Objective To evaluate the significance of t he abnormality of VHL gene and VEGF expression in primary renal cell carcinoma ( RCC). Methods Mutation and abnormal methylation of VHL g ene and the expression of VEGF were dtected by means of multiplex PCR,PCR-SSCP and immunohistochemistry in 42 cases of primary RCC,in 18 specimens adjacent to the neoplastic lesion and in 10 specimens of normal renal tissues. R esults The alteration rate of VHL gene was higher in RCC tissue (61. 9%) than in the normal renal tissue (3.6%,P

Objective:To study the effects of the overexpression of hsa-miR-202 on the proliferation and molecular mechanism of lung cancer A549 cells. Methods:A sequence of hsa-miR-202 with ppG/miR/eGFP/Blasitidin pasmid was directionally connected and a eukaryotic expression vector pmiR-202 of the target hsa-miR-202 gene was constructed. pmiR-202 was transtected to A549 cell with Lipofectamine 2000. The WST assay was used to detect the cell proliferation rate, and RT-PCR was used to detect the relative gene expression levels. Western blot analysis was used to detect the IL-10 protein expression levels. The interaction between miR-202 and IL-10 was examined using a luciferase reporter assay. Results:The design from the DNA sequencing results shows that a eukaryot-ic expression vector of miR-202 was successfully constructed. The proliferation inhibition rates of A549 cells by Pmir-202 were 12%, 38%, and 52%. The differences in the treatment group compared with the blank control and negative control groups were statistically significant. The RT-PCR results showed that the relative expression levels of miR-202 increased after transfection with pmiR-202. Over-expression of miR-202 can downregulate the relative gene and protein expression levels of IL-10, and the relative levels were 25%and 0.75, respectively. Compared with the blank control and the negative control groups, the difference was statistically significant (P<0.05). The fluorescent activity was reduced when transfection was performed with miR-202 mimics, and IL-10-3'UTR plasmid was cloned. Conclusion: pmiR-202 effectively inhibited the proliferation of A549 cells and exhibited a time-effect relationship with miR-202 by targeted combination with IL-10 3'UTR to downregulate IL-10 expression in A549 cells.

Objective To investigate the effect of hypoxia on the expression of vascular endothelial growth factor (VEGF) in villous explants, and the effect of hypoxia-inducible factor 2? (HIF-2?) on the VEGF expression. Methods The villous were obtained from women who underwent suction and curettage for termination at 6-8 gestational weeks. Four groups were divided: normoxia group; hypoxia group; sense HIF-2? oligonucleotide group and anti-sense HIF-2? oligonucleotide group. All villous were cultured in vitro. In anti-sense and sense group, villous were incubated under hypoxia with HIF-2? oligonucleotide. The villous explants were collected after 48 h of cultivation. The protein level of HIF-2? was detected by Western blot and the mRNA of HIF-2? and VEGF were examined by real-time PCR. Results The expression of HIF-2? mRNA and VEGF mRNA as well as the protein level of HIF-2? were increased in hypoxic group comparing with the normoxia group. Compared with the sense group, the expression of HIF-2? protein and HIF-2?, VEGF mRNA showed significant decrease in anti-sense group. Conclusions Hypoxia may result in the increased expression of VEGF in villous explants during the early trimester, which might be mediated by HIF-2?.

OBJECTIVE: To optimize the extraction process of Fructus Schisandrae Chinensis. METHODS: Taking the contents of schizandrin, schisantherin A, deoxyschizandrin, r-schizandrin as the indexes, the influence on the extraction process by extract solvent and medicinal material particle diameter was examined, and the extraction parameters were optimized. RESULTS: Active components of Fructus Schisandrae Chinensis, after the medicinal material being crushed, can be effectively utilized through extracting by alcohol. The optimized extraction parameters were as follows: the medical material being ground into 40 meshes, extracted twice by 70% alcohol, 2 hours per time. CONCLUSION: The optimized extracting process works better than current technique adopted for formulated granule preparation, and can ensure quality to the highest degree.

OBJECTIVE: To optimize the process for extraction and purification of Schisandrin in Fructus Schisandra Chinensis. METHODS: The orthogonal test L9( 34) was adopted to optimize the water extraction process using Schisandrin and its extracts as indicators. The parallel test was used to optimize the parameters of the alcohol precipitation technique. RESULTS: The optimized process for extraction and purification of Schisandrin was as follows: extracting three times using 10- fold water, 1. 5h each time, with filter liquids concentrated to 1. 4g of crude drug mL- 1 then precipitated to 80% ; filtering alcohol liquids, and adjusting pH value to 7. CONCLUSIONS: This method can be used as a reference for the extraction of Fructus Schisandra Chinensis.

Objective To observe enhancing effect of nerve regeneration on peripheral nerve defect models bridged by a new PRGD/PDLLA/VPA composite conduit.Methods In this study from February,2012 to March,2014,PRGD/PDLLA/VPA nerve conduits were tested in the rat sciatic nerve transection model.At different periods after operation,its ability to promote nerve regeneration was evaluated by sciatic functional index(SFI),electrophysiology (CMAPs,NCVs) and histologic assessment.Forty rats were randomly divided into 4 groups (n =10),group A:PRGD/PDLLA/VPA,group B:PDLLA/VPA,group C:PRGD/VPA and group D:autograft.Results At 12 weeks after surgery,the SFI value of group A (-45 ± 3.19)and group D (-42 ± 3.01)were significantly higher than those of group B(-79 ± 3.06) and group C(-72 ± 2.07)(P ＜ 0.05);The CMAPs of group A (24.89 ± 5.01) and group D (25.39 ± 5.63) were significantly higher than those of group B(14.88 ± 3.11) and C(15.00 ± 5.54);the NCVs of group A (31.42 ± 2.43) and group D (31.50 ± 2.16) were significantly higher than those of group B (20.11 ± 2.39) and group C(21.00 ± 2.13)(P ＜ 0.05).At 12 weeks after surgery,the numbers of regenerated nerve in the tube of group A (258 ± 6.18) and D(259 ± 5.59) were significantly higher than those of group B (231 ± 5.00) and group C(230 ± 5.07)(P ＜ 0.05).There was no significant difference between groups A and D(P ＞ 0.05).Conclusion These results illustrated that this new PRGD/PDLLA/VPA conduit could significantly facilitate the regeneration of short nerve defect and recovery of motor nerve,which provides a new thought for treatment of peripheral nerve injury.

Objective To investigate the changes and clinical significance of calcitonin gene related peptide (CGRP) and interleu-kin 1α(IL-1α) before and after warm acupuncture therapy in the patients with lumbar disc herniation (LDH ) .Methods Serum CGRP and IL-1αcontents were detected before and after the warm acupuncture therapy in 60 cases of lumbar disc herniation and 15 healthy control cases by ELISA method for conducting the statistical analysis .The changes of the McGill pain scores and serum CGRP and IL-1αconcentrations before and after the warm acupuncture therapy were observed .Results The McGill pain scores af-ter the warm acupuncture therapy in the LDH patients were significantly decreased ,the difference was statistically significant (P<0 .01) ,and serum CGRP and IL-1α concentrations were significantly decreased ,the difference was statistically significant (P<0 .01) .Conclusion CGRP and IL-1αcan be used as the observation index of the effects for the warm acupuncture therapy in trea-ting LDH ,which can provide the basis for the conservative treatment of LDH .

Abstract: Objective　To construct a mouse model of Schistosoma japonicum liver disease induced by direct injection of Schistosoma japonicum eggs through the portal vein and evaluate its effectiveness, in order to provide a new animal model for schistosomiasis liver disease research. Methods　Fifteen 8-week-old C57BL/6 male mice were randomly divided into control group and egg injection group, with 5 in the control group and 10 in the egg injection group. On day -14, 5 000 live eggs were injected into the abdominal cavity of mice, and on day 0, the mice were anesthetized and the abdominal cavity was opened. 5 000 live eggs were injected through the portal vein, and the control group was injected with equal volume of phosphate buffer (PBS). 5 mice in the egg group were killed on day 10 and 30, respectively. The control group mice were killed on day 10, and their serum and liver tissue were collected. Hematoxylin eosin staining (HE) and Masson staining were performed, and alanine aminotransferase (ALT), aspartate aminotransferase (AST), and liver hydroxyproline (HYP) content were detected using a microplate spectrophotometer. Liver fibrosis-related genes, Th1 and Th2 type immune response-related genes were analyzed by real-time fluorescence quantitative PCR (qPCR). Liver injury, egg granuloma and fibrosis, and adaptive immune response were detected to evaluate the effect of portal vein injection of eggs while inducing mouse model of schistosomiasis liver disease. Results　The results showed that significant egg granulomas appeared in the liver of mice after injection of eggs into the portal vein for 10 and 30 days. There was no statistically significant difference in the area of egg granulomas between the 10-day group and the 30-day group (t=0.975, P=0.332). Masson staining and liver hydroxyproline content detection showed significant fibrosis in the liver. The qPCR results showed that, compared with the control group, the expression levels of fibrosis marker genes, such as α⁃Sma (alpha smooth muscle actin), Col1a1 (collagen type Ⅰ alpha 1), and Tgfb1 (transforming growth factor beta 1), were significantly increased (t=6.380, 7.533, 5.314; P=0.002, 0.001, 0.007), and then decreased on the 30th day, with no statistical difference compared to the control group (t=0.940, 1.529, 1.746; P=0.778, 0.543, 0.457). At the same time, the expression levels of Th1 type immune response-related genes, such as tumor necrosis factor alpha (Tnf), interferon gamma (Ifng), and Th2 type immune response-related genes, such as interleukin-5 (Il5), interleukin-13 (Il13), significantly increased 10 days after eggs injection (t=6.163, 4.589, 5.651, 5.367; P=0.003, 0.018, 0.020, 0.009). In addition, there was no significant change in the levels of AST and ALT in the serum of each group of mice (t=0.982, 3.450; P=0.771, 0.074. t=1.164, 0.564; P=0.697, 0.917). Conclusions　A mouse model of schistosomiasis liver disease induced by portal vein injection of worm eggs was constructed. The study provides a new modeling method for studying the mechanism of liver fibrosis in schistosomiasis..

Objective: To establish the fingerprint chromatogram of Compound Renshen Injection (CRSI). Methods: HPLC with ZORBAX SB-C18[4.6(i.d)?250mm] column was used, the acetonitrile-water (gradient elution) as a mobile phase and detection wavelength at 203nm. Results: Indicating 27 peaks on the HPLC-fingerprint of CRSI. Conclusion: The method is simple and accurate with a good reproducibility and can be used as a quality control method for CRSI.

ObjectiveTo investigate the role of CD4+ CD25+ regulatory T cells(Treg) in peripheral blood and intestinal endotoxin (ET) in liver injury of rat severe acute pancreatitis (SAP).MethodsSixty male Wistar rats were randomly allocated into sham operation group(SO group) and severe acute pancreatitis group(SAP group).The rats in SAP group recevied the injection of Sodium Taurocholate(NaTc) into their far-end of bile-pancreatic duct and were sacrificed in 3-,6-and 12-hour,serum amylase(AMY) and alanine aminotransferase(ALT) determined by full-automatic biochemistry instrument,limulus reagent method is used to determine ET content in plasma,the proportion of Treg among peripheral lymphocytes was determined by Flow Cytometry(FCM),HE stain is used to observe pathological changes in liver and pancreas,the protein of Foxp3 activity was evaluated by immunohistochemistry staining,and the relationships between these indicators were analyzed using Pearson correlation analysis test.ResultsHistopathologic examination and the level of ALT revealed the occurrence of pancreatic inflammation and pathological changes of liver in SAP group.The percentage of Treg in SAP groups significantly increased at 3 h(2.26％ ±0.32％),6 h(2.36 ％ ±0.48％)and 12 h(2.80％ ±0.35％) comparing to the SO groups(P＜0.01) ; plasma ET levels compared with the SO group was statistically significant (P＜ 0,01 ),and 12 h (0.85 ± 0.11) compared to,3 h (0.74±0.11) and 6 h (0.78-±-0.07) was no significant difference (P＞0.05).The expression of Foxp3 protein on the livers were upregulated markedly.Pearson correlation analysis teat showed that quantities of Treg were positively proportional to the levels of ET(r=0.89,P＜0.01) after liver injury of SAP.ConclusionsSAP may lead to liver injury and the high plasma levels of ET and Treg in peripheral blood may play an important role in liver injury of SAP.