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2008, Volume 39 Issue 5
06 October 2008 Previous Issue    Next Issue
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ULTRASTRUCTURE ANALYSIS OF HUMAN NEURAL STEM CELLS/PROGENITORS CULTIVATED EM>IN VITRO/EM> 2008, 39 (5):  615-619.  doi: Abstract ( )   Objective Neural stem cells have become a major concern in the current research of neuroscience, for they are involved in the neural injuries and recoveries, the origin of neural tumors, as well as other fields. The study on their ultrastructures, which is still limited at present, is indispensible. To offer more information is the aim of this paper. Methods Neural stem cells/progenitors from human fetal brain tissue were cultivated EM>in vitro/EM> and observed under a scanning electron microscope (SEM) and a transmission electron microscope (TEM). Results Neural stem cells/progenitors presented to be neurospherelike after days of culture EM>in vitro/EM>. The neruospheres were made up of neural stem cells/progenitors and nonfixiform material inside, and cells in neruospheres could be divided into lucent and dark ones according to electron densities. Between adjacent cells as well as on the cytoplasmic sides of the apposed plasma membranes, there were vesiclelike structures. Cell membrane fusions were also observed between some adjacent cells. Single neural stem cell /progenitor was spherical with rough surface under SEM. Many kinds of organellas, e.g. Golgi’s complex and endocytoplasmic reticulum, were underdeveloped in neural stem cells/progenitors, which generally had big, nuclei and scanty cytoplasm. The numbers, types and maturities of cellular organs in different cells were not always identical, which showed their heterogeneities. For instance, both neurofilaments and microtubules could only be observed in a few neural stem cells/progenitors； lysosomes were very abundant in some, but even hardly founded in others. What’s more, autophagosomes at different stages and in differernt formations could be seen in most cells. The nuclei, frenquently containing huge amounts of euchromatin and a small quantity of heterochromatin, mostly were globular, sometimes reniform or lobulated; most neural stem cells/progenitors had only one chromatospherite, seldom two or more, and sometimes no obvious chromatospherite could be seen. Conclusion Developed autophagosomes, vesiclelike structures between adjacent cells as well as on the cytoplasmic sides of the apposed plasma membranes and cellular membrane fusions could be seen in the human embryooriginated neural stem cells/progenitors and Related Articles | Metrics

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EXPLORATION OF THE MECHANISM FOR THE AGE-RELATED CHANGE OF THE UNMYELINATED NERVE FIBERS IN RAT WHITE MATTER 2008, 39 (5):  620-625.  doi: Abstract ( )   Objective To investigate agerelated changes in rat white matter with the new stereological techniques in order to figure out the exact reason for the decline of myelinated nerve fibers in aged white matter and explore the effect of sex on the agerelated change of brain white matter. Methods Nine adult Long-Evans rats (6-8 months) and 8 aged Long-Evans rats (18 months) were used. The volume density, length density, total volume and total length of unmyelinated fibers in white matter were estimated with the stereological techniques and transmission electron microscope technique. Results In the male and female rats, there was no significant decrease in the total length of unmyelinated fibers in white matter of young and aged rats. There was a significant decrease in the total volume of unmyelinated fibers in white matter of aged female rats compared with that of young female rats. The total volume of unmyelinated fibers in white matter of aged male rats was decreased by 31% compared with that of young male rats even though the difference was not statistically significant. In young and old rats, there was no significant differences of all unmyelinated fiber parameters between male and female rats. Conclusion In aged rat white matter, there was a significant decrease in the unmyelinated fibers with large diameters. Meanwhile, the myelinolysis of the myelinated fibers with small diameters in aged white matter made the age-related decrease of the unmyelinated fibers with small diameters in white matter unnoticeable. Related Articles | Metrics

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DIFFERENT DOMAINS OF α-SYNUCLEIN INVOLVED IN MITOCHONDRIAL OF MN9D CELLS 2008, 39 (5):  626-630.  doi: Abstract ( )   Objective To investigate the interaction of different domains of α-synuclein gene with mitochondrial by over-expressing N-terminal, NAC and C-terminal of α-synuclein in MN9D cell. Methods N-terminal of α-synuclein/1-65 (amino acid, aa), NAC of α-synuclein/61-95aa and C-terminal of α-synuclein/96-140aa were generated by PCR amplification and these cDNA fragments were then subcloned into retroviral pLNCX2 vectors. The reconstructed plasmids were transfected into PT67 packaging cell line by Lipofectin and viral particles were selected. After using the viral particle to infect the MN9D cells, the cell viability was evaluated by Cell Counting Kit-8 assay (CCK-8). Gene expressing level was detected by real Time RT-PCR. The different domains of α-synuclein distribution and co-localization of target protein with mitochondrial were measured by immunofluorescence and the mitochondrial membrane potential was determined by flow cytometry. Results N-terminal of α-synuclein was found to be located together with mitochondrial and to make the membrane potential of mitochondrial decline. α-synuclein/NAC expressed mainly in nuclear while α-synuclein/C was chiefly found in cytoplasm. Conclusion N-terminal of α-synuclein may interact with mitochondrial and interfere its function by depolarizing its membrane potential. α-synuclein Related Articles | Metrics

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THE MORPHOLOGICAL ANALYSIS OF APOPTOSIS IN HIPPOCAMPUS OF POSTTRAUMATIC STRESS DISORDER RATS AND THE EXPRESSION OF APOPTOSISRELATED GENES 2008, 39 (5):  631-635.  doi: Abstract ( )   Objective To investigate relationships between the expression of Bcl-2, Bax protein and neuronal apoptosis in hippocampus of posttraumatic stress disorder (PTSD) rats. Methods The SPSmethod was used to set up the rat PTSD models. There were five groups：after SPS 1d，4d，7d，14d groups and control group. Apoptotic cells were detected by electron microscopy and TUNEL method. The expressions of Bcl-2 and Bax proteins were detected with immunohistochemistry, double fluorescent, confocal laser scanning microscopy and Western blotting. Results Apoptotic cells were present in hippocampus of PTSD rats. The number of apoptotic cells increased with the development of PTSD and peaked on the 7th day after SPS, then decreased gradually. The expression of Bcl-2 protein peaked on the 4th day after SPS and Bax protein peaked on the 7th day after SPS, then decreased gradually. The ratio of Bcl-2/Bax increased at the beginning and then gradually decreased. Conclusion The neuronal apoptosis in hippocampus of PTSD rats may be one of reasons inducing hippocampus atrophy. The increase of Bcl-2 and Bax protein and the change of Bcl-2/Bax ratio would play an important role in regulating the hippocampal neuronal survival Related Articles | Metrics

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EXPRESSION OF FASCICULATION AND ELONGATION PROTEIN ZETA1 IN TYPE-1 AND TYPE-2 ASTROCYTES 2008, 39 (5):  636-639.  doi: Abstract ( )   Objective To observe and analyze the expression of fasciculation and elongation protein zeta-1 (FEZ1) gene in purified type-1 astrocytes (T1A) and type-2 astrocytes (T2A). Methods By the technique of gene-chip, some related genes in the different gene expression profiles of purified T1A and T2A were compared and analyzed. Increased mRNA levels of FEZ1 in T2A was further confirmed by RT-PCR. Results 138 different genes were differentially expressed in T1A and T2A, 60 of which were highly expressed in T1A and 78 in T2A. The expression of FEZ1 was up-regulated in T2A, and the expression of FEZ1 was detected in the different parts (cerebral cortex, olfactory bulb and spinal cord) of the central nervous system of P1 rats.Conclusion FEZ1 gene is among several genes that expressed differently in T1A and T2A. The expression of FEZ1 is found in central nervous system of P1 rats. FEZ1 may play an important role in the development and regeneration of central nervous system. Related Articles | Metrics

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THE PROMOTION OF NEUROGENESIS IN SUBVENTRICULAR ZONE BY BONE MARROW STROMAL CELLS AFTER PERMANENT FOCAL CEREBRAL ISCHEMIA IN RATS 2008, 39 (5):  640-644.  doi: Abstract ( )   Objective To investigate the effect of transplanted bone marrow stromal cells on the cells in the subventricular zone of the rats with permanent focal cerebral ischemia and analyze cell types. Methods Permanent focal cerebral ischemia models were established with middle cerebral artery occlusion (MCAO) and divided into three groups: MCAO alone, intravenous infusion of 1ml PBS at 24 hours after MCAO, and intravenous infusion of 2×10SUP>6/SUP> BMSCs 24 hours after MCAO. Then, the groups were subdivided into 7-day and 14-day groups after MACO. Neurological functions were detected by Zausinger evaluation; meanwhile, 5-bromodeoxyuridine was injected to label the proliferating cells in the subventricular zone, and double-immunofluscent technologies were used to identify the cell type. Results On the 7th day and the 14th day after MACO, neurological functional scores of BMSCstreated group were higher than those of the other two groups (EM>P/EM><0.05). On the 14th day after MCAO, BrdU-positive cells in SVZ of ipsilateral ischemia of BMSCstreated group were more than those in the two controls (EM>P/EM><0.05); double-immunofluorescence label demonstrated that BrdU-positive were collocated with markers of neuron and astrocyte. Conclusion BMSCs can promote the neurological function of the rats with permanent focal cerebral ischemia and this action may be related with the neurogenesis in the subventric Related Articles | Metrics

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BARREL CORTEX CHANGES IN YOUNGER AND ADULT SPRAGUEDAWLEY RATS 2008, 39 (5):  645-648.  doi: Abstract ( )   Objective To study the development and changes of the Barrel Cortex in younger and adult SD rats. Methods Cytochrome oxydase histochemistry and Nissl’s counterstaining were applied to locate the encephalic region of Barrel in adult rats, and Barrel cortex changes in different ages were analyzed(postnatal 20 days and postnatal 3 months). Results During the development of Barrel cortex, an obvious area change was found（EM>P/EM><0.01）. The space between Barrel rows shrinked and the number of neurons in septa decreased. Compared with that of younger Sprague-Dawley rats, the Barrel cortex of adult Sprague-Dawley rats had more cytochrome oxydase highly active neurons（EM>P/EM><0.01）.Conclusion Subtle changes exist during the development in the fine assignment and structure o Related Articles | Metrics

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DISTRIBUTION AND EFFECTS OF PERIPHERAL NERVE AXOLEMMA ION CHANNELS IN EXPERIMENTAL ALLERGIC NEURITIS 2008, 39 (5):  649-655.  doi: Abstract ( )   Objective To reveal the distribution and effects of peripheral nerve axolemma ion channels in experimental allergic neuritis (EAN). Methods The alteration of Na+ and K+ channels on peripheral nerves (PNs) in the course of EAN was observed and the relationship between the channels and nerve conduction properties was analyzed by assessing histology and electrophysiology of PNs as well as clinical severity. Results Na+ and K+ channels obviously decreased on day 9 post immunization (p.i.), a time point of disease onset, and quickly became undetectable in next two weeks. Undergoing a slow and incomplete regeneration, neither of the channels regained the normal appearance on day 85 p.i. even if the clinical symptom disappeared several weeks before. Na+ and K+ channels had a synchronous development during disease course and remained a close correlation with the alteration of paranodal myelin. Electrophysiological abnormality kept consistent with the disturbance of PNs just in the acute period of EAN (923d p.i.) and the compound muscle action potential (CMAP) amplitude partly reflected the distribution of axolemma ion channels. Conclusion Loss of axolemma ion channels of PNs might be one of the reasons directly leading to the early symptoms of EAN. As a structural component, the channels were liable to damage and difficult to restore. The clustering and maint Related Articles | Metrics

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EFFECT OF EphB/Ephrin-B1 SIGNAL ON THE CURRENTS OF α7-nAChRs ON CILIARY GANGLION NEURONS 2008, 39 (5):  656-660.  doi: Abstract ( )   Objective Recent researches showed that Eph/Ephrin tyrosine kinase family plays an important role in the development and functional maintanence of nervous system, but it is unclear if it affects sympathetic nervous system.The study is to detect the effect of EphB/Ephrin-B1 signal on the currents of nicotinic acetylcholine receptor (α7-nAChRs) on ciliary ganglion neurons. Methods Currents of α7-nAChRs on cultured and acute-associated ciliary ganglion neurons were recorded respectively. Cells were randomly divided into four groups: the control group, the EphrinB1Fc treated group (cells stimulated with different concentrations of Ephrin-B1Fc which is a recombination of IgG and Ephrin-B1), IgG treated group (cells stimulated with IgG at the same concentration of IgG needed for Ephrin-B1Fc recombination), Ephrin-B1 treated group (cells stimulated with Ephrin-B1 at the same concentration of Ephrin-B1 needed for Ephrin-B1Fc recombination). Results There was no difference between the control, IgG and Ephrin-B1 groups, but Ephrin-B1Fc could inhibit α7-nAChRs currents of both cultured and acute-associated CG neurons in a concentration-dependent manner. The inhibition happened quickly and was reversible. ICSUB>50/SUB> of ISUB>peak/SUB>/CSUB>m/SUB> of cultured and acuteassociated cells was 0.032mg/L and 1.4mg/L respectively. Conclusion The results showed that EphB/Ephrin-B1 signal can affect currents of CG α7-nAChRs. Is suggests that Related Articles | Metrics

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THE SILENCE OF SHORT HAIRPIN RNAs INDUCED Smad2 IN NIH/3T3 FIBROBLAST CELLS 2008, 39 (5):  661-665.  doi: Abstract ( )   Objective To construct five shRNA-expression plasmids and to investigate the expression of EM>Smad2/EM> in TGFβ/ Smads signal transduction treated with shRNA-expression plasmid. Methods Five shRNA-EM>Smad2/EM> DNA sequences from mRNA sequence of mouse EM>Smad2/EM> gene were designed and synthesized. DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively. Five shRNAEM>Smad2/EM> expression plasmids were obtained and then transfected into NIH/3T3 cells. The suppressed expression of EM>Smad2/EM> was assessed by RT-PCR and Westernblotting. Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of EM>Smad2/EM>. The suppression effect of the RNAipool composed of four different plasmids was more obvious than that of any single. Conclusion The shRNAexpression plasmids were successfully constructed, which could specifically and effectively suppress the expression of EM>Smad2 Related Articles | Metrics

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THE RELATIONSHIP OF THE PROLIFERATION AND PHENOTYPIC MODULATION OF THE STROMAL CELLS AND THE EXPRESSION OF ESTROGEN RECEPTOR α IN BENIGN PROSTATIC HYPERPLASIA 2008, 39 (5):  666-669.  doi: Abstract ( )   Objective To investigate the different expression patterns of the phenotypic marker protein for stromal cell, proliferating cell nuclear antigen(PCNA) and estrogen receptor α(ERα) in normal human prostate (NP) and benign prostatic hyperplasia (BPH) tissue, and to locate cells stained positively with the marker protein for stromal cell and PCNA or ERα at the same position. Methods Serial sections of 4 NP and 8 BPH specimens were subjected to immunohistochemistry (IHC) staining for vimentin, α-smooth muscle actin (α-SMA), myosin, PCNA and ERα. Results Compared with that in the NP tissues, the number of α-SMA positive cells increased significantly in BPH tissue, and the number of vimentin positive cells increased moderately in the stroma and prominently surrounding the acinus and in the basal layer. In BPH tissue, the myosin and ERα staining signal was lost in the stromal cells surrounding the acinus, and the positive staining cells gathered into bunches in the stroma far away from the acinus, while the positive cells were sporadically distributed in the NP tissue. The PCNA positive cells increased moderately in the stroma and increased significantly in the basal layer. The serial sections staining results showed that there was a colocation of PCNA, vimentin and ERα in basal cells, and a colocation of myosin and ERα in stromal cells. Conclusion There are apparent phenotypic modulations of the stromal cells in the BPH specimen co Related Articles | Metrics

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THE EFFECTS OF COMBINATION OF CHINESE MEDICINE EFFECTIVE CONSTITUENTS ON CONFIGURATION AND COMPONENTS OF NUCLEAR MATRIX OF THE OSTEOSARCOMA MG-63 CELLS 2008, 39 (5):  670-676.  doi: Abstract ( )   Objective To explore the alteration of nuclear matrix proteins and the configurational changes of nuclear matrix (NM) system during differentiation of the osteosarcoma MG-63 cells EM>in vitro/EM> after being induced by the combination of ginsenoside Rg1, cinnamic acid and tanshinoneⅡA (RCT). Methods Cell cycle was investigated by flow cytometry and the growth curve was drawn with MTT method. The cells treated with or without RCT were subjected to selective extraction method and prepared for whole mount electron microscopy observation. The nuclear matrixintermediate samples were examined under light and transmission electron microscopes respectively. The nuclear matrix proteins were subjected to analysis of proteomic technique. Results The proliferstion of the MG-63 cells treated with RCT were repressed markedly; the rate of repression was 72.37％ on the seventh day，and the cell cycle was blocked at G0/G1 phase. The typical malignant phenotype of nuclear matrix of MG-63 tumor cell were reversed to normal cells. The expression of many functional proteins of nuclear matrix were changed in the MG-63 cells after the treatment with RCT. Conclusion RCT can repress markedly the proliferation of MG-63 cells, block cell cycle, induce a restorational change to the nuclear matrix architecture similar to that of normal cells, and change the const Related Articles | Metrics

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A STUDY ON THE CORRELATION BETWEEN THE CHANGES OF AQUAPORIN4 EXPRESSION AND INJURIES IN RAT RETINA UNDER DIFFERENT ACUTE OCULAR HYPERTENSION 2008, 39 (5):  677-682.  doi: Abstract ( )   Objective To investigate the changes of aquaporin 4(AQP4) and its mRNA expression in rat retina under different acute ocular hypertension ( AOH )，so as to study the role of AQP4 in some diseases related with the alteration of ocular pressure. Methods The AOH rat models were established by perfusing isotonic salt solution into the anterior chamber of rat eyes through a connected reservoir, which was situated at 75cm,125cm and 150cm heights to produce appropriate intraocular pressure. HE staining was performed to check the thickness and morphological changes of the retinal inner layers, and Nissl staining was used to count the number of ganglion cells in the ganglion cells layer (GCL). The changse of AQP4 and its mRNA expression in rat retina under different AOH were detected by immunofluorescence staining and confocal microscope，in situ hybridization and RTPCR respectively. Results The retina was swelling and the thickness of retina inner layer was improved under different AOH, and the number of ganglion cells in GCL was decreased after the perfusion stopped. The expressions of AQP4 and its mRNA in rat retina under different AOH were significantly elevated compared with the normal control groups. This upregulation expression had a strong correlation with the thickness changes of retinal inner layer and the decrease in the cell number of the GCL under different AOH. Conclusion The upregulation of AQP4 and its mRNA expression in rat retina was correlated with the increased intraocular pressure. The results indicate that AQP4 may play an important role in the pathophysiol Related Articles | Metrics

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STUDY ON POLYMORPHISMS OF 15 STR LOCI IN MIAO ETHNIC GROUP OF RONGSHUI COUNTY IN GUANGXI PROVINCE 2008, 39 (5):  683-687.  doi: Abstract ( )   P>Objective To investigate the distribution patterns of 15 short tandem repeats (STR) loci (TPOX，TH01，CSF1PO，D19S433，vWA，D18S51，D5S818，FGA，D8S1179，D21S11，D7S820，D3S1358，D13S317，D16S539，D2S1338 ) in Miao ethnic group of Rongshui County in Guangxi Province. Methods The sodium citratedblood specimens were collected from 208 healthy unrelated Miao individuals (man: 102, female: 106) in Rongshui County in Guangxi Province, and then the DNAs from the samples were extracted by phynolchloroform technique. The DNAs were amplified by using AmpFlSTR IdentifilerTM PCR Amplification Kit, and finally the data were detected with 3100 Genetic Analyzer. Result Altogether 5.20 alleles and 11.62 genotypes of 15 STR were found in 208 healthy unrelated Miao individuals of Rongshui County in Guangxi Province. The allele frequency and genotype frequency were 0.004 8-0.466 3 and 0.004 8-0.317 3 respectively. Total discrimination power wa Related Articles | Metrics

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THE DISTRIBUTION OF CD2ASSOCIATED PROTEIN IN RENAL CELLS AND ITS POSSIBLE ROLE IN PODOCYTES 2008, 39 (5):  688-692.  doi: Abstract ( )   Objective To study the distribution of CD2-associated protein (CD2AP) in normal renal cell lines and its interaction with nephrin and F-actin in podocytes. Methods The human mesangial cells (HMC) and HK-2 were cultured in DMEM. Conditionally immortalized murine podocyte cell line was cultured in RPMI 1640 The expressions of CD2AP and nephrin in podocytes were examined by RT-PCR and Western blotting. The distribution of CD2AP in HMC, HK-2, differentiated and undifferentiated podocytes was observed by laser scanning confocal microscopy. The coexistence of CD2AP with nephrin and F-actin in undifferentiated podocytes were also detected. Results CD2AP was distributed within the cytoplasm and perinulcear region of HK-2 and undifferentiated podocytes, but was absent in HMC cells. Its distribution profile changed and presented as peripheral accumulation when podocytes were put into differentiationpermissive conditions. CD2AP was located together with nephrin and F-actin in podocytes. Conclusion CD2AP can be detected in epithelial-originated renal cells. The alteration of distribution profile of CD2AP indicates it may participate in the process of podocytes differentiation and be involved in the regulation of slit diaphragm and cytoskeleton. Related Articles | Metrics

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INHIBITION OF GLUCOCORTICOID ON ORNITHINE DECARBOXYLASE DURING RAT LIVER REGENERATION 2008, 39 (5):  693-698.  doi: Abstract ( )   Objective The regulation of ornithine decarboxylase (ODC) gene expression and enzyme activity by corticosterone, the main glucocorticoid in rat, during rat liver regeneration induced by partial hepatectomy (PH) was evaluated. Methods Bilateral adrenalectomies (ADX) and sham-ADX were performed on ether-anesthetized rats 3 days before PH. Corticosterone in sesame oil was injected subcutaneously to adrenalectomied rats. ODC mRNA, ODC protein and enzyme activity were detected by RT-PCR, Western blotting and high performance liquid chromatography (HPLC), respectively. Results The ODC mRNA levels, protein accumulation and enzyme activity were lower in the intact liver compared to the regenerating liver. After PH, mRNA levels were remarkably enhanced in all groups (EM>n/EM>=6 in each group) and peaked at 5 hours postPH. Till 7 hours, the contents in all groups from high to low were ADX group,control group (Sham-ADX group), ADX treated with 10mg/kg and 40mg/kg body weight corticosterone group, respectively. ODC protein accumulation in ADX rats was higher than that in control rats (EM>n/EM>=13, the same below), but it decreased in corticosteronetreated (10mg/kg) rats until 24 hours post-PH, with a strong decline seen in 40mg/kg corticosteronetreated rats. ODC activity was rapidly promoted, and the highest levels were observed at 6 hours after PH in all groups (EM>n/EM>=6 in each group). After corticosterone treatment, the activities declined significantly at 6 hours post-PH, with the lowest value found in the 40mg/kg group. Conclusion Corticosterone treatment results in dose-dependent decreases in ODC mRNA and enzyme protein both in the intact liver and the regenerating liver. The change in ODC activity is partia Related Articles | Metrics

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ESTABLISHMENT OF TRANSGENIC MICE EXPRESSING SV40T IN PARIETAL CELLS 2008, 39 (5):  699-702.  doi: Abstract ( )   Objective SV40 T antigen had been used widely in establishment of transgenic tumor animal models, which could induce cellular immortalization and carcinogenesis. The objective of the study was to construct the eukaryotic specific expression vector of SV40 large T antigen in gastric parietal cells and to establish transgenic mouse models with microinjection technique. Methods 38kb DNA fragment HSUP>+/SUP>KSUP>+/SUP>ATPase β promoter/SV40T was released from specific expression vector of SV40 large T antigen in gastric parietal cells pcDNA31(-)/HKSV and injected into oocytes to establish transgenic mice. Transgenic positive mice were detected by PCR and Southern blotting. Gene expression was measured by RT-PCR. Results 422 injected eggs were transferred to 16 pseudopregnant female mice and 77 offspring were born, the frequency of transplant was 18.2%. In 77 liveborn mice, 2SUP># /SUP>, 4SUP>#/SUP> , 8SUP>#/SUP> , 16SUP>#/SUP> , 24SUP>#/SUP> , 51SUP>#/SUP> , 57SUP>#/SUP> , 61# , 68# , 73# were proved to be positive founder mice by PCR. 68# founder was sterile, and 99 F1 mice were borne by the other nine founders. None F1 positive mouse was found in 8# offspring, 31 F1 mice were proved to be positive by PCR and Southern blotting in the other eight pedigrees. The rate of positive mice was Related Articles | Metrics

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CORRELATION OF ZAC WITH SOMATOSTATIN AND ITS RECEPTOR EXPRESSION IN GASTRIC CANCER TISSUES 2008, 39 (5):  703-707.  doi: Abstract ( )   Objective To study correlation of zinc finger protein which regulates apoptosis and cell cycle arrest(ZAC) with somatostatin and its receptor expression in gastric cancer tissues, and explore its relationship with clinicopathological factors of gastric cancer. Methods Immunohistochemistry and RT-PCR techniques were used to detect expression of ZAC, somatostatin and its receptor in 30 cases of gastric cancer, adjacent cancer and normal gastric mucosa tissues. Results The positive rate of ZAC in gastric cancer, adjacent cancer and normal gastric mucosa tissues was 33.3%(10/30)、73.3%(22/30)and 66.7%(20/30) respectively; that of SST was 36.7％(11/30)、53.3％(16/30) and 73.3％(22/30) respectively. The positive rates of both ZAC and SST in gastric cancer tissues were significantly lower than those in normal gastric mucosa tissues. Five SSTR subtypes were expressed in gastric cancer, adjacent cancer and normal gastric mucosa tissues. Compared with normal gastric mucosa tissues, positive rate of SSTR3 decreased significantly, and those of SSTR2 and SSTR5 increased significantly in gastric cancer tissues. In gastric cancer tissues, the expression tendency of ZAC was consistent with those of SST and SSTR3 Co-expression of ZAC and SST was obviously related to differentiation type and infiltration. However, no relation was found between co-expression of ZAC and SST to lymph node metastasis. Co-expression of ZAC and SSTR3 was significantly related to differentiation type of gastric cancer. Conclusion ZAC might be a downstream gene of somatostat in in inhibition of development of gastric cancer. Somatostatin effects on ZAC expression via binding with SSTR3 to inhi Related Articles | Metrics

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THE RELATIONSHIP BETWEEN THE INSTABILITY OF EM>PINX1/EM> GENE AND GASTRIC CARCINOMA 2008, 39 (5):  708-712.  doi: Abstract ( )   Objective To examine loss of heterozygosity (LOH) and microsatellite instability (MSI) of locus D8S277 on chromosome 8 and their influence on the expression of PINX1 in the gastric carcinoma,which may provide an experimental basis for the mechanism of PINX1 gene and tumor development. Methods DNA was extracted from formalinfixed paraffinembedded tissues. Polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and ordinary silver staining were used to study LOH and MSI of locus D8S277. Envision immunohistochemistry and LeicaQwin computer imaging techniques were used to assess the expression of PINX1. Results The frequency of LOH was significantly higher in the cases with lymph node metastasis than in those without metastasis (21.15% vs 0, P<0.05),and was significantly higher in the cases at TNM stage Ⅲ＋Ⅳ than in those at stage Ⅰ＋Ⅱ(24.39% vs 3.33%, P<0.05). The positive rate of PINX1 protein was significantly higher in the cases without lymph node metastasis than in those with lymph node metastasis(78.95% vs 48.08%, P<0.05);and was significantly higher in the cases at TNM stage Ⅰ＋Ⅱ than those at stage Ⅲ＋Ⅳ(73.33% vs 43.90%, P< Related Articles | Metrics

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INFLUENCE OF HAISHENGSU ON CELL GROWTH AND EXPRESSION OF APOPTOSIS GENE IN K562 CELLS 2008, 39 (5):  713-716.  doi: Abstract ( )   Objective To study the influence of Haishengsu on cell growth and the expression of apoptosis gene in leukemia K562 cells by Haishengsu. Methods K562 cells were cultured in RPMI1640 containing 10% fetal bovine serum at 37℃ in a humidified atmosphere with 5% CO2. The experiment was performed on three groups: the control group and two Haishengsu groups (10mg/L and 20mg/L).The cell cycle, growth and the expression of Bcl-2 and bax were evaluated respectively by means of flow cytometer(FCM), thiazoyl blue tetrazolium bromide(MTT)assay and immunocytochemistry to determine Haishengsu’s influence. Results Both the Haishengsu (10mg/L and 20mg/L), restrained cell cycle in G1/S phase. The 20mg/L inhibited the growth of K562 cells and the survival of cells declined along with the time in this concentration. It was observed under light microscope that the number of positive cells increased in the two Haishengsu groups(20mg/L and 40mg/L).It was analyzed by image analysis instrument that the expression of Bcl-2 proportion decreased to (58.78±4.65)% and (26.57±2.13)% respectively in the Haishengsu groups (20mg/L and 40mg/L) from (90.98±8.66)% in the control group and the expression of bax proportion increased to (77.69±3.55)% and (90.60±3.73)% respectively in the Haishengsu groups (20mg/L and 40mg/L) from (10.88±6.57)% in the control group.Haishengsu (20mg/L and 40mg/L) could downregulate the expression of Bcl-2 and upregulate the expression of bax in K562 cells. Conclusion Haishengsu could significally restrain the cell cycle in G1/S phase and inhibit the growth of K562 cells. Haishengsu could reduce the expression of Bcl-2 and increase th Related Articles | Metrics

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SIRNA FOR SNAIL REFRAINS FROM MIGRATING OF MESENCHYMAL STEM CELLS TRIGGERED BY TRAMSFORMING GROWTH FACTOR β1 2008, 39 (5):  717-722.  doi: Abstract ( )   Objective To assess the effect of transforming growth factorβ1(TGF-β1) on cell proliferation，cell migration and Snail，MMP-2 expression of mesenchymal stem cells(MSCs)，and to further investigate its mechanisms. Methods Density gradient centrifugalization combined with adherence method were used to segregate and cultivate rat MSCs.MSCs cultivated to the 3rd passage were characterized using immunofluorescence technique.The effects of different concentrations of TGF-β1 on the migration was detected by the modified Transwell chambers.The effects of TGF-β1 at different time points on the expression of Snail and MMP-2 mRNA were measured by RT-PCR，and Snail expression was detected by immunofluorescence technology.siRNA for Snail was synthesized and transfected into MSCs by liposomel before TGF-β1 were treated.The expression of Snail and MMP-2 before and after the transfection were measured by Western blotting. Results Density gradient centrifugalization combined with adherence method could segregate and purify rat MSCs effectively.The results of immunofluorescence staining showed that CD29 and CD44 expressions were positive while CD34 and CD45 expression was negative for MSCs.The TGF-β1 induced a dose-dependent increase in cell migration,which peaked at 2μg/L.The expression level of Snail mRNA with TGF-β1 treatment for 6 hours and the expression level of MMP-2 mRNA for 12 hours were significantly higher than the expression level of those at other time points.24 hours later,the expression of Snail in TGF-β1treated group was significantly higher than t Related Articles | Metrics

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EXPRESSION OF BONE MORPHOGENETIC PROTEIN 4 DURING THE HUMAN EMBRYONIC YOLK SAC HEMATOPOIESIS 2008, 39 (5):  723-727.  doi: Abstract ( )   Objective To further explore the mechanism by which bone morphogenetic protein 4（BMP-4）might be involved in hematopoietic differentiation of the yolk sac. We observed the expression of BMP-4,CD34, CD133 and tyrosine kinase receptors（KDR） in the blood island of the yolk sac at embryonic 3 to 8 weeks. Methods Gene expression was analyzed by RT-PCR and the presence of BMP-4, CD34, CD133 and KDR proteins was confirmed by immunohistochemistry in 57 human embryos. Results In the human yolk sac, we found that BMP-4 was expressed at high levels from the 16th day to the 7th week, and decreased quickly after week 7 The results showed that KDR, CD133 and CD34 largely appeared on the 21st and 30th day, then increased at the 6th week, and decreased quickly after week 7. Furthermore, Ihh，SCl, GATA-1, GATA-2 and PU.1 mRNAs showed that PU.1 was not expressed on the 16th day; however, other factors were expressed all the time. Conclusion The distribution of BMP-4,KDR,CD34,CD133 and transcription factors expression highly suggested that BMP-4 was secreted from the yolk sac which might exert its effects on the specification of human hemangioblast and hematopoietic ste Related Articles | Metrics

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EFFECT OF DIFFERENT CONCENTRATION OF TRICHOSTATIN A ON CHICKEN WING BUD DEVELOPMENT 2008, 39 (5):  728-733.  doi: Abstract ( )   P>Objective To investigate the reaction of the limbs treated with different concentration of Trichostatin A（75μmol/L，750 μmol/L，1.5mmol/L），a histone deacetylase (HDAC)inhibitor. This may be useful to improve our understanding of the role of chromatin remodelling and epigenetic control of gene expression patterns and ultimately the development of drugs against cancer. Methods Using the chicken embryonic limb as an experimental model. The embryos received grafts of TSA soaked beads or PBS control beads into the right forelimb buds. Then they were submitted to EM>in situ/EM> hybridization with probes and apoptosis test. Results TSA could modulate the expression of some myogenesis related genes, MyoD and Myf5 during chicken myogenesis. Using apoptosis staining methods, there was no significant apoptosis in the TSA (75μmol/L) treated embryos. However the induction of morphological changes and apoptosis at specific stage was possible with high concentration of TSA. Conclusion TSA (75μmol/L) regulates certain important transcriptional targets and strongly control muscle differentiation. Increasing th Related Articles | Metrics

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INFLUENCE OF QUERCETIN AND BORNYL ACETATE ON CD4SUP>+ /SUP>/CD8SUP>+/SUP>T LYMPHOCYTES IN THE UTERUS FOLLOWING LPSINDUCED ABORTION IN PREGNANT MICE 2008, 39 (5):  734-737.  doi: Abstract ( )   Objective To investigate the significance of CD4SUP>+ /SUP>and CD8SUP>+/SUP> T lymphocytes in the uterus in early embryo loss (or resorption), and to elucidate the anti-bortive effect and the immunological modulation of the maternalfetal interaction with quercetin and bornyl acetate. Methods Lipopolysaccharide (LPS)（0.10μg/mouse）was injected via the tail vein in order to induce abortion in 7-day-gestation mice which also received quercetin and bornyl acetate at days 4-7 of gestation. Uterine CD4SUP>+/SUP> and CD8SUP>+/SUP> T lymphocytes of each group (EM>n/EM>=10) were detected by immunohistochemistry. Results The amount of CD4EM>SUP>+ /SUP>/EM>T lymphocytes in the uterus of LPSinduced abortion mice was much higher (EM>P/EM><0.01) than that of the control mice, but there was no significant difference in the amount of CD4SUP>+ /SUP>T lymphocytes (EM>P/EM>>0.05). Also, the rate of CD4SUP>+/SUP> /CD8SUP>+/SUP> was increased significantly (P<0.01). When quercetin and bornyl acetate was used to prevent LPSinduced abortion, fewer CD4SUP>+/SUP> T cells were counted. The effect of quercetin combined with bornyl on antiLPSinduced abortion was more significant, and the rate of CD4SUP>+ /SUP>/ Related Articles | Metrics

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COPULATION ABSTINENCE DOWNREGULATED ESTRADIOL LEVEL AND ERα EXPRESSION IN MAMMARY GLAND TISSUE OF ADULT FEMALE RATS 2008, 39 (5):  738-741.  doi: Abstract ( )   Objective To investigate the effects of copulation abstinence on estradiol levels and estrogen receptor-α(ERα) expression in the mammary gland of adult female rats. Methods Twelve 8-week-old female SpragueDawley (SD) rats were randomly divided into two groups: control(copulation) and copulation abstinence. Histological changes of the mammary gland tissue samples was assessed by light and electron microscopic methods and ERα protein was detected by immunohistochemistry and Western blotting analysis. The estradiol levels in serum and mammary gland tissue were measured by radioimmunoassay. Results After a 60-day abstinence period, mammary estradiol levels were reduced (EM>P/EM><0.05). Moreover, ERα protein in the mammary gland tissue was downregulated (EM>P/EM><0.05). No morphological changes was observed in the mammary tissue sections. Conclusion Our results indicate sexual abstinence induced dec Related Articles | Metrics

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CAPABILITY OF MATURATION，FERTILIZATION AND FURTHER DEVELOPMENT OF OOCYTES RETRIEVED FROM RABBIT OVARIAN TISSUE TREATED WITH CRYOPRESERVATION AND AUTOLOGOUS TRANSPLANTATION 2008, 39 (5):  742-746.  doi: Abstract ( )   Objective To investigate the effects of transplantation of frozenthawed ovarian tissues on the maturation, fertilization and further developmental potency of oocytes retrieved from grafts. Methods Twenty five New Zealand white female rabbits were divided into three groups randomly, group 1(EM>n/EM>=5), control group; group 2 (EM>n/EM>=10), fresh ovarian tissues were autologously transplanted into the mesometrium; and group 3(EM>n/EM>=10), frozenthawed ovarian tissues were autologously transplanted into the mesometrium. Three months after the transplantation, rabbits were stimulated with folliclestimulating hormone and oocytes were retrieved from antral follicles 13 hours after human chorionic gonadotropin injection. In vitro maturation (IVM) and intracytoplasmic sperm injection (ICSI) were performed to evaluate the fertility potency of the oocytes from frozen ovarian grafts. Results The number of retrieved oocytes in group 2 and 3 were lower than those of the control group (P<0.05); But no significant differences were observed between group 2 and group 3 (EM>P/EM>>0.05); There were no significant differences both in the percentage of immature oocytes and the maturation rate after IVM, among the 3 groups (EM>P/EM>>0.05); Also, among the 3 groups or in each group, the fertilization rate, cleavage rate and blastocyst formation rate showed no difference, no matter the oocytes matured EM>in vivo/EM> or EM>in vitro/EM> (EM>P/EM>>0.05); The blastocyst formation rate derived from oocytes that matured in vitro was significantly lower than oocytes that matured EM>in vivo/EM> (EM>P/EM><0.05). Conclusion Related Articles | Metrics

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COMPARISON OF ESTRADIOL RECEPTOR β EXPRESSION IN THE OVARY OF MICE AND PIGS 2008, 39 (5):  747-750.  doi: Abstract ( )   Objective To study the diversities of expression of estradiol receptor β（ERβ）in the ovary of mice and pigs. Methods Immunohistochemistry was used in the present study in order to detect the expression and distribution of estradiol receptor β in tissue section taken from both the mouse and pig. Results In the ovary of mouse, there was a higher intensity of immunohistochemical staining for ERβ in the corpora lutea and the cytoplasm of oocytes in all sizes of growth follicles, and a weaker intensity of immunohistochemical staining for ERβ in the granulose cells and thecal cells in all sizes of follicle. In addition, there was also a weaker intensity of immunohistochemical staining for ERβ in the interstitial cells and germinal epithelium. In the ovary of pig, there were highly intensity of immunohistochemical staining of ERβ in the thecal cells of follicles. Staining was also detected in the granulose cells in the follicles before the formation of an antrum, but some of the granulose cells in the antrum follicle expressed ERβ, at the same time the others did not expressed it. There was no obvious expression in the interstitial cells or germinal epithelium. Conclusion There are great differences in the expression and distr Related Articles | Metrics

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IMMUNOHISTOCHEMICAL STUDIES ON ENDOCRINE CELLS IN THE DIGESTIVE TRACT OF EREMIAS ARGUS 2008, 39 (5):  751-755.  doi: Abstract ( )   Objective To study the localization and morphology of 5hydroxytryptamine(5-HT),somatostatin(SS),gastrin(Gas),glucagon(Glu),substance P(SP), pancreatic polypeptide(PP) endocrine cells in the digestive tract of Eremias argus. Methods The ABC (avidinbiotin compex method) immunohistochemical method was used. Results 5-HT cells distributed more widely than the other five kinds of endocrine cells in the digestive tract, and they were observed throughout the digestive tract, from the esophagus to the rectum. In addition, the density of 5HT cells was the highest in the jejunum. SS cells were not found in the esophagus or the rectum. The distributive density of SS cells was the highest in the stomach. Gas cells and PP cells distributed in the pyloricus and the small intestine with the highest density in the duodenum. Glu cells distributed mainly in the pyloricus，duodenum and jejunum, the distributive density in the pyloricus was higher than other part obviously. We did not find any SP cell in the digestive tract of Eremias argus. Conclusion The five kinds of endocrine cells which we studied in this paper were mainly round and shuttledshape, which widely lied between epithelial cells, between glandular epithelial cells and at the b Related Articles | Metrics

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EXPRESSIONS AND CLINICAL SIGNIFICANCE OF THYROID STIMULATING HORMONE RECEPTOR AND NaSUP>+/SUP>ISUP>-/SUP> SYMPORTER IN HUMAN THYROID CARCINOMA 2008, 39 (5):  756-759.  doi: Abstract ( )   Objective To investigate the expressions of thyroid stimulating hormone receptor (TSHR) and NaSUP>+/SUP>ISUP>-/SUP> symporter (NIS) in thyroid carcinoma and their role in the process of treatment of thyroid carcinoma. Methods The expressions of TSHR and NIS in 45 normal thyroid tissues, 45 papillary thyroid carcinoma (PTC) tissues and 21 anaplastic thyroid carcinoma (ATC) tissues were detected by immunohistochemical SABC technique respectively. Results The expressions of TSHR and NIS in normal thyroid tissues were higher than those in PTC tissues and ATC tissues. TSHR was all expressed on the membrane of follicular epithelial cells of normal thyroid tissues, which were all expressed in the cytoplasm of ATC cells. In PTC the percentage of expressions in cytomembrane and cytoplasm were 71.1% and 15.6% respectively. NIS was all expressed on the membrane of follicular epithelial cells of normal thyroid tissues, which were all expressed in the cytoplasm of PTC cells and ATC cells. Conclusion The expression location and the expression level of TSHR and NIS are closely related with the differentiation degrees of thyroid carcinoma, which may become the effect indication of TSH suppression therapy and radioiodin Related Articles | Metrics

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SECTIONAL AND IMAGING ANATOMY OF THE RIGHT HEMILIVER INTRAHEPATIC PORTAL VEIN 2008, 39 (5):  760-764.  doi: Abstract ( )   Objective The purpose of this study was to evaluate the course, distribution characteristic of the right hemiliver intrahepatic portal vein. We also investigated the relationships between hepatic veins and portal hepatic fissures, including sectional anatomic data for the division of hepatic segments. Methods Using 30 successive sections of the upper abdomen of the adult cadavers (10 transverse, 20 coronal) and multi-slice computed tomography enhanced images and threedimensional images of 20 patients, we investigated the branch pattern and common variation of right hemiliver intrahepatic portal veins and the relationship between the hepatic veins and intrahepatic portal veins. Results The intrahepatic portal vein of the right hemiliver can be divided into the anteriorsuperior and posteriorinferior group branches in all 50 specimens and images. In 38 (76%) of the 50 specimen and CT images, the branches of the right anterior portal vein supplied the area that was posterior to the right hepatic vein. In 20 (40%) of 50 specimen and CT images the first branch arising from the initial part of the right posterior portal vein towards the caudal supply the part area where anterior to the right hepatic vein. The bloodsupply to the area of the right anterior portal vein expands to the left of the middle hepatic vein, especially in 15 patients whose right portal vein trunks was absent. There was no obvious transverse fissure in the right anterior lobe of liver. Further subdivision of the portal segments showed marked individual variability, with no prevailing branching pattern. Conclusion The right hemiliver could be divided into right anteriorsuperior and right posteriorinferior lobes, and there was a curved fissure between them. There was a constant longitudinal fissure in the right anteriorsuperior lobe of liver. In addition, the right hepatic vein was not an accurate indicator of the position of the right interlobar fissure especially in the superior part and inferior part. The middle hepatic vein was not an accurat Related Articles | Metrics

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RELATIONSHIP BETWEEN DIGIT RATIO AND CORONARY HEART DISEASE 2008, 39 (5):  765-768.  doi: Abstract ( )   Objective To investigate the relationship between the digit ratio and coronary heart disease. Methods Use anthropometry, digit length of both hands in each person was measured. Digit ratios were counted and compared between right hands and left hands, for coronary heart disease patients and controls respectively. Results (1) Our results showed the trend as 2D∶3D<2D∶4D<3D∶4D<2D∶5D<4D∶5D<3D∶5D in both groups. (2)The mean values for digit ratio was higher in the patient group than that of the control group. Significant differences in digit ratio for 2D∶3D (left, EM>P/EM><0.05) and 2D∶4D(left, EM>P/EM><0.01: right, EM>P/EM><0.05) were abserved between patients and controls. (3) We have shown a negative relation between the digit ratio and the onset age of coronary heart disease in Ningxia Han males(EM>P/EM><0.001). Conclusion Digit rati Related Articles | Metrics

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SOMATOTYPE OF GANSU YUGU ADOLESCENTS BY THE HEATHCARTER METHOD 2008, 39 (5):  769-772.  doi: Abstract ( )   Objective To study the characteristics and regularities of somatotype growth of Yugu adolescents. Methods The somatotype growth of 989 Yugu adolescents（male:512,female:477）in Sunan was evaluated by the Heath-Carter method. Results The average somatotype of YuGu adolescents in males was mesomorphic ectomorph (3.0-3.6-3.7), and in females, the average somatotype was ectomorphic endomorph (3.8-2.9-3.6). The somatotypes develop from central,endomorphic ectomorph to mesomorphic ectomorph in the male, however, in the female from central, ectomorphic endomorph,endomorphic ectomorph, to mesomorphic endomorph.Conclusion The somatotypes of Yugu adolescents are very different between males and females. In the male group, the somatotypes of the 7-12 yearold group of Yugu adolescents are similar to the Mongolia，Han ethnic,Zhuang ethnic and Hungary. The somatotypes of 13-17 year-old group are similar to Tibetan，Zhuang ethnic,Han ethnic and Daur. However for the female group ,the somatotypes of the 79 yearold group are simila Related Articles | Metrics

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THE EXPRESSION OF HISTAMINE IN THE CARDIAC SYMPATHETIC FIBERS FROM THE GUINEA PIG SUPERIOR CERVICAL GANGLION 2008, 39 (5):  773-775.  doi: Abstract ( )   Objective To investigate the expression of histamine in the cardiac sympathetic fibers from the guinea pig superior cervical ganglion and its coexistence with norepinephrine so as to provide morphological evidence for histamine as a cardiac sympathetic neurotransmitter. Methods Biotinylated dextranamine (BDA) anterograde tracing and immunofluorescence histochemical staining for histamine/norepinephrine were applied. Results After injection of BDA into the superior cervical ganglion, BDA labeled sympathetic fibers in the left and right atria and ventricle were observed. Meanwhile, the tracing fibers proved histamine-like immunoreactive or both histamine and norepinephrine-like immunoreactive. Conclusion Histamine is expressed in the cardiac sympathetic fibers from the guinea pig superior cervical ganglion and coexisted with norepinephrine. Related Articles | Metrics

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RELATIONSHIP BETWEEN 5-LIPOXYGENASE AND LUNG INFECTION OF MICE 2008, 39 (5):  776-778.  doi: Abstract ( )   Objective To investigate the relationship between the 5-lipoxygenase(5-LO) in mouse lung and during infection in the respiratory system. Methods Investigate the expression of 5-LO in the lung of a mouse by immunocytochemistry. Results The intensity of the immune staining against 5-LO antibody was stronger in infected mice than that in noninfected rats. A weaker immuneoreation was observed in the treated group of animals. Conclusion Changes in the expression of 5-LO in pulmonary tisssues could be designed to detect the treatment efficacy during the lung infection. Related Articles | Metrics

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AN IMPROVED COX METHOD FOR NEURON STAINING BY USING VIBRATOME SECTION 2008, 39 (5):  779-781.  doi: Abstract ( )   Objective To explore a quicker and simpler method for the staining of cerebral pyramidal cells and cereberullar Purkinje cells. Methods Blocks of the cerebrums and cerebellums of the cat, rabbit, rat and guinea pig were removed, fixed and immersingstained with a mixture of mercuric chloride, potassium bichromate and potassium chromate. Some of fixed blocks were embedded in collodion and sectioned on a sliding microtome. The sections were prepared by traditional Cox method. The other fixed blocks were cut on a vibratome without embedding, and the sections were prepared by the improved Cox method. Results Compared with Cox method, the improved method by using vibratome section had less procedure and took less time. Moreover, the staining of neurons by the improved method was more even, showing more complete processes and more details of small processes, with a lighter background and no or fewer if any, precipitations of staining reagents in the sections.Conclusion The improved Cox method by using vibratome section is quick and simple with higher successful rate and is superior in terms of neuron staining compared with the traditional Cox method. Related Articles | Metrics