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2010, Volume 41 Issue 1
06 February 2010 Previous Issue    Next Issue
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bFGF induced dedifferentiation of glial cells in injured rat optic nerve 2010, 41 (1):  1-8.  doi: 10.3969/j.issn.0529-1356.2010.01.001 Abstract ( )   Objective To investigate the mechanism that bFGF promotes the regeneration of injured optic nerve and induces dedifferentiation of glial cells in it. Methods Fifty-five adult male SD rats were randomly divided into 3 groups as normal control group, injury group and bFGF group. At day 7 post operation, optic nerves from injury group and bFGF group were detected by gene chip and real-time PCR. At day 7, 14 post operation, optic nerves were harvested and detected by HE staining and immunohistochemistry. Results Compared with the injury group, there were 645 genes expression up-regulated and 458 genes down-regulated including genes related neural stem cell or precursor cell neural development, proliferation, apoptosis, chromatin configuration, transcription regulation, signal transduction, neural growth and so on in the bFGF group. There were bigger nuclei, more cells, more immunoreactivity of nestin, extracellular signal-regulated kinase(Erk1/2), glial fibrillary acidic protein(GFAP), and myelin basic protein(MBP) in the distal optic nerves and more immunoreactivity of neurofilament(NF) in the proximal optic nerves in the bFGF group than that in the injury group.Conclusion bFGF could promote the proliferation of neuroglia cells, dedifferentiation of neural glias and improve the microenvironment to favour the regeneration of injured optic nerve. Related Articles | Metrics

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Interference effect of picrosideⅡon cerebral ischemia reperfusion injury in rats 2010, 41 (1):  9-12.  doi: 10.3969/j.issn.0529-1356.2010.01.002 Abstract ( )   Objective To investigate the neuroprotective effects of picrodideⅡ on cerebral ischemic reperfusion injury in rats. Methods Intraluminal thread methods were applied to establish the left middle cerebral artery occlusion reperfusion models (MCAO/R) in rats. PicrodideⅡ (10mg/kg) and salvianic acid A sodium (10mg/kg) were injected from tail vein for treatment. The neurological behavioral function was evaluated with Bederson’s test. The cerebral infarction volume was observed with tetrazolium chloride (TTC) staining. The structure of cells was observed with histopathology. The apoptosis positive cells were counted by terminal deoxynucleotidyl transferase midiated dUTP nick end labeling (TUNEL). Results The neurological behavioral malfunction appeared in all rats with MCAO/R. The infarction focus showed in the ischemic hemisphere following cerebral ischemia reperfusion injury. In the picrodideⅡ and salvianic acid A sodium treatment groups, the number of apoptosis positive cells decreased and the cerebral infarction volume reduced, while the neurological behavioral function was significantly improved than those in the model control group (EM>P/EM><0.05). The cerebral infarction volume in the picrodideⅡ group was smaller than that in the salvianic acid A sodium group (EM>P/EM><0.05).Conclusion PicrodideⅡ might reduce cerebral infarction volume and improve the neurological behavioral function through inhibit Related Articles | Metrics

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Effects and injury mechanism of reactive oxygen species after spinal cord injury 2010, 41 (1):  13-17.  doi: 10.3969/j.issn.0529-1356.2010.01.003 Abstract ( )   Objective To explore the effect and injury mechanism of reactive oxygen species (ROS) after spinal cord injury (SCI) through detecting the dynamic changes of malonyldialdehyed （MDA）content in spinal cord and observing neurocyte apoptosis and correlation apoptosis factor expression after SCI. Methods Totally 132 adult SD male rats were randomly divided into three groups: sham group, SCI group, methylprednisolone (MPSS) group. The SCI of SD rats was performed by Allen’s weight dropping way to impact on the posteriors of spinal cord T10. The contents of MDA were determined by chromatometry, the expression of Caspase-3 and Bcl-2 family in the injured spinal cord was detected by immunohistochemical staining; Apoptotic cells were detected by using fluorometric terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (fluorometric TUNEL) staining. Results The content of MDA in the injured cord increased significantly after SCI, reached the peak at 6 hours and 3 days post-injury, then dropped down gradually, then was back to the normal level after 7 days. The number of TUNEL labeling positive cells of SCI group increased at 6 hours post-injury, reached the peak at 3 days, then dropped down gradually; Bcl-2, Bax protein began to increase at 6 hours post-injury, reached the peak at 5 days after injury, then dropped down gradually. Caspase-3 protein began to increase at 6 hours post-injury, reached the peak at 3 days after injury, then dropped down gradually. The content of MDA, the number of TUNEL labeling positive cells, the expression of Caspase-3 and Bax of MPSS group decreased significantly than that of SCI group at the same time, respectively, while Bcl-2 protein was up-regulated after administration of MPSS.Conclusion ROS could promote the expression of Caspase-3 and degrade the ratio of Bcl-2/Bax to induce apoptosis of neurocyte, which might play significantly role in the process of secondary SCI. In addition, MPSS exerts neuroprotective effects against ROS toxicity, which might be of importance and might contribute to their clinical efficacy for the treat Related Articles | Metrics

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Construction and function of the recombinant vector expressing human glutamic acid decarboxylase 65 2010, 41 (1):  18-21.  doi: 10.3969/j.issn.0529-1356.2010.01.004 Abstract ( )   Objective To construct the recombinant rAAV2-hGAD65 vector and detect its function both in vitro and in vivo. Methods The cDNA of human glutamic acid decarboxylase 2 (hGAD65) gene, which was one of gamma-aminobutyric acid (GABA) synthetase, cloned by the method of RT-PCR, was subcloned into the adeno-associated virus (AAV) vector and formed the recombinant vector of AAV-hGAD65 (rAAV2-hGAD65). The recombinant vector was packaged by the AAV Helper-Free System and its titer was detected. The primary fibroblast, cultured from the rat lung, was transfected by the rAAV2-hGAD65. The expression of the hGAD65 in fibroblast was detected by immunohistochemical method and the level of GABA in the media was assayed by high performance liquid chromatograph (HPLC). EM>In vivo/EM>, the hGAD65 was detected by immunohistochemical method in STN and the concentration of the GABA in the reticular part of substantia nigra (SNr) was assayed by HPLC after the rAAV2-hGAD65 was injected into the subthalamic nucleus (STN) by the stereotaxic method. Results The sequence of hGAD65 cDNA was in accordance with that in the Genebank. The amino acid sequence of hGAD65 had no mutation and the titer of rAAV2-hGAD65 was reached 4.5 ×10SUP>11 /SUP>per milliliter. The efficiency of infection to the rat primary firoblasts was 80%, and the concentration of GABA in the media of fibroblasts was (45.66±6.07)nmol/L per liter. EM>In vivo/EM>, hGAD65 could be detected in STN, and the concentrateion of the GABA in the SNr was increased from (5.66±1.07)nmol/g to (12.66±2.59)nmol/g.Conclusion The cDNA of hGAD65 was cloned by RT-PCR and the recombinant vector of rAAV2-hGAD65 was constructed. The AAV can infect the primary fibroblast in vitro and the hGAD65 can catalyse the glutamic acic to GABA. In vivo, the concentration of GABA in the SNr was heighten after the rAAV2-hGA Related Articles | Metrics

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Mechanism of cyclin-dependent inhibitor p27Kip1 in regulating the differentiation of immortalized human neural progenitor cells 2010, 41 (1):  22-26.  doi: 10.3969/j.issn.0529-1356.2010.01.005 Abstract ( )   Objective To investigate whether there is any functional link between p27Kip1 function and all-trans retinoic acid (RA) in the control of neuronal differentiation of immortalized human neural progenitor cells (hSN12W-TERT cells). To investigate the mechanism by which p27Kip1 regulates the differentiation of immortalized human neural progenitor cells. Methods hSN12W-TERT cells were derived from the striatums of human embryos at 12 weeks gestation and cultured with serum-free medium in presence of EGF and bFGF. At the appropriated time, hSN12W-TERT cells were exposed to 1μmol/L RA for 3, 5, 7 days respectively. The experiment was repeated there times. Cell cycle analysis was performed by flow cytometry analysis (FACS). The expression of p27Kip1, p21Cip1, cyclin-dependent kinase 2 (cdk2）, p-cdk2 and S-phase kinase-associated protein 2 (skp2) in hSN12W-TERT cells before and after RA treatment cells were determined by using Western blotting analysis. Results FACS result showed that 77.25% of proliferating hSN12W-TERT cells were in the G1/G0-phase while 9.38% of cells in the S-phase. Following RA treatment, cell growth was arrested, and 85.68% of cells accumulated in G1/G0-phase while 8.57% of cells in the S-phase. Western blotting analysis demonstrated that the levels of p27SUP>Kip1/SUP> in the hSN12W-TERT cells increased following 3 days’ treatment with RA compared with those of normal untreated cells, with a peak at 5 days (EM>P/EM><0.05). The similar results were acquired both in nuclear proteins and in cytoplasm proteins of hSN12W-TERT cells. The expression level of p21SUP>Cip1/SUP> decreased in response to RA treatment. RA did not affect the expression of cdk2, but the expression of p-cdk2, which represented the activity of cdk2, was markedly decreased in response to RA treatment. Skp2, which was required for the ubiquitin-mediated degradation of p27SUP>Kip1/SUP>, was detected in proliferating hSN12W-TERT cells. The expression of skp2 reduced dramatically in response to RA treatment in a time-dependent manner.Conclusion There is a functional link between RA and p27SUP>Kip1/SUP> function in the control of neuronal differentiation in hSN12W-TERT cells. p27SUP>Kip1/SUP> plays a key role during neuronal differentiation. Moreover, high levels of p27SUP>Kip1/SUP> are associated with its degradation inhibiting through reducing proteasome-dependent Related Articles | Metrics

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Relation of ethanol treatment with dopaminergic system in rat brain 2010, 41 (1):  27-31.  doi: 10.3969/j.issn.0529-1356.2010.01.006 Abstract ( )   Objective To study the effect of ethonal on the dopaminergic system by analyzing the altered expression of tyrosine hydroxylase (TH) and dopamine transporter (DAT) in the brain of ethanol-treated rats. Methods Sixty Wistar rats were selected and divided into control group and ethanol-treated group, 30 per group, the ethanol-treated rats were treated with 20% ethanol for 6 months. Immunohistochemistry, flow cytometry and Western blotting were used to analyze the altered expression of TH and DAT in the DA energic system in different brain regions of the ethanol treated rats. Results 1. Immunohistochemistry showed the mean gray value of TH in substantia nigra(SN)-ventrotegmental area (VTA), caudae putamen (CPu) and nucleus accumbens (NACC), DAT in CPu and NACC of the ethanol were smaller than those in control (P<0.05). 2. Flow cytometry showed the expression of TH in middle brain of the ethanol-treated rats increased significantly compared with the control(EM>P/EM><0.05). 3. Western blotting showed the ratio of IA of TH/β-actin and DAT/β-actin in different brain regions of the ethanol-treated rats were larger than those in control(EM>P/EM><0.05).Conclusion Ethanol treatment increases the expression of TH and DAT in rat brain. Related Articles | Metrics

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Up-regulation of β-amyloid precursor protein cleavage enzyme 1 and β-amyloid protein in brain with diabetic rats 2010, 41 (1):  32-36.  doi: 10.3969/j.issn.0529-1356.2010.01.007 Abstract ( )   Objective To investigate expression of β-site APP-cleaving enzymel(BACE1) and Aβ in brain of diabetes mellitus of Wistar rats，to study pathophysiological mechanism of Alzheimer disease from diabetic metabolic disorder. Methods Animal model of diabetes mellitus was established by streptozocin with intraperitoneal injection. Wistar rats were randomly divided into normal group (N), sham-operation group (S), 4 weeks diabetes mellitus model group (M4), 6 weeks diabetes mellitus model group (M6) and 8 weeks diabetes mellitus model group (M8). Behaviour was tested with Morris water maze and shuttle box test. Expression of Aβ was measured by enzyme linked immunosorbent assay and BACE1 by immunohistochemistry, enzyme linked immunosorbent assay, Western blotting and RT-PCR. The absorbance value was measured by imaging analysis. Results The electric times and latancy of memory and study were more increased in model group than that in N and S group but the times of escape more decreased(EM>P/EM><0.01). The expression of Aβ1SUB>40/SUB> increased from (64.13±6.76)pg/mg in normal group to (86.43±7.03)pg/mg in model group by ELISA(EM>P/EM><0.001) and Aβ1SUB>42/SUB> from (67.43±5.12 )pg/mg in normal group to (89.45±5.28) pg/mg (EM>P/EM><0.001) in model group. The expression of BACE1 increased from (116.46±8.10)pg/mg in normal group to (158.73±6.24)pg/mg in model group by ELISA and from 0.61±0.11 in normal group to 1.52±0.16 by Western blotting absorbance valule and from 1.62±0.26 in normal group to 3.61±0.32 by RT-PCR absorbance valule and from 0.81±0.21 in normal group to 2.01±0.36 by immunohistochemistry absorbance valule (EM>P/EM><0.001). The expr Related Articles | Metrics

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Effects of exogenous estrogen on the distribution of nitric oxide synthase positive neurons in supraoptic nuclus of hypothalamus of ovariectomized female rats 2010, 41 (1):  37-40.  doi: 10.3969/j.issn.0529-1356.2010.01.008 Abstract ( )   Objective To observe how exogenous estrogen influences the distribution and the expression of NOS positive neurons in the supraoptic nuclei of hypothalamus in the overiectomized female rats. Methods The 2-3-month-old female rats(EM>n/EM>=40) were selected as the healthy and nulli-copulatory experimental animals. Rats were divided into following groups: normal control group(EM>n/EM>=10), ovariectomized control group(EM>n/EM>=10), and two experimental groups that have been injected with estrogen for post-operative 40 days(EM>n/EM>=10) and for post-operative 70 days(n=10). Finally, all the animals were infused and the brains were removed. Immunohistochemical (SABC) method was adopted to count the number of NOS poitive neurons and observed the NOS poitive neuronal morphology under the light microscope. The image analysis system was used to test the average gray value of immunoreactivity in NOS positive neurons. Results In the ovariectomized control group, the density of NOS positive neurons in supraoptic nucleus was significantly increased and their shapes were bigger than that of the normal control group(EM>P/EM><0.05). The density and the form of the NOS positive neurons in supraoptic nucleons had no apparent difference between the estrogen for post-operative 40 days group and the ovariectomizeed control group(EM>P/EM>>0.05).In the group after estrogen-injection 2 months compared with the normal control group, and the ovariectomized control group, both of the NOS positive neurons’ density and the size become significantly decreased, and the staining of cells was lesser in the group injected with estrogen for post-operation 70 days. Conclusion The present results suggest that exogenous estrogen may influence the distribution and the expression of NOS positive neurons in supraoptic nucleus of hypothalamus of ovariectomized female rats. Related Articles | Metrics

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Fibrin scaffold promoted the differentiation of neural stem cells to the neurons and inhibited the proliferation of astrocytes 2010, 41 (1):  41-47.  doi: 10.3969/j.issn.0529-1356.2010.01.009 Abstract ( )   Objective To investigate the effects of the fibrin scaffold on the differentiation and the proliferation of neural stem cells and astrocytes. Methods Neural stem cells and the gliocytes derived from spinal cord were cultured in vitro respectively. The purified neural stem cells or gliocytes were seeded separately onto the fibrin scaffolds as experimental group and the glass slides modified with poly-L-lysine（PLL）as control group. At different time in culture the neural stem cells were immunofluorescence stained with antibodies against the marker of neurons i.e. neurofilament(NF).The length of NF-positive neuritis was masured and the average value was calculated in the culture well (EM>n/EM>=4). The gliocytes were immunofluorescence stained with antibodies against the marker of astrocytes i.e. glial fibrillary acidic protein (GFAP ). The total number of the cells and the GFAP-positive cells were counted from 5 different fields of vision in the culture well (EM>n/EM>=4), then the average ratio of GFAP-positive cells was calculated. The differentiation of neural stem cells, the extension of neurites and the proliferation of astrocytes on the fibrin scaffolds were compared with those on the slides. The protein of GFAP was detected by Western blotting to analyse the mature degree of astrocytes. All above experiments were repeated 3 times respectively. Results Immunofluorescence staining showed that the NF-positive neurites in the fibrin scaffold group were longer than those in the control group, whereas GFAP-positive cells were fewer than those in the control group. The expression of GFAP in the cells on the scaffold was lower than that in the control group.Conclusion The fibrin scaffold could promote differentiation of the neural stem cells to neurons and extension of the neurites. Meanwhile, the scaffold could inhibit proliferation and mature of the astrocytes. Related Articles | Metrics

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Effect of adenovirus-mediated IL-24 gene on the topoisomeraseⅡα and Caspase-3 expression in glioma cell line U251 2010, 41 (1):  48-52.  doi: 10.3969/j.issn.0529-1356.2010.01.010 Abstract ( )   Objective The present study is to investigate IL-24 gene(Ad5F35-hIL-24) effect on the topoisommeraseⅡα(topoⅡα) and Caspase-3 expression in glioma cell line U251. Methods After transfected the U251 glioma cells with the Ad5F35-hIL-24, the methyl thiazolyl tetrazolium (MTT) was used to analyse the inhibition rate of Ad5F35-hIL-24 on the cells. Hoechst 33258 fluorescent staining and flow cytometric assay were used to detect apoptosis. The immunohistochemistry assay was used to detect topoⅡα expression, and Western blotting was applied to detect the protein expression of topoⅡα and Caspase-3. Transwell experiment was used to test the invasiveness of the cells. Results It was found that the Ad5F35-hIL-24 could inhibit U251 cell proliferation and induce apoptosis in a dose dependent manner compared with the control groups. It showed that Ad5F35-hIL-24 could inhibit topoⅡα expression reveled by immunohistochemistry and Westeren blotting, while it increased Caspase-3 protein expression. The Transwell experiment showed that the Ad5F35-hIL-24 could reduce the invasiveness of the U251 glioma cells.Conclusion The exogenous IL-24 gene can inhibit the cell proliferation and induce apoptosis of U251 glioma cells. The to Related Articles | Metrics

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Effect of pineal gland and melatonin on CD4SUP>+/SUP>CD25SUP>+/SUP>Treg cells development and Foxp3 expression of rat thymus 2010, 41 (1):  53-59.  doi: 10.3969/j.issn.0529-1356.2010.01.011 Abstract ( )   Objective To investigate the influence of pinealectomy（Px）and melatonin (MLT) supplementation on CD4SUP>+/SUP>CD25SUP>+/SUP>Treg cells development and Foxp3 expression in rat thymus. Methods One hundred and twenty clean-grade male SD rats were divided into five groups: normal control group, sham pinealectomized group, pinealectomized group, Px +7.5 mg/(kgI>&#/I>8226;d) MLT group, Px +15 mg/(kgI>&#/I>8226;d) MLT group, and the thymus were taken out at the 4th week and the 8th week; Flow cytometric analysis was used to analyse the number of double positive cells in the thymus and peripheral blood; Semiquantitative RT-PCR and immunohistochemistry were applied to analyse Foxp3 expression of rat thymus. Results There was no difference of the number of CD4SUP>+/SUP>CD25SUP>+/SUP> Treg cells in rat thymus between Px and normal/sham group, and the number increased dependently on time and dose after MLT supplementation. In rat peripheral blood the double positive cells significantly increased in models of Px at the 4th week, and then backed to normal level after MLT supplementation. The results showed no significant changes in all groups at the 8th weeek; At the 4th weeks, Foxp3 expression increased in the thymus of Px rats compared to nomal/sham group, which returned to normal level after MLT supplementation. On the other hand, Foxp3 expression showed no significant difference in all groups at the 8th weeek.Conclusion Px made no difference between the development of CD4+CD25+Treg cells in rat thymus, but resulted in significant increase of thymic output and Foxp3 expression. All the effects disappeared at the 8th week. MLT supplementation could reverse the abnormity. Related Articles | Metrics

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Analysis of proteins secreted by bone marrow stromal cells using shotgun mass spectrometry 2010, 41 (1):  60-64.  doi: 10.3969/j.issn.0529-1356.2010.01.012 Abstract ( )   Objective Using shotgun mass spectrometry to detect proteins probably contained in bone marrow stromal cells(BMSCs) conditioned medium. Methods Mixed with BMSCs conditioned medium was divided into two parts which is(>5kD and <5kD) by means of ultrafiltration. The two parts were used to culture neural stem cells(NSCs) separately, and the proportions of neurons, astrocytes and oligodendrocytes in the offsprings of NSCs were calculated, then the effective part that could regulate the differentiation of NSCs was detected by Shotgun mass spectrometry. Results The BMSCs conditioned medium which is >5kD could promote the NSCs differentiate into more neurons and oligodendrocytes. The SDS-PAGE of this part showed that the most proteins were above 14kD, then the protein bands were enzymed. In total, 456 proteins were identified by Shotgun mass spectrometry after all the protein bands were enzymed, there were 154 similar proteins, 17 hypothetical proteins and 56 unknown proteins. And in the rest of 229 proteins, most of them were cytoskeletal proteins, secreted proteins, signal transduction proteins, enzymes, transporter and so on. Conclusion Many proteins secreted by BMSCs could regulate the differentiation of NSCs so as to prove the protein components probably existed in the BMSCs conditioned medium. Related Articles | Metrics

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Clinical significance of thrombospondin 4 expression in gastric carcinoma 2010, 41 (1):  65-69.  doi: 10.3969/j.issn.0529-1356.2010.01.013 Abstract ( )   Objective The aim of this study was to measure quantitatively the TSP-4 mRNA expression and its significance in gastric carcinoma was evaluated by correlating its expression with clinicopathological features, microvessel density(MVD) and MMP-9.Methods Eighty-two patients with gastric carcinoma were recruited in this study. TSP-4 mRNA expression in gastric carcinoma and adjacent tissue was detected by real-time PCR. CD34 and MMP-9 protein expression were examined by immunohistochemistry. MVD was determined according to CD34-positive tubular structures. Results The expression level of TSP-4mRNA in gastric carcinoma was obviously higher than those of adjacent tissue(EM>P/EM>=0.03) and it was associated with tumor size, histological type, lymph node metastasis, MVD and MMP-9(EM>P/EM>=0.002, P=0.031， EM>P/EM>=0.014,EM>r/EM>＝0.67 EM>P/EM><0.01, EM>P/EM>=0.008),but not sex, age and depth of invasion. （EM>P/EM>=0.53，EM>P/EM>=0.57，EM>P/EM>=0.15）.Conclusion TSP-4 may play an important role in occurrence and progression of gastric carcinoma and that the effect of TSP-4 on tumor growth and metastasis may be exerted through reg Related Articles | Metrics

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Differentiation of rat bone marrow stromal stem cells into neuron-like cells induced by breviscapine 2010, 41 (1):  70-74.  doi: 10.3969/j.issn.0529-1356.2010.01.014 Abstract ( )   P>Objective To explore the feasibility of differentiation into neural cells from bone marrow stromal cells(BMSCs) in vitro by breviscapine,and to offer reference for the applying of BMSCs. Methods The bone marrow stromal cells of SD rat were separated and purified by passsage culture.After phenotype characterization,the 4th passage of BMSCs was exposed to breviscapine. Differentiation was observed under phase contrast microscope every 6 hours and the cells were stained immunocytochemically with neuronspecific enolase（NSE）and glial fibrillary acidic protein（GFAP）.The induced cells activity was detected by MTT. NSE and GFAP were determined by flow cytometry and RT-PCR. Results The BMSCs were CD44SUP>+/SUP> ，CD54SUP>+/SUP> ，CD34SUP>- /SUP>. After 18 hours of induction, cytoplasm of BMSCs partly contracted with protruding;The immunocytochemical staining was performed on cells after induction for 24 hours，the rates of NSE and GFAP staining positive were（48.7±3.4）％ and（56.8±4.2）%.By using flow cytometer, expressions of NSE and GFAP showed much higher in induced gro Related Articles | Metrics

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Expression of NF-κB and regulated upon activation normal T cell expressed and secreted chemokine in experimental abscending aortic aneurysm rat model 2010, 41 (1):  75-79.  doi: 10.3969/j.issn.0529-1356.2010.01.015 Abstract ( )   Objective To investigate the expression of nuclear factor kappaB(NF-κB) and regulated upon activation normal T cell expressed and secreted chemokine(RANTES) during the formatiom of ascending aortic aneurysm. Methods Forty Wistar rats were randomly divided into the control group(n=20) and the experimental group(n=20).The rat models were made by ligating the ascending aorta. The ascending aortas were taken after ligation for 3months. Immunohistochemistry staining was performed to detect the protein expression of NF-κB and RANTES. The expression of NF-κB and RANTES mRNA were detected by RT-PCR. Results Immunohistochemisry staining results showed NF-κB and RANTES expression significantly increased in aneurysm, while there was a little positive staining in the control group. RT-PCR results indicated that the expression levels of NF-κB and RANTES in the aneurysm were stonger than that of the control group. The expression of NF-κB and RANTES mRNA were remarkably correlated. Conclusion The expression of NF-κB and RANTES in ascendin aortic aneurysm are stronger than that in Related Articles | Metrics

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Development of the outflow tract ridge in the embryonic mouse heart 2010, 41 (1):  80-86.  doi: 10.3969/j.issn.0529-1356.2010.01.016 Abstract ( )   Objective To investigate the origin of α-SMA positive cells in the outflow tract ridge of the embyonic mouse heart and to explore the ultrastructure change of the mesenchymal cells during the ridges fusion. Methods Sections of embryonic day 10(E10d) to E14d mouse embryonic hearts were stained by immunohistochemistry assay with monoclonal antibodies against α-smooth muscle actin (α-SMA), α-sarcomeric actin(α-SCA) and in situ hybridization method with PlexinA2 probe. The outflow tract ridges fusion was observed by transmission electron microscopy at E12.5d. Results From E10d to E11d, PlexinA2 positive cells were seen in the neural tube with mesenchymes around it, the aortic sac and aortic arch. These cells also migrated into the outflow tract ridge along the aortic sac wall and part of them expressed α-SMA. At E12d, PlexinA2 was expressed in the spinal ganglia, the pharyngeal mesenchyme, the aortopulmonary septum and the ascending aorta and pulmonary trunk. The septum showed α-SMA strongly positive. But only a few of α-SMA positive cells were observed in the ascending aorta and pulmonary trunk. At E12.5d, two clusters of condensed mesenchymal cells in the outflow tract ridges formed and began to express PlexinA2 weakly and α-SMA strongly. When the ridges began to fuse, the endothelial cells formed a cellular seam, which rapidly broke into pieces and disappeared and were replaced by the sparsed mesenchymal cells containing a few of microfilaments. Two clusters of condensed mesenchymal cells gradully moved to merge. It was noted that some mesenchymal cells contained plenty of microfilament bundles, mitochondria and focal contacts between them. Some mesenchymal cells only had a few of microfilaments and plasma membrane fusion was seen between them. Conclusion α-SMA positive cells in the outflow tract cushion may be derived from cardiac neural crest. The endothelial cells might participate in the fusion of the outflow tract ridges by endothelial-mesenchymal trans Related Articles | Metrics

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Anti-aging effect of allografting adipose-derived stem cells in rats 2010, 41 (1):  87-92.  doi: 10.3969/j.issn.0529-1356.2010.01.017 Abstract ( )   ObjectiveTo observe the effect of transplanting the adipose-dervived stem cells(ADSCs) on free radical metabolism and immune function of rat aging model induced by D-galactose from fasiaology perspective; to explore a new method for anti-aging. MethodsThe ADSCs were cultured in vitro. Thirty male SD rats were randomly divided into 3 groups: normal control group(A), aging model group(B) and treat group(C). Ten rats in each group. Rats in B and C groups were injected D-galactose continually into make the sub-acute aging model rats.After 8-week injections of D-galactose, rats in group C were injected ADSCs(3×106/ml) through caudal vein. After 2-week transplantions of ADSCs, T-SOD, CuZn-SOD, MDA, NO, IL-2 and spleen index levels in serums of each group were detected and compared among the three groups. ResultsCompared with the A group, the SOD, NO, IL-2 level and spleen index in serum in group B decreased significantly, while the contents of MDA increased significantly. Compared with group B, the SOD, NO, IL-2 level and spleen index in serum in group C had been improved, and the contents of MDA decreased significantly. ConclusionTransplanting ADSCs can improve the antioxidant ability and strengthen the cellular immune function of aging rats.Further more, it can delay the ageing procedure induced by D-galactose in rats. Related Articles | Metrics

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Salvia miltiorrhiza promotes vascular endothelial growth factor expression in frozen-thawed mouse ovarian in an early stage after transplantation 2010, 41 (1):  93-99.  doi: 10.3969/j.issn.0529-1356.2010.01.018 Abstract ( )   Objective To investigate the angiogenesis effects of Salvia miltiorrhiza in heterotopically grafted frozen-thawed mouse ovaries. Methods The ovaries thawed after cryopreservation were xenografted into the donated kidney capsules of 8- to 12-week adult male mice. The mice were divided into two groups, saline and Salvia miltiorrhiza groups, the mice either in the saline or in Salvia miltiorrhiza groups were administered i.p. daily either saline(0.5ml) or Salvia miltiorrhiza(0.5g)respectively, from the day prior transplantation. The two groups were sacrificed 1 day,2 days and 7 days after transplantation respectively, the grafts from thawed,1 day,2 days,7 days were removed for follicle counting, immunohistochemical studying and detecting of the mRNA expression of vascular endothelial growth factor(VEGF). Results The number of follicles and survival rates in grafts after transplantation of Salvia miltiorrhiza group were more than that of saline group (EM>P/EM><0.05); the expression of VEGF increased after transplantation,peaked on day 7,there was no difference between the two groups (EM>P/EM>>0.05); the apoptosis index of Salvia miltiorrhiza group was less than that of saline group (P<0.05), the mRNA expression of VEGF188 and VEGF164 of Salvia miltiorrhiza group was more than that of saline group on 48 hours after transplantation(EM>P/EM><0.05). Conclusion Salvia miltiorrhiza may provide benefits for folliculogensis and decreasing the apoptosis index of follicles. Nevertheless,a increase in the VEGF188 and VEGF164 isoform in the Salvia miltiorrhiza group may suggest the positive effect of exogenous Salvia miltiorrhiza therapy in the early stage of angiogenesis. Related Articles | Metrics

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Effects of heroin dependence on rat’s calcitonin positive cells in thyroid gland and serum FT3, FT4, TSH 2010, 41 (1):  100-103.  doi: 10.3969/j.issn.0529-1356.2010.01.019 Abstract ( )   Objective To investigate the effect of the heroin addicts on rat’s thyroid C cells, the secretion of calcitonin(CT) and serum FT3, FT4, TSH. Methods Adulthood 55 male SD rats were randomly divided into normal control group(NCG), saline control group(SCG) and heroin dependence group(HDG). Heroin-dependent rat models were set up by the way of subcutaneous injection of heroin and the thyroid glands were excised on the 10th, 17th, 24th, 31st, and 38th days after models were set up. Tissues were assessed by immunohistochemical SABC method, cell counting, image analysis method and chemiluminescence techniques. Result Compared with the normal control group(NCG)and saliane control group( SCG), the immune staining intensity was weakened; the number of CT positive cells was reduced(EM>P/EM><0.05); the average gray value was significantly higher(EM>P/EM><0.05) and showed a gradual increase in the CT positive cells of heroin dependence group(HGD). The concentration of TSH in HDG was not statistically significant as compared with the NCG. The concentration of FT3 in HDG decreased gradually on the 10th day and the 17th day as compared with the NCG.(EM>P/EM><0.05). The concentration of FT4 in HDG increased gradually on the 24th day and the 38th day as compared with the NCG.(EM>P/EM><0.05). Conclusion During heroin dependence period, the number and functional of CT positive cells has decreased. The changes of concentration of FT3 and FT4 suggest that the function of thyroid gland has changed. Related Articles | Metrics

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Expression of endothelin-1 in cerebral vessels of scalded rats 2010, 41 (1):  104-108.  doi: 10.3969/j.issn.0529-1356.2010.01.020 Abstract ( )   Objective To investigate the expression and effects of endothelin-1 (ET-1) in cerebral vessels of scalded rats. Method Histological method was used to observe the alteration of histological structure of basilar artery in 8 scalded rats. Reverse transcription polymerase chain reaction(RT-PCR) was performed to examine expression of ET-1 mRNA; Western blotting to analyse expression of endothelin-1; radioimmunoassay to measure ET-1 levels, respectively, of cerebral basilar artery in 48 scalded rats. Result Vascular pathohistological changes were observed in cerebral basilar artery after rats were scalded; ET-1, ET-1 mRNA expression and ET-1 levels of basilar artery in scald 6 hours, 12 hours and 24 hours group were obviously higher than that of control group. Conclusion Scald may cause an increases of ET-1 expression in basilar artery, and elevated ET-1 expression may relate to the pathogenesis of cerebral vascular injury in scalded rats. Related Articles | Metrics

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Expression of hepatic growth factor and C-met in reserved liver tissue after partial hepatectomy of hepatic fibrosis 2010, 41 (1):  109-113.  doi: 10.3969/j.issn.0529-1356.2010.01.021 Abstract ( )   Objective To study the expression of hepatic growth factor(HGF) and C-met in reserved liver tissue after partial hepatectomy of rats with hepatic fibrosis. Methods Totally 130 SD rats were randomly divided into four groups: normal group （n=7）, group of normal rats with partial hepatectomy(EM>n/EM>=50)，hepatic fibrotic group(EM>n/EM>=7), and group of hepatic fibrotic rats with partial hepatectomy(EM>n/EM>=66). Rats were killed after operation 12 hours, 1 day, 3 days, 5 days, 7 days and 14 days separately, then HGF and C-met of reserved liver tissues were detected with immunohistochemistry staining and Western blotting. Results In the group of normal rats with partial hepatectomy, immunohistochemistry staining indicated that the expression of HGF and C-met increased to get the peak point after partialhepatectomy 12 hours and 3 days respectively, and HGF maintained at the high level to the 7th day, then decreased gradually, finaly approched to the level of pro-operation at 14th day, but C-met fell sharply，and declined to the the level of pro-operation at the 14th day. In the group of hepatic fibrotic rats with partial hepatectomy, the expression of HGF and C-met decreased sharply after operation 12 hours, next HGF increased to get the peak point at the 1st day, and then declined speedily, and decreased to the bottom at the 14th day, but C-met declined to the bottom at the 3rd day, then increased slightly till the 7th day, affter that increased sharply to the summit at the 14th day. Western blotting analysis showed the results of HGF and C-met coincided with that of immunohistochemistry. Conclusion The high isochronous expression of HGF and C-met in hepatic tissue is propitious to hepatocellular division, Which indicates that the expresson out of step of HGF and C-met might be the key reason of hard regeneration of fi Related Articles | Metrics

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Expression and ultrastructure location of PPAR-γ and COX-2 in endometrial carcinoma 2010, 41 (1):  114-118.  doi: 10.3969/j.issn.0529-1356.2010.01.022 Abstract ( )   Objective To investigate the development and prognosis of peroxisome proliferators-activated receptor-γ(PPAR-γ) and cyclooxygenase-2(COX-2) in endometrial carcinoma. Methods The electron microscopic immunohistochemical technique was used to observe the ultrastructure location of COX-2 protein labeled by colloidal gold in the endometrial carcinoma. Results 1.There were negative immunohistochemical staining signals of PPAR-γ in both the normal endometrium and hyperplasia endometrium, whereas, was positive staining in the endometrial carcinoma; 2.The intensities of the COX-2 immunohistochemical staining were significantly statistical difference among the normal endometrium, the hyperplasia endometriu and the endometrial carcinoma(EM>P/EM><0.01); 3. The COX-2 labelled by colloidal gold granules was observed in the endoplasmic reticulum, nuclear membrane and nuclei in the endometrial carcinoma. Conclusion Both PPAR-γ and the COX-2 might play an important role in the development Related Articles | Metrics

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Electron microscopic observations on changes of cytoplasmic organelles in the oocytes of Cervus nippon hortulorun 2010, 41 (1):  119-123.  doi: 10.3969/j.issn.0529-1356.2010.01.023 Abstract ( )   Objective Revealing the developmental regulation of Cervus nippon’s oocyte and organelles. Methods In the experiment,follicle systems during both estrum and non- estrum were divided into the primordial follicle,growing follicle and mature follicle according to the Cervus nippon’s follicle diameter size,formation of zonapellucida,appearance time of follicular cavity.At the same time,observations on cytoplasmic organelles in development of oocytes were conducted with electron microscopic,eyepiece micrometer and photomicrographic technique(The number of every oocyte observed is 6-8). Results In the primordial follicle and early growing follicle phase,the quantity of mitochondria,golgi apparatus,smooth endoplasmic reticulum and cortical granules increased gradually and all organelles moved to the cortical area.However,in the late growing follicle and mature follicle phase,Golgi apparatus and rough endoplasmic reticulum disappeared,cortical granules began to arrange themselves in line beneath the plasma membrane of the oocyte,mitochondrias dispersed toward the central region of cytoplasm,and almost all the round mitochondria with rare cristae turned into hooded ones, and nucleus compaction occurred.In addition,the short and thick microvilli began to appear from the primary follicle ovocyte,become intensive and slender when secondary follicle’s ovocyte;It’s until tertiary follicle’s ovocyte,microvilli started to shorten and become coarse,and even parts of them contract from the zona pellucida gradually.Conclusion In the development of oocytes, the changes of type,quantity and distribution on mitochondria has a close relation with cells at proliferation,differentiation and metabolism level.Cortical granule has no association with golgi apparatus basically,but smooth endoplasmic reticulum(SER).The nuclei are the sites of RNA synthesis and warehouses,and its d Related Articles | Metrics

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Progeny of 2-cell embryo blastomeres distribute in mouse blastocyst randomly 2010, 41 (1):  124-127.  doi: 10.3969/j.issn.0529-1356.2010.01.024 Abstract ( )   Objective Kunming strain(KM) mice were used as animal models. Nontoxic dextran conjugated with tetramethylrhodamine（TMR）and fluorecein isothiocyante（FITC）was microinjected to two of the 2-cell blastomere as molecular probe to trace the development fate of the blastomere ,in order to figure out the mechanisms of the formation of Em-Ab axis. Methods FITC- dextran was injected to zygote in order to make sure if it is noxious. Two blastomeres of 2-cell embryo were injected FITC- dextran and TMR- dextran respectively. Results When labeled embryo develeped to blastocyst, distribution of progeny of 2-cell embryo blastomeres can be detected.Conclusion The cells of blastomere randomly distributed either embryonic parts or extra Related Articles | Metrics

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Expression of estrogen receptor alpha and beta in the uvea tissue of female rats 2010, 41 (1):  128-131.  doi: 10.3969/j.issn.0529-1356.2010.01.025 Abstract ( )   Objective To research the expression of the estrogen receptor alpha ( ERα) and estrogen receptor beta ( ERβ) in uvea tissues of the female rats, and to provide molecular biology data for further studies of the relation estrogen to uvea diseases. Methods Twenty-two adolescent SD female rats were selected. All rats were killed by dislocation of cervical vertebra, the eyeballs were in paraffin imbedding and made to a series of sections, using immunohistochemical method; ERα and ERβ distribution were investigated in uvea tissue of rats; and quantitied by Tanaka scores analytical method. The uteri of rats was used as positive control and PBS as negative control. The level of estradiol in serum of the rats were examined by radioimmunoassay. Results The expression level of ERβ was moderate or highter in stroma cell, anterior pigment epithelium as well as pasterior pigment epithelium of the iris, unpigmented epithelium, pigmented ciliary epithelium and vascular endocemet of the choroid layers. But ERα was not obviously expressed in uvea tissues. The expression rate of ERβ was higher than ERα in these tissues（EM>P/EM>0.05). Immnoreactivity positive substance was granule, which was distributed in the cytoplasm or in the nucleus. The level of estradiol in serum of the rats was (2 Related Articles | Metrics

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Expression of p16SUP>INK4A /SUP>in mouse endometrium during the estrus cycle and the early pregnancy period 2010, 41 (1):  132-136.  doi: 10.3969/j.issn.0529-1356.2010.01.026 Abstract ( )   Objective To investigate the expression of tumor suppressor gene p16SUP>INK4A/SUP> in mouse endometrium during the estrus cycle and early pregnancy and its possible role in blastocyst implantation. Methods Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of p16INK4A mRNA,immunohistochemistry and Western blotting were applied to detect p16SUP>INK4A/SUP> protein in mouse endometrium tissues during the estrus cycle and early pregnancy.　 Results The intensity of p16SUP>INK4A/SUP> mRNA expression in mouse early pregnancy was higher than that in the estrus cycle.Compared with the other 3 stages, the level of p16SUP>INK4A/SUP> mRNA expression at estrus was obviously higher. During the early pregnancy, the level of p16SUP>INK4A/SUP> mRNA expression increased steadily from day 2 to day 5,reaching the maximal level on day 5,then decreasing. Both immunohistochemical and Western blotting analysis showed the same results in expression patterns of p16SUP>INK4A/SUP> protein for mouse endometrium tissues as those results of RT-PCR.Conclusion p16SUP>INK4A/SUP> is involved in the embryos penetrating into the endometrial barrier. Related Articles | Metrics

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Radiologic measurement and sexual diagnosis of metacarpal in Kazakh nationality by discriminant analysis 2010, 41 (1):  137-140.  doi: 10.3969/j.issn.0529-1356.2010.01.027 Abstract ( )   Objective To study the sex difference and discriminating formulas by measuring both metacarpal bones on X-ray and body height of Kazakh nationality adults,providing anatomical data for physical anthropology and forensic medicine. Methods The metacarpal bones length on Xray films and body height from 200 adults of Kazakh nationality，including 100 male and 100 females were measured. The obtained date were statistically dealt by SPSS15.0 software. Results The length of the 4th metacarpal bone in male and the 5th in female were statistical significance beside, 4 discriminating formulas were obtained as follows: Y＝2.824r1＋2.563r4-0.654r5＋0.614l7-0.039l9＋2.452 l10-212.186，Y＝2.350r1＋2.377r4-0.995r5＋0.445l7＋0.046l Related Articles | Metrics

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Microanatomy and hemodynamic numerical simulation of the cerebral bridging veins entering superior sagittal sinus 2010, 41 (1):  141-146.  doi: 10.3969/j.issn.0529-1356.2010.01.028 Abstract ( )   Objective To simulate the hemodynamic feature in cerebral bridging veins (BVs), in order to provide a morphologic basis for the pathogenesis explanation and imaging diagnosis of cerebral venous thrombosis (CVT). Methods Totally 6 human cadavers (12 sides) were examined in this study. Each head of the cadavers was injected with blue-coloured latex via the superior sagittal sinus (SSS) and internal jugular veins. The diamter and the angle of BVs entering SSS were measured. Based on the data of cadavers and computational fluid dynamics software pack, the hemodynamic models were established. The wall shear stress (WSS) was carefully studied and compared between different models. Results The total of 137 BVs formed two clusters along the SSS: anterior group and posterior group. Compared with anterior group BVs, the diameter of posterior group BVs was large, and the angle was smaller. In 137 models，when the diameter of a BV was more than 1.2mm, and the angle was between 65 and 105 degree, the local WSS decreased in the downstream wall of SSS. When the diameter of a BV was more than 1.2mm, and the angle was less than 65 degree, the local WSS decreased in the downstream wall of SSS and the upstream wall of BVs. The minimum WSS in BVs was 63% of the minimum WSS in SSS. Compared with the anterior group BVs, the minimum WSS in the wall of posterior group BVs was samller, and the distance from the minimum WSS to the dural entrance was longer. Conclusion CVT occurs easily when the diamter of a BV is more than 1.2mm and the angle is less than 65 degree. The embolus forms early in the upstream wall of BVs entering the posterior part of SSS. Related Articles | Metrics

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Comparison of human mesenteric artery multislice spiral CT images with anatomy 2010, 41 (1):  147-152.  doi: 10.3969/j.issn.0529-1356.2010.01.029 Abstract ( )   Objective To measure and evaluate mesenteric artery by comparing the multi-slice spiral CT mesenteric artery images with autopsy specimens. Methods Totally 230 normal subjects were selected to undergo abdominal multi-slice spiral CT and enhanced CT. We processed the images, reconstructed 3D images, analyzed and compare the mesenteric artery images obtained by multi-slice spiral CT with autopsy specimens. Results 1. Diameters of mesenteric artery obtained by vivo image were significantly larger than that of autopsy specimens (EM>P/EM><0.05); 2. Start locations, branch types and running directions of both superior mesenteric artery and inferior mesenteric artery were much different between traditional autopsy specimens information and our results.3. Different reconstruction methods had different advantages. Especially, STS-MIP method could present the level of mesenteric artery better. Conclusion The method for mesenteric artery study using multi-slice spiral CT can enhance scanning and 3D reconstruction with workstation has been approved to work well, and it is superior to traditional autopsy specimen method. It is also convenient for mesenteric artery scientific evaluation. The result data of this method are reliable. Moreover, this method is available to research with large number of specimen. Related Articles | Metrics

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Optimal computer aided measure for screw internal fixation in the cavitas glenoidalis through human coracoid process of scapula 2010, 41 (1):  153-156.  doi: 10.3969/j.issn.0529-1356.2010.01.030 Abstract ( )   Objective To investigate a special optimization technique for computer aided measure, and provide anatomical basis for screw internal fixation in the cavitas glenoidalis through the coracoid process of scapula. Methods Thirty accurate scapula models were reconstructed from CT data sets. First, special optimization objective function was designed for single screw internal fixation configuration, and the optimal placement of screw was found automatically under constraints. Then, the placements of double screws internal fixation configuration were searched taking advantage of principal component analysis. Finally, statistical measure data were provided according to new anatomical reference landmarks for clinical use. Results For single screw internal fixation configuration, the distance from the optimal screw entry point P to the acromion process point X was (39.15±2.28) mm, to the coracoid process point Y was (28.66±2.68) mm, to the angulus superior point Z was (61.13±6.57) mm; The angle was (81.27±7.15)° between PX and PY, and (133.27±6.84)° between PX and PZ. The mean inclination of the lag screw was (104.08±4.41)° for the angle with line PX, (101.29±3.51)° with line PY, and (76.23±5.03)° with line PZ. For double screws configuration, the distance from the original single screw entry point P to the screw entry point E was (5.12±1.37)mm，to the screw entry point F was (3.88±0.94)mm. The angle between the long axis of coracoid process and li Related Articles | Metrics

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Measuring the displacement and deformation under the internal fixation with compressing steel plate for femoral fractures by digital speckle method 2010, 41 (1):  157-159.  doi: 10.3969/j.issn.0529-1356.2010.01.031 Abstract ( )   Objective Measuring the displacement and deformation of the fifth screw under different tensile force before or after adultal internal fixation with compressing steel plate for femoral fractuer by digital speckle correlation method.And analysis of the displacement and deformation after reducing the number of screws.Aimed at providing a theoretical basis for clinical femoral internal fixation with compressing steel plate about the number of screws. Methods Taking the antisepsis femoral specimen of grown-ups, embedding and fixing with tooth powder after removing the soft tissue, producing a fixed model in the 1/2 femur.Fixing steel plate below the lateral femur subperiosteal with five screws through the fourth floor of cortical bone on both sides of the fractures, samples were divided into 10 states, group A-J were measured by orders, under the rally of 400 N,500N , measuring the displayment of the fifth screw with CSS-44100-e-universal test machine for loading rally experiment ,and automatic data processing. Results For analysis of variance, the mean displacement measurements of the fifth screw under rally of 400N,500N, B-J group and A group have significant differences（EM>P/EM><0.05）.The mean deformation measurements of the fifth screw - under rally of 400N,500N, group A and group F-J group have significant differences（EM>P/EM><0.05）; group A and B-E group have no significant differences.Conclusion In a certain range, the fracture displacement and the numuber of the screws show negative correlation. The screws near the fracture line for femoral compression plate internal fixation of femoral fractuer is easily broken because Related Articles | Metrics

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Application of hydrodynamics-based transgene to gene transfection in rat fatty liver 2010, 41 (1):  160-164.  doi: 10.3969/j.issn.0529-1356.2010.01.032 Abstract ( )   Objective To study the condition and method of hydrodynamics-based transgene(HDT) in rat fatty liver. Methods Inject different dosages and concentrations of green fluorescent protein plasmid pEGFP-C1 at different speeds, then collect 4 rats’ liver leaves at different time points after injection and prepare their frozen section, finally observe and quantify the GFP expression with fluo rescence microscope at 488 nm excitation wavelength. Results Plasmid pEGFP-C1 concentration 33mg/L, injection speed 2ml/s, injection volume 8.5%of rat body weight, injected plasmid. After 6 hours of injection, GFP-positive cells rate of pedicel leaf is about 18%, left leaf about 14%, middle leaf about 12.5%, right leaf about 10% and tail leaf about 8%. GFP begin to gradually reduce since 24 hours, until 72 hours almost no GFP-positive cells were checked in all liver leaves. Conclusion Hydrodynamics-based transgene can be applied to rat fatty liver, the appropriate conditions of this method are 33mg/L plasmid concentration, 8.5% rat avoirdupois, 2ml/s injection speed, and the suitable time to observe the proportion of GFP-positive cells is 6-24 hours after gene injection. Related Articles | Metrics

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Improvement of mouse spermatogonial cells transplantation method 2010, 41 (1):  165-168.  doi: 10.3969/j.issn.0529-1356.2010.01.033 Abstract ( )   Objective To improve the current method of spermatogonial cells transplantation, and make the new method become more operative and easy to carry out. Methods The spermatogonial cells in rC57BL6/tg14 (act-EGFO-Osb Y01) mice that expresse GFP protein were collected as the donor cells, and by using a modified syringe, and then were injected into the seminiferous tubules of pretreated wild type GFP-null mice through testicular efferent duct. The transplantation outcome was evaluated by trypan blue stainting and fluorescent microscopic examination and pathohistological analysis of transplanted testis. Results The transplanted testis of recipient mice showed green fluorescence signal, and the signal of seminiferous tubules was found significantly higher than that of the surrounding tissues. The transplanted GFP-positive cells generated colonies and spermatogenesis but pretreated wild type GFP-null mice were not found. Conclusion The expressed GFP protein spermatogonial cells in rC57BL6/tg14 mice were successfully transplanted in the wild type GFP-null recipient mice, fourthermore the improved transplantation method simplified the triditional one and achieved the same transplantation results. Related Articles | Metrics