Objective To evaluate the role of 5-HT5A receptors (5-HT5A R) in activation of astroglia in the spinal dorsal horn in a rat model of neuropathic pain induced by vincristine. Methods Forty adult male SD rats weighing 180-200 g were randomly divided into 4 groups ( n = 10 each): control group (group C);neuropathic pain group (group P);Ad-X-HK group (group B) and Ad-5-HT5A-siRNA group (group S). Neuropathic pain was induced by repeated intraperitoneal (IP) injection of vincristine 0.1 mg/kg according to the method described by Weng et al in group P, B and S. On the 2nd day after the last IP injection, the animals received artificial cerebrospinal fluid, Ad-X-HK and Ad-5-HT5A-siRNA 25 μl administered intrathecally (IT) in group P, B and S respectively. Paw withdrawal threshold to mechanical stimulus was measured before and on the 7th day after IT administration. The animals were then sacrificed. The lumbar segment ( L4.5 ) of the spinal cord was removed for determination of 5-HT5A R and GFAP expression. Results Body weight and paw withdrawal threshold were significantly decreased after repeated IP vincristine administration in group P compared with group C. IT Ad-5-HT5A-siRNA reduced pain threshold further in group S compared with group P. Repeated IP vincristine significantly increased the expression of 5-HT5A R and GFAP in spinal dorsal horn, and IT Ad-5-HT5A-siRNA significantly decreased the expression of 5-HT5A R while increased the expression of GFAP in spinal dorsal horn in group S compared with group P. Conclusion 5-HT5AR is involved in the inhibition of astrocyte activation, resulting in reduction of vincristineinduced neuropathic pain.

Objective To evaluate the efficacy and adverse effect of brachial plexus block with morphine and ropivacaine for postoperative analgesia in children. Methods Forty-five children undergone upper extremity surgery were randomly divided into three groups (n = 15) , ie, Croup A (brachial plexus block with ropivacaine), Croup B (brachial plexus block with ropivacaine and subcutaneous injection of morphine) and Croup C (brachial plexus block with morphine and ropivacaine). The analgesic effect and the analgesia maintenance time were recorded by FLACC method at 1, 2, 4, 6, 8, 12, 24 hours after operation. The adverse reactions were observed 48 hours after operation. Results There was no statistical difference between Croup A and Croup B in the analgesia maintenance time and the number of cases without analgesic therapy at each time point (P > 0. 05). Compared with Croup A or Croup B, the analgesia maintenance time in Croup C were significantly extended and the number of cases without analgesic therapy at 4, 6, 8, 12, 24 hours after operation were significantly increased (P < 0.05). The adverse effect was fairly light in three groups. Conclusion Brachial plexus block with morphine and ropivacaine may extend analgesia maintenance time effectively and safely in children undergone upper extremity surgery.

Objective To investigate the pathogenesis of age-related cortex and nuclear cataract.Methods The levels of transforming growth factor-?_2 (TGF-?_2)mRNA,proliferating cell nuclear antigen (PCNA),ratio of Bcl-2/Bax,fibronection (FN),vimentin and the density of lens epithelial cells (LECs) were measured and compared among age-related nuclear cataract,cortex cataract and normal LECs.Results The ratio of TGF-?_2 mRNA expression/?-actin expression,the levels of PCNA protein,Bax and Bcl-2 in age-related nuclear cataract(0.16?0.02,16.01,15.97, 16.97)and in cortex cataract (0.27?0.01,1.98,16.93,1.44) were significantly different from those in normal LECs(0.15+0.02,2.43,16.84,6.12),all P

Lentiviral vectors have drawn considerable attention recently and show great promise to become important delivery vehicles for future gene transfer manipulation. In the present study we have optimized a protocol for preparation of human immunodeficiency virus type-1 (HIV-1)-based defective lentiviral vectors (DLV) and characterized these vectors in terms of their transduction of different cells. Transient co-transfection of 293T packaging cells with DNA plasmids encoding lentiviral vector constituents resulted in production of high-titer DLV (0.5-1.2 × 107IU/mL), which can be further concentrated over 100-fold through a single step ultracentrifugation. These vectors were capable of transducing a variety of cells from both primate and non-primate sources and high transduction efficiency was achieved using concentrated vectors. Assessment of potential generation of RCV revealed no detection of infection by infectious particles in DLV-transduced CEM, SupT-1 and MT-2 cells. Long-term culture of transduced cells showed a stable expression of transgenes without apparent alteration in cellular morphology and growth kinetics. Vector mobilization to untransduced cells mediated by wild-type HIV-1 infection was confirmed in this test. Challenge of transduced human T-lymphocytes with wild-type HIV-1 showed these cells are totally resistant to the viral infection. Considering the effective gene transfer and stable gene expression, safety and anti-HIV activity, these DLV vectors warrant further exploration for their potential use as a gene transfer vehicle in the development of gene therapy protocols.

Objective To analyze the risk factors for failures of intramedullary and extramedullary internal fixation in surgery of femoral intertrochanteric fractures in elderly patients.Methods A retrospective study was conducted of the 205 elderly patients with osteoporotic femoral intertrochanteric fracture who had accepted closed reduction together with intramedullary and extramedullary internal fixation between September 2005 and August 2014.They were 89 men and 116 women,from 65 to 98 year of age (average,78.8 years).By AO classification,137 cases were of types AI.1-A2.1 (stable fractures),and 68 of type A2.2-A3.3(unstable fractures).The incidence of internal fixation failure and Harris scores at the last follow-up were recorded.The factors possibly contributing to the failure were analyzed using the univariate analysis and multivariate logistic regression analysis.Results Of the patients,192 obtained a mean follow-up of 39 months (from 14 to 60 months),but 13 were lost after a 10-month follow-up.Of the 205 patients,internal fixation failure occurred in 12 (incidence of 5.9％).Five failed cases received DHS fixation and 7 accepted PENA fixation.At the last follow-up when the 13 cases were lost after 10-month follow-up,the mean Harris hip score was 81.6 (from 57 to 92),and the excellent to good rate was 84.9％ (29 excellent cases,145 good ones,10 fair ones and 21 poor ones).The multivariate regression analysis revealed that tip-apex distance (TAD) ＞ 25 mm(OR ＝ 333.33),severe osteoporosis (OR =267.44),AO types A2.2-A3.3 (OR ＝ 22.24),functional reduction of fracture (OR =20.79),and concomitant medical diseases (OR =4.59) were independent risk factors for failures of internal fixation.Conclusions DHS and PFNA fixations are effective treatments for elderly patients with femoral intertrochanteric fractures.TAD＞ 25 mm,severe osteoporosis,unstable factures,functional reduction of fracture,and concomitant medical diseases may lead to internal fixation failure in surgery of femoral intertrochanteric fractures in elderly patients.

Numerous epidemiological studies have studied the association of adiponectin (ADIPOQ) gene and adiponectin receptor (ADIPOR) gene polymorphisms with risk of colorectal cancer (CRC),but the outcomes were incomplete and inconsistent.Therefore,we conducted a meta-analysis to assess the associations systematically.All eligible case-control studies published up to Jan.2015 were searched from PubMed,the Cochrane library,Elsevier,Wiley Online library,China National Knowledge Infrastructure,WanFang data and Chongqing VIP.Effect sizes of odds ratio (OR) and 95％ confidence interval (95％CI) were calculated by using a fixed-or random-effect model.Twelve case-control studies including 6141 cases and 7398 controls were selected.Significant differences in the distributions of allele frequency with CRC risk were directly present in ADIPOQ variants rs2241766,rs1501299 and ADIPOR variant rs1342387.In stratified analysis for different populations,significant differences were present in ADIPOQ variant rs822396 for Ashkenazi Jewish,in ADIPOQ variant rs1501299 and ADIPOR variant rs1342387 for Chinese and in ADIPOQ variant rs 2241766 for Ashkenazi Jewish and Chinese.In addition,the factors correlated with insulin resistance had synergistic effect with ADIPOQ variants rs2241766 T/G and rs1501299 G/T on risk of CRC.ADIPOQ variants rs2241766 T/G,rs1501299 G/T and ADIPOR variant ADIPOR rs1342387 G/A had a population specific correlation with CRC risk,which may be mediated by insulin resistance.And large well-designed studies are still needed for further evaluation of rs822396 and rs1063538,especially for their interaction and combined effect in the correlation with CRC risk.

BACKGROUND:Development of new coronary thrombolytic agents is hot in the market. A new drug, mutated recombinant tissue-type plasminogen activator (rtPAm), is the product of mutation of tPA by changing binding loci with plasminogen activator inhibitor (PAI)-1 to reduce the degradation. In vitro test has demonstrated that the activity of rtPAm is much higher than rtPA in the absence of PAI. The present study is to observe the efficacy of mutated recombinant tissue-type plasminogen activator (rtPAm) in coronary thrombolytic therapy. METHODS:A total of 30 adult beagles were equally divided into 5 groups after thrombi:vehicle group, urokinase group, rtPAm low-dose group, rtPAm medium-dose group, and rtPAm high-dose group. Thrombolytic effect and myocardial infarction were observed after thrombolytic therapy. RESULTS:In the urokinase group, time to reperfusion was (15.8±3.8) minutes. TIMI 2 flow was demonstrated in 4 beagles, TIMI 3 flow in 2, and re-occlusion in 4 after 90 minutes respectively. In the low-dose rtPAm group, time to reperfusion was (15±4.5) minutes; TIMI 2 flow was demonstrated in 2 beagles, TIMI 3 flow in 4, and re-occlusion in 2 after 90 minutes. In the high-dose rtPAm group, time to reperfusion was (7.5±2.6) minutes. None of the beagles showed re-occlusion after 90 minutes. The infarction areas were (2.1+0.9)％ in the medium-dose rtPAm group and (0.7+0.4)％ in the high-dose rtPAm group, which decreased significantly than those in the low-dose rtPAm group. The aggregation rate in the medium-dose and high-dose rtPAm groups decreased significantly than that in the urokinase group. CONCLUSION:rtPAm may serve as a thrombolytic agent with platelet-targeted fibrinolysis and antiplatelet aggregation activities.

Objective To analyze the related factors with prognosis in patients with serous ovarian adenocarcinoma and to set up a prognostic model of serous ovarian adenocarcinoma.Methods The clinical, pathological and follow-up data of 104 cases with serous ovarian adenocarcinoma were retrospectively analyzed.Kaplan-meier univariate analysis was used to screen the prognostic factors;COX univariate and multivariate analyses were used to determine the risk coefficient of each factors and different layers in each factor.Pearson rank correlation was used to reject the influence of different factors with each other.And the prognostic model of serous ovarian adenocarcinoma was set up based on the result of the above study,which could be used to deduce the survival probability of patients with serous ovarian adenocarcinoma.Results International Federation of Gynecology and Obstetrics(FIGO)stage(P=0.0029),histological grade(P= 0.0054),residual disease(P=0.0000),metastasis of lymph nodes(P=0.0000)and chemotherapy(P= 0.0000)were the related factors of prognosis in patients with serous ovarian adenocareinoma,of which FIGO stage was the most important one,followed sequentially by histological grade,metastasis of lymph node, residual disease and chemotherapy(the independent risk coefficient of each factor was 1.3392,0.9206, 0.7071,0.6004,0.4985 in sequence).We set up a prognosis model according to the prognostic index of each factors.The effect of chemotherapy and residual disease on prognosis could be quantified by this model, and the higher the score,the lower the survival probability of patients.Condusions FIGO stage, histological grade,residual disease,metastasis of lymph nodes and chemotherapy are important prognostic factors of serous ovarian adenoearcinoma.This model can be used to estimate the prognosis of patients with serous ovarian adenoearcinoma,and the effect of both chemotherapy and residual disease on the prognosis could be quantified by the model.

Full-length NS5A gene of the hepatitis C virus was amplified by PCR using plasmid pBAC25 containing HCV nonstructural gene as template. The amplified fragment (about 1.34 kb) was cloned into plasmid pQE32, and the recombinant plasmid pQENS5A was expressed in JM109 strain. The NS5A protein was purified by NiSO4 metal chelating resin, and characterized by Western-blot. Its antigenecity was determined by ELISA. The positive detection rate of anti-NS5A was 75% (69/92) in ninety-two clinic sera. The positive rate of anti-NS5A was 82.5% (33/40) in fourty positive standand sera, and the negative rate of anti-NS5A was 100% (40/40) in fourty negative standand sera. The results showed that the Full-length NS5A proteinn had the higher sensitivity and specificity in the detection of HCV antibody in sera, we suggested that NS5A protein was a useful antigen for blood screening.

SARS-CoV is a newly discovery pathogen causing severe acute respiratory problems.It has been established that the S protein in this pathogen plays an important rule in the adsorption and penetration of SARS-CoV into the host cell by interaction with the ACE2 receptor.To determinant which functional motif of the S protein was involved in the interaction with ACE2,seven truncated S proteins deleted from the N or C terminal were obtained by an E.coli expression system and purified by column chromatography to homogeneity.Each truncated S protein was fixed on to the well of an ELISA plate and an interaction was initiated with the ACE2 protein.The adsorption were quantified by ELISA,and the results indicated that amino acids from 388 to 496 of the S protein was responsible for the interaction with the ACE2 receptor,and the interaction could be completely disrupted by an antibody specific to these amino acids.Deletions adjacent to this domain did not appear to have a significant impact on the interaction with ACE2,suggesting that the S protein of SARS-CoV could be developed as a vaccine to prevent the spread of SARS-CoV.