Abstract: Objective To express and purify MPT83 protein of Mycobacterium tuberculosis and evaluate its application value in immunological diagnosis of tuberculosis (TB) using clinical samples. Methods Using Mycobacterium tuberculosis (Mtb) H37Rv genome as the template, Mtb mpt83 gene was amplified by PCR and connected to PET-21a (+) to construct prokaryotic expression vector, and then transferred into E.coli DH5α. The positive colonies were picked out and retained. The recombinant plasmid pET-mpt83 of the strain with positive colony PCR was extracted, identified by double digestion, and the samples of the positive colonies were sent for sequencing. The correctly sequenced plasmids then were transferred into BL21 competent cells for induction, expression and purification with nickel column affinity chromatography. The purified products were identified by 12 alkyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and Western blotting. Mouse polyclonal antiserum was prepared by immunizing mice with purified protein. 8 patients clinically diagnosed as tuberculosis pleural effusion (TB group) and 8 adenocarcinomas patients (CA group) were enrolled and their pleural effusion and plasma were collected. 8 healthy people (HC group) were enrolled as the control group and their plasma were collected. An indirect ELISA was used to detect the level of specific antibodies recognizing MPT83 protein in the samples. Results Mtb MPT83 protein was successfully expressed and purified. The serum titer of MPT83 mouse polyclonal antibody was as high as 1∶1 280 000. The plasma levels of MPT83 antigen specific antibodies in TB group were significantly higher than those in HC group (P<0.05), while the plasma levels of MPT83 antigen specific antibodies in CA group were not significantly different from those in HC group (P>0.05). Compared with the HC group, there was no significant difference in pleural fluid in both the TB and CA groups (P>0.05). The ROC curve was used to analyze the OD values of plasma in TB group and HC group, and the area under the curve was greater than 0.7, showing high diagnostic efficacy. Conclusion　MPT83 protein has high antigen specificity and immunogenicity, which has great application value in the immunological diagnosis of tuberculosis.

Objective To evaluate the effect of acupressure on chemotherapy-induced digestive tract reaction of malignant tumor patients.Methods We searched Pubmed,Medline,Embase,AMED,Cochrane Library,Cancerlit and Cinabl,China Biology Medicine disc (CBM),China National Knowledge Infrastructure (CNKI),VIP database (VIP)and Wanfang database,the databases were searched from the establishment of the database to March 2014.We identified randomized controlled trials (RCTs) and of acupressure on chemotherapy-induced digestive tract reaction for malignant tumor patients.Results In total,8 RCT,959 patients were included.Meta-analysis showed that compared with the no implementation of acupressure,acupressure could effectively reduce the frequency,duration and severity of chemotherapy-induced nausea,vomiting and retching.Meanwhile,acupressure could decrease anti-emetic medication dosage,but could not increase quality of life.Conclusions Acupressure can improve the digestive tract reaction of malignant tumor patients,alleviate the suffering of patients.Meanwhile acupressure couldn't increase the medical burden and mental burden of patients.More attention should be paid to the effect of acupressure on baseline level of state anxiety,functional state and more items.

Aim To investigate the effect of sphingo-sine kinase 1 (SphK1 )on unilateral ureteral obstruc-tion(UUO)-induced tubulointerstitial fibrosis and ex-plore the possible mechanism.Methods The CD-1 mice were randomly divided into four groups:sham-op-eration group(Sham),PF-543 treatment control group (Sham +PF-543),model group(UUO)and PF-543 treatment group(UUO +PF-543).On 1 ,3,7 and 1 4 d after operation,eight mice were selected randomly from each group and sacrificed.The protein expressions of SphK1 ,mature TGF-β1 ,FN,ColⅠ,LC3,Beclin1 ,Atg5 and Atg1 2 were observed by Western blot.The histo-logical changes were examined by Masson′s trichrome stain.Immunhistochemistry was performed to measure the levels of expression of SphK1 ,FN and Col Ⅰ. Transmission electron microscope was used to observe the autophagic body.Results SphK1 expression and autophagy were both upregulated in a mouse model of kidney fibrosis induced by UUO. Meanwhile, in-creased mature TGF-β1 and deposition of extracellular matrix(ECM)were observed in tubulointerstitial areas compared with sham-operated mice.After intraperito-neal injection with the SphK1 specific inhibitor PF-543 in UUO mice,enhanced expression of SphK1 and acti-vated autophagy were significantly abrogated.Howev-er,aggravation of renal fibrosis was detected when SphK1 inhibitor PF-543 was applied to suppress SphK1 expression in UUO mice.Conclusion SphK1 activa-tion is renoprotective through the induction of autoph-agy in the pathogenesis of kidney fibrosis.

ObjectiveTo develop a transgenic mouse model for N.gonorrhoeae researches.Methods Human carcinoembryonic antigen-related cellular adhesion molecules 1 (hCEACAM1) eukaryotic expression vector,pCDPGICAM1,was used to generate transgenic mice by microinjection.The funder mice were screened by PCR,sequence analysis,Western blot and fluorescence-activated cell sorting analysis,respectively.The transgenic mice expressing hCEACAM1 were inoculated with N.gonorrhoeae intravaginally.Adhesion and infection of gonococci to mice were analyzed by bacteria culture and microscopy.Results Four (lines 50,53,54,and 59) of the 22 F0 generation transgenic mice were found to carry the transgene.The hCEACAM1 protein was expressed on the cell membrane of various tissues in the line 53 transgenic mouse.Compared with normal mice,N.gonorrhoeae can successfully infect and cause inflammation in the transgenic mice.Conclusion The hCEACAM1 transgenic mouse can be used as an animal model for gonococcal infections.

AIMTo investigate the effects of NPPB, a chloride channel blocker, on proliferation of mesangial cells.

METHODSCell proliferation was determined by measuring cell number and 3H-thymidine incorporation. The LDH activity released from these cells was measured as evaluation of cell viability. The phase of cell cycle was detected by flow cytometry.

RESULTSCell proliferation assays showed that treatment with both NPPB (50 and 25 micromol x L(-1)) and in hypertonic media (100% increased osmolarity with D-mannitol ) significantly reduced the number of human MC and 3H-thymidine incorporation in a dose-dependent manner. But the LDH activity was not significantly altered in the treatment with 50 micromol x L(-1) NPPB. Flow cytometry experiments showed that 50 and 25 micromol x L(-1) NPPB arrested (84.2 +/- 2.4) % and (80.8 +/- 2.9) % of cells at G0/G1 stage, versus (70.5 +/- 1.4) % of control cells. Conclusion NPPB suppresses cell proliferation and produces growth arrest at G0/G1 phase in human MC by a mechanism probably associated with changes in cell volume.

Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chloride Channels ; antagonists & inhibitors ; Dose-Response Relationship, Drug ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Mesangial Cells ; cytology ; metabolism ; Nitrobenzoates ; administration & dosage ; pharmacology

Objective To test the effectiveness of an integrated intervention to improve control and management of type 2 diabetic patients.Methods The study was designed as a randomized and multi-center clinic trial for 24 weeks.150 overweight patients from 2 companies and one hospital in Shanghai were recruited.The principal objective of the project was to compare the effectiveness of a structured diabetes management program consisting of 200 kcal breakfast as a meal replacement with low glycemic index (GI),scheduled blood glucose monitoring(6 times per week),low GI diet consulting,monitoring and medical check up and nutritional education (diet consulting,health education on nutrition and healthy lifestyle) in overweight individuals with type 2 diabetes.Results At 12th week,the level of fasting blood glucose(FBG) in both groups decreased by 16.5% and 10.6%,and at 24th week,the FBG also decreased by 25.0% in intervention group,but in control group FGB increased 2.8% (P＜0.01).Meanwhile,there were 8.2% and 11.1% reductions in HbA1C respectively at 12th week and at 24th week in intervention group,while the values were increased slightly in control group(P＜0.01).Compared with control group,waist circumference,hip circumference,and blood pressure were significantly reduced after treatment for 24 weeks in intervention group (P＜0.01).Conclusions Integrated intervention is an effective approach in managing FBG,HbA1C blood pressure,and weight control in overweight diabetic patients.

Nine proteins encoded by Mycobacterium tuberculosis RD1 region are important protective antigens that become absent in long passaging of Mycobacterium tuberculosis. They only exist in pathogenic Mycobacteria and are absent in Bacille Calmette-Guerin and environmental Mycobacteria. With good immunogenicities, they may play an important role in the diagnosis and prevention of Mycobacterium tuberculosis. This article reviews recent studies on using RD1-encoded proteins as antigens in the diagnosis of active tuberculosis and tuberculous pleurisy.
Antigens, Bacterial ; isolation & purification ; metabolism ; Humans ; Mycobacterium tuberculosis ; physiology ; Tuberculosis ; diagnosis

OBJECTIVETo investigate the relationship between condylar marrow signal abnormalities and temporomandibular joint internal derangement (TMJID).

METHODSOblique sagittal T1 weighted MR imaging at closed and open mouth and Oblique sagittal T2 weighted MR imaging at closed mouth were obtained from 88 joints of 44 patients suffering from TMD. Condylar marrow signal abnormalities were reviewed and classified into bone marrow edema pattern (hypointense T1, hyperintense T2), sclerosis pattern (hypointense T1 and hypointense T2) and combined edema and sclerosis pattern.

RESULTSOf 88 joints, 13 (14.8%) joints showed condylar marrow signal abnomalities, among which 11 belonged to edema pattern and, 1 was sclerosis pattern and the other was the combined patten. Of 13 joints with condylar marrow signal abnomalities, 11 (84.6%) had TMJID. Of 75 joints with normal marrow signal, 25 (33.3%) joints had TMJID. There was significant correlation between condylar marrow signal abnormalities and TMJID (P < 0.05).

CONCLUSIONDisc displacement is one of the factors inducing condylar marrow signal abnormalities. The pathological process from disc displacement to osteonecrosis requires further study.

Adolescent ; Adult ; Aged ; Child ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Mandibular Condyle ; pathology ; Middle Aged ; Temporomandibular Joint Disc ; pathology ; Temporomandibular Joint Disorders ; diagnosis ; pathology

Objective To explore the characteristic of the genetic frequency distribution of human leukocyte antigen(HLA)-DR alleles in children with bronchial asthma in Guangxi area.Methods Eighty-four unrelated asthmatic individuals and 168 healthy people without asthma and atopy living in Nanning region of Guangxi as control group were involved in the study.All asthmatics had their serum total IgE levels measured with Pharmacia UniCAP system,and skin-prick test with 10 kinds of inhalant allergens were taken,and pulmonary function were measured among the asthmatic.HLA oligonucleotide array was used to 21 gene frequencies of HLA-DR.Results The frequencies of HLA-DR B1*070X allele and HLA-DR B1*11XX allele among the asthmatic(2.98% and 13.69%)were significantly higher than those in control group(0.3% and 5.95%)(?2 =6.915,9.478 P

Objective To investigate the current distribution of hepatitis C virus (HCV) genotype in Southern China and to understand the HCV transmission and to infer its transmitting trend.Methods The HCV gene subtypes of 3 524 specimens from Southern China were detected and analyzed by polyonerase chain reaction (PCR)-fluorescence probe method or sequencing.The regular nested PCR and sequencing were used for the phylogenetic tree analysis when the fluorescence PCR inefficiently identifying virus isolates.Results Among 3 524 specimens,there were 2 922 cases from Guangdong,78 cases from Fujian,152 cases from Hainan and 372 cases from Guangxi.Genotype 1b comprised the majority (1 808/ 3 524,51.3％),followed by genotype 6a (925/3 524,26.2％),2a (298/3 524,8.46％),3a (246/ 3 524,6.98％),3b (200/3 524,5.68％) and 1a (27/3 524,0.77％).In addition,1 case wasgenotype 6e,1 case was genotype 6q,1 case was genotype 6r,3 case were genotype 6w,2 case were genotype 6xa,2 case were genotype 6n,and 1 case was genotype 6 with unclassified subtype.The genotype 1b accounted for the majority in most areas of 21 cities and counties in Guangdong Province,followed by genotype 6a.But in some areas,the major genotype was genotype 6a,followed by 1b.Genotype 4,genotype 5 and genotype 7 were not found in this study.Conclusions In the past two years,genotype 1b and 6a are still the epidemic genotypes in Guangdong,Guangxi and Hainan provinces.However,genotype 6a has replaced 1b as the dominant one in some areas in Guangdong Province.The distributions of HCV genotypes do not change significantly in Guangxi and Fujian provinces.