Objective To construct recombinent retroviral vectors containsins the expression sequence of hIL-4,and detect the expression of target genes.Methods Design the primer with restrictive enzyme spot and amplify target gene by PCR from plasmid including hIL-4 gene.Clone the target gene into retroviral vector pLXSN,and identify the aquired plasmid by sequencing.Transfect human synoviocytes with acquired virus in vitro.Detect the protein expresssion by western blotting.Results We successfully constructed the recombinant retrovirus-rRV-hIL-4.The hIL-4 gene was transduced into human synoviocytes by recombinant retrovirus in vitro.The protein expression of genes was detected by Western blotting.Conclusion Recombinant retrovirus rRV-hIL-4 is constructed successfully.The hIL-4 gene is transduced successfully into the human synoviocytes in vitro and the transduced synoviocytes can express hIL-4 protein.

Acupuncture Therapy ; Adult ; Humans ; Male ; Myopathies, Structural, Congenital ; therapy

Tyrosine phosphorylation is a key post-translational mechanism that regulates cellular processes and maintains homeostasis.Aberrant changes in tyrosine phosphorylation are often associated with disease states such as metabolicdisorders,cancer and cardiovascular disease.Protein tyrosine phosphatases (PTPs) are the enzyme family that regulates protein phosphorylation level of tyrosine residues in the cellular processes and signaling ways.So far,scientists have discovered 112 kinds of human PTPs.Among them,PTP1B is widely and clearly studied.Recently,as an enzyme that play a role in oncogenesis,PTP1B has been wildey studied by scientists.Here,we highlight the relationship between protein tyrosine phosphatase 1B and hematologic neoplasms.

Objective To investigate the expression and the roles of Angiopoietin 1(ang-1) and Angiopoietin 2(ang-2) in the endometrium of women with abnormal bleeding after medical abortion. Methods Analyzing endometrial pathological change of 1087 patients with abnormal bleeding after medical abortion in early pregnancy,and the endometrial specimens from 40 patients were randomly chosen for the study. The endometrial specimens from 20 women without abnormal bleeding after medical abortion were used as control group.Immunohistochemistry was used to detect the levels of Ang-1 and Ang-2 proteins in endometria. Results The percentage of patients with both the residul decidua and villus was 80.5%; positive immunoreactive signals of Ang-1 and Ang-2 were found in the endometrial glandular epithelium, stromal cells and the endothelial cells of vessels; the expression rate of Ang-1 and Ang-2 in the patients with abnormal bleeding were higher than that in the control group(P

0.05). The mortality in group A was obviously higher than that in group AI ( P

Objective:To investigate the effect of mycophenolate mofetil(MMF)on proliferation and apoptosis of cyst-lining epithelial cells in patients with autosomal dominant polycystic kidney disease(ADPKD),and to compare its effect with that of rapamycin(RAPA)in vitro.Methods: Primary cultured cyst-lining epithelial cells were treated with MMF and RAPA at different concentrations(0,0.005,0.05,0.5,5 ?g/ml)for 48 h or 72 h.The inhibitory effects of them on the cells were evaluated by MTT assay;the cell cycle distribution and apoptotic ratio were determined by flow cytometry.The morphological changes of cyst-lining epithelial cells were observed under transmission electron microscope.Results: Both MMF and RAPA significantly inhibited the proliferation of cyst-lining epithelial cells in a dose-and time-dependent manner.After 48 h treatment,the cells were blocked at S phase by MMF and at G0/G1 phase by RAPA.Both drugs induced cell apoptosis,with the maximal apoptotic rate being(5.53?0.27)% for MMF and(4.36?0.10)% for PAPA.Typical morphological changes of apoptotic cells were observed under electron microscope.Conclusion: MMF can effectively inhibit proliferation and induce apoptosis of cyst-lining epithelial cells,but its inhibitory effect is weaker than that of RAPA.

Objective To investigate the effect of rosiglitazone on p38 mitogen-activated protein kinase (p38MAPK) pathway in polycystic kidney cyst-lining epithelial cells. Methods The cyst-lining epithelial cells (PKD cells) from human polycystic kidney were treated with rosiglitazone (10 μmol/L), peroxisome proliferator-activated receptor-γ (PPARγ) inhibitor GW9662 (10 μmol/L), rosiglitazone (10 μmol/L) +GW9662 (10 μmol/L), p38MAPK specific inhibitor SB203580 (10 μmol/L), SB203580 (10 μmol/L)+ rosiglitazone(10 μmol/L) for 2 hours followed by epidermal growth factor (EGF) stimulation. Protein expressions of p38, phuspho-p38 (p-p38) and proliferating cell nuclear antigen (PCNA) were detected by Western blot. p38 mRNA was examined by RT-PCR. Expression of c-fos and c-jun was observed by immunocytochemistry. Results (1) EGF markedly up-regulated the expressions of p38, p-p38, PCNA, c-fos anti c-jun compared with control group (P<0.01). (2) Compared with EGF treated group, rosiglitazone significantly reduced p38 activation and mRNA expression (P<0.01, respectively). Rosiglitazone, rosiglitazone+SB203580 could significantly down-regulated p-p38, PCNA, c-fos and c-jun expression (P<0.01, respectively) with no significant difference between these two groups. (3) GW9662 partially reversed the reduction effect of rosiglitazone. Conclusions Rosiglitazone can inhibit proliferation of autosomal dominant polycystic kidney disease cyst-lining epithelial cells partially through down-regulating p38 activation and reducing c-fos, c-jun and PCNA expression. The above effect of rosiglitazone is in part PPARγ-independcnt.

Objective To observe the effect of jiaweibugan decoction on VEGF expression of sciatic nerve in experimental diabetic rat and explore the preventive and therapeutic effects of iiaweibugan decoction on peripheral neumpathy in~xperimental diabetic rats.Methotis The diabetic rat model was established by streptozotocin(STZ).The rats were killed on the 4th or 8th week from the beginning of treatment respectively,and VEGF mRNA of sciatic nerve was detected by reverse-transcriptase polymerase chain reaction(RT-PCR).Resuits The results of RT-PCR showed that VEGF expression ofthe model group and treated group on the4th or8th week WaS higherthan that of the normal control group(P＜0.05).Compared with the model group,VEGF expression in the diabetic rats treated byjiaweibugan decoction increased markedly at the end of4th week(P＜0.05),while decreaSed significantly at the end of8th week(P＜0.05).Conclusion Jiaweibugan decoction has a proventive and therapeutic effect on peripheral neumpathy in experimental diabetic rats.

Objective　To construct an anti-LPS single chain phage antibody library in mice for further biomedical works. Methods　Total RNA was extracted from splenic cells for reverse transcription after BALB/C mice had been immunized with pure LPS for 4 weeks. The designed primers were used to amplify the variable region genes of both heavy and light chain (VH,VL) with polymerase chain reaction. The VH and VL were then conjugated to form a single chain of variable fragment (ScFv) by a linker. The ScFv was cut by NotⅠ and SfiⅠ and then ligated into a pCANTAB5E phagemid vector which was digested with the same restriction enzymes. The ligated vector was then tranfected into the competent E.coli TG1 cells. Four TG1 clones were randomly selected to detect the exotic DNA. Results　The titer of anti-LPS in murine sera was 1∶12 800. The concentration of total RNA was 12.3813 μg/ml. The length of the fragments were 340 bp for VH, 320 bp for VL and 800 bp for ScFv respectively. 1.9×107 clones were grown after transfection and one from the four randomly selected clones was identified to contain the exotic DNA. Conclusion　A 4.75×106 murine anti-LPS single chain phage antibody library was successfully constructed.

Objective To study appliction value of quantitative dynamic contrast enhanced MRI(T1-DCE MRI)in preoperative grading of brain glioma.Methods 80 patients who were pathologically confirmed with a tumor grade (WHO grade Ⅰ 20 cases, grade Ⅱ 20 cases,grade Ⅲ 20 cases and grade Ⅳ 20 cases).All patients were examined with MR enhancements and T1-DCE MRI. The original perfusions imaging datas were analyzed using the GE Omni Kinetic software,which produced the transfer constant (Ktrans )map,the rate constant (Kep )map and fractional volume (Ve )map.Choose ROI and get values of Ktrans ,Kep and Ve .Pearson correlation was carried out to analyze the correlation between values of Ktrans ,Kep ,Ve of different grades of gliomas and pathology classifications.The Ktrans ,Kep and Ve values of the different grade gliomas were statistically analyzed using an ANOVA .Receiver operator characteristics (ROC)curve was used to analyze sensitivity and specificity of permeability parameters.Results The Ktrans ,Kep and Ve values of each levels has a strong correlation with pathological grading (r=0.95 1,0.804,0.766).There was obviously statistically significant difference between different grade groups(P < 0.01 )by Ktrans .Kep values have statistically difference between different grades except grade Ⅱ and Ⅲ.Ve values were different between different grades except grade I andⅡand grade Ⅲ and IV.Accord-ing to ROC curve,Ktrans seemed to be a better parameter for evaluating the tumor grade with the highest sensitivity and specificity. With the cutoff thresholds of Ktrans of 0.1 60,0.420 and 0.935,different grades of glioma can be differentiated with sensitivities of 90%,95%,95% and specificities of 95%,95%,85% respectively.Conclusion Quantitative analysis of microcirculation perfusion status of different grade gliomas by Ktrans values obtained from T1-DCE MRI can assessment the degree of the destruction of the blood brain barrier and evaluate the grade of gliomas more accurately before operation.