AIM: To investigate the anticoagulant effect of pravastatin and low molecular weight heparin (LMWH), as well as their combination, over time, in a rat model of experimental nephrotic syndrome. METHODS: Healthy SD male rats were chosen randomly to perform nephrotic syndrome models by single injection of adriamycin via tail vein, the matched normal control rats received single injection of equivalent 0.9% sodium chloride instead. After 14 days, the models were set up and randomized into model control group, pravastatin group (pravastatin 2 mg·kg-1·d-1 gavage once a day), LMWH group(LMWH 200 U·kg-1·d-1 intraperitoneal injection once a day) and combined treatment group(pravastatin 2 mg·kg-1·d-1 gavage+ LMWH 200 μ·kg-1·d-1 intraperitoneal injection), then all rats underwent measuring proteinuria every two weeks and fibrinogen, antithrombinⅢ(ATⅢ), D-dimer, platelet count, serum total protein and serum albumin after 4 weeks of treatment. RESULTS: The concentration of fibrinogen and D-dimer in model group was higher significantly than that in control group, and the level of ATⅢ was lower remarkably than that in control group, but platelet count had no obvious change; Compared with model group, pravastatin could increase the level of ATⅢ and decrease the concentration of D-dimer, but the concentration of fibrinogen and platelet count did not change obviously; LMWH and combined treatment could also decrease level of D-dimer, but had no great effects on ATⅢ, fibrinogen and platelet count; all treatment group had no obvious change of serum total protein and serum albumin. CONCLUSION: Adriamycin-induced nephrotic syndrome rat models have hypercoagulability and pravastatin can increase the level of ATⅢ.

Human cytomegalovirus (HCMV) is the most common cause of congenital infection, resulting in birth defects such as microcephaly. In this study, RT-PCR and Western Blotting were performed to quantify the regulation of endogenic nerve growth factor expression in neuroglia cells by HCMV infection. The results showed that basal, endogenous NGF expression in U251 was unchanged during early HCMV infection. NGF expression is strongly down-regulated during the latent phase of infection. These results suggest that HCMV can depress the NGF expression in U251 cells.

To investigate the inhibitory effects of Ginsenoside Rb1 (GRb1) on apoptosis caused by Herpes Simplex Virus-1 (HSV-1) in Human Glioma Cells (U251),U251 cells were infected by HSV-1 at a multiplicity of infection of 5 and GRb1,GRb1+HSV-1,HSV-1 and control groups.MTT and cell apoptosis assays were used to detect the inhibitory effects of GRbl on the apoptosis of U251 cells that caused by HSV-1 infection for various concentrations of drug and virus treatments by MTT assay.We found that in the 400 μg/mL GRbl and 400 μg/mL GRbl+HSV-1 groups,MTT values were higher than control group at all times (P＜0.05).Moreover,the apoptosis rate in the 400 μg/mL GRb1+HSV-1 group was lower than the HSV-1 group (P＜0.05).These results confirmed that,at appropriate concentrations,GRb 1 could inhibit nerve cell apoptosis in HSV-1 infections.

Objective:To construct a recombinant eukaryotic expressive vector of pEGFP-N3-M-IL-2(88 Arg,125 Ala),and to study the expression of this gene in the Glioma cell line U 87,and to detect its antitumor activities of the fusion protein M-IL-2(88 Arg,125 Ala).Methods:The target fusion gene M-IL-2 (88 Arg,125 Ala) was amplified by PCR from pPICZαA/M-IL-2 (88 Arg,125 Ala) and cloned into pEGFP-N3 vector after digestion to construct recombinant eukaryotic expressive vector pEGFP -N3-M-IL-2( 88 Arg,125 Ala).And then recombinant plasmid pEGFP-N3-M-IL-2(88Arg,125Ala) was transfected into Glioma cell U87 by LipofectamineTM2000 immediately after it was confirmed by restrictive enzyme analysis and sequencing .RT-PCR and Western blot were used to confirm expression of the fusion gene in the U87.Prohibitory effect of recombinant M-IL-2(88Arg,125Ala) protein on U87 was assessed by CCK-8 assay.Results:Restrictive analysis and sequence analysis revealed that M-IL-2(88Arg,125Ala) fusion gene was cloned into the vector pEGFP-N3 suc-cessfully,fusion gene M-IL-2(88Arg,125Ala) could express in U87 cells and could inhibit the growth of U87 cells.Conclusion:The eu-karyotic expression plasmid pEGFP-N3-M-IL-2( 88 Arg,125 Ala) was constructed and expressed in U 87 cells successfully ,the fusion protein could inhibit the growth of U87 cells.We laid a foundation for further research of gene M-IL-2(88 Arg,125 Ala).

OBJECTIVE To develop a macroarray method to detect pathogens in cerebrospinal fluid.METHODS According to the bacterial 16S rRNA genes,designed 10 kinds of specific probes and a pair of universal primers that can amplify rRNA gene of all bacteria.The tailed probes were spotted onto a nylon membrane.DNA was isolated from each pathogen,and subjected to UP-PCR to amplify target fragments,which were labeled with bio-16-dUTP at the same time.All those denatured fragments were hybridized to the probes on nylon membrane and visualized by AKP labeled avidin.The sensitivity and specificity of the system were detected.A total of 32 CSF samples,which were verified the bacterial infection by the routine method,were tested by this method.RESULTS It was sensitive to 10 CFU/ml when detecting Escherichia coli.Every kind of pathogens only reacted to its corresponding probes fixed on nylon membranes,which showed high specificity.The result of identifying 32 CSF clinical specimens accorded with that of routine method.CONCLUSIONS The method can screen out common pathogens in CSF sensitively and exactly.

Objective To detect specific polymorphisms in Toll-interleukin 1 receptor domain containing adaptor protein(TIRAP) coding region for Chinese Han population, and verify whether they are associated with susceptibility to tuberculosis. Methods Search TIRAP polymorphisms by sequencing in small sample; detect single nucleotide polymorphism(SNP) by ligase detection reaction technique in large sample; analyze whether polymorphisms are related to tuberculosis by statistic methods. Results Four polymorphisms were present in the TIRAP coding region. 394A had higher frequencies in the tuberculosis(TB)group than the control. But allelic and genotypic analysis showed that there were no significant difference in statistic between TB patients and controls(P>0.05). The SNP G164A mutation related with TB patient's condition. Comparing to controls, retreatment patients' allelic frequencies had significant difference in statistic(P<0.05), sputum positive patients and lung cavitation patients had lower 164A frequencies. Conclusion TIRAP coding region polymorphisms may be risk factors for TB occurrence and development in Chinese Han population.

Objective To observe the influence of human umbilical cord mesenchymal stem cells (hUCMSC) transplanted into the tail vein of diabetic rats on apoptosis of retinal neurons and the retinal expression level of glial fibrillary acidic protein (GFAP).Methods Seventy clean male Sprague-Dawley rats were randomly divided into the normal control group (group A),diabetes mellitus (DM) only group (group B),DM + balanced salt solution (BSS) group (group C),DM + hUCMSC group (group D),with 10 rats in each group.DM rats were induced by intraperitoneal injection of streptozotocin.Apoptosis of retinal cells was assayed by dUTP nick end labeling.Immunohistochemistry and Western blot was performed to detect the retinal expressions of GFAP in rats.Results Compared with group A,large numbers of apoptotic cells could be found in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) of group B and group C,however the apoptotic cells in group D were significantly reduced than group B and C.The expression of GFAP was mainly located in the retinal GCL and retinal nerve fibre layer (RNFL) in group A,throughout the inner plexiform layer (IPL) in group B and C,only distributed in RNFL and GCL in group D.It was obvious that the expression of GFAP in group B and C was higher than group A.Compared with group B and C,the expression of GFAP in group D was significantly reduced.The difference of GFAP expression among the 4 groups was significant (F=79.635,P＜0.05).Conclusion hUCMSC could inhibit the apoptosis of retinal cells and activation of glial cells in early DM rats.

Objective To find out the composite risk factors associated with early neurological deterioration (END) in acute ischemic stroke,to reveal the mechanism of END,and to provide the data base for the early prediction and prevention.Methods Five hundred and fifty-eight patients with cerebral infarction admitted to our hospital from October 2009 to December 2012,were screened.Among them,107 patients met the diagnostic criteria of END in acute ischemic stroke,451 patients met the diagnostic standard of early acute ischemic stroke without END.Neurological function scale and other variables included 58 related factors of 9 categories were selected.Association rule mining methods were used to analyze relations between END in acute ischemic stroke and risk factors sets.Results The results of association rule mining discovered that there were 2 individual risk factors,3 double-factor combinations,7 triple-factor combinations and 15 four-factor combinations related with END in the early stage of acute ischemic stroke;the more the composite factors,the higher the probability of neurological deterioration;the composite factors were mostly the combination of variables of different categories,involving neurological function scale scores,infection condition,dysphagia,personal life history (smoking and drinking),infarction location,age,and levels of electrolyte,C reactive protein,and homocysteine.Besides some independent risk factors which had been reported in the literatures,the results of this study found that heart rate and time interval from onset to hospitalization also related with END in early acute ischemic stroke.Conclusion END in acute ischemic stroke may be attributed to the combination effect of variable factors;all risk factors should be considered and a variety of targeted measures should be taken to prevent and treat the patients with END in early acute ischemic stroke.

OBJECTIVETo investigate the morphology and proliferation effects of adenovirus containing bone morphogenetic protein-2(BMP-2) on tongue cancer Tca8113 cells.

METHODSTca8113 cells were transfected with the Ad-BMP-2 of 0, 50, 100 multiplicity of infection (MOI) respectively. Inverted fluorescence microscope was used to evaluate the morphological changes of these cells. Western blot analysis was performed to determine the expression levels of BMP-2 in the transfected cells. Methyl thiazolyl tetrazolium (MTT) assay was performed to monitor the proliferative activity of the infected Tca8113 cells and then the growth curve was made.

RESULTSThe transfection efficiency reached the highest when the MOI was 100. Moreover, the expression of BMP-2 was detected in Tca8113 cells by Western blot. There were no obvious morphological changes of the Tca8113 cells before and after transfection. And the proliferation of transfected Tca8113 cells decreased compared with control.

CONCLUSIONAd-BMP-2 gene can inhibit the proliferation of Tca8113 cells in vitro.