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2009, Volume 40 Issue 1
06 January 2009 Previous Issue    Next Issue
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综述
肿瘤基础研究专题报道
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Neurochip’s advancement research: neurons and silicon get intimate by microcontact printing 2009, 40 (1):  1-6.  doi: 10.3969/j.issn.0529-1356.2009.01.001 Abstract ( )   Neurochip is a kind of biochip by using microelectrode array to achieve information transmission between neural cells and electronic devices, as well as an interface of artificial neural compensation device. By connecting living neurons to silicon circuits, long-term recording and stimulating are completed non-invasively. At present in this field, the key point is how to make neural cells adhere on the chips, direct cell bodies to electrodes, establish the communication links and promote the signal to noise ratio between the neuron and substrate. Micro-fabrication technology can be expected to achieve this goal in which microcontact printing technique is the most widely used. By the application of polydimethylsiloxane as stamps, biological macromolecules or organic polymer as “inks", the fine patterns are transferred to the silicon from the template in order to achieve surface modification of materials and structural micro-fabrication. In this review, we introduce the advancement of neural chip research briefly, describe the current research of microcontact printing technology in detail and discuss the application of this approach in the field of neural chip nowadays. Related Articles | Metrics

肿瘤基础研究专题报道

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Preparation of magnetic gemcitabine stealth nanoliposomes and its antitumor effects EM>in vitro/EM> 2009, 40 (1):  7-12.  doi: 10.3969/j.issn.0529-1356.2009.01.002 Abstract ( )   P>Objective To explore a new approach to prepare magnetic gemcitabine stealth nanoliposomes（MGSL）， and evaluate its quality and its antitumor effects EM>in vitro/EM>.  Methods MGSL was prepared by reverse-phase evaporation method. The coated rate, shape, diameter, distribution of MGSL were examined by high performance liquid chromatography(HPLC), microscopy and laser light scattering. Then the stability of MGSLs was also evaluated. The antitumor effect on breast cancer cell line MCF-7 EM>in vitro/EM> was investigated. Electron microscopy was employed to observe morphological features and ultrastructure changes of apoptosis of MCF-7 cells. The effect of MGSL on proliferation inhibitory rate of MCF-7 cells was measured with MTT method. The FCM analysis was carried out to examine the cell cycle distribution and cell apoptotic rate. Results The electron microscope and atomic force microscope photographs of MGSL showed that the shape of liposome was round or elliptical. It was showed that 97.62 percent of the liposome optimized was 206.6nm with above the coated rate 81.7%. Moreover, it possessed a good magnetic response and a high stability. MCF-7 cell line was sensitive to MGSL and the cytotoxity was correlated with the loaded drug dose. After treated for 2h, the effect of MGSL on proliferation inhibitory rate of MCF-7 cells was lower than that of gemcitabine group（EM>P/EM><0.05）, while treated for 48h, the effect on proliferation inhibitory rate was similar（EM>P/EM>>0.05）. Moreover, the effect on proliferation was also similar between MGSL group and MGL group（EM>P/EM>>0.05）.The effect of apoptosis of MGSL on MCF-7 was obvious and the rate of apoptosis was 51 Related Articles | Metrics

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Effects of the combination of ginsenoside Rg1,cinnamic acid and tanshinone IIA on the morphology and terminal differentiation of MG-63 cells EM>in vitro/EM> 2009, 40 (1):  13-19.  doi: 10.3969/j.issn.0529-1356.2009.01.003 Abstract ( )   Objective This study explored the effects of the effective component of Chinese herbs on the morphology,ultrastructure and terminal differentiation phenotype of human osteosarcoma,and in addition identified its inducing function of terminal differentiation of MG-63 cell. Methods MG-63 cells were treated with the combination of 33 mg/L ginsenoside Rg1,2mmol/L cinnamic acid and 0.3 mg/L tanshinoneIIA(RCT) for 7 days and three samples of each groups were subjected to immuncocytochemical assay,and light and electromicroscopy. The MG-63 cells treated by hexamethylene bisacetamide(HMBA) were investigated as a positive control of induced differentiation. Results Immunocytochemistry analysis revealed that the expression of the molecular biomarkers of terminal differentiation,including type Ⅰ collagen，osteocalcin and osteonectin,were notabley positive,and more calcified glycogen particles and classic bone nodules were obsreved in the RCT treated cells.Using light and electromicroscopy, we found that the morphology and ultrastructure of MG-63 cells had undergone restorational changes after RCT treatments similar to that of normal cells: the cells were regular and inclined to have the same size, the size increased, the nucleo-cytoplasm ratio lessened, the number and size of nucleous decreased, organelles tended to be well-developed, and were typical and polarized. Conclusion From these observations we have drawn the conclusion that RCT could reverse the malignant phenotypes of tumor cells, promote the expression of terminal differentiational biomarkers, and remarkably induced MG-63 Related Articles | Metrics

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The effects of Thy-1 expression on cell motility and regulated genes of lung adenocarcinama cell line A549 cells 2009, 40 (1):  20-24.  doi: 10.3969/j.issn.0529-1356.2009.01.004 Abstract ( )   Objective To investigate the function of Thy-1 gene, especially its possible roles in the migration of A549 cells as well as the possible mechanisms by which it works. Methods Construct recombinant expression plasmid vector of pcDNA3.0-Thy-1 and transfected it into A549 cells. These cell lines were further studied for the exogenous Thy-1 mRNA expression by RT-PCR and expression of endogenous Thy-1 protein by immunofluorescence assay. Then the transwell cell migrations were assayed. In order to study the possible mechanisms by which Thy-1 influences the invasiveness potential, microarray were used to screen out cell migrations associated genes. Results Eukaryotic expression vector pcDNA3.0-Thy-1 was constructed; Stable Thy-1 expressing cell line were established; Transwell assay showed that over expression of Thy-1 in A549 cell line resulted in reduced ability of cell migration (EM>P/EM><0.0001); and microarray suggested that the Thy-1 gene could influenced the expression level of 25 cell motility-related genes. In these genes， 11 genes expressions were up regulated, and 14 genes were down regulated. These results could be consistent when microarray and RT-PCR methods were used to detect the difference of gene expression.Conclusion Overexpression of Thy-1 leads to a less invasive phenotype of A549 cells and changes the expression level of many cell motility-related genes. This suggested that Thy-1 expression patterns in lung cancer tissue may be associated with the invasiveness of non-small cell lung cancer. Related Articles | Metrics

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The expression of BHD gene in human lung adenocarcinoma A549 cells and its effect of BHD gene on the metastasis of A549 cells 2009, 40 (1):  25-31.  doi: Abstract ( )   Objective This study was designed to investigate the BHD gene expression location in human lung cancer cells and its effect on the invasion ability of lung cancer cells. Methods The BHD vector and the small interfer RNA targeting BHD was constructed and transfected into the cell line A549. The expression of BHD was examined by immunofluorescence. Transwell test was used to detect the invasion ability of A549 cells. Results After transfected BHD vector into A549 cells, immunofluorescence showed the expression of Folliculin located in cellular plasma. The invasion ability of A549 cells decreased after transfected with BHD vector, and the invasion ability of A549 cell increased after transfected with BHD siRNA. Conclusion In the lung cancer cell line A549, Folliculin is expressed in the cytoplasm and BHD gene is a metastasis suppressor gene. Related Articles | Metrics

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Radix Actinidiae extractive inhibits the growth of transplanted tumor of esophagus cancer cell EC109 in nude mice 2009, 40 (1):  32-36.  doi: Abstract ( )   Objective To study the effect of acetidin extractive of Radix Actinidiae on transplantation tumor of esophagus cancer EC109 cells in nude mice,and to explore its mechanism of action. Methods Establish the model of transplanted tumor bearing esophagus cancer EC109 cells in nude mice and to detect the effect of acetidin extractive of Radix Actinidiae EM>in vivo/EM>. Control group and treatment groups (high-dose group and low-dose group)were set for comparison,there was six nude mice in each group．The transplanted tumor growth curve was drawn according to transplanted tumor volume, and inhibition ratio was calculated according to the final tumor weight. Terminal deoxynucleotidyl transferase mediated UTP nick end labeling (TUNEL) was used to examine the apoptosis index of the transplanted tumor.Immunohistochemistry (SP)was used to examine the expression of Survivin protein and Ki-67 antigen in the transplanted tumor.RT-PCR was used to examine the expression of Survivin mRNA. Results The growth of transplanted tumor in treatment groups became slow after taking drug.The final tumor volume and tumor weight of treatment groups were smaller than that of control group. The tumor weight inhibiting rates of highdose group and lowdose group were 64.57% and 46.78% individually. The apoptosis index of the transplanted tumor of treatment groups was higher than that of control group. The expression of Survivin mRNA and protein,Ki-67 antigen in treatment groups were lower than that of control group. And high-dose group was more differently. The statistically significant difference was found between the index of each groups that have mentioned (P<0.01).Conclusion The acetidin extractive of Radix Actinidiae has the inhibitory effect on the growth of transplanted tumor of esophagus cancer EC109 cell in nude mice.To inhibit proliferation activity of transplantation tumor,to degrade the expression of Survivin protein and mRNA,and to induce apoptosis of cancer cells might b Related Articles | Metrics

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Fluctuating asymmetry of dermatoglyphy in breast cancer patients 2009, 40 (1):  37-40.  doi: Abstract ( )   Objective To investigate the characteristics of dermatoglyphic parameters of healthy controls and breast cancers in women of Han ethnic from Ningxia, and analyze their fluctuating asymmetry (FA). Methods Fingerprints and palmar prints of both hands were collected with cyclostyling ink　(breast cancer patients: 112,control: 112). Results Breast cancers had significant difference in right FRCⅠand atd （EM>P/EM><0.05）of both hands. There was a significant difference in FAⅤ（atd angle,EM>P/EM>0.01）,FAⅩ（Ⅳ ridge count,EM>P/EM>0.05）between two groups, breast cancers had higher FA. And there was also a significant difference in |R-L|≥7 on distribution of atd angle bet Related Articles | Metrics

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Significance of alterations of mitochondrial DNA copy number in breast carcinoma 2009, 40 (1):  41-43.  doi: Abstract ( )   Objective To explore the effect in human breast cancer occurring by analyzing the difference of mtDNA copy number in breast cancers and paracancerous tissues. Methods Polymerase chain reaction（PCR）was applied to detect the HV1 and HV2 of mtDNA D-loop in 20 cases invasive ductual breast cancer tissues and 20 cases peritumoral tissues, meantime β-actin was served as a quantitative standard marker, it compared the difference between breast cancer and paracancerous tissues by polyacrylamide gel electrophoresis(PAGE) and analyzing nucleic acid in the Gel-Pro Express system. Results The bands of HV1 and HV2 in breast cancer tissues was weaker than that in peritumoral tissues. There was difference between two groups by HV1 (EM>P/EM><0.05); There was difference between two groups by HV2 (P<0.01). It was detected that the HV1 and HV2 were positively correlated by statistics analyzing (P<0.01). There was no statistic relationship between the difference of mtDNA copy number and age and diameter, but the mtDNA copy number in lymph node metastasis group was higher than that of no lymph node metastasis group, there was difference between two groups (P<0.05). The mtDNA copy number in phase Ⅰof TNM stage was higher than that of phase Ⅱand Ⅲ, and there was difference between phase Ⅱ and phase Ⅲ (EM>P/EM><0.05), there was difference between phase Ⅰ and phase EM>Ⅲ(P/EM><0.05). Conclusion The mtDNA number is descended in i Related Articles | Metrics

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Association of expression of vascular endothelial growth factor D, lymphangiogenesis and lymph node metastasis in human breast cancer 2009, 40 (1):  44-47.  doi: Abstract ( )   Objective To investigate lymphangiogenesis, lymphovascular invasion (LVI) and the expression of vascular endothelial growth factor D (VEGF-D) and their correlations in different regions of human breast cancer, and to evaluate their associations with axillary lymph node metastasis. Methods Tissue samples of primary tumors from 79 patients undergoing curative surgical resections for breast cancer were divided into four regions: tumor region, peritumoral region, nearertumor region and farthertumor region. Samples from the different regious were immunohistochemically examined for D2-40 and VEGF-D to evaluate lymphatic vessel density (LVD), LVI and VEGF-D expression in different regions. Results LVD in the peritumoral region was highest 20.25±2.03, and the positive rates of VEGF-D expression and LVI in tumor region were also highest (87.34% and 63.29%, respectively). In the tumor region, there was no significant correlation (P>0.05) between LVD and VEGF-D expression, between LVD and LVI, and between VEGF-D expression and LVI; however, there were significantly positive linear correlations (P<0.05) in other regions. Axillary lymph node metastasis significantly correlated with LVD in the peritumoral region, and significantly correlated with VEGF-D expression and LVI in the peritumoral and nearertumor regions (EM>P/EM><0.05). Conclusion The expression of VEGF-D might promote lymphangiogenesis and increase the chance of lymphovascular invasion in human breast cancer. The increase of LVD easily promotes LVI and axillary lymph node metastasis. The peritumoral and nearertumor regions may play more important roles in the study of lymphangiogenesis, lymphatic spread of breast cancer and in th Related Articles | Metrics

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Association of expression of vascular endothelial growth factor D, lymphangiogenesis and lymph node metastasis in human breast cancer 2009, 40 (1):  44-47.  doi: Abstract ( )   Objective To investigate lymphangiogenesis, lymphovascular invasion (LVI) and the expression of vascular endothelial growth factor D (VEGF-D) and their correlations in different regions of human breast cancer, and to evaluate their associations with axillary lymph node metastasis. Methods Tissue samples of primary tumors from 79 patients undergoing curative surgical resections for breast cancer were divided into four regions: tumor region, peritumoral region, nearertumor region and farthertumor region. Samples from the different regious were immunohistochemically examined for D2-40 and VEGF-D to evaluate lymphatic vessel density (LVD), LVI and VEGF-D expression in different regions. Results LVD in the peritumoral region was highest 20.25±2.03, and the positive rates of VEGF-D expression and LVI in tumor region were also highest (87.34% and 63.29%, respectively). In the tumor region, there was no significant correlation (P>0.05) between LVD and VEGF-D expression, between LVD and LVI, and between VEGF-D expression and LVI; however, there were significantly positive linear correlations (P<0.05) in other regions. Axillary lymph node metastasis significantly correlated with LVD in the peritumoral region, and significantly correlated with VEGF-D expression and LVI in the peritumoral and nearertumor regions (EM>P/EM><0.05). Conclusion The expression of VEGF-D might promote lymphangiogenesis and increase the chance of lymphovascular invasion in human breast cancer. The increase of LVD easily promotes LVI and axillary lymph node metastasis. The peritumoral and nearertumor regions may play more important roles in the study of lymphangiogenesis, lymphatic spread of breast cancer and in th Related Articles | Metrics

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Influence of low expression of annexin A5 on the proliferation and apoptosis of HeLa cells 2009, 40 (1):  48-51.  doi: Abstract ( )   Objective To observe the change of proliferation and apoptosis of HeLa uterine cervical carcinoma cells when expression of annexin A5 protein was suppressed. Methods RNA interference method was employed. The pair of siRNAs which can supress the expression of annexin A5 protein best were screened out by Western blotting and then the siRNAs were transfected into HeLa cells; 48 hours later, cells were collected. MTT method was used to observe the change of proliferation of the HeLa cells and the apoptosis examining box was used to examine cell apoptosis. Results The proliferation speed of the HeLa cells in which annexin A5 protein was low expressed had no significant difference with those of the cells untransfected while apoptosis of the cells were significantly decreased. Conclusion Proliferation of HeLa cells was not identified to be influnenced when the expression of annexin A5 protein was suppressed while cell apoptosis was affected significantly. Related Articles | Metrics

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Construction of recombinant plasmid pcDNA3.1-ANXA5 and the affection to the proliferation of the SiHa cells by overexpressed annexin A5 2009, 40 (1):  52-56.  doi: Abstract ( )   Objective To construct the recombinant plasmid pcDNA3.1-ANXA5 so as to study the affection of the overexpressed annexin A5(ANXA5) protein to the proliferation of uterine cervical carcinomal SiHa cells. Methods The ANXA5 cDNA fragment was cloned into pcDNA3.1 vector to construct the recombinant plasmid pcDNA3.1-ANXA5; The pcDNA3.1-ANXA5 plasmid and pcDNA3.1 vector were transfected into the uterine cervical carcinoma SiHa cells respectively and screened by G418. Western blotting was used to identify cell clones which overexpress ANXA5 and immunohistochemistry was applied to identify cells transfected with pcDNA3.1 vector which has a His-tag. Cells overexpressed ANXA5, cells transfected with pcDNA3.1 vector and cells without any treatment were tested by MTT and CCK-8 method to examine whether overexpressed ANXA5 protein can affect the proliferation of the SiHa cells. Results The recombinant plasmid pcDNA3.1-ANXA5 was successfully constructed and the ANXA5 gene sequence was proved correct; The proliferation of the SiHa cells without treatment and those transfected with pcDNA3.1 vector was fast and there was no difference（P>0.05）between the two kinds of cells；while the proliferation of the SiHa cells overexpressed ANXA5 was very slow. The difference from the other two kinds of cells was significant (P<0.05). Conclusion The ANXA5 protein maybe affect the Related Articles | Metrics

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Role of cord blood-derived T lymphocytes transplanting for BALB/c mice with erythroleukemia 2009, 40 (1):  57-62.  doi: Abstract ( )   Objective To study the survival and infiltration condition of cord blood derived-T lymphocytes in mice and its therapeutic action for erythroleukemic (EL) mice; and to provide the experimental and theoretical basis for studying the biological characteristics and antileukemia effects of cord bloodderived T cells. Methods Fourty-eight BALB/c mice were randomly divided into three groups, the control group(CG),the experimental group 1(EG 1)and the experimental group 2(EG 2). Cord blood derivedT lymphocytes were separated and prepared, and transplanted it into EL mice via the tail vein or peritoneal injection after being signed to their group. This was done in order to observe the survival time and infiltrate condition of the cord bloodderived T cells in mice and its effect on the morphosis of EL mice spleen by immunofluorescence (IMF), immunohistochemistry (IMC) and transmission electron microscope (TEM). Results Most activated cells were CD3SUP>+ /SUP>T lymphocytes by PHA, and the positive rate was (83.42±1.26)%; human CD3SUP>+/SUP> cells and BrduSUP>+/SUP> cells were detected in peripheral blood, spleens and liver of all mice at the 3rd and 7th day after cord blood derived-T lymphocytes were transplanting; CD25SUP>+/SUP> cells could be detected in the liver and spleen of EL mice in both experimental groups; and the average survival time of EL mice in the two experimental groups was (27.25±7.06) days and (24.74±2.93) days respectively (this was statistical significance compared to control group, P<0.05). The results of Hoechst 33258 staining showed: 1.Nuclear fluorescence was uniform in the control group and there were few apoptotic cells; 2. In the experimental group, some of the leukemia cells appeared to have morphological changes consistent with apoptosis; chromatin had a high degree of cohesion; dense concentration,or splited into massives, edge was smooth and clear; Under TEM, leukemic cells infiltrate was generally observed in the spleens of the control group mice. Parts of the leukemic cells showed typical apoptosis morphological changes in the experimental groups: nuclear chromatin concentration and edge accumulation was seen；nuclei fragmentation and formation of apoptotic bodies; the expansion of RER, and the vacuolar or medullar degeneration of mitochondria could be found. Conclusion The cord bloodderived T lymph Related Articles | Metrics

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Raf-1 is possibly related with the multidrug resistance in oral squamous cell carcinoma 2009, 40 (1):  63-66.  doi: Abstract ( )   Objective Multidrug resistance is a major barrier for most patients with oral squamous cell carcinoma. In this study, we investigated the difference of the expression of Raf-1 mRNA and protein between multidrug resistant (KBV) and sensitive (KB) human oral squamous carcinoma cells to explore the effect of Raf-1 on the multidrug resistance in the human oral squamous carcinoma. Methods The changes of the mRNA and protein levels of Raf-1 in both KBV cells and KB cells were determined by RT-PCR,Western blotting assay and immunofluorescent method. By MTT and flow cytometry (FCM), ICSUB>50/SUB> and apoptosis rates in both KBV and KB cells were detected. Results The findings indicated that expressions of Raf-1 mRNA and protein levels in KB cells were lower than those in KBV cells statistically (P<0.01). Immunofluorescent image showed that Raf-1 protein expression in KBV cells were much stronger than that in KB cells. The ICSUB>50/SUB> in KB cells was (21.33±2.25)μg/L and that in KBV cells was (352.68±12.36)μg/L.The resistance index (RI) was 16.5The apoptosis rate in KB cells was (98.8±1.2)% and that in KBV cells was (23.5±2.1)%.These results showed that differences in the chemosensitivity to VCR between KBV cells and KB cells were significant statistically (P<0.01). Conclusion Related Articles | Metrics

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Study of the age-related changes of the white matter and the myelinated nerve fibers in the white matter of the left hemisphere and right hemisphere of male rats 2009, 40 (1):  67-72.  doi: Abstract ( )   Objective To explore if there are significant differences in the white matter and the myelinated fibers between left and right hemispheres of male rats and if the age-related changes of the white matter and the myelinated fibers in the white matter are same for left hemisphere and right hemisphere of male rats. Methods The white matter and the myelinated fibers in the white matter from 5 male young Long-Evans rats (6-8 month old) and 4 male aged Long-Evans rats (18 month old) were quantitatively investigated with the stereological techniques and transmission electronic microscope technique. Results There are no significant differences of the white matter and the mylinated nerve fibers in the white matter between the left and right hemispheres in both young and aged groups. Although the total volume of the white matter and the total length and the total volume of the myelinated nerve fibers in the white matter were reduced in both left hemisphere and right hemisphere, only the white matter volume and the total volume of the myelinated fibers in the white matter of right hemisphere and the total length of the myelinated fibers in the white matter of left hemisphere were significantly decreased by 32.9%, 28.6% and 49.3%, respectively. Conclusion There are no significant differences in the white matter and myelinated nerve fiber in the white matter between left and right hemispheres in both young and aged male Long-Evans rats. The white matter volume and the total volume of the mylinaed fibers in the white matter of right hemisphere and the total length of the myelinated fibers in the white matter of left hemisphere in the aged male rats are significantly decreased when compared to young rats. Related Articles | Metrics

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Relationship of the epilepsy induced by IL-2 and JAK/STATs signal transduction pathway 2009, 40 (1):  73-77.  doi: Abstract ( )   Objective To explore the relationship of the epilepsy induced by interleukin2(IL-2) and the Janus kinase(Jak)/ signal transducers and activators of transcription(Stats）signal transduction pathway. Methods Immunocytochemistry was applied to study the expression changes of tyrosine kinase Jak1 in brain after IL-2 administration to induce epilepsy, and the changes of N-methyl-D-aspartate receptor-1(NMDAR-1) and glutamic acid(Glu) when preusing Jak1 inhibitor and glucocorticoid prior to IL-2 Results After injecting IL-2 into the lateral ventricle, the rats displayed obvious seizures and the expression of Jak1 was significantly stronger comparing with the control group(P<0.01), immunosuppressants such as cyclosporin A(CsA) could partly inhibit this effect. Jak1 inhibitor genistein and immunosuppressant glucocorticoid could restrain the epilepsy behaviors, and the NMDAR-1 and Glu expressions significantly decreased conferring to the IL-2 group（P<0.05）. Conclusion Jak1 plays an important role in signal tr Related Articles | Metrics

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Changes in somatostatin-immunopositive neurons of the hippocampal formation in the Meynertlesioned rat 2009, 40 (1):  78-82.  doi: Abstract ( )   Objective To investigate the changes of somatostatin(SS)immunopositive neurons in the hippocampal formation in Meynert(NBM)-lesioned rat and to make a preliminary analysis of the mechanism of SS participating in learning and memory. Methods In this study, the memory dysfunction model was established by the microinjection of kainic acid (KA) into the bilateral NBM. The passive avoidance task was used to estimate the degree of memory dysfunction. After 1-4 weeks survival, brain sections were stained by immunohistochemical technique to show the SS-immunopositive neurons. Then the SS-immunopositive neurons were counted with an optic microscope. Results Lesions of NBM could remarkably disrupt memory continuance compared with the normal group. There was a highly significant difference between the NBM-lesioned group and the normal group in the number of SS-positive neurons of hippocampal formation(P0.01). There were a few changes of lesion in cellular morphology which present as the slightly illegible cell outline and the shortening or disappeanig processes. Conclusion SS-positive neurons in the hippocampal formation decreased significantly in the NBM-destroyed rat and it may have a role in the memory dysfunction to a certain extent. Related Articles | Metrics

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Method for expansion and induction of porcine fetus islet stem cell 2009, 40 (1):  83-86.  doi: Abstract ( )   Objective To find a method for proliferating porcine fetus islet stem cells. Methods Cells collected from porcine fetus pancreas were digested with trypsine and cultured in medium RPMI1640 with 10% FBS. Some cells survived after seven days, these cells could be passed for 5 to 6 times. Then, most of those cells were trend to die and float in the medium, only were a few little cells attached on culture plate. These little cells proliferated quickly when medium was changed to RPMI 1640 with 15% FBS and each of the little cells can formed cells’ clone in 4 days. The cell’s character of islet stem cell was detected by immunochemical method. Results Cells obtained by the method posses strong proliferation ability and it has been sub cultured over 64 passages till now. It expressed islet stem cell’s mark such as PDX-1，GLUT-2，nestin，vimentin et.al. After induction, it expressed insulin, glucagon, somatostatin and polypeptide. Conclusion Cells obtained by this method were determined as islet stem cells. They can differentiate into islet endocrine cell EM>in vitro/EM>. This study can provide reference for isolation and expansion of isl Related Articles | Metrics

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Effects of recombinant human endostatin on intracellular CaSUP>2+/SUP> homeostasis in fibroblast-like synoviocytes in rats with adjuvant arthritis 2009, 40 (1):  87-91.  doi: Abstract ( )   Objective To observe the effects of recombinant human endostatin(rhEndostatin) on intracellular CaSUP>2+/SUP> homeostasis in fibroblastlike synoviocytes(FLSs) in rats with adjuvant arthritis(AA), approach ionic mechanisms of AA FLSs apoptosis induced by rhEndostatin, and provide experimental data for possible drug treatment and their targets in rheumatoid arthritis (RA). Methods Twelve male SpragueDawley(SD) rats weighing 140160 g were used for these studies. The rats were divided at random into a normal group (EM>n/EM>=3) and an AA model group (EM>n/EM>=9). Adjuvant arthritis was induced in rats in the AA model group and the FLSs obtained from AA rats were cultured EM>in vitro/EM>. The cultured AA FLSs were incubated with the calcium ionsensitive fluorescent indicator fluo3/AM. A confocal laser scanning microscope was used to measure changes of fluorescence intensity of intracellular CaSUP>2+/SUP> in the cells caused by rhEndostatin when the cells were immersed in buffer (with or without CaSUP>2+/SUP>), and to investigate the effect of rhEndostatin on intracellular free calcium concentration {［CaSUP>2+/SUP>］i} in AA FLS. Results Recombinant human endostatin could cause rapid ［CaSUP>2+/SUP>i elevation in static AA FLSs in CaSUP>2+/SUP>containing buffer. Within 10s after treatment with rhEndostatin, the elevated levels of ［CaSUP>2+/SUP>］i ran rapidly to peaked and was followed by a plateau period. The elevated ［CaSUP>2+/S Related Articles | Metrics

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The induction of SMMC-7721 cells into polyploid giant cells using triazole schiff base derivative EM>in vitro/EM> 2009, 40 (1):  92-97.  doi: Abstract ( )   Objective To analyze the features of polyploid giant cells that come from SMMC-7721 cells after being induced by triazole Schiff base derivative(LH-37) EM>in vitro/EM>. Methods The SMMC-7721 cells（1×10SUP>4/SUP>/ml） were cultivated in medium containing of 1×10SUP>-5/SUP> mol/L LH-37 for 72 hours, and their DNA content was analyzed by flow cytometry. The Co-staining with Hoechst 33258 and propidium iodide were used to observe cell death. Behavior of the microtubule(MTs) and laminB configuration apparatus in cells were examined by immunofluorescence, counterstained with Hoest33258 staining，and the ultrastructure of giant tumor cells was observed by JEM 100CX-II electron microscopes.Results Increased the proportion of mono-nucleated or multi-nucleated giant cells (MOGC and MNGC) after SMMC-7721 cells were cultured in the presence of LH-37 at 72 hours. The proportion of the giant cells in MOGC and MNGC was 12.32%and 0.92% respectively. Arrangement and radial segregation of subnuclei, subnuclei were united through MTs which radically emerged and embraced them from a single microtubule organizing centre (MTOC),thick microtubule tracks reached individual nucleoli; sub-nuclei of this radial arrangement was encircled by the intact lamin; and most of the MNGCs died by apoptotic crisis. MTs of MOGC uniformly spread throughout the cytoplasm from the nuclear envelope and usually accumulated in the perinuclear region. Abnormal mutipolar mitosis, and spontaneous chromosome condensation were observed. In addition electron micrographs demonstrated features of autophagy during apoptosis of some MOGCs. Conclusion The data presented indicate th Related Articles | Metrics

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Biological characteristics of human bone marrow mesenchymal stem cells and differentiation to neural precursor cells 2009, 40 (1):  98-102.  doi: Abstract ( )   Objective To investigate biological characteristics of cultured human bone marrow mesenchymal stem cells (hMSCs) EM>in vitro/EM>, and differentiation potential to neural precursor cells (NPCs). Methods hMSCs were isolated with density gradient centrifugation and cell adhesion from adult bone marrow. The cell morphology, growth, surface markers were investigated, and osteogenic, chondrogenic, adipogenic differentiation potential were analyzed as well. At the 3 passage, hMSCs were expanded by embryonic stem cell maintenance medium, then these cells were exposed to neural induction medium, that contained 5-aza-2-deoxycytidine (5azadc) and Trichostatin A (TSA). After 7 days, half of the samples were detected the markers for multipotent NPCs, Nestin and Sox2 by immunofluocence and RT-PCR. Further, another half of the samples were grown an additional week in Neurobasal/B-27，to observe to differentiate into neural phenotypes. Results For acquired hMSCs, the positive ratio of CD-29，CD-44 were all above 90%; The osteogenic, chondrogenic and adipogenic differentiation potential were detected obviously; hMSCs induced by 5azadc and TSA could be identified by the positive staining for Nestin and Sox2, RT-PCR assessment showed that the differentiated cells expressed Nestin and Sox2. And after 1 week in Neurobasal/B-27, hMSCs differentiated into NF-L immunopositive cells with similar neuronal morphology. Conclusion hMSCs can be cultured and expanded EM>in vitro/EM>; after treated with epigenetic modifiers agents in neural environment, Related Articles | Metrics

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Application of hydrodynamics-based transgent togene transfection in rat liver 2009, 40 (1):  103-107.  doi: Abstract ( )   Objective To study the condition and method of hydrodynamics-based transgene when applied to gene transfection in rat liver. Methods Inject different dosages and concentrations of green fluorescent protein (GFP) plasmid (pEGFP-C1) at different speeds, then collect 4 rats’ liver leaves at different time points after injection for frozen section, and to observe and quantify the GFP expression with a fluorescence microscope at 488 nm excitation wavelength. Results With one-off injection(the optimal conditions), plasmid concentration is 30mg/L, injection speed is 2ml/s, injection volume is 9％ of rat body weight. At 6 hours after injection, GFP-positive cells take on the highest proportion of above 20％; GFP begin to gradually reduce since 24 hours, until 72 hours almost no GFP-positive cells are seen in any liver leaf. Briefly(under the above optimal conditions), GFP expression in each liver leaf is described as follows: about 30％ for pedicel leaf, about 20％ for all of left, middle and right leaves,and about 10％for tail leaf.Conclusion Conclusion Hydrodynamics-based gene delivery procedure can be well applied to transgene in the rat liver, with the appropriate conditions of this method being 30mg/L plasmid concentration, 9％rat avoirdupois, 2ml/s injection speed. The most suita Related Articles | Metrics

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The effects of heroin dependence on expression of islet amyloid polypeptide and glucose transporter 2 in pancreas islet β cells of rats 2009, 40 (1):  108-112.  doi: Abstract ( )   P>Objective To explore the effects of heroin dependence on expression of islet amyloid polypeptide (IAPP)and glucose transporter 2(Glut2) in islet β cells of rats, and to study the possible, mechanism of expression changes. Methods Sixty-six normal SD rats were divided into a normal control group (NCG), a saline control group (SCG) and a heroin dependence group (HDG). We established the heroin dependence model in rats by subcutaneous injection of heroin, and excision of the pancreas tails on 10 days, 17days, 24days, 31days and 38days. Tissues were assessed by immunohistochemical SABC, image analysis and morphometric methods in the study. Results The results of immunohistochemical staining showed that: 1. IAPP immunoreaction was localized in the cytoplasm of β cells,whereas Glut2 immunostaining was observed on the membranes of β cells; 2.As compared with the NCG and SCG, the immunoreaction of IAPP immunoreactive(IR) positive cells in HDG increased and the immunostaining of cells turned intensive, the numerical density on area also increased after the 24th day HDG (P<0.05). Meanwhile, the results of image analysis showed that the mean grey degree of IAPP-IR cells in HDG decreased as compared with NCG and SCG(P<0.05),especially on the 38th day; 3.The immunoreaction of Glut2 positive cells in HDG increased obviously and the results of image analysis showed that the mean grey degree of Glut2-IR cells in HDG. Decreased as compared with NCG and SCG (P<0.05), with the lowest degree on the 38th day. Conclusion By enhancing the synthesis and secretion of IAPP and Glut2, β cells of islets are involved in the regulation of body metabolic and carbohydrate metabolic processes Related Articles | Metrics

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Experimental observation on angiogenesis of the frozen-thawed human fetal ovaries after xenotransplantation 2009, 40 (1):  113-117.  doi: Abstract ( )   Objective To observe the angiogenesis in the human fetal ovarian grafts one month after heterotransplantation. Methods Ovaries from 3 inevitable abortion human fetuses 22 gestational weeks of age were frozen- thawed, xenografted into the immunodeficient nu/nu mice(EM>n/EM>=18), and recovered on 2, 7, and 28 days after transplantation respectively. These grafts, together with the fresh and the frozen-thawed ovarian tissue were examined for blood vessels by immunolocalization of CD34, ultrastructure, and detection of the mRNA expression of vascular endothelial growth factor (VEGF)， Angiopientin2(Ang-2). The untransplanted tissues of the fresh and the frozenthawed ovarian were investigated exactly the same as the grafts. Results The density of the microvessel was significantly increased in the grafts on day 2, peaked on day 7, and slightly reduced on day 28, together with distended microlvessel on day 2 which turned to normal on day 7 under the light Microscope. Transmission electron microscope (TEM) showed the majority primordial follicles were well preserved. The grafts of day 2 showed some edema in stroma and vessels filled with red blood cells. As they disappeared, the microvessel was growing in the counterparts on day 7 and day 28. The mRNA levels of VEGF and Ang-2 showed the similar temporary rising on day 2 and decreasing since. Conclusion The angiogenesis of the frozen-thawed ovarian tissue from human fetus completed within one month after transplantation. The first week after transplantation was the critical time to achieve prolonged graft functionality and increased the yield Related Articles | Metrics

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Apoptosis and expression of apoptosis-related proteins in lymphocytes during the post embryonic development of the duck bursa 2009, 40 (1):  118-121.  doi: Abstract ( )   Objective The apoptosis and expression of Caspase-3,Bcl-2 proteins in lymphocytes throughout the post embryonic development of the Tian Fu meet duck bursa of fabricius were studied in order to explore the mechanism of B-lymphocytes apoptosis. Methods The Tian Fu meet ducks from 0-29 weeks posthatching period were divided into 11 groups, 5 ducks per group.The routine HE, terminal deoxynucleotidyl transferase mediated dUTP-biotin nick endlabeling(TUNEL)，and immunohistochemical methods were used. Results From 0-3 weeks posthatching period, the apoptosis index of lymphocytes in the medullar of follicle rose, whereas that in the cortex of follicle remained the same. The apoptosis indexes of lymphocytes in the medullar and cortex of follicle decreased at the 5th week and maintained at this level until the 14th week, and then increased at the 17th week, reaching their plateaus at the 29th week. During 0-14 weeks, the apoptosis indexes of lymphocytes in the medullar of follicle in each group was higher than that in the cortex, and lower than that in cortex from 17-29 weeks. The changing pattern of Caspase-3 expression in lymphocytes was paralleled positively with that of apoptosis index, while that of Bcl-2 expression in lymphocytes was reversely with that of apoptosis index (except 5-14 weeks). Conclusion The lymphocyte apoptosis exists universally in the duck bursa during the post embryonic development period which undergoes obvious age-related changes. The expression patterns of Caspase-3 and Bcl-2 proteins in lymphocytes might be one of the crucial mechanisms in such age-related changes of lymphocytes apoptosis. Related Articles | Metrics

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An evaluation on the response of the mouse bone marrow to magnesium alloy EM>in vivo/EM> 2009, 40 (1):  122-126.  doi: Abstract ( )   Objective To investigate the degradation mechanism at the bone-implant interface of degrading magnesium alloys in bone and to determine their effect on the surrounding bone. Methods Sample rods of magnesium alloys were implanted intramedullary into the femora of rats received a subcutaneous injection of gentamycin as an antibiotic prophylaxis and weekly subcutaneous injections of calcein green. After 6, 9, 15, 26 and 80 weeks, decalcified sections were maken for histomorphologic analysis. The bone-implant interface was characterized in calcified sections by optical microscopy, scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS). Results The metallic implants made of magnesium alloys were degraded entirly in vivo with the elapse of time.The corrosion layer was indirectly contact with the surrounding bone through basement membrane containing C, N and O elements. Besides,the results further showed an increased bone mass around the magnesium rods, while no bone was induced in the surrounding soft tissue.Conclusion The results of this study suggest that high magnesium ion concentration with Zn and Mn could lead to bone cell activation, and might be a suitable material for bone-implanting. Related Articles | Metrics

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Histological architecture of reticular fibers and age-related changes of myocardial tissue in rats 2009, 40 (1):  127-129.  doi: Abstract ( )   Objective To observe the histological architecture of reticular fibers and agerelated changes during development of rats, in order to establish abase for probing myocardial tissue remodeling characteristics. Methods Using technique of paraffin section, Gomori silver staining, the myocardial tissue of fetal rat (pregnant about 19d), newborn rat (9d), young rat (8m) and old rat (14m) were observed with light microscopy. The expression differences of Ⅲ type collagen were detected by Western blotting. The reticular fibers was quantitated with image analysis system, and the data were processed with SPSS statistical software. Results The reticular fiber could be divided to thin and thick fibers that encircled myocytes. The reticular fibers around myocyte showed thin string or absence in fetal rat. As for the newborn rat, the reticular fibers around myocytes gradually increased and formed continuous membranes, and some spaces that were full of branch of reticular fibers appeared between myocytes. The area percentage of reticular fibers was gradually changed. Ⅲ type collagen (reticular fiber) expression changed with aging, and sharply increased from newborn to young rat.Conclusion The reticular fibers are composed of thick or thin fibra sheaths around myocytes, and the branchs of reticular fibers are full of the space between myocytes or between myocytes and interstitium, and the expression of Ⅲ type Related Articles | Metrics

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Effect of VEGF and AR activity upon femoral head structure and micrangium morphology of diabetic rats 2009, 40 (1):  130-136.  doi: Abstract ( )   Objective To observe the distribution and morphologic variation of micrangium in diabetic rat’s femoral head and to probe into the difference of the expression of VEGF mRNA at the different time and the effect of the variation of the aldose reductase(AR) activity on the femoral head structure as well as micrangium lesion. Methods To set up the diabetic rat’s model with STZ, we divided the rats into three normal groups: CON1, CON2 and CON3 and three diabetic groups：DM1, DM2 and DM3, ten rats in each group. The structure of femoral heads of different group rats was observed under light microscope. The expression of coagulation factor Ⅷ was detected with immunohistochemistry. The density of micrangium was measured by infusion with ink. We also observed the expressive intenstiy of VEGF mRNA EM>in situ/EM> hybridization and analysised the activity of AR by making biochemical detection of the tissue homogenate of femoral head of the rat in each group. Results The diabetic rats’ femoral heads developed with the course of illness，the bone trabecula became thin and few,the marrow cavities enlarged.The expression of coagulation factor Ⅷ rose，the microvessel density enlarged.The expression of VEGF mRNA in vascular of the marrow cavities endothelial cells rose，and the activity of AR was enhancement（EM>P/EM><0.05）.Conclusion In the early period of diabetic rat’s femoral head t Related Articles | Metrics

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Quantitative analysis on the relationship between ureteric bud and nephron number in developing mouse kidney 2009, 40 (1):  137-140.  doi: Abstract ( )   Objective To investigate the growth pattern of ureteric bud (UB) and nephron number induced by UB in developing mouse kidney. Methods Immunohistochemical technology and stereological method were used to observe and measure UB and nephron at different developmental stages of embryonic and postnatal mice. Results Immunohistochemical examination showed that ureteric buds surrounded by metanephrogenic tissue were observed in embryonic day 12, and each UB tip branched dichotomisously. In embryonic day 14, nephrons induced by UB emerged. From embryonic day 16 to postnatal day 5, the number of UB tip and nephrons were increased. In postnatal 7, UB disappeared and nephron number was not changed. Stereological examination showed that from embryonic day 14 to 18, each UB tip induced one nephron . From postnatal day 1 to 5, each UB tip induced four nephrons. Those data indicated that a total of 10-11 UB branching events occurred during mouse kidney morphogenesis.Conclusion It is well established that nephron number is proportional to the extent of UB branching that occurs during developing kidney, but it is not constant. Related Articles | Metrics

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Expression of LPA-R3 in uterine endometrium during the period of peri-implantation of mouse embryo 2009, 40 (1):  141-145.  doi: Abstract ( )   Objective To investigate the expression and distribution of LPA-R3 during the period of mouse embryo implantation. Methods Seventyeight mice were randomly divided into pregnancy group（EM>n/EM>=36）,pseudopregnancy group(EM>n/EM>=36) and control group(EM>n/EM>=6).Immunohistochemistry and reverse transcription polymerase chain reation (RT-PCR) and Western blotting were employed to analyze the expression and distribution of LPA-R3 in the uterine endometrium during mouse embryo implantation. Results LPA-R3 was mainly expressed in cytoplasm of glandular and luminal epithelium and part of stromal cells of mouse uterus.The LPA-R3 mRNA and protein level in the uteri increased during early pregnancy, increased at 3 days post coitus (3 d.p.c.), peaking around 4 d.p.c., periimplantation period, then returned to the basal levels on 5 d.p.c. This temporal profile of LPA-R3 expression in the uterus of pseudopregnant mice was essentially similar to that observed in normal pregnant mice. The expression level of LPA-R3 receptor protein and mRNA had no significant difference between pseudopregnant mice and normal pregnant mice(EM>P/EM>>0.05).Conclusion LPA-R3 may participate in the period Related Articles | Metrics

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Comparison between EM>Canis lupus/EM> and EM>Canis familiaris/EM> in morphological and cytological characteristics of their digestive tract 2009, 40 (1):  146-152.  doi: 10.3969/j.issn.0529-1356.2009.01.030 Abstract ( )   Objective To investigate the anatomic and histologic characteristics of the digestive tract of EM>Canis lupus/EM>and EM>Canis familiaris/EM> and types, regional distributions and cell densities of gastrointestinal endocrine cells in various parts of their digestive tract, in order to clarify morphology and cytohistology mechanism of their different feeding habits. Methods Six wolves and six domestic dogs have been tested, using morphological observation and paraffin sectioning to compare the anatomic and histologic characteristics of the digestive tract; using immunohistochemical techniques to compare the types, regional distributions and cell densities of gastrointestinal endocrine cells in various parts of their digestive tract. Results From the comparison of the anatomic and histologic characteristics of the digestive tract between EM>Canis lupus/EM> and EM>Canis familiaris/EM> we found that wolves have shorter in total length and lighter in total weight than that of domestic dogs, especially colon. They also possessed much thinner in cardiac glandular region and oesophagus and cardiac mucosa but thicker in fundus gland region, jejunum and colon canal wall and large intestine mucosa. From the comparison of the characters of endocrine cells we could find that, most of their endocrine cells have the similar regional distributions except pancreatic polypeptide (PP) positive cells and substance P (SP) positive cells.Conclusion The comparison between EM>Canis lupus/EM> and EM>Canis familiaris/EM> indicate that different feeding habits have caused several anatomical and histometical changes of their digestive tract, but Related Articles | Metrics

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Pattern influence on the fingers of Hui and Han ethnics in Ningxia 2009, 40 (1):  153-158.  doi: 10.3969/j.issn.0529-1356.2009.01.031 Abstract ( )   Objective To analyze the occurrence and probability of the manifestation of finger pattern types in Han and Hui ethnics from Ningxia. Methods In this study, dermatoglyphic traits in Hui 262(male 129, female 146) and Han 352(male 206, female 146) were analyzed. Results In the entire sample, both Hui and Han were located around the ulnar loop and the whorl. Whereas this arrangement was significantly and typically different in pattern influence cases. When the arch occurred on one of the fingers, this pattern type was also much more frequent on the other nine fingers, whereas whorls were much less common. When there was an ulnar loop on one of the fingers, the frequency of the ulnar loop on the other nine fingers was higher, whereas the frequency of the whorl was lower, than in the entire sample. The frequency of the arch was not influenced by the occurrence of the ulnar loop on other fingers. However, the tendency was the opposite when there was a whorl on one of the fingers.Conclusion The formation and occurrence of pattern types on certain fingers is closely related to the pattern type on the other fingers, which is pattern influence. The pattern influence shows that if a given pattern type is present on one of the fingers, then, in these cases, the frequency of pattern types on the other nine fingers typically is different from that in the entire sample. Pattern influence was found in both Hui and Han nationalities, and in both sexes. Related Articles | Metrics

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Thin transverse sectional anatomy of the sellar region with corresponding MRI 2009, 40 (1):  159-163.  doi: 10.3969/j.issn.0529-1356.2009.01.032 Abstract ( )   Objective To explore the anatomy of the sellar region and its adjacent structures so as to provide intimate morphological data for clinical image diagnosis and surgical operation. Methods Two Chinese adult male cadavers were examined along canthomeatal line with a 15T magnetic resonance(MR) imaging unit by using a spin echo sequence. After being embedded with 5% gelatin and frozen to -40℃ for a week, the specimens were sliced into 0.1mm continuous sections on transverse plane along canthomeatal line from down to up with SKC500 computerized freezing milling machine (milling accuracy of 0.001mm) in a -15℃ laboratory,the slices of the sellar region were as thin as 005mm, subsequently, the serial crosssections were photographed with high-resolution digital camera and imported into an animation computer. Typical slices were selected to investigate important structures with the corresponding MR images. Results Forty and forty-one T1 and T2 MR images, 304 and 310 thin sectional images were obtained separately. Comparing the MRI with thin sectional images, we investigated the sectional anatomy of the sellar region: 1. The internal carotid artery run sinuously in the cavernous sinus and formed some gaps with it. 2. The Ⅲ、Ⅳ、Ⅵ cranial nerves and trigeminal branches ophthalmic nerve, maxillary nerve displayed one by one from anterior to posterior in the lateral wall of cavernous sinus. In the section through the optic chiasm we could observe the stalk of hypophysis crossing the diaphragma sellae and connecting with the pituitary body which could seldom be shown in the thick sections. Conclusion Combination of slices obtained from computerized freezing milling technique and magnetic resonance images offers a better understanding of the complex anatomy structures and provid Related Articles | Metrics

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Comparison of the geometric morphology and compliance of common bile duct between human and pigs 2009, 40 (1):  164-167.  doi: 10.3969/j.issn.0529-1356.2009.01.033 Abstract ( )   Objective To explore the relationship of the geometric morphology and compliance of the common bile duct (CBD) between human and pig and to provide theoretical basis for pigtohuman liver xenotransplantation. Methods The CBDs of 5 adult corpses and 50 Hubei white pigs aged between 3-12 months were obtained. The pressurediameter relationship of the CBD was measured in the biomechanical testing system for soft tissues; the compliance was calculated then. The materials were extracted through transverse frozen sections, and then stained with HE. The diameter and wall thickness of CBD were measured by computer image analysis system. Results The diameter and the wall thickness of CBD of pigs aged between 36 months were significantly shorter or thinner respectively than that of adult human (P<0.01), while the results of pigs aged between 7-10 months are similar to that of the adult human (P>0.05). The compliance of the CBD of pigs aged between 3-6 months and 11-12 months was significantly smaller than that of adult human (P<0.01), while the results of pigs aged between 7-10 months were similar to that of the adult human (P>0.05).Conclusion The geometric morphology and compliance of the CBD of pigs aged between 7-10 months are similar to that of the adult human, suggesting that it is possible for the liver of pigs aged between 7-10 months to be used as the donor for liver xenotransplantati Related Articles | Metrics