Human gut microbiota plays a key role in the development of obesity. Intestinal flora can regulate energy absorption and nutrition metabolism, increasing the energy harvesting from diet. Alteration of gut flora produces excessive lipopolysaccharide, which, when absorbed into the blood, can induce inflammatory reactions and promote the high-fat diet-associated obesity and metabolic syndrome. Intestinal flora increase visceral fat deposition by lowering the expression of Fiaf in intestinal mucosa. Different immune status also affects the intestinal flora.The gut microbiota is hypothesized to be an environmental factor that contributes to obesity; by interacting with factors such as host and diet, it adjusts the energy metabolism. Antibiotics or probiotics may alter the composition of intestinal microflora and improve the metabolic syndrome, and thus provides new treatment options.
Animals ; Diet, High-Fat ; Gastrointestinal Tract ; microbiology ; Humans ; Inflammation ; etiology ; Mice ; Obesity ; microbiology ; therapy ; Probiotics ; therapeutic use

OBJECTIVETo study the mechanism of hepatitis B virus infected patients who is negative for HbsAg.

METHODSDNA sequences of 46 patients were analyzed. In these patients, HBsAg was negative but HBV DNA was positive and six new HBsAg variants were identified. Four of the six variants were combined point mutants and two were insertion variants. These S genes were subcloned into eukaryotic expression vector EBO-plpp, and the recombinant eukaryotic expression plasmids were transfected into COS7 cells. Cell lines expressing mutant type HBsAg were obtained. The supernatants were detected by ELISA and RIA.

RESULTSOnly the two-amino acid-insertion variants could be detected and the others failed to react with polyclonal and monoclonal antibodies against HbsAg.

CONCLUSIONThe results indicated that the point mutations and insertions may result in a conformational change of the S gene, which affect HBsAg antigenicity, suggesting a possible relationship between the variants and the negative conversion of HBsAg of the patients.

Animals ; Antigenic Variation ; COS Cells ; Cercopithecus aethiops ; Hepatitis B Surface Antigens ; genetics ; immunology ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; immunology ; virology ; Humans ; Plasmids ; genetics ; Point Mutation ; Transfection

BACKGROUNDThe extended spectrum β-lactamase (ESBL)-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are the major pathogens causing pneumonia and have a significant impact on the clinical course. Limited data exist on molecular characterization of ESBL-producing E. coli and K. pneumoniae that cause pneumonia. The aim of this study was to investigate the comprehensive multilevel characteristics of E. coli and K. pneumoniae causing pneumonia in China for the first time.

METHODSE. coli (17) and K. pneumoniae (21) isolates responsible for pneumonia were isolated from 1270 specimens collected in a prospective multi-center study in eight teaching hospitals in China from June to December in 2007. The susceptibilities, ESBL confirmation, sequence typing, blaCTX-M and blaSHV genes, their genetic environment and plasmid Inc/rep types were determined.

RESULTSSixteen E. coli (94.1%) and eleven K. pneumoniae (52.4%) isolates were ESBL producers. About 77.8% and 66.7% of them were resistance to ciprofloxacin and levofloxacin, and 100% were susceptible to imipenem. The most prevalent ESBL gene was CTX-M-14, followed by SHV-2, CTX-M-15, CTX-M-3, CTX-M-65, SHV-12, SHV-26 and SHV-28. SHV-1 and SHV-11 were also detected and coexisted with blaCTX-Ms in five strains, and three strains contained only SHV-1. All CTX-M-14 were detected ISEcp1 upstream and nine were found IS903 downstream and the majority of them (64.3%) were carried by IncF plasmids. All blaSHV were flanked by recF and deoR, located on IncF, IncN, IncX and IncH plasmids. Two SHV-2, one SHV-1 and the only SHV-28 were further preceded by IS26. Genes lacY and lacZ were detected at further upstream of two blaSHV-1. The K. pneumoniae carrying SHV-28 was susceptible to β-lactams, and no mutations or deletions in gene or promoter sequences were identified to account for susceptibility. Multilocus sequence typing experiments showed the ESBL-producing strains were genetically diverse.

CONCLUSIONSThe rate of occurrence of blaESBL in E. coli and K. pneumoniae causing pneumonia was high, and blaCTX-M-14 was dominant and probably mobilized by ISEcp1 mainly on IncF plasmids. Importantly, unexpressed blaESBL genes may occur in susceptible isolates and hence may have clinical implications.

Blotting, Southern ; Escherichia coli ; enzymology ; genetics ; Klebsiella pneumoniae ; enzymology ; genetics ; Plasmids ; genetics ; Pneumonia ; microbiology ; Promoter Regions, Genetic ; genetics ; Prospective Studies ; beta-Lactams ; metabolism

Thyroid cancer is the one of the most common endocrine tumors. The biological behaviors and prognoses of the thyroid cancer of different histological types remarkably differ. The highly invasive thyroid cancer responds poorly to traditional therapies. Recent research advances in the molecular mechanisms of the pathogenesis of thyroid cancer have revealed the roles of many genetic and epigenetic variations such as gene mutation, abnormal gene amplification, and abnormal gene methylation in the development of thyroid cancer, which provides new insights in the molecular diagnosis, prognosis, and target therapy of the thyroid cancer.
Carcinoma ; genetics ; Humans ; Mutation ; Signal Transduction ; Thyroid Neoplasms ; genetics

Objective To investigate the epidemiology of BDV infection in Yili horses and Yili donkeys and to analyze phylogenetic source of BDV in Yili area, Xinjiang. Methods We established fluo- rescence quantitative nested RT-PCR to detect BDV p24 segment in peripheral blood mononuclear cells (PBMCs) of 518 Yili horses and 206 Yili donkeys in Yili area, Xinjiang. Positive products were validated by detecting BDV p40 segment and plasmid to preclude the contamination, and were sequenced to analyze the homology of gene sequence, amino acid sequence and phylogenetic tree. Results The positive rates of BDV infection in PBMCs of 518 Yili horses and 206 Yili donkeys were 0.97% and 1.94%, respectively. The results of BDV p40 segment verification were positive in all of the samples of BDV p24 positive. All the samples tested were not contaminated by plasmid. There was a homology of the gene sequence of positive PCR samples with strain He/80. And the gene sequence revealed more than 93% identical to H1766 and strain V. Conclusion Our study suggested BDV natural infection in Yili horses and Yili donkeys. The en- demic BDV had a high degree of identity to strain He/80.

Objective To study the biological significance of HBV gene mutation at nucleotide (nt) 1862. Methods The EB virus eukarotic expression vector for HBV pre-C/C gene was constructed using molecular biological method, HBV pre-C/C gene mutation at nt 1862 was induced by way of site-specific mutation technique, and identified by PCR-RFLP and sequencing analysis. The resulted recombinant plasmid containing the HBV variant was subsequently transfected into Cos7 cell line mediated by lipofectin, to observe the expression of HBeAg. The cells transfected with the recombinant plasmid con- taining wild HBV pre-C/C gene fragment served as control. Results HBeAg expression was detected in the cells transfected with wild recombinant plasmid but not in those with HBV variant transfection. Conclusion The success in the construction of eukarotic expression vector for HBV pre-C/C gene mutation at nt 1862 may pave the way for further studying a series of biological changes of HBV resulted from the mutation addressed in this study.

Objective To study the biological significance of HBV gene mutation at nucleotide (nt) 1862. Methods The EB virus eukarotic expression vector for HBV pre-C/C gene was constructed using molecular biological method, HBV pre-C/C gene mutation at nt 1862 was induced by way of site-specific mutation technique, and identified by PCR-RFLP and sequencing analysis. The resulted recombinant plasmid containing the HBV variant was subsequently transfected into Cos7 cell line mediated by lipofectin, to observe the expression of HBeAg. The cells transfected with the recombinant plasmid con- taining wild HBV pre-C/C gene fragment served as control. Results HBeAg expression was detected in the cells transfected with wild recombinant plasmid but not in those with HBV variant transfection. Conclusion The success in the construction of eukarotic expression vector for HBV pre-C/C gene mutation at nt 1862 may pave the way for further studying a series of biological changes of HBV resulted from the mutation addressed in this study.

Background and Objective:The gross tumor volume (GTV) obviously reduces after induction chemotherapy (IC) for primary Iocoregionally advanced nasopharyngeal carcinoma (NPC). This study was to investigate the impact of changing gross tumor volume delineation on the dose distribution and clinical treatment outcome after IC. Methods: From January 2008 to April 2009, 24 patients with Stage Ⅲ-Ⅳb primary locoregionally advanced NPC were treated with TPF regimen IC followed by intensitymodulated radiotherapy (IMRT) with concurrent chemotherapy The primary GTVs were delineated into two parts: the post-IC primary GTV (GTVpost-IC-NP),and the region of pre-IC primary GTV minus GTVpost-IC-NP (GTVprepost-IC-NP). The dose distributions of two plans with GTVpost-IC-NP or pre-IC primary GTV were assessed by analyzing ten cases. The clinical treatment outcome and toxicity of all patients were observed. Results:The post-IC GTV was significantly smaller than the pre-IC GTV(primary GTV 25.5 cm~3 vs.51.1 cm~3,P=0.001;lymph nodes GTV 9.1 cm~3 vs. 31.4 cm~3, P=0.035;primary+lymph nodes GTV 33.2 cm~3 vs. 82.6 cm~3, P=0.004), the overall GTV with an average shrinkage of 61%. The high dose region was also smaller after IC(volumes covered by 64.4 Gy were 422.9 cm~3 vs.457.9cm~3, P=0.003; 274.2 cm~3 vs.334.5 cm~3 by 68 Gy, P=0.041). The complete response rate was 38% after IC,and 100% three month after radiotherapy.The toxicity of following IMRT with concurrent chemotherapy was similar to that of IMRT with concurrent chemotherapy alone. With median follow-up of 9 months, the locoregionally control rate was 100% and only one patient presented metastasis 15 months after treatment. Gonclusions: TPF regimen IC could significantly reduce tumor volume. The following IMRT with GTVpost-IC-NP plan reduced the high dose region,which didn't add toxicity while had excellent short-term treatment outcome.

Background:Pioglitazone may be beneficial in the treatment of psoriasis.However,based on the effectiveness and safety considerations,it has not been widely used.To fully evaluate the strength of evidence supporting psoriasis treatment with pioglitazone,we conducted a meta-analysis of existing published studies.Methods:PubMed,Ovid,Cochrane Library,Google Scholar,and Web of Science databases were systematically searched before February 2019.Randomized controlled trials (RCTs) of pioglitazone administration compared with placebo,administered to patients with psoriasis for at least 10 weeks,and published in English were included.Quality of the included RCTs was identified by the modified Jadad scale.The quality of evidence for each outcome was evaluated using the GRADEpro Guideline Development Tool online software.Primary outcomes were proportion of patents showing psoriasis area and severity index (PASI) score improvement (＞75％) and the mean percent change in PASI score from baseline to the end of treatment.Dichotomous data were analyzed using odds ratios (ORs) corresponding to the 95％ confidence interval (CI),whereas continuous variables,expressed as mean and standard deviation,were analyzed using the mean differences (MD) with the 95％ CI.Results:Six RCTs were analyzed.Meta-analysis showed that pioglitazone reduced the PASI scores in patients with psoriasis compared with the control group when administered at 30 mg per day (P ＜ 0.001,MD =-3.82,95％ CI =-5.70,-1.93) and at 15 mg per day (P =0.04,MD =-3.53,95％ CI =-6.86,-0.20).The PASI-75 of the pioglitazone group was significantly higher than that of the control group at 30 mg per day (P ＜ 0.001,OR =8.30,95％ CI =3.99,17.27) and at 15 mg per day (P =0.03,OR =2.96,95％ CI =1.08,8.06).No statistically significant differences in total adverse events were observed between the groups.There were no significant differences in common adverse reactions such as weight gain and elevated liver enzymes between the two pioglitazone groups.Conclusions:Use of pioglitazone in the current treatment of psoriasis is beneficial.The therapeutic effect of the daily 30 mg dose may be greater than that of the 15 mg dose per day with no significant change in the frequency of adverse reactions.

OBJECTIVETo prepare rat whole-kidney acellular matrix (ACM) scaffolds using fluid perfusion method.

METHODSThe kidneys with ureters and renal vessels were harvested from 12-week-old Wistar rats. Intravenous catheters were inserted through the renal arteries to establish channels for whole-kidney retrograde perfusion successively with heparinized PBS, 1% SDS, deionized water, 1% TritonX-100 and antibiotic-containing PBS under a pressure of 100 cmH2O. After decellularization, the scaffolds were observed under microscope with HE staining, scanning electron microscope, and fluorescence microscope with DAPI fluorescence staining.

RESULTSNo cell residue was found in the scaffolds under microscope. Scanning electron microscope identified reticular structures consisting of basilar membrane and collagen without normal cellular structures in the scaffolds, and no strong fluorescence due to the binding of DAPI to the cell nuclei was observed under fluorescence microscope.

CONCLUSIONFluid perfusion is simple and reliable to prepare rat whole-kidney acellular matrix, which may serve as an ideal cell-free scaffold.

Animals ; Biocompatible Materials ; Cell Separation ; methods ; Extracellular Matrix ; Female ; Kidney ; cytology ; Male ; Perfusion ; Rats ; Rats, Wistar ; Tissue Engineering ; methods ; Tissue Scaffolds