Objective: To learn the prevalence and drug resistance of nontuberculous mycobacteria ( NTM ) in Zhejiang Province, so as to provide evidence for NTM prevention and control. Methods: A total of 2 878 clinical mycobacterium isolates in Zhejiang Province were collected from the drug resistance surveillance in 2008-2009, 2013-2014 and 2018-2019, PNB/TCH growth tests were used to preliminarily identify the NTM in these mycobacterium-positive isolates. 16SrRNA, rpoB, ITS and hsp65 gene sequencing analysis were used to confirm strains initially identified as NTM. Proportional method was applied to detect drug susceptibility of NTM isolates. Results : Finally, 135 strains were confirmed as NTM and the isolation rate was 4.69%. The isolation rates of NTM in 2008-2009, 2013-2014 and 2018-2019 were 1.85%, 4.56% and 7.84%, respectively, with an increasing trend ( P<0.05 ). Thirteen species were identified and the top two species were M. intracellulare ( 82, 60.74% ) and M. kansassi ( 18, 13.33% ). The NTM isolates showed the highest drug resistance rate to isoniazid ( 97.78% ), followed by p-aminosalicylic acid ( 94.87% ) and streptomycin ( 94.81% ). Conclusions The isolation rates of NTM showed an upward trend in the drug resistance surveillance in 2008-2019,2013-2014 and 2018-2019 in Zhejiang Province. M. intracellulare and M. kansassi were the main strains isolated. The NTM isolates showed high resistance against both first and second-line antituberculosis drugs.

Objective To know the application of MPB64 antigen detection of immune colloidal gold method for identification of Mycobacterium.Methods Taking the PCR amplified sequencing method as the 'gold standard',a total of 418 clinical isolated mycobacterium tuberculosis and 2 standard strains were tested by immune colloidal gold method,and was compared with the traditional method of PNB/TCH.Results Compared with sequencing method,the coincidence rate of immune colloidal gold method and PNB/TCH identification test was 95. 93%and 87. 08%respectively.The sensitivity, specificity,positive predictive value (PPV ) and negative predictive value (NPV ) of immune colloidal gold method identification were 88. 41%,99. 64%,99. 19% and 94. 58%respectively.It costs MPB64 immune colloidal gold method 15 minutes to carry out the results,less than 28 days of the traditional method,as well as 3 days of sequencing method. Conclusion Compared with PNB/TCH method,MPB64 immune antigen colloidal gold method has higher specificity and sensitivity,and could be applied in identification of Mycobacterium.

Objective To monitor the drug resistance of tuberculosis in Zhejiang Province and to provide scientific evidence for the control of drug resistant tuberculosis.Methods Thirty counties in Zhejiang Province were selected as sample during July 1,2013 and June 30 2014,and smear positive cases were selected to be monitored during study period. Results The rate of drug resistance was 30.88%,and the rate of multi drug resistance was 5.02%.The rate of the extensively drug -resistant tuberculosis was 0.32%.Among initial treatment cases,the rate of drug resistance was 29.22%,higher than 2008,and the rate of multi drug resistance was 3.21%.Among retreatment cases,the rate of drug resistance was 45.74%,and the rate of multi drug resistance was 21.28%.All kinds of monitored drugs were found resistance phenomenon.The drug resistant rate of SM was highest (15.28%),and aminoglycosides (2.35%)were relatively low.Conclusion The status of retreatment TB drug resistance suggested that we had reduced acquired TB drug resistance through implementation of DOTS strategy and standard short course chemotherapy treatment in Zhejiang Province. But it is still not optimistic to control TB drug resistance,and the status of initial treatment TB resistant suggested that resistant strains spread had not been effectively controlled in Zhejiang Province.So we should strengthen the early detection of drug resistant among TB patients,and to further improve the standard of conventional TB treatment.

OBJECTIVETo clone an A3IP gene and investigate its cellular and histological localization based on previous research which has identified part of A3IP sequence interacting with carboxyl-terminal of ataxin-3.

METHODSBioinformatic and Northern blotting were applied to clone the A3IP gene and detect the expression of its transcripts in various human tissues and brain regions. Western blotting and immunofluorescence staining were applied to detect expression of A3IP protein in cultured cells. Immunohistochemistry staining was applied to study the expression of A3IP protein in various human tissues and brain regions.

RESULTScDNA cloning of A3IP gene's reading frame and its sequence assembly were completed. Three transcripts (1 kb, 1.35 kb and 6 kb, respectively) of A3IP were found to express in various human tissues and brain regions. A3IP pEGFP expresses in cytoplasm of cultured COS-7 cells and various human tissues and brain regions including cerebral cortex, cerebellum, muscle, peripheral nerve, liver and kidney.

CONCLUSIONThe cloned A3IP gene encodes A3IP, a novel ataxin-3 interacting protein. Three transcripts of A3IP are expressed in various human tissues and brain regions. A3IP is a cytosolic protein.

Ataxin-3 ; Base Sequence ; Carrier Proteins ; genetics ; metabolism ; Cloning, Molecular ; Humans ; Molecular Sequence Data ; Nerve Tissue Proteins ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Protein Binding ; Protein Transport ; Repressor Proteins ; genetics ; metabolism

This study was purposed to investigate the B cell-activating factor belonging to the TNF family (BAFF) and a proliferation-inducing ligand (APRIL) levels in bone marrow, and the BAFF receptor expression level on B cells in multiple myeloma (MM) patients, in order to explore the characteristics of B cells in bone marrow of MM patients. MM patients were studied before treatment (newly diagnosed group, 19 patients) and after treatment with improvement (stable group, 17 patients), 10 non-hematologic patients were selected as control (control group). The BAFF receptors (BAFF-R) and transmembrane activator and calcium modulator and cyclophilin ligand interactor (TACI) on B cell (CD19(+)), naive B cell (CD19(+)IgD(+)) and memory B cell (CD19(+)CD27(+)) of bone marrow in all groups were detected by flow cytometry. The BAFF, APRIL level in bone marrow supernatant were tested with ELISA. The results showed that the BAFF-R expression level on CD19(+) cells in newly diagnosed group were higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group and stable group, but BAFF-R expression level on CD19(+)IgD(+) cells in newly diagnosed group was higher than that in control group; the BAFF-R expression level on CD19(+)CD27(+) cells in newly group was higher than that in stable group and control group; there was no significant difference between the BAFF-R expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in stable group and control group. There was no significant difference among the TACI expression level on CD19(+) cells, CD19(+)IgD(+) cells or CD19(+)CD27(+) cells in newly diagnosed group, stable group and control group. The bone marrow supernatant BAFF level in newly diagnosed group was higher than that in stable group and control group, but there was no significant difference between stable group and control group. There was no significant difference among the bone marrow TACI levels in newly diagnosed group, stable group and control group. It is concluded that both the bone marrow BAFF level and the BAFF-R expression level on CD19(+) cell, CD19(+)IgD(+) cells and CD19(+)CD27(+) cells in MM patients increase, which may help to stimulate B cells, thereby may relate with to MM pathogenesis.
B-Cell Activating Factor ; metabolism ; B-Cell Activation Factor Receptor ; metabolism ; Bone Marrow ; metabolism ; Humans ; Middle Aged ; Multiple Myeloma ; metabolism ; pathology

Objective To explore the cut-off value of three dimensional(3D)vena contracta area(VCA)in diagnosing severe tricuspid regrugitation(TR)under different etiologies and its accuracy and practicality in clinical application.Methods From Mar 2019 to May 2021,ninety-two patients with confirmed TR underwent two dimensional(2D)and 3D transthoracic echocardiography.The correlation and consistency between 3D VCA 3D calculated based on the proximal isokinetic surface area(PISA)effective regurgitant orifice area(EROA)was calculated.Comprehensive 2D multi-parameter method was used as a reference method to calculate the cut-off value of the diagnosis of severe TR.Results A total of 85 patients were ultimately included.3D VCA and 3D PISA EROA had similar and acceptable correlations in both primary TR and secondary TR(primary TR:r=0.831,P<0.01;secondary TR:r=0.806,P<0.01).Bland-Altman analysis showed that 3D VCA overestimated TR compared with 3D PISA EROA(62%overestimated in the total patient population,51%overestimated in primary TR,and 74%overestimated in secondary TR).In secondary TR,the cut-off value of 3D VCA for diagnosing severe TR was 0.45 cm2(sensitivity 89%,specificity 82%);combining clinical symptoms,positive 2D PISA EROA results and 3D VCA results for severe TR,the chi-square value was higher than those only included clinical symptoms or incorporated clinical symptoms and positive 2D PISA EROA results(42.168 vs.26.059 and 16.759,P<0.01).Conclusion 3D VCA would overestimate TR,and had high and incremental diagnostic value for evaluating severe TR in secondary TR.

This study design of specific identification primers for the ITS2 sequence of F. ussuriensis. The reaction system and conditions were optimized, and PCR-nucleic acid test strips were constructed to realize the visual detection of F. ussuriensis Bark. Through molecular cloning and sequencing technology, we constructed a positive control for F. ussuriensis DNA and formulated quality standards. The established method was evaluated for sensitivity, specificity and reproducibility, and the authenticity of the commercially available samples was identified. Results demonstrated that based on the ITS2 sequences, F. ussuriensis and its mixed forgeries could be distinguished. The PCR products of the authentic F. ussuriensis on test strips showed two bands in the T and C lines, while the pseudo products and negative control showed only one band in the C line, which was consistent with the results of agarose gel electrophoresis. The specificity was 100%, and the sensitivity of the PCR-nucleic acid test strip was up to 0.1 ng·μL^-1, which was 10 times higher than that of the gel electrophoresis assay. 11 out of 16 commercially available samples of F. ussuriensis were qualified, and 5 were unqualified. Collectively, the PCR-nucleic acid test strip method established in this study is specific, rapid, accurate and visualized, which can provide a new technical idea for the detection of F. ussuriensis.