Objective To investigate the characteristics of human immunodeficiency virus(HIV-1)V3 loop amino acid mutations among AIDS patients in Shenzhen. Methods Fragments of HIV-1 env gene were amplified by RT-nested-polymerase chain reaction(PCR)from plasma of the AIDS patients in Shenzhen.The PCR products were then directly sequenced and the sequences were aligned,translated and analyzed. Results Sequence analysis showed that there were B and CRF01-AE HIV-1 subtypes in Shenzhen.The V3 loop terra peptide GPGQ accounted for about 48%,GPGR about 36%,other V3 loop terra peptide forms were 16%;the V3 loop central motif in the majority(76.9%)of the CRF01-AE strains was GPGQ,the majority(75%)of the B strains was GPGR.We also found some unusual terra peptide compositions:DQDR and DQGQ on the tip of V3 loop,some amino acid mutations at specific of V3 loop associated with SI phenotype were very common in AIDS patients(80%). Conclusions The V3 loop amino acid mutations were very common in AIDS patients,the V3 loop central motif in the majority was GPGQ and GPGR in Shenzhen,the mutations shows the change of HIV-1 phenotype and predicts that disease progress to AIDS.

Objective To investigate the antimicrobial resistance mechanisms and genetic homogeny of Salmonella from community acquired infections in Shenzhen,China.Methods Ninety-three of Salmonella were isolated from 2002 to 2007 at Shenzhen People's Hospital,China.PCR and DNA sequencing were used to investigate the mutation in QRDR of the gyrA,gyrB,parC and parE.Plasmid mediated quinolone resistance genes including qnr and aac(6')-Ib-cr,β-lactamase genes including blaTEM,blaSHV,blaOXA, blaCTX-M, and class 1 integron were detected. All isolates were typed by PFGE. Results S. enterica typhi and S. enterica paratyphi A were susceptible to ampicillin, chloramphenicol, trimethoprim/sulfamethoxazole, ceftriaxone and ciprofloxacin, with the susceptible rate of 96%-100%. Fifty-two percent (13/25) of S. enterica typhi and 95% (61/64) of S. enterica paratyphi A were resistant to nalidixic acid. Twenty-four percent (6/25) of nalidixic acid-resistant S. enterica typhi and 94% (60/64) of nalidixic acid-resistant S. enterica paratyphi A showed decreased susceptibility to ciprofloxacin (MIC of 0. 125-1 μg/ml).All nalidixic acid-resistant (susceptible to ciprofloxacin ) Salmonella (NARS) isolates had a single substitution in the QRDR of GyrA, and 91% (68/75) of these isolates carried the substitution Ser83Phe in GyrA. Two mutations in the QRDR of GyrA were detected in both of two ciprnfloxacin-resistant Salmonella,with the additional one mutation in the QRDR of parC. Plasmid mediated quinolone resistance genes including qnr and aac(6')-lb-cr were not detected in any isolate. The blaCTX-M-14 gene was detected in a ceftriaxoneresistant isolate of S. enterica paratyphi A, with ISEcpl located on the upstream of it. Three muhidrugresistant strains of Salmonella all carried one 1 900 bp classⅠ integron gene cassette dhfrⅫ-orfF-aadA2,with the additional one β-lactamase gene of blaTEM-1, or blaOXA-30. Twenty-two distinct PFGE patterns were observed among twenty-five S. enterica typhi. The PFGE patterns of sixty-four S. enterica paratyphi A showed limited genetic diversity (average similarity of 91% ). Ninety investigated inpatients were infected in the community. Six patients infected by S. enterica paratyphi A had a travel history before infection. Conclusions Nalidixic acid-resistant S. enterica typhi and S. enterica paratyphi A are highly prevalent in Shenzhen,China. The mutation in the QRDR of GyrA is the prevalent mechanism responsible for the resistance to nalidixic acid in Slmonella. The great genetic similarity among S. enterica paratyphi A isolates indicates endemic disease from the presence of a single clone over 6-year period.

Objective: To analyze the mortality trend of bladder cancer among residents in Qidong, Jiangsu Province from 1972 to 2016, so as to provide the basis for the prevention and treatment strategy of bladder cancer in Qidong. Methods: The data of bladder cancer was collected from Qidong Cancer Registry.The crude mortality rate ( CR ), age-standardized rate by Chinese population in 2000 (CASR) and world population in 1960 ( WASR ), truncated rate (35-64 years) and cumulative rate ( 0-74 years ) were calculated. The annual percent change ( APC ) was used to analyze the trend of mortality in bladder cancer. Results: During from 1972 to 2016, There were 1 497 deaths due to bladder cancer in Qidong from 1972 to 2016. The CR, CASR and WASR were 2.96/105, 1.83/105 and 1.80/105, respectively. The APCs in CR, CASR, WASR of bladder cancer were 5.29%, 1.86% and 1.81%, respectively ( P<0.05 ), showing upward trends. The truncated rate, cumulative rate and cumulative risk were 1.47/105, 0.17% and 0.17%, respectively. The CR, CASR and WASR in males were 4.71/105, 2.97/105 and 3.31/105, respectively, which was higher than that of 1.26/105, 0.75/105, and 0.66/105 in females ( P<0.05 ). The APC of CR, CASR and WASR in males were 5.71%, 1.96% and 2.17%, respectively ( P<0.05 ), all showed upward trends. For females, the APC of CR was 4.47% ( P<0.05 ), showing an upward trend, but there was no significant change in CASR and WASR ( P>0.05 ). The CR of bladder cancer was high among people aged more than 55 years. The CR in 55-64-year-old group, 65-74-year-old group and more than 75-year-old group showed upward trends, with APC of 4.50%, 2.22% and 4.51%, respectively ( P<0.05 ). Conclusions From 1972 to 2016, the mortality of bladder cancer in Qidong showed an upward trend, which was relatively high in men and people aged over 55 years.

Antigens, CD ; metabolism ; CD3 Complex ; metabolism ; Celiac Disease ; complications ; Chromosome Aberrations ; Chromosomes, Human, Pair 9 ; Diagnosis, Differential ; Enteropathy-Associated T-Cell Lymphoma ; complications ; genetics ; metabolism ; pathology ; Humans ; Integrin alpha Chains ; metabolism ; Intestinal Mucosa ; pathology ; Ki-1 Antigen ; metabolism ; Lymphoma, Extranodal NK-T-Cell ; pathology ; Lymphoma, Large B-Cell, Diffuse ; pathology

NS2 and NS3 are two post-transcriptional gene silencing suppressors that are encoded by Rice stripe virus. Gene silencing suppressors are always related to the pathogenicity of viruses. In this study, the cDNA of NS2 and NS3 were recombined by overlapping PCR assays, ligated to the RNAi vector, and inserted into the PXQ expression vector using Pst I; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'Nipponbare', using an Agrobacterium-mediated method. Thirty-one T0 transgenic plants were selected by G418 screening. PCR and southern blot analyses confirmed that the target gene was transformed into transgenic rice successfully, and different transgenic plants contained various copies of the gene. The disease resistance assay revealed that T0 transgenic rice had a delayed onset of RSV for approximately 10-20 d, and the accumulation of virus in the transgenic plants was reduced by 30%-50%. This was related to the delayed onset of disease.
Disease Resistance ; Oryza ; genetics ; immunology ; virology ; Plant Diseases ; genetics ; immunology ; virology ; Plants, Genetically Modified ; genetics ; immunology ; virology ; RNA Interference ; Tenuivirus ; genetics ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology

Introduction: Tuberculosis (TB) in internal migrants is one of three threats for TB control in China. To address this threat, a project was launched in eight of the 19 districts of Shanghai in 2007 to provide transportation subsidies and living allowances for all migrant TB cases. This study aims to determine if this project contributed to improved TB control outcomes among migrants in urban Shanghai. Methods: This was a community intervention study. The data were derived from the TB Management Information System in three project districts and three non-project districts in Shanghai between 2006 and 2010. The impact of the project was estimated in a difference-in–difference (DID) analysis framework, and a multivariable binary logistic regression analysis. Results: A total of 1872 pulmonary TB (PTB) cases in internal migrants were included in the study. The treatment success rate (TSR) for migrant smear-positive cases in project districts increased from 59.9% in 2006 to 87.6% in 2010 (P < 0.001). The crude DID improvement of TSR was 18.9%. There was an increased probability of TSR in the project group before and after the project intervention period (coefficient = 1.156, odds ratio = 3.178, 95% confidence interval: 1.305–7.736, P = 0.011). Conclusion: The study showed the project could improve treatment success in migrant PTB cases. This was a short-term programme using special financial subsidies for all migrant PTB cases. It is recommended that project funds be continuously invested by governments with particular focus on the more vulnerable PTB cases among migrants.

Background The injury or surgery of cornea cause the proliferation of corneal stromal cells and scar formation.Recent research showed that cureumin can obviously reduce the degree of fibrosis of tissue.But if curcumm play inhibitory effect on corneal keratocytes fibrosis is rarely reported.Objecttve This studv was to investigate the effect of curcumin on the transformation of corneal keratocytes into fibroblasts in vitro and further explore the antifibrotic effect of curcumin on corneal keratocytes.Methods The murine corneal keratocytes from 150 BALB/c mice were isolated and primary culture in DMEM culture medium containing 10% fetal bovine serum and then divided into blank control group(inducer group,CG),low-dose group(CG+7.5 mg/L curcumin),mediumdose group(CG+10.0 mg/L curcumin),high-dose group(CG+12.5 mg/L curcumin),non-inducer group.Seven days following intervention,the expression of cell markers such as keratocan,aldehyde dehydrogenase(ALDH),decorin and fibronectin-1 in keratocytes were analyzed by RT-PCR.The effect of curcumin on cultured murine corneal keratocytes proliferation was evaluated by MTS technique.The expression of fibronectin-1 in murine cornea was investigated by immunofluorescence assay.Results The primarily cultured keratocytes showed tlIe fusiform-like shape with the abundant cytoplasm and big nuclei.In the presence of curcumin,the mRNA levels of keratocan and ALDH were down-regulated and those of CD90 and decorin were up-regulated,showing the significantly differences with the increase of dose(P<0.05),but the expression pf fibronectin-i was not obviously changed with the alteration of dose of curcumin. MTS showed that the inhibitory rates of curcumin on keratocytes in 10.0 mg/L and 2. 5 mg/L groups were enhanced in comparison with 7.5 mg/L group, showing statistically significant difference among three groups( F = 956.00, P<0.05). The expression of fibronectin-1 was found in the corneal keratocytes with the red fluorescence in stroma. Conclusion Curcumin can inhibit the fibrosis of corneal keratoeytes in a dose-dependent manner. These results offer a preliminary theoretical basis for the application of curcumin in controlling corneal scar formation during wound healing.

Background Researches demonstrated that dendritic cells(DCs) are uniformly immature in the central cornea but mature in the peripheral region of cornea.So an important question is which factor impact the maturation of DCs,especially in terms of corneal transplant rejection and the known roles of DCs in the development and persistence of some corneal diseases.Objective This study aimed to examine whether corneal stroma cells (CSCs) inhibit DCs maturation through secreting transforming growth factor beta 2 (TGF-β2) and prostaglandin E2 (PGE2).Methods DCs,T cells and CSCs were isolated and cultured from clean BALB/c and C57BL/6 mice.The level of PGE2 and TGF-β2in CSCs culture supernatant and the fresh RPMI 1640 medium were then analyzed by enzyme linked immunosorbent assay (ELISA).During the DCs maturation stage,the neutralizing TGF-β2 antibody and the EP2 receptor antagonist AH6809 were added in the CSCs culture supernatant respectively.According to the different treatment,cultured cells were assigned to different groups as follows:control group,CSCs culture supernatant group,AH6809 group,TGF-β2 antibody group,AH6809 +TGF-β2 antibody group.Subsequently,the cellular surface markers for DCs,including CD11c,CD80,CD86,and MHC- Ⅱ,were analyzed by flow cytometry.The capability of stimulating the proliferation of T lymphocytes was evaluated by allogeneic mixed lymphocyte reactions,and the function of endocytosis was assessed by fluorescein isothiocyanate-dextran(FITC) uptake.Results The data of ELISA showed a higher concentration of TGF-β2 and PGE2 in murine CSCs culture supernatant than in the fresh RPMI 1640 medium.Compared with the CSCs culture supernatant group,the expression of CD80,CD86,and MHC- Ⅱ was up-regulated ( P ＜ 0.05 ),the expression of dextran was down-regulated ( P ＜ 0.05 ),and the stimulate index was increased( P＜ 0.05 ) in the TGF-β2 antibody group; the expression of CD86,and MHC-Ⅱ was up-regulated (P＜0.05),the expression of dextran was down-regulated ( F =13.740,P =0.006 ),and the stimulate index was increased(P＜0.05) in the AH6809 group;the expression of MHC-Ⅱ was up-regulated and the stimulate index was increased with statistical difference in interaction(P＜0.05 ) in the AH6809+TGF-β2 antibody group.Compared with the control group,the expression of CD80 and CD86,and the stimulate index was still lower(P＜0.05 ).Conclusions TGF-β2 and PGE2 contribute to the inhibitory effects on DCs maturation mediated by murine CSCs in vitro and further have additive effect on the immunosuppression of DCs.

Objective To analysis the immunological characteristics of patients with severe H1N1 influenza A, and to provide theoretical basis for predicting the prognosis of the disease. Methods A retrospective analysis was conducted. The clinical data of 15 patients diagnosed with severe H1N1 influenza A and admitted to Shanghai General Hospital of Nanjing Medical University from October 2015 to December 2016 were collected. All the patients were divided into survival and death groups according to 28-day survival. Clinical characteristics, treatment algorithm, organ function, inflammatory reaction and immune cell status were compared, and Cox regression was used to decide the risk factors of 28-day death in patients with severe H1N1 infection A. Results All 15 patients with severe H1N1 infection A were enrolled, most of who presented with cough (93.3%), fever (86.7%), sputum production (80.0%), shortness of breath (73.3%), myalgia (40.0%) and fatigue (40.0%). All had been received anti-virus, antibiotics, mechanical ventilation and anti-coagulation therapy; some were treated with prone position, neuromuscular blocker and extracorporeal membrane oxygenation (ECMO). The incidences of acute myocardial and kidney injury were high, and the acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score (14.1±6.1) and sequential organ failure assessment (SOFA) score (9.6±4.1) implicated the critical condition. Of 15 patients, 4 patients died in 28 days, while 11 were cured and discharged. Compared with survival group, the patients in death group had higher levels of APACHE Ⅱscore (22.7±3.8 vs. 11.8±3.8), troponin [cTn (μg/L): 0.52 (0.07, 2.02) vs. 0.15 (0.10, 0.45)] and blood urea nitrogen [BUN (mmol/L): 11.9 (6.7, 29.1) vs. 3.9 (2.7, 6.8)] and a lower level of blood platelets count [PLT (×109/L): 76±33 vs. 146±49, all P < 0.05]. The levels of C-reactive protein (CRP) and interleukin-6 (IL-6) within 24 hours of admission in death group were significantly higher than those of survival group [CRP (mg/L): 172.2±88.5 vs. 74.8±33.1, IL-6 (ng/L):283.3 (140.1, 711.0) vs. 18.5 (12.7, 71.4), both P < 0.01]. Compared with survival group, the expressions of CD3+, CD4+, CD8+ T cells and natural killer cell (NK cell) in death group were significantly decreased (CD3+ T cell: 0.348±0.119 vs. 0.573±0.106, CD4+ T cell: 0.135±0.046 vs. 0.344±0.098, CD8+ T cell: 0.089±0.057 vs. 0.208±0.054, NK cell: 0.124±0.057 vs. 0.252±0.182, all P < 0.05), but there were no significant differences in CD4+/CD8+ ratio and human leucocyte antigen-DR positive (HLA-DR+) T cell between death group and survival group (CD4+/CD8+ ratio:1.57±0.26 vs. 1.83±0.54, HLA-DR+ T cell: 0.035±0.022 vs. 0.062±0.036, both P > 0.05). B lymphocyte in death group was significantly higher than that of survival group (0.477±0.136 vs. 0.229±0.121, P < 0.01). Cox regression analysis revealed that APACHE Ⅱ score [risk ratio (RR) = 20.4, 95% confidence interval (95%CI) = 5.3-31.2, P = 0.017], CD4+ T cell (RR = 11.1, 95%CI = 5.1-20.0, P = 0.048) and CD8+ T cell (RR = 9.1, 95%CI = 4.3-16.7, P = 0.049) were independently risk factors of 28-day survival of patients with severe H1N1 influenza A. Conclusion Immunological paralysis and severe inflammatory response were early complicated with severe H1N1 influenza A, and these were significantly associated with prognosis.

Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals ; Body Fluids ; chemistry ; Chemical Precipitation ; Chemistry Techniques, Analytical ; methods ; Humans ; Liquid-Liquid Extraction ; Proteins ; isolation & purification ; Solid Phase Extraction