Table of Content

25 April 2007, Volume 27 Issue 4

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研究论文

Relation of quantitation of spermatozoan surface wheat germ agglutinin(WGA) Receptor and fertilization rates of metaphase IIoocytes in vitro

2007, 27(4):  361-363. 

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Objection To determine the predictive value of wheat germ agglutinin receptor (WGAR) on human spermatozoa surface, the frequency of abnormitic sperm morphology and human spermatozoa acrosin activities to fertilization rate of IVF-ET.Methods Spermatozoa and oocytes were obtained from both normospermic men and women patients with oviduct obstruction,They came to our center for IVF procedure. Statistics of the quantitation of WGAR on spermatozoa, the frequency of abnormitic sperm morphology, human spermatozoa acrosin activities and the fertilization rate of IVF-ET were made. Result Two groups based on the fertilization rate less than 65% (n=64)and over 65%(n=194) were divided, there was a significant difference about their amount of the WGAR(P<0.01);there was not significantly different about the frequency of abnormitic sperm morphology and human spermatozoa acrosin activities(P>0.01;P<0.05). Conclution The fertilization rate of IVF-ET was affected by the amount of WGAR on spermatozoa, the frequency of abnormitic sperm morphology and human spermatozoa acrosin activities,while the amunt of WGAR on spermatozoa might be more useful to predict fertilization rates in vitro

Proteomic analysis of human nasophryngeal carcioma cdll: the proteins associated with malignant activities

2007, 27(4):  364-371. 

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Objective To find genes associated with malignant activities of human nasopharyngeal carcinoma cell CNE-2L2. Methods Differential display of proteins between the CNE-2L2 cell with high malignant activities (called W cell) and the CNE-2L2 cell with profound reduced malignant activities (called AS cell) was analyzed by proteomic technique. The differential display was validated by real-time PCR or immune fluorescence staining. Expression of CD44 gene was inhibited by means of siRNA. The inhibition was validated by means of Western blotting. Tumorigenesis of cells was examined in nude mice. Results MOLDI-TOF MS showed 12 protein spots differential displayed. They were: Peroxiredoxin 2 isoform b；Tropomyosin 3；TPM4-ALK fusion oncoprotein type 2；MLL protein；CD44；Retinoblastoma binding protein 7；Tubulin, b polypeptide；ATP synthase，H+ transporting, mitochondrial F1 complex, b polypeptide；Vimentin；D21S2056E protein；Proteasome (prosome, macropain) 26S subunit, non-ATPase，4；ENO1 protein。The former 4 were up-regulated and the latter 8 down-regulated in AS cell. The differential display of the 7 proteins with highest scores was confirmed. Tumorigenesis of the CNE-2L2 cell with inhibition of CD44 gene expression caused by siRNA in nude mice was profoundly inhibited. Conclusion Proteomic analysis showed 12 proteins differential displayed between AS cell and W cell. Inhibition of CD44 gene expression resulted in profound reduction of the malignant activities of CNE-2L2 cell.

Association of PTEN with NHERF-1

2007, 27(4):  372-376. 

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Objective To identify PDZ domain containing proteins interacting with PTEN and its characterization with NHERF-1 by proteomic analysis. Methods The interactions between PTEN and PDZ domain containing proteins were screened with PDZ protein array, and the novel one was then identified with GST pull-down and co-immunoprecipitation assay. Results Using a PDZ protein array, we found PTEN　binding with NHERF-1. The interaction of PTEN and NHERF-1 was further characterized by GST pull down assay, and demonstrated that PTEN associated with NHERF-1 via the binding of PTEN carboxyl-terminal with the PDZ domain 1 (PDZ1) of NHERF-1. The last four amino acids (I-T-K-V) of the PTEN were the key determinants of this interaction as mutation of any of the four amino acids to alanine resulted in markedly reducing association of PTEN with NHERF-1. In addition, full-length of PTEN robustly associated with NHERF-1 was also determined by co-immunoprecipitation experiment in cos-7 cells. Conclusion The interaction of PTEN and NHERF-1 was identified both in vitro and in cells. PTEN/NHERF-1 association was mediated via the binding of PTEN carboxyl-terminal with the PDZ1 of NHERF-1, and the last four amino acids of the PTEN carboxyl-terminal were important for PTEN/NHERF-1 interaction.

Hypoxia enhanced the differentiation of human bone marrow-derived mesenchymal stem cells into dopaminergic neurons

2007, 27(4):  377-381. 

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Objective The aim of the study is to investigate the effect of hypoxia on the differentiation of human bone marrow-derived mesenchymal stem cell (hMSCs) into the dopaminerigic neurons. Methods The numbers of differentiated cells were tested by the methods of immunocytochemistry and flow cytometry, and the content of dopamine and its metabolits are measured by HPLC-ECD. Result The number of TH positive cells differentiated from BME-induced hMSCs under hypoxic condition increases about 3 times as compared with BME-induced hMSCs under normoxic condition. Furthermore, the content of dopamine (DA) and its derivation homovanillic acid (HVA) synthesized by differentiated cells in hypoxia group are significantly higher than normoxia group (P<0.01 and P<0.05 respectively). Conclusion Hypoxia could improve the differentiation of human bone marrow-derived mesenchymal stem cell into dopaminergic neurons; this provides a new clue for clinical treatment of Parkison's Disease with hMSCs.

Assesment of low quantitative ultrasound values of calcaneus in chinese mainland women

2007, 27(4):  382-385. 

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Objectives To study the rule of ultrasonic bone mineral density of calcaneus with age, height and body weight, and establish the normal reference value of stiffness index (SI) of females in Chinese southern region and provide scientific foundation for osteoporotic diagnosis and prevention. Methods SI for calcaneus in 2498 healthy people 10-87 years old were measured with Achilles Express ultrasound apparatus made in USA. They were divided into groups according to sex and age. One group for 10 years each and the records beyond 69 years were classified into one group, total 7 groups. Results The SI exhibited a characteristic mild rise-then-fall pattern with increasing age. And the peak value of SI presented in 20～29 age group. Pearson correlation analysis showed negative correlation between SI and age and positive correlation between SI and height and weight. The prevalence of osteoporosis increased gradually with age. Conclusion There were significant correlations in SI with age, height and weight; the values of SI will provide an important data reference to establish the normal values and diagnostic standard of osteoporosis.

The culture and phenotypic examination of human fetal hepatocytes in vitro

2007, 27(4):  386-390. 

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Objective: Determining the characteristics of human fetal hepatocytes in vitro. Methods: Isolating and culturing human fetal hepatocytes by stepwise trypsinization of liver fragment in vitro; collecting the culture medium to determine the quantity of AFP, ALB and the functional enzymes (including ALT, AST, GGT, ALP and LDH) of different generations in culture; determining the expression of cytochrome C by immunohistochemistry; testing the effects of sodium byturate on human fetal hepatocytes. Results: Human fetal hepatocytes were polygonal epithelial cells in DMEM medium. They could be maintained for 5.5 months (about 30 passages) in vitro. They secreted ALB and functional enzymes all over their cultivation. Conclusion: Human fetal hepatocytes can be maintained and keep function in vitro for several months.

Research on the SMAD4-independent pathway of TGF-β1 in human pancreatic cancer by suppression subtractive hybridization

2007, 27(4):  391-397. 

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Objective：To elucidate the smad4-independent signaling pathway of TGF-β 1in the pancreatic cell carcinogenesis Methods: We transfected pancreatic cancer cell line, BxPC3 which carries homozygosity loss of SMAD4 , with TGF-β1 and pcDNA3 as a control. Cell growth was detected by flow cytometry (FCM) and expression of TGF-β1 detected by western blot. By the technique of suppression subtractive hybridization (SSH), we constructed a substracted cDNA library of TGF-β1 related genes. Amplification of the library was carried out with the E. coli strain JM109. Reverse northern blot was used to confirm the genes differentially expressed after TGF-β 1 functioned. Results: The growth of BxPC3 was slightly inhibited after transfected with TGF-β and overexpression of this cytokine was convinced by western blot. TGF-β 1 related subtractive library with high subtractive efficiency was set up successfully. The amplified library contained 300 positive clones. Reverse northern blot showed that 32 clones were actually differentially expressed. After sequencing and blastn searching, we found 10 genes up-regulated and 12 down-regulated,13 of which habour known function and 9 unknown. Conclusions: With this subtracted cDNA library, it is easy for us to reveal the molecular mechanism in the pancreatic carcinogenesis.

The specificity of airway epithelial cells transfected by elafin on bacterial biofilm

2007, 27(4):  398-401. 

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Objective To explore the different effect of elafin incubated by different bacteria on P.aeruginosa(Pa) bioflim. Methods To cultivate the A549 cells in vitro, the pEGFP-N1-elafin eukaryotic expression vectors have been transfected to the cells by LipofectamineTM 2000. Elafin transfected cells incubated by the supernatant of S. epidermidis (S.epidermidis group), Pa (Pa group) and E.coli (E.coli group) respectively 24 hours，the A549 cells were transfected the vector without elafin gene as normal group. Then the levels of elafin were detected by ELISA and Western Blot. To establish the Pa biofilm model in vitro，a rapid silver nitrate staining procedure and scanning electronic microscope (SEM)demonstrated bacterial biofilm. After biofilm carriers put into each group and incubated 8 hours, we measured the proportion of bacteria biofilm by silver nitrate staining in light mocroscope and observed the structure of biofilm by SEM in each group. Results The Pa and E. coli groups（especially Pa） induced the content of elafin in cells and the level of secretion increasing compared to the normal group, while the S. epidermidis group have no increase. Both silver nitrate staining procedure and SEM demonstrated bacterial biofilms. The the proportion of bacteria biofilm and the structure of BF in Pa and E. coli groups were changed, especially in Pa group. Conclusions There was specificity of different bacteria to induce the express of elafin. The inducing effect of Pa were exceed that of E.coli, S. epidermidis had厮 no obvious effect. Elafin could clean the bacteria of biofilm.

Safety and feasibility of autologous bone marrow mesenchymal stem cell transplantation for acute myocardial infarction in pig

2007, 27(4):  402-407. 

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Objective To test the feasibility and safety of autologous bone marrow mesenchymal stem cell (MSC) transplantation by intracoronary injection in acute myocardial infarcted pig (AMI). Methods Coronary occlusion with balloon was used to produce AMI.Labeled MSCs were implanted by intracoronary injection.Four weeks after AMI,single photon emission computed tomography was used to evaluate the relative infarct size,myocardial perfusion score and ejection fraction (EF) of the animals. Left ventricular systolic pressure( LVSP), left end-diastolic pressure (LVEDP), peak rate of pressure rise(+dP/dt) and peak rate of pressure fall (-dP/dt) were assessed by catheterization. Immunofluorescence was performed to evaluate MSC engrafment and differentiation. Results Compared with control group, heart function parameters were all improved in pigs receiving MSCs transplantation, including EF [（44.1±4.3）% vs (39.0±5.2)%, p<0.05] perfusion score （30.0±5.3 vs 41.8±8.8, p<0.01）, relative infarct size[（28.6±4.7）% vs (33.5±3.4)%, p<0.05], LVSP（87.1±8.7mmHg vs 78.0±4.4 mmHg, p<0.05）, +dP/dt（1818.1±116.7mmHg/s vs 1610.6±114.5 mmHg/s, p<0.01） , -dP/dt（1800.9±88.5mmHg/s vs 1618.4±65.7 mmHg/s, p<0.01） and LVEDP（9.4±1.7mmHg vs 12.6±2.8 mmHg, p<0.05）. MSCs implanted in myocardium were stained positively for myosin heavy chain, connexin 43, smooth muscle actin and von Willebrand factor. Conclusions Transplantation of MSCs could improve cardiac function in AMI pig, and that the improvement might result from myocardial regeneration and angiogenesis by MSC differentiation.

The effects of NGF on the caspasecascade reaction induces by caspase-12 in cerebral ischemia/reperfusion injury

2007, 27(4):  408-411. 

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Objective To observe the effect of NGF on the caspase-12 in cerebral ischemia-reperfusion, and elucidate the protective mechanism of NGF on cerebral tissue underwent ischemia-reperfusion in rabbit. METHODS Twenty-six healthy male white rabbits weighting （2.6±0.2）kg were randomly divided into sham operation group, ischemia-reperfusion group and NGF group. The tissues of ischemia-reperfusion region were taken to detect the expression of caspase-12, caspase-3 by immunohistochemistry and to analyze them with Motic 6.0 image analysis system, and to detect the I/R cerebral apoptosis by flow cytometry and TUNEL staining methods. RESULTS The caspase-12 and caspase-3 expresses in a low level, a few apoptotic cells were detected by FCM and the TUNEL staining in sham operation group. As compared with that in sham operation group, the expression of caspase-12 and caspase-3 in I/R group and NGF group were higher (P<0.01); the apoptotic cells detected by FCM and TUNEL methods were significantly increased, as compared with those of sham operation group(P<0.01). As compares with those of I/R group, the levels of caspase-12 and caspase-3 in NGF group were lower than that of I/R group, the percentage of apoptotic cells was lower than that of I/R group (P<0.01). CONCLUSION NGF can decrease the number of apoptotic cells of the cerebral ischemia-reperfusion, inhibit the caspase cascade reaction induced by caspase-12, which is one of the protective mechanisms of NGF.

Effect of 5-aza-2'-deoxycytidine on cell proliferation and apoptosis in ovarian cancer cell line and expression of hMLH1 and hMSH2

2007, 27(4):  412-416. 

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Objective To observe the effects of 5-aza-2'-deoxycytidine(5-Aza-CdR)on cell proliferation and apoptosis in ovarian cancer cell line SKOV3 and the expression of mismatch repair gene HMLH1 and HMSH2, and to investigate the potential mechanism of its antitumorigenesis. Methods Human ovarian cancer cell line SKOV3 was treated with 5-Aza-CdR(0.5 , 5 and 50μmol/L), a specific demethylation agent for 3d, and then cultured in RPMI-1640 medium for 7d.The growth of cells was observed by MTT assay. The apoptosis was analyzed by flow cytometry. The expression of HMLH1 and HMSH2 mRNA was observed by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). Results SKOV3 cells treated with 5-Aza-CdR displayed a slowed growth rate in comparison with that of the control cells, The apoptosis rate of each group were 10.59%±1.57%、17.52%±1.72%、34.10%±1.45% ,respectively ，which were markedly higher than that of control（P<0.01）.The apoptosis was also notably correlated with 5-Aza-CdR concentration（F=227.6，P<0.01）. After 0.5, 5 and 50μmol/L 5-Aza-CdR treatment, the level of HMLH1 and HMSH2 mRNA expression was increased differently, and it was significantly correlated with the concentration of 5-Aza-CdR. Conclusions 5-Aza-CdR can inhibit the proliferation and partly induce the apoptosis of human ovarian cancer cell line SKOV3 by inducing the expression of HMLH1 and HMSH2 genes ，which may be inactivated by hypermethylation. Methylation of mismatch repair genes plays an important role in the carcinogenesis and progression of ovarian cancer.

Focal adhesion kinase prompoted human pulmonary smooth cells enhncement

2007, 27(4):  417-420. 

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Obiective:To study whether focal adhesion kinase (FAK) prompte human pulmonary artery smooth cells (HPASMCs) proliferation .Method: Cultured HPASMCs stimulated by fibronectin (40mg/L) were passively transfected with sense -FAK oligonucleotides(ODNs), FAK activity was measured by immunoprecipitation and expression of FAK protein was detected by Western blots .Meanwhile, The change of cell proliferation were measured by MTT and 3H-TdR absorbation experiment. Results: The changes of FAK activity and FAK protein content exhibited dose and time dependence at diferential concentration and time passively transfected with sense-FAK ODNs in Cultured HPASMCs. At the same time, sense-FAK ODNs prompoted HPASMCs proliferation and 3H-TdR absorbation .Conlusion: FAK can faciliate HPASMCs proliferation multiplication,which may play an important function in pulmonary artery hypertension development.

Renal protective effect of mycophenolate mofetil in diabetic rats

2007, 27(4):  421-425. 

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Objective To assess the renal protective effects of mycophenolate mofetil(MMF) in diabetic model rats and explore its mechanism. Methods SD rats were divided randomly into there groups: control group,diabetic model group and diabetic group treated with MMF. Observed twelve weeks, blood glucose(BG), blood urea nitrogen(BUN), serum creatinine(Scr), index number of kidney hypertrophy(kidey weight to body weight, KW/BW),creatinine clearance(Ccr) and 24- hour urinary protein (24Upro) were detected. Kidney tissure were examined by light microscopy and electron microscopy. The protein exprssion of intercellular adhesion molecule-1 (ICAM-1) and proliferating cell nuclear antigen(PCNA) in kidney tissure were determined by immunohistochemical technique. The mRNA expression of ICAM-1 in kidney tissure were semi-quantitatively determined with reverse transcription-polymerase chain reaction(RT--PCR). Results Compared with control group, BG ﹑KW/BW﹑24Upro﹑BUN﹑Scr and Ccr were significantly increased in diabetic model rats(P<0.05 or 0.01), the density of mesangium and the depth of glomerular basement membrane were significantly enlarged(P<0.01), the protein expression of ICAM-1 and PCNA and the mRNA expression of ICAM-1 were significantly up-regulated (P<0.05or 0.01). Except blood glucose, the above mentioned parameters in MMF treated group were all significantly inhibited(P<0.05 or 0.01). Conclusion MMF showed the protective effect for diabetic nephropathy. The mechanism may be correlated with that it down-regulate the exprssion of ICAM-1 and PCNA.

Exploring the relationship of angiogenesis and GH-Secreting Pituitary Adenoma

2007, 27(4):  426-431. 

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The aim of this study was to investigate MVD, TSP-1, TGF-β1 expression in GH-Secreting Pituitary Adenomas by immunohistochemistry, and to correlate data with clinical characteristics. Method: The protein expression of TSP-1,TGF-β1 in 48 surgical specimens (21invasive cases; 27 noninvasive cases)of pituitary adenomas was measured using immunohistochemical method，and explored the relationship between the expression and invasion of adenomas. MVD were measured by immunohistochemical method with detecting CD34. Results: Compared with the noninvasive group, no differences of expression of CD34 (t=2.257; P=0.083) was observed in the study. The expression of TSP-1 in invasive group was lower. The expression of TGF-β1 was stronger in invasive cases than in noninvasive ones. The expression of TGF-β1 had a positive correlations with MVD; but there was no correlations between the expression of CD34 and the invasion of pituitary adenomas. In addition, MVD count was not associated with the expression of TSP-1. Size, sex or rate of recurrence did not influence MVD and TSP-1 expression. Conclusion: These results suggest that evaluation MVD values do not necessarily represent angiogenesis in pituitary adenomas.However,TGF-β1 maybe involved in upregulating the expression of MVD, and TSP-1 do not influence MVD in pituitary adenomas and maybe regulate angiogenesis by other pathway.

Effects of fluvastatin on intercellar adhesion molecules-1 expression induced by C reactive protein in human ujmbilical vein endothelial cells

2007, 27(4):  432-435. 

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Objective To study the effects of Fluvastatin on the expression of intercellular adhesion molecule-1 (ICAM-1) in human umbilical vein endothelial cells (HUVECs) induced by C-reactive protein (CRP). Method The HUVECs were primary cultured . HUVECs of the third to fifth generations were stimulated in different concentration CRP of 5,10,50 and 100mg/L and different time of 6, 12 and 24h. Then the cells were treated with Fluvastatin in different concentration of 10-7,10-6 and 10-5mol/L. The content of ICAM-1 protein was detected with ELISA, the mRNA expression of ICAM-1 was evaluated by RT-PCR. Results In the control group, HUVECs produce ICAM-1 protein and mRNA in low concentrations. In the CRP group, the content of ICAM-1 protein was increased significantly (p<0.01) and the mRNA expression of ICAM-1 was increased significantly (p <0.01). The increasing effect of ICAM-1 protein was concentration -dependent and time- dependent (p<0.01). In the Fluvastatin group, the content of ICAM-1 protein and the mRNA expression of ICAM-1 was decreased significantly (p<0.01). The inhibitory effects of fluvastatin on ICAM-1 protein was concentration -dependent (p<0.05). Conclusion Fluvastatin maybe exert the function of anti- atherosclerosis by inhibiting the expression of ICAM-1 induced by CRP in HUVECs.

Effect of transforming growth factor β1 on expression of vascular endothelial growth factor on hepatoma cells induced by hypoxia

2007, 27(4):  436-439. 

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Objective: To observe the effect of Vascular Endothelial Growth Factor(VEGF) expression in hepatoma cells affected by TGF-?1 and hopxia . Methods: HepG2 cells were cultured? invitro, and treated with different doses of TGF-?1 and Cobalt chloride hexahydrate(CoCl2), or treated with TGF-?1 and CoCl2. The changes of VEGF protein and mRNA were detected by immunohistochemistry and semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). Results: Immunohistochemistry and RT-PCR showed that the expression of VEGF protein and mRNA were significantly higher in TGF-β1 groups or CoCl2 groups than that of control group(p<0.01), they increased the expression of VEGF in concentrate-dependently manner. The addition of TGF-β1 with CoCl2 results in significantly more VEGF expression than either stimulation alone(p<0.05). Conclusion: TGF-?1 can increase the expression of VEGFmRNA in Hepatoma cells；TGF-?1 can coordinate with hypoxia induced by CoCl2 to increase the expression of VEGFmRNA.

Effect of singal transducer and activator of transcription -1 on pulmonary inflammatory response in septiv rats

2007, 27(4):  440-444. 

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Objective The purposes of the present study were to assess ①the effect of signal transducer and activator of transcription （STAT） on pulmonary injury induced by cecal ligation puncture (CLP) in septic rats; ②the significance of interleukin (IL)-6 and IL-10 expression in pulmonary injury mediated by STAT in septic rats. Methods Sepsis of rats was induced by CLP. 56 male Wistar rats were randomly divided into normal control (n=8), CLP group (n=24), and inhibitor (rapamycin, RPM) of STAT pretreatment group (n=24). At serial time points in each group, animals were sacrificed. Then, pulmonary tissue and serum samples were harvested to determine IL-6 and IL-10 mRNA expression levels by reverse transcription polymerase chain reaction (RT-PCR) and protein expression levels by enzyme-linked immunosorbent assay (ELISA). Meanwhile, pulmonary STAT1 DNA-binding activity was detected by electrophoretic mobility shift assay (EMSA) and activity of myeloperoxidase (MPO) as well as histopathology were also evaluated. Results Compared to normal control, pulmonary STAT1 activity at 6h, 24h and 48h following CLP significantly elevated (P<0.01). In the meantime, pulmonary MPO, histopathologic scoring as well as IL-6 and IL-10 mRNA and protein expression levels in lung or serum markedly increased respectively. Following pretreatment with RPM, pulmonary STAT1 activity, MPO and histopathologic scoring decreased compared to CLP group, IL-10 expression significantly elevated, IL-6 levels without alteration at the same time. Conclusion These data suggest that severe intra-abdoment infection elevates pulmonary STAT1 activity and induces injury of remote lung, and inhibiting the activation of STAT1 attenuates pulmonary tissue damage, however contributions from STAT1 driven expression of IL-10 as well as IL-6 may be needed in a balanced fashion to maximize the animals' ability to survive septic challenge.

技术与方法

A study on mochanism of dermal fibroblasts as feeder layers to support the growth of human keratinocyes

2007, 27(4):  445-448. 

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Objective To study the mechanism of dermal fibroblasts as feeder layer to support the growth of human keratinocytes. Methods Human dermis fibroblasts were isolated and cultured and then treated with mitomycin-C and the expression of type Ⅰand type Ⅲ precollagen mRNA and relevant protein in feeder layer were examined by RT-PCR and Immunohistochemistry. KCs were cultured both on FB and NIH3T3 feed layer as control， the adhering numbers and the time of fusion were observed. Results RT-PCR revealed an increase of type Ⅰprecollagen mRNA in FB feeder layer compared with that of normal fibroblasts （P＜0.05）and no change was seen in type Ⅲ precollagen. Positive stains of collagen type Ⅰ and Ⅲ were shown in the FB feeder layer and those KCs grew and differentiated well on the FB feeder layer seeded at appropriate densities. There was no statistical difference in the adhering numbers and the time of fusion in two different culture feeder layer. Conclusions Human dermal fibroblasts as the feeder layer could secrete collagen type Ⅰ and Ⅲ which will contribute to multi-layered sheets of cultured keratinocytes.

Compraison of different extraction methods of alternaria alternate allergens

2007, 27(4):  449-452. 

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objective In order to find one of the best methods to make the extraction of alternaria alternate, in this study, we made different methods to compare the influence on the extraction allergen. Methods Alternaria alternate was gotten from the air and identified by Chinese Academy of Sciences, cultured under 26℃ for four weeks. Protein extracted by different methods and its concentration was determined using the Bradford assay， its bioactivity was tested by RAST inhibition, and residual proteins were analyzed by SDS-PAGE. Result The extraction methods, ranging from grinder（a）, grinder plus ultrasonic(b), ground with liquid nitrogen(c) and liquid nitrogen plus ultrasonic(d). The concentration of protein extractions were as follows （g/ml）: a was 0.44±0.04；b was 0.75±0.03；c was 0.72±0.03 and d was 1.29 ±0.05. From SDS-PAGE, we could find that we got the most component in the extraction by liquid nitrogen plus ultrasonic. From RAST, 50% inhibition as follows(μg/ml): a was 8.5, b was 9.4, c was 7.0,d was 3.7. Conclusion: Ground with liquid nitrogen plus disintegration by ultrasonic was the most effective method.

Establishment of artery transpantation model of chronic rejection in rat

2007, 27(4):  453-454. 

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The culture characters of neural stem derived from spinal cord of rats in different time after birth

2007, 27(4):  455-456. 

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临床园地

Reversded island flap pedicled with accompanying vesses of the cutaneous nerves in the liwer leg anatomy and design

2007, 27(4):  462-465. 

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Objective To study the blood supply of the flap accompanying vessels of the cutaneous nerves in the lower leg, and design the reversed flap for clinical reference. Methods Anatomic observation was performed on 30 adults' lower extremity specimens perfused via pressure with red latex through femoral arteries. Results Superficial peroneal nerves, sural nerves and saphenous nerves are all nourished by their accompanying arteries which, anastomosed with the cutaneous perforating branches of other arteries, also nourish the corresponding skin areas. Conclusion The blood supply of the reversed island flap accompanying vessels of the cutaneous nerves in the lower leg proves reliable and makes it possible to design long flaps along the cutaneous nervous axis.

研究短文

The study on the effect of L-arginine in pulmonary hypertension therapy

2007, 27(4):  466-467. 

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HOXA-11 gene and female reproductive reguation

2007, 27(4):  470-471. 

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协和大查房

Fever,hypocytosis and osteodynia

2007, 27(4):  472-475. 

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讲座

The discrimination and explanation of statistical errors in the expression and description of statistical data

2007, 27(4):  476-480. 

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