Objective To establish a rapid method of real-time fluorescence RT-PCR to quantify Soul virus.Methods The pro-fessional software was adopted to design the primer and the TaqMan-BHQ probe.With artificially synthesized L gene segment as the template of Soul virus,the real-time RT-PCR for detecting Soul virus was researched.Results The Ct value of templates had a good linear relationship with the log value of the template diluted concentration.The standard curve was Y =-3.607X +41.84, r2 =0.998,the PCR amplification efficiency was 108.1%,its lowest detection limit was 53.2 copies/μL.Conclusion Applying the real-time fluorescence RT-PCR by the TaqMan-BHQ probe for detecting nucleic acid of Seoul virus has the characteristics of short time-consuming and high sensitivity.

Objective To compare the accuracy of nucleic acid detection and antibody detection for diagnosing human immunode‐ficiency virus(HIV) infection .Methods Retrospectively analysed data of nucleic acid detection and antibody detection from 124 ca‐ses of patients diagnosed with HIV infection from 2005 to 2014 .The positive rates of the two methods were compared respectively in patients with early‐stage of HIV infection(76 cases) and patients with intermediate and advanced stage of HIV infection (48 ca‐ses) .Results In patients with early‐stage of HIV infectionn ,the positive rate of nucleic acid detection (94 .74% ) was higher than that of antibody detection (84 .21% );while in patients with intermediate and advanced stage of HIV infection ,the positive rate of antibody detection(97 .92% ) was higher than that of nucleic acid detection (81 .25% );both had statistically significant difference (P<0 .05) .Conclusion On the early stage of HIV infection ,the accuracy of nucleic acid detection is higher than that of antibody detection ;while on the intermediate and advanced stage of HIV infection ,antibody detection shows better accuracy .

Objective To determine the diagnostic value and clinical significance of sonographically detected fetal dysplastic kidney with normal amniotic fluid volume. Methods At the 2nd or 3rd trimester of gestation,the fetuses with unilateral or bilateral renal anomalies (ahnormal size,echo,shape or cyst of the kidney) and normal amniotic fluid volume received systemic ultrasound examination,autopsy or follow-up until after birth. The fetus with only dilated renal pelvis was not included. Results Eleven fetuses of dysplastic renal anomalies with normal amniotic fluid volume were identified by prenatal ultrasound. Among the five fetuses affected by unilateral multicystic kidney dysplasia (MCKD),the renal anomaly was isolated in four fetuses,and the other one was complicated with absence of the ipsilateral hand. One of the two fetuses of unilateral renal agenesis had no other associated anomaly and the other one was complicated with hydrocephalus,spina bifida,ipsilateral absent radius and single umbilical artery,correspongding to the VACTERL syndrome. Two fetuses of pelvic kidney and horseshoe kidney respectively was proved by postnatal ultrasound. One fetus was diagnosed as autosomal dominant polycystic kidney disease(ADPKD)on the basis of multiple renal cysts and a positive family history,the fetus also had cardiac rhabdomyoma. One fetus of bilateral normal sized hyperechoic kidneys was proved to be renal dysplasia by autopsy. Conclusions Unilateral MCKD is the most common type of fetal renal dysplasia which can be detected by prenatal ultrasound with normal amniotic fluid volume. Based on the sonographic characteristics and the family history,most of the dysplastic renal anomalies can be diagnosed prenatally and the prognosis can be predicted.

Objective To evaluate the effect of hydrogen peroxide (H2O2) on autophagy in melanocytes,and to explore its possible regulatory mechanisms.Methods Normal human melanocytes at exponential growth phase were divided into several groups:blank control group receiving no treatment,positive control group treated with 100 nmol/L sirolimus solution,and experiment groups treated with H2O2 solution at different volume fractions of 10-7-10-3 respectively.After 4-hour treatment,cell counting kit-8 (CCK-8) assay and flow cytometry were performed to evaluate the cellular proliferative activity and detect apoptosis of melanocytes respectively.Acridine orange staining was performed to detect autophagosome formation,transmission electron microscopy to observe ultrastructural changes of autophagosomes,and Western blot analysis to measure the expression of autophagy-specific protein Beclin 1 and microtubuleassociated protein 1 light chain 3B (LC3B).A total of 84 autophagy-related genes were analyzed by RT2 Profiler PCR Array,so as to screen differentially expressed autophagy-related genes.Results After the treatment with H2O2 at different volume fractions of 10-3,5 × 10-4,10-4,5 × 10-5,10-5,5 × 10-6 and 10-6,experiment groups showed significantly decreased cellular proliferative activity,but significantly increased apoptosis rate compared with the blank control group (F =286.95,301.23,respectively,both P ＜ 0.05).With the increase in volume fractions of H2O2,the cellular proliferative activity was significantly gradually decreased (P ＜ 0.05),while the apoptosis rate showed an opposite trend (P ＜ 0.05),except that the 5 ×10-6 H2O2 group showed no significant differences in the apoptosis rate compared with the 10-5 H2O2 group and 10-6 H2O2 group.Acridine orange staining and electron microscopy showed autophagosome formation in the 10-5 H2O2 group,10-6 H2O2 group and positive control group.Western blot analysis revealed that Beclin1 expression and LC3B-Ⅱ/LC3B-Ⅰ ratio were significantly higher in the 10-5 H2O2 group,10-6 H2O2 group and positive control group than in the blank control group (all P ＜ 0.05).RT2 Profiler PCR Array showed significant up-regulation of ATG12,ATG3,ULK1,PIK3CG,PTEN and PIK3C3 genes and significant downregulation of EIF2AK3 gene in the 10-5 H2O2 group,10-6 H2O2 group and positive control group compared with the blank control group.In the 10-5 H2O2 group and positive control group,the mTOR gene was significantly up-regulated,and the ULK2 gene was significantly down-regulated.The 10-6 H2O2 group showed no obvious changes in the expression of mTOR gene,but significant up-regulation of AMPK and JNK1 genes.Conclusion H2O2 at volume fractions of 10-5 and 10-6 can induce autophagy in melanocytes,likely by influencing the expression of some related signaling molecules.

Objective To evaluate the efficacy of systemic glucocorticoids and intravenous immunoglobulin (IVIG)for the treatment of toxic epidermal necrolysis (TEN). Methods Clinical data on TEN inpatients treated with systemic glucocorticoids alone or in combination with IVIG were collected from the Department of Dermatology, First Affiliated Hospital of Nanjing Medical University from January 2006 to December 2012. Therapeutic outcomes were evaluated in these patients. Statistical analysis was carried out by using a multiple linear regression analysis, binary logistic regression analysis and Cox regression analysis with the SPSS 16.0 software. Results A total of 48 inpatients with TEN were included in this study. Multiple linear regression analysis showed that the maximum daily dose of glucocorticoids for disease control was decreased gradually over years (β=-0.461, P=0.004). However, binary logistic regression analysis revealed no obvious changes in the frequency of administration of IVIG over years. Cox regression analysis showed that both hospitalization duration (RR=0.351, 95.0%CI:0.150-0.825)and the time required for the control of skin lesions (RR=0.492, 95.0%CI:0.245-0.986)decreased with the increase in the frequency of IVIG administration. In addition, with the increase in the maximum daily dose of glucocorticoids for disease control, the time required for the control of skin lesions was also shortened (RR=0.997, 95.0%CI:0.994 -1.000), while no obvious changes were observed in hospitalization duration. Conclusions IVIG shows superiority in controlling lesions, reducing complications and improving the prognosis of TEN. Compared with systemic glucocorticoids, IVIG shows better therapeutic efficacy and less adverse effects, and may be preferentially selected.

Objective:To investigate the clinical and pathological features of breast cancer of young women.Methods:The clinical and pathological characteristics of breast carcinoma were compared between young women(≤35 years old,181 cases) and elder women(≥60 years old,186 cases).Results:More tumors were found in left breast,and located in the upper outer quadrant.The major pathological type was invasive ductal carcinoma.There were significant differences in tumor size,histological grading,TNM staging and axillary lymph node metastases(P

Objective To clone express and purify Schistosoma japonicum fructose?1 6?bisphosphate aldolase SjFBPA in E. coli and observe its expression in different developmental stages of S. japonicum. Methods FBPA gene was amplified from S. japonicum adult worm cDNA by using PCR. The amplified product was recombined into pET28a plasmid and inducibly expressed with IPTG in E. coli BL21. SDS?PAGE and Western blotting were employed to analyze and identify the recombinant protein SjFBPA rSjFBPA . Then rSjFBPA was purified by chromatographic purification and its purity was analyzed by SDS?PAGE. The protein concentration of rSjFBPA purified was measured by the BCA method. Furthermore SjFBPA mRNA was ana?lyzed in different developmental stages of S. japonicum by RT?PCR. Results SjFBPA was successfully amplified by using PCR and identified by restriction enzyme digestion and sequencing. The Western blotting analysis confirmed that the recombinant pro?tein could specifically reactive to the anti?His?tag monoclonal antibody. The concentration of the purified recombinant protein was about 4 mg/ml. The result of RT?PCR showed that SjFBPA mRNA was expressed in cercaria schistosomulum adult worm and egg of S. japonicum. Conclusion SjFBPA is successfully recombined and expressed in a prokaryotic system and SjFBPA mRNA is expressed in cercaria schistosomulum adult worm and egg of S. japonicum.

To investigate the cellular immunity abnormalities of patients with rheumatoid arthritis(RA), peripheral blood lymphocytes subpopulations, interleukin 2 (IL-2) production and natural killer (NK) cell activity were determined in 9 patients with RA. The results showed that there were a remarkable decrease in NK cell activity and poor response to IL-2 stimlulation. IL-2 production and.the expression of membrane-bound IL-2 receptor were increased in RA patients. The percentage of T4 positive cells and the ratio of T4/T8 were also increased in patients with RA. The results indicate that there are severe cellular immunity abnormalities in patients with RA.

Resting-state functional magnetic resonance imaging (rs-fMRI) can be applied to study the resting-state network, and it is easier than task-related fMRI. Imaging principle, research methods of rs-fMRI, as well as its application in cognitive impairment were reviewed in this article.

[Objectives]To investigate the protective effects of recombinant Trichinella spiralis excretory-secretory 53ku pro-tein(rTsP53)on acute lung injuries in mice.[Methods]Thirty Balb/c mice were randomly divided into normal group. ALI group and rTsP53 group(n=10,respectively). Macrophages were harvested by bronchoalveolar lavage. Mortality in 72 hours was counted and compared. Pathological damage of lung tissues was observed by HE staining and graded by Smith score. Wet/dry ratio was measured. The bronchoalveolar lavage fluid concertration of IL-6 and IL-4 was measured by ELISA. The mRNA expression of TNF-α,iNOS, IL-10 and Arg-1 in alveolar lavage macrophages was detected by RT-PCR.[Results]72 h mortality of ALI mice was 70%,which was reduced to 30% in mice received rTsP53 treatment. Compared with ALI mice,the pathological damage of in rTsP53 treated-mice was improved and Smith score was declined ,combined with descending W/D ratio. IL-6 level of alveolar lavage fluid was elevated in ALI mice compared with normal group. And alveolar lavage macrophage was polarized to M2 sub-type,appeared as higher mRNA expression of TNF-α and iNOS and lower level of IL-10 and Arg-1. Bronchoalveolar lavage fluid concentration of IL-6 was declined and IL-4 was elevated in rTsP53-treated mice compared with ALI group. The macrophages of alveolar wash had higher mRNA expression of IL-10 and Arg-1,while lower level of TNF-α and iNOS,manifesting M2 polarization characteristics.[Conclusion]Recombinant T.spiralis P53 protein could protect mice from acute lung injuries induced by LPS via modulating M2 macrophage polarization,which play a role in depression of inflammatory reaction and tissue repairment.