Objective To investigate the features of immune response of human immunodeficiency virus type-1 (HIV-1) antigen specific T lymphocytes in HIV-1 monoinfected or HIV 1/hepatitis C virus (HCV) coinfected individuals.Methods Twenty-six HIV-1 monoinfected and 23 HIV-1/HCV coinfected individuals were enrolled.Immunomagnetic microbeads were used to isolate T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from human peripheral blood mononuclear cells (PBMC).Frequencies of interferon-γ (IFN-γ) secreting cells of CD4+,CD8+ T lymphocytes and PBMC stimulated by a peptide pool containing 12 overlapping peptides in HIV-1 P24 from 49 patients were assessed by enzyme-linked immunospot (ELISPOT) assay.HIV-1 RNA levels of these patients were also detected by real-time fluorescence quantitative polymerase chain reaction.The data were compared by one-way ANOVA and Mann-Whitney U test,and Spearman test was used for correlation analysis.Results Frequencies of HIV-1 antigen specific CD4+ T lymphocytes [median =25 spot-forming cells (SFC)/106 cells] were significantly lower than those of CD8+T lymphocytes (median=38SFC/106 cells,F=4.592,P=0.037) and PBMC (median=53 SFC/106 cells,F=5.436,P=0.025) in HIV-1 monoinfected group.Frequencies of HIV-1 antigen specific CD4+ T lymphocytes (median=5 SFC/106 cells,Z=-2.432,P=0.015),CD8+T lymphocytes (median=5 SFC/106 cells,Z=-1.996,P=0.046) and PBMC (median=10 SFC/106 cells,Z=-2.306,P=0.021) in HIV-1/HCV coinfected group were significantly lower than those in HIV-1 monoinfected group.Conclusions In HIV-1 infection,antigen specific immune response of CD4+ T cells can be activated,but weaker than that of CD8+ T cells.Co-infection with HCV might down-regulate the responses of HIV-1 antigen specific T lymphocytes in HIV-1 infected individuals.

Objective To investigate the effect of isoflurane preconditioning on the nuclear factor kappa B (NF-kB)activity of leukocytes in patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Twenty ASAⅡorⅢpatients of both sexes aged 20-60 yrs undergoing cardiac valve replacement under CPB were randomly divided into 2 groups(n=10 each):control group(C)and isoflurane preconditioning group(I).Anesthesia was induced with midazolam 0.08-0.12 mg?kg~(-1),fentanyl 5-10?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1).The patients were mechanically ventilated(FiO_2=100%)after tracheal intubation.Anesthesia was maintained with intermittent i.v.boluses of fentanyl and midazolam in group C or with 2 MAC isoflurane and intermittent i.v.boluses of fentanyl and midazolam in group I before CPB.Systolic BP was kept between 90-120 mm Hg,diastolic BP between 50-80 mm Hg and HR between 60-90 bpm in both groups. Isoflurane was discontinued at the initiation of CPB.Arterial blood samples were taken after tracheal intubation and before inhalation of isoflurane(T_0)at 30 min(T_1),1 h(T_2)and 2 h(T_3)after aortic unclamping for determination of NF-kB activity of leukocytes using electrophoretic mobility shift assay(EMSA).The amount of fentanyl,midazolam,dopamine and sodium nitroprusside(SNP)consumed during operation and the rate of recovery of spontaneous heart beat in both groups were recorded.Results The NF-kB activity was significantly increased after aortic unclamping in C group but there was no significant difference in NF-kB activity before CPB (T_0)and after aortic unclamping(T_(1-3))in I group.The NF-kB activity was significant lower at T_(1-3) in group I than in group C.The total amount of fentanyl consumed was 40-60?g?kg~(-1) in C group and 20-30?g?kg~(-1) in group I. Significantly less amount of dopamine was used in group I than in group C.There was no significant difference in SNP consumption between the 2 groups.The rate of recovery of spontaneous heart beat was significantly higher in group I than in group C(P＜0.01).The amount of dopamine consumed was positively correlated with the highest level of NF-kB activity in both group[r=0.962 in group C;r=0.908 in group I(P＜0.01)].Conclusion Isoflurane preconditioning can attenuate the NF-kB activity of leulocytes in patients undergoing cardiac valve replacement under CPB.

Objective To investigate the major risk factors for posterior circulation stroke and the clinical and imaging features of posterior circulation stroke patients with diabetes.Methods The patients with acute cerebral infarction were enrolled.The clinical data of patients with posterior circulation and anterior circulation stroke were compared.The patients with posterior circulation stroke were further divided into either a diabetic group or a non-diabetic group,and the vascular risk factors and imaging features of both groups were compared.The patients with posterior circulation stroke were divided into proximal segment,middle segment and distal segment and mixed groups according to the distribution of vascular lesions.The correlations between diabetes and each group and the imaging features were analyzed.Results A total of 328 patients with posterior circulation stroke (male 194,the diabetic group 108) and 336 patients with anterior circulation stroke (male 214,the diabetes group 59)were enrolled.The proportions of patients with diabetes (32.9％ vs.21.7％ ; x2 =10.501,P =0.001),hyperlipidemia (60.1％ vs.47.9％ ;x2 =9.852,P =0.002),previous stroke or transient ischemic attack (TIA) (29.0％ vs.22.0％ ;x2 =4.213,P =0.040) in the posterior circulation ischemic stroke group were significantly higher than those in the anterior circulation ischemic stroke group,and the proportion of smoking patients was significantly lower than that in the anterior circulation ischemic stroke group (18.3％ vs.26.2％ ; x2 =5.977,P =0.014).The levels of total cholesterol (4.72 ±1.07 mmol/L vs.4.56 ± 0.98 mmol/L; t =2.079,P =0.038),triglycerides (1.54 ± 1.07 mmol/L vs.1.33±0.71 mmol/L; t=3.085,P=0.002) and low-density lipoprotein cholesterol (2.91±0.90 mmol/L vs.2.75 ±0.80 mmol/L; t =2.373,P =0.018) were significantly higher than those in the anterior circulation ischemic stroke group,and the level of high-density lipoprotein cholesterol was significantly lower than that in the anterior circulation ischemic stroke group (1.13 ± 0.31 mmol/L vs.1.18 ±0.32 mmol/L; t =2.045,P=0.041).Multivariate logistic regression analysis showed that diabetes (odds ratio [OR] 1.560,95％ confidence interval [CI] 1.086-2.239; P =0.016) and previous stroke or TIA history (OR 1.455,95％ CI 1.013-2.090; P =0.042) were the independent risk factors for posterior circulation ischemic stroke.In patients with posterior circulation ischemic stroke,the patient's proportions of hyperllpidemia (66.7％ vs.55.5％ ;x2 =5.069,P =0.024) and drinking (13.0％ vs.4.5％;x2 =7.568,P=0.006) in the diabetic group (n =108) were significantly higher than those in the non-diabetic group (n =220); the proportion of atrial fibrillation patients was significantly lower than that in the non-diabetic group (3.7％ vs.11.4％ ;x2 =5.274,P =0.022).The levels of triglycerides (1.70 ± 0.93 rnmol/L vs.1.45 ± 1.11 mmol/L; t =1.989,P =0.048),fasting glucose (8.46 ± 2.96) mmol/L vs.5.30± 0.96 mmol/L; t=10.706,P=0.000) and glycosylated hemoglobin (8.36％ ± 1.94％ vs.6.07％ ± 0.55％ ; t =10.576,P =0.000) in the diabetic group were significantly higher than those in the non-diabetic group.The proportion of patients with large artery atherosclerosis stroke in the diabetic group was significantly higher than that in the non-diabetic group (73.1％ vs.60.0％; x2=5.457,P=0.019); the proportion of the patients with cardioembolism was significantly lower than that of the non-diabetic group (2.8％ vs.9.1％;x2 =4.428,P =0.035).The proportion of patients with posterior circulation middle segment infarction in the diabetic group was significantly higher than that of the non-diabetic group (49.1％ vs.31.4％ ;x2 =9.726,P =0.002).The proportions of the patients with brainstem infarction (60.2％ vs.48.2％ ;x2 =4.182,P =0.041) and single brainstem infarction (55.6％ vs.30.5％ ;x2 =19.235,P =0.000) in the diabetic group were significantly higher than those in the non-diabetic group.In patients with single brainstem infarction,the proportions of the patients with pontine infarction (43.5％ vs.25.9％ ;x2 =10.374,P =0.001) and medulla oblongata infarction (7.4％ vs.1.8％ ; P =0.023) in the diabetic group were significantly higher than those in the non-diabetic group.Conclusions Diabetes and previous stroke or TIA history are the independent risk factor for posterior circulation stroke.Diabetes is closely associated with brainstem infarction,and it is more likely to result in pontine infarction.

AIM:To point the susceptible gene in Avellino corneal dystrophy family with autosomal dominant inheritance.
●METHODS: Genomic DNA was extracted from the peripheral blood samples of all individuals of the pedigree. Several microsatellite makers were selected for gene scan in the hot regions of mutation. Linkage analysis was carried out using a Linkage software package. The haplotype data were processed using Cyrillic software to define the region of the disease gene.
●RESULTS: ln our pedigree, significant evidence of linkage was obtained at marker D5S396 and D5S393 [LOD score (Z)=3. 01, recombination fraction (θ)=0. 00]. The haplotype analysis of our pedigree was located between the microsatellite markers D5S808 and D5S638.
●CONCLUSION:The pathogenic gene of the Avellino corneal dystrophy pedigree is traced to a 11. 2 cM region in the chromosome 5q.

The spores suspension of Streptomyces roseosporus D-38 irritated with 20mW He-Ne laser for 20 min were incubated on G1 medium plates containing 1. 9?g/mL of streptomycin. Ten percent of mutants increased the potency of daptomycin by streptomycin-resistance method, including the mutant LC-54, which could produce daptomycin 81. 2 mg/L, which was 39% higher than that of the beginning strain by flask fermentation.

Objective To investigate the possible mechanism of glomerular injury in diabetes mellitus by determining whether epithelial-mesenchymal transition (EMT) is caused by high glucose in mice podocytes. Methods Using mice glomerular podocyte cell line as an in vitro system, podocytes were incubated with glucose(12.5 mmol/L, 25 mmol/L, 50 mmol/L) and mannitol (50 mmol/L) for 36 hours. Then the cells were collected and expression of alpha-smooth muscle actin(α-SMA), fibronectin (FN), CD2 associated protein (CD2AP) and Wilms' tumor 1 gene (WT-1) was detected by Western blot and indirect immunofluorescence staining. Results Under low glucose (5.6 mmol/L) and mannitol (50 mmol/L) condition, there were high expression of CD2AP and WT-1, and low expression of α-SMA and FN in mice podocytes. After 36 hours treatment with high glucose (12.5 mmol/L), the expression of α-SMA and FN in podocytes was significantly increased, and the expression of α-SMA and FN was further up-regulated with the increase of glucose dosage (25, 50 mmol/L). The indirect immunofluorescence staining revealed the similar result, and the percentage of positive α-SMA cells was also increased compared with low glucose and mannital group (P<0.05). Meanwhile, Western blot showed that high glucose could down-regulate the expressions of CD2AP and WT-1 in a dose-dependent manner. Conclusion EMT may be a potential pathway leading to podocyte dysfunction and glomerular injury under high glucose conditions.

Objective:To express rationally engineered antibodies against EGFR and assess their affinity to EGFR and anti-tumor cell migration effect. Methods:L and V_H genes of humanized antibodies against EGFR were designed and synthesized. Genes encoding V_H and C_H were connected and then cloned into a pIRES based bicistronic expression vector. Gene encoding the corresponding L gene was also cloned into the same vector. 293T cells were transfected with the recombinant plasmid and the antibody expression was confirmed by Western blotting. The antibodies were purified by protein A based affinity chromatography. Binding of the humanized antibody to the EGFR was assessed by Surface Plasmon Renainance with Biacore3000, and the biological activity of the humanized antibody was determined by tumor cell invasion test.Results:Three expression vectors were constructed and the humanized anti-EGFR antibodies were expressed and purified successfully. In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25×10~3and 50×10~3, respectively. Western blot assay showed that the humanized antibodies had recognition specificity to goat-human IgG antiserum. Biacore assay revealed that the humanized antibody C3 binds to EGFR with high affinity(6.13×10~(-10)M). Cell migration test showed that C2,C3 and C5 could suppress growth and migration of tumor cells.Conclusion:Three anti-EGFR humanized antibodies (C2,C3 and C5) have been constructed and expressed successfully, and the C3 antibody retained high affinity for EGFR and showed improved inhibitory effect on tumor cell growth and migration.

Objective To assess the effectiveness of the combined treatment model for Wilms'tumor and to improve treatment results.Methods Fifty-five patients diagnosed with Wilms' tumor between July 1981 to June 2010 were analyzed retrospectively.Eighteen patients were diagnosed by preoperative ultrasound-guided fine needle biopsy,and 53 patients were confirmed by postoperative pathology results.Seven cases were in clinical stage Ⅰ,19 cases in clinical stage Ⅱ,21 cases in stage Ⅲ,six cases in stage Ⅳ and two cases in stage Ⅴ.Thirty-five cases had histopathological subtype,30 cases had the favorable type,and five cases had the unfavorable type.Among the 55 patients,kidney tumor resection was performed on 48 cases,wide edge partial nephrectomy was performed on two cases,tumor enucleation was performed on one bilateral renal tumor case,kidney tumor resection with pulmonary metastasectomy was performed on two cases,and two cases had no surgical procedures.Eighteen cases received preoperative chemotherapy,40 cases received postoperative chemotherapy,and 12 cases received postoperative radiotherapy.Patients were grouped according to age,stage,histological type,treatment model,treatment course and whether or not they had radiotherapy.The Kaplan-Meier method was used in the evaluation and comparison of over survival (OS),disease free survival (DFS) and relapse free survival (RFS) of the different groups to reveal the relationship between different grouping factors with the prognosis of Wilms' tumor. ResultsThe median of follow-up was 34 mon ( ranging from 3 to 355 mon).The 3-year OS,5-year OS and 2-year DFS were 77.6％,69.0％ and 52.4％,respectively.The differences of OS in different stages ( P =0.006 ),DFS between pure operation group and combined therapy group ( P =0.004 ) and RFS between radiotherapy group and no radiotherapy group ( P =0.03 ) were significant,P ＜ 0.05.ConclusionsThe normative multi-disciplinary treatment model for patients with Wilms' tumor can achieve good results and is well tolerated.

Objective To build a high quality diagnosis system for schistosomiasis surveillance in the situation of low infec-tion in Jianglin County. Methods The network laboratory for schistosomiasis diagnosis was built according to the national crite-ria in Jianglin County in 2012. Results The network laboratory for schistosomiasis diagnosis was established successfully and the operation was quiet well. Conclusion The establishment and operation of the laboratory play an important role in the real-ization of schistosomiasis elimination.

Objective To investigate the effect induced by specific inducible nitric oxide synthase (iNOS) inhibitor 1 400 W on nasopharyngeal carcinoma CNE-2 cells lines and its mechanism.Methods CNE-2 cells were treated by different concentrations of 1 400 W,diamminedichloroplatinum (DDP),and both chemicals.Cell counting kit-8 (CCK-8) was used to examine the viability of cells.Quantitative realtime polymerase chain reaction (qRT-PCR) was applied to detect the iNOS mRNA expression.Results The expression of iNOS mRNA was down-regulated by 1400W and was positively correlated with inhibitionrate of cell proliferation.1 400 W inhibits proliferation of CNE-2 cell in a concentration-dependent manner.The proliferation inhibition rate of CNE-2 cells treated by 1 400 W combined with DDP was not enhanced.Conclusions Specific inducible nitric oxide synthase inhibitor 1 400 W can exerts anti-tumor effect though inhibiting the expression of iNOS mRNA;The mechanism of chemosensitization induced by iNOS inhibitor on CNE-2 cells may be closely related level of down-regulation of iNOS expression.