To identify the species of Mycobacterium clinical isolates by molecular biology techniques,six clinical isolates which were preliminarily recognized as Mycobacterium tuberculosis by the TCH and PNB culture methods were selected in this study.PCR was applied to amplify the oxyR-ahpC interval resistant-zone(intergenic region) and the length of PCR product in four strains was the same as that of H37Rv,while the length in the other two strains was different from that of H37Rv but same as Mycobacterium intracellulare 95002.With sequencing and on-line homology comparison with H37Rv,U18263 and U71061,the DNA sequence of oxyR-ahpC intergenic region displayed a 99% homology with Mycobacterium intracellulare U71061 and an 84% homology with Mycobacterium tuberculosis H37Rv.In addition,the results of hsp65 PCR-restriction fragment length polymorphism analysis and multi-locus PCR amplification in the two strains were identical with those of Mycobacterium intracellulare 95002.These two clinical isolates which were preliminarily recognized as Mycobacterium tuberculosis by PNB and TCH culture methods were finally identified as Mycobacterium intracellulare.Results predicted that the application associated with various techniques of molecular biology would provide a faster,easier and more correct way for the species identification of Mycobacterium.

This study was aimed to explore the mechanism of Xin-Feng capsule (XFC) on pulmonary function of Sjogren's syndrome (SS) rat model based on the TGF-β1-ERK1 signal pathway. A total of 50 SD rats were ran-domly divided into the normal control (NC) group, model control (MC) group, hydroxychloroquine sulfate (HCQ) group, radix paeoniae alba (total glucosides of paeony capsules, TGP) group, and the XFC treatment group, with 10 rats in each group. Except the NC group, the complete Freund's adjuvant (CFA) and the same rat sub-mandibular gland antigen induced method were applied to each rat after metatarsus injection on two feet and CFA after sufficient emulsification of 0.2 mL submandibular gland protein mixed antigen induced SS. The water drink-ing quantity, body weight change, pulmonary function, ERK1, TGF-β1 protein expression and serum cell factor (IL-17, IL-4) changes were detected in each group. The results showed that compared with the NC group, the body quality and serum IL-4 of MC rats were obviously decreased, and the water drinking quantity, submandibu-lar gland / lung index, submandibular gland pathological score, ERK1, TGF-β1 integral and IL-17 were increased (P < 0.01 or P < 0.05), and the pulmonary function parameter was decreased (P < 0.01 or P < 0.05). Compared with the MC group, the body quality, pulmonary function parameters FEF50 and MMF of the XFC group were in-creased, the IL-4 expression was increased, the water drinking quantity, submandibular gland / lung index, sub-mandibular gland pathological score, serum IL-17 expression, ERK1 and TGF-β1 integral were decreased (P <0.01 or P < 0.05). Compared with the HCQ group, the body quality and IL-17 were significantly decreased in the XFC group (P < 0.01), FEF25, FEF75 and MMF were increased (P < 0.01 or P < 0.05). Compared with the TGP group, the lung index and IL-17 were decreased in the XFC group (P < 0.01 or P < 0.05). It was conclud-ed that the lung function of SS rats was declined, which is closely related to the activation of TGF-β1-ERK1 sig-nal pathway. Traditional Chinese medicine of XFC is able to downregulate the TGF-β1 expression and to restrain epithelium-mesenchymal cell transformation, in order to inhibit ERK1 phosphorylation activation and proliferation, decrease immune inflammation, in order to improve SS and lung function.

The effect of PubMed on the development of biomedical journals and the role of its Linkout function in extending the effect of journals were analyzed , followed by a description of the technologies and methods for the full-text seamless link of journal Websites and PubMed using its Linkout function with Biomedical and Environmental Sciences as an example .

Objective To investigate and evaluate the PCR-RFLP method for identification of Mycobacterium abscessus (M.abscessus) group.Methods 46 clinical acid-fast bacilli (AFB) isolates from Pulmonary Hospital of Fujian Province,Center for Disease Control and Prevention of Zhaoyang District in Beijing and Beijing People Liberation Army General Hospital were collected in 2009 -2010 and identified by traditional bacteriological characteristics according to clinical laboratory handbook of mycobacteria (2004).The PCR-RFLP method was used for species identification of M.abscessus group using hsp65 (441 bp) or rpoB (380 bp) gene fragment as specific target,while the direct DNA sequencing was performed as a control method.Results Of 46 AFB isolates,30 strains were identified as Mycobacterium tuberculosis by its traditional bacteriological characteristics and 16 strains were identified as non-tuberculosis mycobacterium (NTM).10 strains of the NTM strains had identical bacteriological characteristics with the reference strain M.abscessus ATCC 19977.Identified by hsp65 PCR-RFLP,9 of these 10 strains got the same pattern of 235bp and 200 bp(BstE Ⅱ )/145 bp,70 bp,60 bp,55 bp,50 bp and 40 bp(Hae Ⅲ ),and 1 got pattern of 235 bp and 200 bp(BstE Ⅱ )/200 bp,70 bp,60 bp and 50 bp(Hae Ⅲ ).While identified by rpoB PCRRFLP,all 10 strains got the same pattern of 105 bp,95 bp and 80 bp( Msp Ⅰ )/130 bp,100 bp and 90 bp (Hae Ⅲ ).By analysis of the DNA sequence,hsp65 and rpoB sequence of these 9 strains showed 100％similarity with those of M.abscessus,while the hsp65 and rpoB sequence of the other one strain showed 100％similarity with those of M.massiliense.Conclusion PCR-RFLP is a rapid and effective method for species identification of M.abscessus group,and hsp65 PCR-RFLP is more effective than rpoB PCR-PFLP in the species identification of M.abscessus group.

The Mycobacterium chelonae/abscessus (M.chelonae/abscessus) complex belongs to the rapidly growing genus Mycobacterium (RGM).It is one of the most important pathogenic members of Mycobacterium leading to nosocomial infections and outbreaks.It includes members of M.chelonae,M.immnunogenum,M.abscessus,M.massiliense,and M.bolletii.In order to investigate the epidemiological characteristics of the M.chelonae/abscessus complex in China and to conduct the molecular methods for species identification of M.chelonae/abscessus,we collected clinical M.chelonae/abscessus complex strains identified by phenotypic tests.Members were verified by sequencing of 16S rRNA,Species and subspecies were identified by hsp65 and rpoB PCR RFLP methods.In total,27 clinical specimens were identified as Mycobacterium chelonae/abscessus complex by phenotypic tests.16s rRNA gene sequence analysis of all 27 clinical samples shared over 99.7％ similarity with M.chelonae and M.abscessus.Species identification with hsp65 PCR-RFLP and rpoB PCR-RFLP revealed that 18 specimens were M.abscessus and 4 were M.absecces.The remaining 5 samples displayed a pattern that failed to match any previously reported pattern.Thus,this might represent a novel species that is part of the Mycobacterium chelonae/abscessus complex.We identified that a majority of the chronic lung infection in China is caused by the M.chelonae/abscessus complex.Specifically,the M.abscessus species might be the most infectious,while other species in the complex can still cause infection.Interestingly,there may be a novel or previously unidentified species that is a part of the complex.Finally,we show that species identification can be carried out more accurately by combined use of hsp65 and rpoB PCR-RFLP.

Objective To observe the changes of lung parameters,the ratios of B and T lymphocyte attenuator(BTLA) and serum cytokines in patients with knee osteoarthritis (KOA),and explore possible molecular mechanism of them.Methods Forty-seven cases of knee osteoarthritis from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from 2011 October to 2012 July were analyzed in this study.Pulmonary parameters were detected by spirometer; B and T lymphocyte attenuator(BTLA) was detected by flow cytometry ; Interleukin (IL)-1β,IL-10,matrix metalloproteinase-9 (MMP-9),tissue inhibitor of matrix metalloprotease-1 (TIMP-1) were detected by ELISA;ESR was determined by Westergren method ; hs-CRP was determined by the automatic biochemistry analyzer.Results (1) Compared with NC group,levels of FVC,FEV1,FEV1/FVC,PEF,MEF25.75,MEF50,MEF25,CD3 + BTLA+ T cell,CD4+ BTLA+ Tcell,IL-10,TIMP1 were significantly decreased,IL-1 β,MMP9 were significantly increased.(2)Correlation analysis showed FVC was negatively correlated to Lequesne MG,symptom classify quantization scores,course,MMP9,while positively related to CD3+ BTLA+T cell,IL-10,TIMP1 ;FEV1 was positively correlated to CD3 + BTLA+T cell,CD4+ BTLA+T cell,TIMP1,while negatively correlated to course ; MEF50 was positively related to CD3+BTLA+T cell,CD4+ BTLA+T cell.Conclusion While articular cartilage lesions occurred in KOA patients,the lung function was also declined.The mechanism may be associated with the declination of expression of BTLA,which can cause up-regulating of IL-1 β,MMP9 and down-regulating of IL-10,TIMP1,thus leading to immune dysfunction and abnormal immune response.Those may induce airway injuries and result in lung function decline finally.

Objective To discuss the effects of multimodal combination dialysis on Klotho protein, fibroblast growth factor-23 (FGF-23) and brain natriuretic peptide (BNP) in patients with maintenance hemodialysis (MHD). Methods A randomized controlled trial (RCT) was conducted. 120 patients who was diagnosed with chronic renal failure (CRF) uremia receiving MHD over 3 months admitted to Blood Purification Centre of Department of Nephrology of the Second People's Hospital of Guiyang from December 2015 to December 2016 were enrolled, who were randomly divided into hemodialysis (HD) group (HD for 8 times a month), HD + hemofiltration (HF) group (HD for 8 times a month + HF once a month), and HD + HF + hemoperfusion (HP) group (HD for 8 times a month + HF for 4 times a month + HP once a month), with 40 patients in each group. Before and after treatment for 6 months and 12 months, blood was taken from venous circuit tube, the serum Klotho protein and FGF-23 levels were determined by enzyme linked immunosorbent assay (ELISA), and the serum BNP level was determined by electrochemiluminescence. Results 120 patients with MHD were enrolled in the final analysis without withdrawal. There were no significant differences in the levels of Klotho protein, FGF-23, or BNP before enrollment among the three groups (all P > 0.05). Compared with those before enrollment, the levels of serum Klotho protein after enrollment in three groups showed a sustained upward tendency, which were higher in HD + HF + HP group than in HD + HF group and HD group (μg/L: 2.59±0.61, 1.63±0.35, 1.13±0.26 at 6 months, F = 119.374, P = 0.000; 6.98±1.21, 3.57±1.03, 2.12±0.43 at 12 months, F = 275.675, P = 0.000); the levels of FGF-23 showed a sustained downward tendency, which were lower in HD + HF + HP group than in HD + HF group and HD group (ng/L: 69.22±38.26, 132.28±61.18, 178.50±74.64 at 6 months, F = 33.509, P = 0.000; 32.81±17.32, 87.93±43.27, 146.33±69.28 at 12 months, F = 55.466, P = 0.000);the BNP showed a similar tendency as FGF-23 (ng/L: 4083.39±2864.53, 7245.69±4643.81, 7969.12±5360.85 at 6 months, F = 8.758, P = 0.000; 1521.86±894.63, 4554.32±1969.84, 5013.89±2033.64 at 12 months, F = 49.003, P = 0.000). Conclusion Multimodal combination dialysis can increase the Klotho protein level, and decrease the levels of FGF-23 and BNP in MHD patients with CRF uremia.

ObjectiveTo investigate the value of shear wave elastography (SWE) in the evaluation of liver damage in patients with Budd-Chiari syndrome (BCS).MethodsSixteen patients with BCS which were all conifrmed by interventional therapy or inferior vena cava angiography were enrolled in the study. And iffty healthy volunteers who were conifrmed with no liver disease history, no surgical history and no metabolic disease such as diabetes were included in this study. All the subjects underwent conventional ultrasound and no obvious abnormality was detected. All the BCS patients and the volunteers underwent SWE. The elastic graph and the quantitative elastic value, including the average value, the maximum value and the minimum value, were acquired and compared between BCS patients and the volunteers.ResultsThe elastic graphs for the healthy volunteers were uniform light blue while the elastic graphs for BCS patients were hybrid images of blue, green and yellow. The average value, the maximum value and the minimum value of BCS patients were (18.1±9.4) kPa, (20.6±10.5) kPa and (15.5±8.5) kPa, which were statistically higher than those of healthy volunteers [(6.2±1.0) kPa, (7.5±1.2) kPa and (4.8±1.1) kPa,t=7.69, 8.94 and 7.83, allP<0.01].ConclusionsSWE can reflect the liver damage of patients with BCS before the change of the two-dimensional ultrasound. It can play an important role in the accurate selection of treatment and the accurate evaluation of treatment effect and the prognosis.

Objective:To assess the efficacy of different non-drug smoking cessation interventions on smoking cessation among the high-risk populations of lung cancer screening by network meta-analysis.Methods:PubMed, MEDLINE (Ovid), PsychINFO, CNKI, WanFang and VIP databases were searched for randomized controlled clinical trials published from January 2010 to September 2020.According to the eligibility criteria, the retrieval literature was screened, the quality evaluation and data extraction were conducted, then, the statistical analysis was performed by using the Stata 14.0 software.Results:A total of 28 trials were included, including 34 640 cases of high-risk population, involving intervention measures, including 5R short quit smoking intervention, cognitive behavior therapy, award model quit smoking intervention, motivational interview, network intervention, telephone intervention, incentive mechanism plus telephone intervention, network plus telephone intervention, conventional treatment and blank group.The results of network meta-analysis showed that on the 7-day follow-up of 6 months, according to the score of SUCRA, the ranking of smoking cessation rate was 5R short smoking cessation intervention (0.88), cognitive behavior therapy (0.85), AWARD model smoking cessation intervention (0.80), incentive mechanism plus telephone intervention (0.73), motivational interview (0.53), Internet plus telephone intervention (0.40), Internet intervention (0.37), telephone intervention (0.31), conventional treatment (0.13) and blank group (0.03). However, on the 7-day smoking cessation rate of 12-month follow-up, the ranking of smoking cessation rate was 5R short smoking cessation intervention (0.94), AWARD model smoking cessation intervention (0.81), motivational interview (0.51), network intervention (0.40), telephone intervention (0.19) and conventional treatment (0.14).Conclusions:The existing evidence shows that no matter the length of follow-up, 5R short smoking cessation intervention has the best effect on smoking cessation in high-risk population of lung cancer screening.

Objective To prepare specific mouse monoclonal antibodies against Homo sapiens dynein,axonemal, heavy chain 2 (DNAH2). Methods Firstly, recombinant plasmid encoding His tagged immunogen, targeting N-terminal sequence of DNAH2 protein (1-300 aa), in E. coli was constructed. IPTG was used to induce the expression of His-immunogen, which was then purified and immunized in BALB/c mice. Hybridoma cells were obtained through the fusion between myeloma cells and splenocytes isolated from BALB/c mice. Finally, ELISA and Western blot assays were performed to screen the positive hybridoma. Results IPTG was used efficiently to induce the expression of DNAH2 immunogen in E. coli. DNAH2 protein bands were detected in screened positive hybridoma. Conclusion Mouse monoclonal anti-DNAH2 antibody is prepared successfully.