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Hu S.,Xiamen University | Li G.,309th Hospital of Chinese PLA | Li H.,U.S. Center for Disease Control and Prevention | Liu X.,Shenzhen Center for Chronic Disease Control | And 6 more authors.
Journal of Clinical Microbiology | Year: 2014

The MeltPro TB/INH assay, recently approved by the Chinese Food and Drug Administration, is a closed-tube, dual-color, melting curve analysis-based, real-time PCR test specially designed to detect 30 isoniazid (INH) resistance mutations in katG position 315 (katG 315), the inhA promoter (positions-17 to -8), inhA position 94, and the ahpC promoter (positions-44 to -30 and -15 to 3) of Mycobacterium tuberculosis. Here we evaluated both the analytical performance and clinical performance of this assay. Analytical studies with corresponding panels demonstrated that the accuracy for detection of different mutation types (10 wild-type samples and 12 mutant type samples), the limit of detection (2 × 103 to 2 × 104 bacilli/ml), reproducibility (standard deviation [SD], <0.4°C), and the lowest heteroresistance level (40%) all met the parameters preset by the kit. The assay could be run on five types of real-time PCR machines, with the shortest running time (105 min) obtained with the LightCycler 480 II. Clinical studies enrolled 1,096 clinical isolates collected from three geographically different tuberculosis centers, including 437 INH-resistant isolates and 659 INH-susceptible isolates characterized by traditional drug susceptibility testing on Löwenstein- Jensen solid medium. The clinical sensitivity and specificity of the MeltPro TB/INH assay were 90.8% and 96.4%, respectively. DNA sequencing analysis showed that, except for the 5 mutants outside the detection range of the MeltPro assay, a concordance rate between the two methods of 99.1% (457/461) was obtained. Among the 26 mutation types detected, katG S315T (AGC→ACC), inhA - 15C→T, katG S315N (AGC→AAC), and ahpC promoter - 10C→T accounted for more than 90%. Overall, the MeltPro TB/INH assay represents a reliable and rapid tool for the detection of INH resistance in clinical isolates. © 2014, American Society for Microbiology. All Rights Reserved.

Wang J.,General Hospital of PLA | Si Y.,309th Hospital of Chinese PLA | Wu C.,General Hospital of PLA | Sun L.,General Hospital of PLA | And 3 more authors.
Lipids in Health and Disease | Year: 2012

Background: Atherosclerosis is a chronic degenerative disease of the arteries and is thought to be one of the most common causes of death globally. In recent years, the functions of adventitial fibroblasts in the development of atherosclerosis and tissue repair have gained increased interests. LPS can increase the morbidity and mortality of atherosclerosis-associated cardiovascular disease. Although LPS increases neointimal via TLR4 activation has been reported, how LPS augments atherogenesis through acting on adventitial fibroblasts is still unknown. Here we explored lipid deposition within adventitial fibroblasts mediated by lipopolysaccharide (LPS) to imitate inflammatory conditions. Results: In our study, LPS enhanced lipid deposition by the up-regulated expression of adipose differentiation-related protein (ADRP) as the silencing of ADRP abrogated lipid deposition in LPS-activated adventitial fibroblasts. In addition, pre-treatment with anti-Toll-like receptor 4 (TLR4) antibody diminished the LPS-induced lipid deposition and ADRP expression. Moreover, LPS induced translocation of nuclear factor-kB (NF-kB), which could markedly up-regulate lipid deposition as pre-treatment with the NF-kB inhibitor, PDTC, significantly reduced lipid droplets. In addition, the lowering lipid accumulation was accompanied with the decreased ADRP expression. Furthermore, LPS-induced adventitial fibroblasts secreted more monocyte chemoattractant protein (MCP-1), compared with transforming growth factor-β1 (TGF-β1). Conclusions: Taken together, these results suggest that LPS promotes lipid accumulation via the up-regulation of ADRP expression through TLR4 activated downstream of NF-kB in adventitial fibroblasts. Increased levels of MCP-1 released from LPS-activated adventitial fibroblasts and lipid accumulation may accelerate monocytes recruitment and lipid-laden macrophage foam cells formation. Here, our study provides a new explanation as to how bacterial infection contributes to the pathological process of atherosclerosis. © 2012 Wang et al.; licensee BioMed Central Ltd.

Liu H.-R.,Chinese PLA General Hospital | Liu H.-R.,309th Hospital of Chinese PLA | Li W.-M.,309th Hospital of Chinese PLA
IUBMB Life | Year: 2015

In certain unique conditions like viral infections of the liver like hepatitis B (HBV) and hepatitis C (HCV), activation of Tregs may be associated with chronicity of the viral infections and subsequent predisposition to development of hepatocellular carcinoma (HCC) by the integrated viral genome. In parallel, potential persistence of Tregs activity may lead to immune evasion of cancerous cells and thus persistence of the carcinomatous conditions. In this study, we hypothesized that although the relative proportions of Tregs may remain unaltered in HCC, persistence of activity of Tregs may lead to immune evasion in advanced stages of HCC. To examine the issue of activation of Treg in liver cancer pathogenesis, we obtained liver biopsy and peripheral blood samples from patients with advanced grades of HCC, isolated Tregs, and examined the methylation status of "Treg-specific demethylated region" (TSDR), a key region whose methylation suppresses Treg activity and demethylation stimulates its genomic activity. This study provides evidence of demethylation of TSDR, increased gene expression examined by luciferase assays, and nuclear translocation of key transcription factors that function as gene enhancers in CD4+CD25+FoxP3 regulatory T cells in advanced grades of HCC. © 2015 IUBMB Life, 2015 67(5):355-360, 2015 © 2015 International Union of Biochemistry and Molecular Biology.

Song L.,309th Hospital of Chinese PLA | Wu X.,309th Hospital of Chinese PLA
International Journal of Antimicrobial Agents | Year: 2016

Resistance and tolerance to antituberculosis (anti-TB) drugs, especially the first-line drugs, has become a serious problem in anti-TB therapy. Efflux of antimicrobial agents via bacterial efflux pumps is one of the main reasons for drug resistance. Efflux pump inhibitors (EPIs) bind to efflux pumps to inhibit drug efflux and thus enhance the drug effect and reduce drug resistance. Studies on EPIs targeting the efflux pumps of Mycobacterium tuberculosis (Mtb) help to understand Mtb resistance and to identify the potential drug target and are of significance in guiding the development of new anti-TB drugs and optimal combinations. Currently, there are many potential EPIs under study, but none of them has been used clinically for anti-TB therapy. In this article, we will provide an overview on the current development of EPIs targeting the efflux pumps of Mtb and discuss their potential clinical applications. © 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.

Zhang Y.,309th Hospital of Chinese PLA | Hao H.,309th Hospital of Chinese PLA
Biochemical and Biophysical Research Communications | Year: 2013

Mutations in gap junction (GJ) family of proteins, especially in the connexin (Cx) 26, are responsible for causing severe congenital hearing loss in a significant portion of patients (30-50% in various ethnic groups). Substitution of glycine at the position 45 of Cx26 to glutamic acid (p.G45E mutation) causes the Keratitis-ichthyosis-deafness (KID) syndrome. Previous studies have suggested that this point mutation caused a gain-of-function defect. However, the molecular mechanism of KID syndrome remains unclear. Since glycine at this position is conserved in many Cxs expressed in the cochlea, we tested the hypothesis that glycine at position 45 is an important component of the sensor regulating the Ca2+ gating of GJ hemichannels. Using reconstituted Cx30, 32 and 43 expressed in the HEK 293 cells, we compared the functions of wild type and p.G45E mutant Cxs. We found that G45E in Cx30 resulted in similar deleterious cellular effects as Cx26 did. Cell death occurred within 24h of transfection, which was rescued by increasing extracellular Ca2+ concentration ([Ca2+]o). Dye loading assay showed that Cx30 G45E, similar to Cx26 G45E, had leaky hemichannels at physiological [Ca2+]o (1.2mM). Higher [Ca2+]o reduced the dye loading in a dose-dependent manner. Whole cell membrane current recordings also indicated that G45E caused increased hemichannel activities. p.G45E mutations of Cx32 and 43 also resulted in leaky hemichannels compared to their respective wild types in lower [Ca2+]o. Our data in this study provided further support for the hypothesis that glycine at position 45 is a conserved Ca2+ sensor for the gating of GJ hemichannels among multiple Cx subtypes expressed in the cochlea. © 2013 Elsevier Inc.

OBJECTIVE: To investigate the association of eNOS 894G-->T, -786T-->C gene polymorphisms with disease severity and outcome in septic patients. METHODS: A total of 117 patients with severe sepsis were randomly selected from ICUs at 9 academic hospitals in Beijing during April 2007 to May 2009. PCR-RFLP and PCR-SSCP were used to analyze the alleles and genotypes in eNOS 894G-->T and -786T--> C gene polymorphisms. Recorded clinical data included demographics, pathogens, APACHE II score within 24 hours and SOFA score within 7 days after ICU admission, percentage of shock patients, days to shock onset (from infection to shock onset), duration of shock and the mortality at Days 7 and 28. RESULTS: In comparison with genotype GT carriers, the patients with genotype GT in eNOS 894G-->T polymorphism had a incremental trend in frequency of shock (87% vs 68.1%, P = 0.071) and a significantly shortened days to shock onset [1.0 (0.1 - 6.5) vs 2.0 (0.10 - 27.0) days, median (range), P < 0.05]. Those patients had been shown to have a significantly high APACHE II score (23.61 +/- 7.00 vs 19.50 +/- 6.99, P < 0.05), SOFA score (9.43 +/- 3.42 vs 5.26 +/- 2.94, P < 0.001) and mortality at Day 7 (34.8% vs 0%, P < 0.001) and Day 28 (78.3% vs 23.4%, P < 0.001). Multivariate analyses revealed that age in years, SOFA score and genotype GT in eNOS 894G-->T polymorphism were independent high-risk factors for the outcome in septic patients. However, eNOS -786T-->C gene polymorphism was not associated with disease severity and outcome in septic patients. CONCLUSION: Carriage of genotype GT in eNOS 894G-->T polymorphism is associated with the occurrence of shock and impaired organ function.

Chen G.,Chongqing Medical University | Zhang L.,Chongqing Medical University | Chen L.,309th Hospital of Chinese PLA | Wang H.,Chongqing Medical University | And 2 more authors.
Transplantation | Year: 2013

BACKGROUND: Biliary fibrosis has been referred to as the "final common pathway" of acute and chronic bile duct injury after orthotopic liver transplantation (OLT). We studied the role of integrin αvβ6 in the pathogenesis of ischemia-related biliary fibrosis after OLT. METHODS: The mouse nonarterialized OLT model with prolonged cold ischemia time was used in this study. A total of 54 FVB/N mice were divided into three groups: sham-operated group (sham, n=18), OLT group that was given the blocking antibody to integrin αvβ6 (OLT+antibody, n=18), and OLT group that was given the isotype control immunoglobulin G (OLT+vehicle, n=18). The expression of αvβ6 and major fibrosis-related genes were studied by real-time polymerase chain reaction and immunohistochemistry. Serum and bile were collected and analyzed biochemically. The histopathologic evaluation was performed to determine the severity of biliary fibrosis and bile duct injury. RESULTS: Integrin αvβ6 was highly expressed on newly formed bile ducts because of cholangiocyte proliferation and was gradually upregulated with the progression of biliary fibrosis after liver transplantation. αvβ6 transcripts closely correlated with fibrosis stages but not bile duct injury severity. Inhibition of αvβ6 attenuated peribiliary collagen deposition remarkably, induced significant downregulation of fibrogenic genes, and improved hepatic function. CONCLUSIONS: Integrin αvβ6 is strongly induced de novo in newly formed bile ducts because of cholangiocyte proliferation during ischemia-related biliary fibrogenesis after liver transplantation. Inhibition of αvβ6 could retard the progression of biliary fibrosis of liver allograft significantly, suggesting that αvβ6 is a potential target for the treatment of ischemic biliary complications. Copyright © 2013 Lippincott Williams & Wilkins.

Zheng S.-J.,309th Hospital of Chinese PLA | Jia C.-Y.,309th Hospital of Chinese PLA
Chinese Journal of Burns | Year: 2012

The delayed healing of diabetic ulcer has been haunting the surgeons and researchers for a long time. Although we have been researching and exploring the effective therapies for many years, the progress has been limited. Bone marrow-derived mesenchymal stem cells (BMSCs) have gradually won worldwide attention for their characteristics of differentiating into tissue repair cells and secreting multiple cytokines as well as growth factors. In recent years, the role of BMSCs in the treatment of diabetic ulcer has been drawing more and more attention. This article reviewed the advancement in the research of BMSCs in promoting the healing of diabetic ulcer. Through a discussion of the treatment of diabetic ulcer, the related research in BMSCs, as well as its role in diabetic ulcer treatment, the mechanism of BMSCs in promoting healing of diabetic ulcers is discussed. We expect through further research, unified criteria for the quality of BMSCs, application approach and dosage of BMSCs could be established.

Bao D.,309th Hospital of Chinese PLA
Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery | Year: 2011

To study the clinical application of Mobi-C prosthesis in treatment of anterior cervical discectomy and artificial disc replacement (ADR). Between January 2009 and June 2009, 20 cases of degenerative cervical disease were treated with anterior discectomy and ADR by Mobi-C prosthesis, including 13 cases of cervical disc herniation and 7 cases of cervical spondylotic radiculopathy, and 25 Mobi-C prosthesis were implanted. There were 8 males and 12 females, aged 29-54 years (mean, 45.2 years). The disease duration was from 4 days to 5 years (mean, 1.2 years). Affected segments of process included C3, 4 in 1 case, C4, 5 in 2 cases, Cs, 6 in 7 cases, C6, 7 in 5 cases, C4, 5 and C5, 6 in 2 cases, and C5, 6 and C6, 7 in 3 cases. Radiographs were taken regularly, and cervical range of motion (ROM) on segments of disc replacements were measured. The functions of cervical spinal cord were evaluated by "40 score" system (COA) preoperatively, immediately postoperatively, and at follow-up. The quality of life was evaluated by neck disability index (NDI) and visual analogue scale (VAS) score. All incisions healed by first intention. No perioperative complication was found. All cases were followed up 16.5 months on average (range, 14-18 months). There was no significant difference in cervical ROM of operated segment between preoperation and follow-up duration (t = 0.808, P = 0.440). No heterotopic ossification was found at follow-up. COA score at last follow-up (38.20 +/- 1.14) was significantly higher than preoperative one (32.10 +/- 2.96), (t = 9.278, P = 0.000), and the improvement rate at last follow-up was 77.2% +/- 5.4%. VAS score at last follow-up (3.20 +/- 1.23) had significant difference when compared with preoperative one (5.10 +/- 1.29), (t = 10.585, P = 0.000). NDI score at last follow-up (29.40 +/- 4.55) had significant difference when compared with preoperative one (39.20 +/- 3.80), (t = 16.039, P = 0.000). A satisfactory short-term curative effect can be obtained by using Mobi-C prosthesis in treatment of anterior cervical discectomy and ADR.

Huang X.W.,309th Hospital of Chinese PLA
Zhongguo gu shang = China journal of orthopaedics and traumatology | Year: 2013

To study the clinical efficacy of double bundle posterior cruciate ligament (PCL) reconstruction with remnant preservation. From January 2007 to November 2011, 50 patients with PCL rupture met the inclusion criteria were divided into two groups: remnant preservation group (RP group) and remnant resection group (RR group). There were 19 males and 7 females in the RP group, ranging in age from 18 to 55 years, with a mean of (32.250 +/- 11.085) years old. The duration from injury to operation ranged from 2 to 66 months, with an average of (17.481 +/- 3.568) months. Among the RR group, 17 patients were male and 7 patients were female, ranging in age from 20 to 54 years old, with an average of (31.458 +/- 9.569) years. The duration from injury to operation ranged from 3 to 72 months, with a mean of (19.354 +/- 3.950) months. The patients in both groups suffered from instability of knee joint, got a positive result of posterior drawer test. In the RP group, the intercondylar notch remnant fiber, scar tissue and synovial were preserved in operation, only the free ligament in the intercondylar notch was resected. In the RR group, the remnant fiber, scar tissue and synovial tissue of adhesive parts were resected. In both groups, autologous semitendinosus and gracilis tendon double-bundle PCL reconstruction were carried out, the tibia was fixed with an absorbable interference screw with post-tie fixation, and the femur side was compositely fixed with absorbable interference screws and suspending fixation. Each patient received both subjective assessment (IKDC subjective evaluation, Lysholm scoring and Cincinnati rating) and objective clinical assessment (IKDC objective evaluation and Kneelax 3 tibia backward measurement) before operation and two years after operation. IKDC subjective evaluation: 92.167 +/- 4.177 in the RP group,which was higher than 87.542 +/- 5.687 in the RR group (P = 0.010). Lysholm scores: 90.917 +/- 4.413 in the RP group, which was higher than 87.083 +/- 5.149 in the RR group (P = 0.027). Cincinnati knee scores: 92.125 +/- 4.003 in the RP group, which was higher than 87.791 +/- 6.665 in the RR group (P = 0.027). IKDC objective evaluation:no significant statistical differences between RP group and RR group. Kneelax 3 assessment : tibia backward test with Kneelax 3 under 132 N showed no significant statistical difference between RP group and RR group, which were (3.958 +/- 0.693) mm and (4.029 +/- 0.846) mm respectively (P = 0.795). The study shows a significant advantage of remnant fiber preservation than remnant fiber resection in double-bundle PCL construction in terms of subjective knee function recovery after operation. There is no significant difference in postoperative knee stability.

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