Objective To Study the expressions and significance of nitric oxide (NO) and forkhead box protein 3 (FOXP3) in patients with pulmonary tuberculosis .Methods Serum NO levels were measured by Griess colorimetric reaction .Flow cytometry was used to determine the number of CD4+CD25+ T cells in peripheral blood .The expression of FOXP3 mRNA was measured by real‐time polymerase chain reaction .Results Serum NO level in patients was (15 .71 ± 1 .26)μmol/L ,higher than the (5 .45 ± 0 .98)μmol/L of healthy controls (P<0 .05) .CD4+ CD25+ T cells comprised (4 .57 ± 0 .85)% of CD4+ T cells in patients ,higher than the (1 .83 ± 0 .49)% in healthy controls (P<0 .05) .CD4+ CD25+ T cells in the peripheral blood of patients with pulmonary tuberculosis highly expressed FOXP3 .Conclusion Patients with pulmonary tuberculosis could be with an increased level of NO and FOXP3 ,which might have important role in the pathogenesis of pulmonary tuberculosis .

OBJECTIVE:To study the effects of Astragalus granules on the expression of Caveolin-3(Cav-3)and Smad family member 3 (Smad3) in the myocardial cells of rats with viral myocarditis. METHODS:90 rats were randomly divided into a nor-mal group,a model group,a Shenmai injection group [positive drug,0.2 g/(kg·d)] and the groups of low,medium and high-dose Astragalus granules [0.42,0.84,1.68 g/(kg·d)],with 15 rats in each group. The rats in all groups except for the normal group were given CVB3 ip for the establishment of viral myocarditis model. Meanwhile,the rats in the drug administration groups were given corresponding drugs ig,while those in the normal group and the model group were given normal saline ig,for 15 consecu-tive days. 5 rats were selected from each group respectively on the 3rd,9th and 15th days of drug use to take an experiment. For the rats,the pathological change of the cardiac muscle tissue was observed and scored,and the mRNA and protein expression of Cav-3 and Smad3 in the myocardial cells were detected. RESULTS:After 15 days of drug use,compared to the normal group,the rats of the model group had hyperplasia of a large number of cardiac muscle fibers,obvious lesions at cardiac muscle fibers, and significantly higher pathological score and levels of the mRNA and protein expression of Cav-3 and Smad3 in the myocardial cells (P<0.05). Compared to the model group,the rats in the drug administration groups had cardiac muscle tissue lesions improved and had obviously lower pathological score and levels of the mRNA and protein expression of Cav-3 and Smad3 in the myocardial cells(P<0.05). CONCLUSIONS:Astragalus granules can markedly downregulate the gene expression of Cav-3 and Smad3 in the myocardial cells of rats with viral myocarditis,which is inferred as a prevention and treatment mechanism of viral myocarditis.

Objective To establish a TaqMan probe-based real-time fluorescent quantitative PCR ( real-time PCR) for the quantitative and rapid detection of viral reservoir in peripheral blood mononuclear cells (PBMCs) isolated from rhesus monkeys with simian immunodeficiency virus (SIV) infection and to evaluate its preliminary application. Methods A pair of primers and one TaqMan probe were designed ac-cording to the conserved sequence of SIVmac239 strain for real-time PCR amplification. A length of 2 090 bp of nucleotide fragment was digested from the plasmid p239SpSp5 containing 5′-end long segments of SIV-mac239 strain by restriction enzymes EcoRⅠand SpeⅠ. The standards used for quantitative detection of SIV DNA in peripheral blood samples were prepared by a 10-fold serial dilution and used for graphing the stand-ard curve. The numbers of SIV DNA ( copies per 106 PBMCs) in rhesus monkeys during acute and chronic phases of SIVmac239 infection were determined and the virological characteristics of SIV DNA at different phages of infection were analyzed. Results A linear positive correlation between cycle threshold ( Ct) val-ues and concentrations (10 copies/μl to 109 copies/μl) of the standards was found. High levels of SIV DNA were monitored in SIV-infected monkeys 14 to 22 days after acute infection. The levels of SIV DNA in the acute phase of infection were about 1 to 2 logs higher than those in the chronic phase of infection. The num-bers of SIV DNA ( copies per 106 PBMCs) were 1 log lower than the SIV viral load in peripheral blood of the same monkey. The ratios of SIV DNA load to SIV RNA load ( DNA/RNA) in chronic phase of infection were higher than those in the acute phase. Conclusion The established TaqMan probe-based real-time fluorescent quantitative PCR was a highly sensitive and specific assay for the detection of SIV DNA with an advantage of wide linear range. It could be used for the quantitative evaluation of latent reservoirs of SIV.

Objective To study the diagnostic and distinguishing diagnostic value in primary hepatic carcinoma (PHC)and metastatic hepatic carcinoma (MHC)by the serum sialic acid (SA)detection.Methods During January 2012 to June 2014, 100 cases of patients with PHC,91 cases of patients with MHC,155 cases of benign liver disease patients,and 139 healthy people in Wuyi Traditional Chinese Medicine Hospital were included into the study.The concentration of serum SA and AFP were detected by chemical enzymatic method and chemiluminescence method,SPSS1 9.0 was used to analysis the results.Re-sults The concentration of serum SA in PHC patients (701.08±189.33 mg/L)were significantly higher than benign liver disease patients (588.38±98.51 mg/L)and healthy people (572.37±89.13 mg/L),there was statistical significance (P=0.000),the significantly statistical differences were also in MHC (790.20±162.29 mg/L)and PHC patients (P=0.027). Serum SA in the diagnosis of MHC sensitivity,specificity and AUC were 84.6%,85.2% and 0.895,compared with serum AFP (sensitivity 22.2%,specificity 29.6% and AUC 0.301)had statistically significance (P=0.000).Conclusion The se-rum SA has important clinical significance in the diagnosis and distinguishing diagnosis of PHC and MHC.

Objective:To investigate the methylation of CpG island of metallothionein-3 (MT-3) gene in esophageal cancer tissues and normal tissues in middle and south area of Hebei Province and Chaoshan area of Guangdong Province and compared the results with those in low risk area of esophageal cancer. Methods:The blood samples from 10 normal volunteers, 10 embryonic esophageal tissues, 20 esophageal mucosa tissues from normal subjects in low risk area as well as 30 fresh surgical specimens of esophageal cancer and 30 normal marginal tissues in the high risk middle-south Hebei Province and Chaoshan area were collected. Methylation-specific polymerase chain reaction (MSP) was used to detect the methylation status of the CpG island of MT-3 gene in these samples. Its relationship with clinicopatho-logical features was analyzed. Results:There were 20 (33.3%) cases with MT-3 methylation in the marginal tissues of esophageal cancer from high-risk area, which was higher than that in the normal mucosa from low-risk area (P=0.013). And there were 49 (81.7%) cases with MT-3 methylation in esophageal cancer tissues, which was higher than that in normal marginal tissues (P<0.001). But there was no significant difference in the methylation degree between middle-south of Hebei Province and Chaoshan area (P=0.739). Conclusion:MT-3 methylation widely exists in esophageal mucosa and carcinoma tissues. Acquired stimulus may be the main cause of these methylations.

Objective To assess the role of promyelocytic leukemia protein (PML)and P53 in the progression of esophageal squamous cell carcinoma(ESCC)and its precursor lesions. Methods Different expression patterns of PML and P53 of 241 cases of ESCC combined with adjacent precursors were analyzed by tissue array and immunohistochemistry and correlated with clinicopathological parameters. Results In ESCC and its precursor lesions, PML and P53 displayed positive or strong positive, while in normal esophageal epithelia, these proteins showednegative or stained positive only in parabasal cell layer. The expression level of PML was correlated with the depth of invasion of esophageal carcinomas (X2=29.461,P<0.001),lymph metastasis status(X2=15.226,P<0.001)and pTNMs(x2=26.956,P

Objective To investigate the protein expression of the canonical transient receptor potential(TRPC)channel in the hippocampus of amyloidβprotein(Aβ)?induced Alzheimer’s disease(AD)mice. Methods A total of 36 ICR mice were randomly divided into AD group and control group,with 18 rats in each group. AD mice models were established by Aβ1?42 microinjection into the lateral intracerebroventricular. Learning and memory abilities of the mice were determined using Morris water maze. All TRPC1?TRPC7 mRNA levels in the hippocampus of the mice were detected using reverse transcriptase polymerase chain reaction(RT?PCR). Hippocampal TRPC4 protein expression was examined using immunofluorescence and Western blotting methods. Results Water maze test results showed that the escape latency of AD group were significant?ly longer than that of the control group(P<0.01),and that the target quadrant occupancy of AD group was significantly shortened(P<0.01)and the frequency of platform crossing of AD group was significantly reduced(P<0.01). RT?PCR results showed that all TRPC(TRPC1?TRPC7) mRNA were expressed in the hippocampal of both AD group and control group. Among these channels ,only TRPC4 mRNA levels of AD group was higher than that of the control group(P<0.01). Immunofluorescence images showed that TRPC4 expressed on the membrane of neurons and the intensities of the immunofluorescence of TRPC4 in AD group were stronger than that of control group. Western blotting results showed that the TRPC4 protein expression of AD group was higher than that of control group(P<0.05). Conclusion The increase of TRPC4 protein expression in the hippocampus of mice after intracerebroventricular injection of Aβ1?42 oligomers suggests that TRPC4 may be involved in the pathogenesis of AD induced by calcium homeostasis.

Objective To investigate the association between chromosome variations,abnormalities and male reproductive hor-mones level with spermatogenesis.Methods The chromosome karyotype,serum reproductive hormone including FSH,LH,T,PRL and E2,and semen were detected in 147 patients with male infertility or recurrent sponotaneous abortion.The results were per-formed the comparative analysis.Results Serum FSH,LH level and the incidence rate of azoospermia in the chromosome abnormal-ity group were significantly higher than those in the chromosome variation group and the normal group(P <0.05,P <0.01),serum T level was significantly lower than that in the chromosome variation group and the normal group(P <0.05).Serum FSH and the incidence rate of oligospermia in the Y chromosome variation group were significantly higher than those in the autosomal variation group(P <0.05).The incidence rate of azoospermia had no statistically significant difference between the Y chromosome variation and the autosomal variation group(P >0.05).Serum FSH,LH level and the incidence rate of azoospermia in the sex chromosome abnormality group were obviously higher than those in the autosomal abnormality group(P <0.05),the serum T level was signifi-cantly lower than that in the autosomal abnormality group(P <0.05).Conclusion The chromosome variation and abnormality are closely related with the reproductive hormones disorder and spermatogenetic function disorder.The obvious increase of serum FSH, LH level and obvious decrease of T level caused by sex chromosome variation and abnormality is one of the pathogenesis of oligo-spermia and azoospermia.

Objective To investigate influence of Qi-strengthening,blood-activating and toxin-removing therapy on plasma and colonic P-selectin in ulcerative colitis(UC) rats.Methods Trinitrobenzene sulfonic acid(TNBS) was used for the establishment of SD rat models of UC.UC rats were randomized into the model group,western medicine group(gastric gavage of suspension of Olsalazine Sodium Capsules 0.266 g?kg-1?d-1),and herbal medicine group(gastric gavage of Kuijie Fufa Recipe 20 g?kg-1?d-1).Meanwhile,a normal control group was set up.After medication for 10 and 30 days,and after drug withdrawal for 10 days,plasma P-selectin level was detected by enzyme-linked immunosorbent assay,and colonic P-selectin expression was detected by immunohistochemical assay.Results Plasma P-selectin level was increased in the model group in different time(P