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@#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.
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As a cellular bulk degradation and survival mechanism, autophagy is implicated in diverse biological processes. Genome-wide association studies have revealed the link between autophagy gene polymorphisms and susceptibility of autoimmune diseases including systemic lupus erythematosus (SLE) and inflammatory bowel disease (IBD), indicating that autophagy dysregulation may be involved in the development of autoimmune diseases. A series of autophagy modulators have displayed protective effects on autoimmune disease models, highlighting the emerging role of autophagy modulators in treating autoimmune diseases. This review explores the roles of autophagy in the autoimmune diseases, with emphasis on four major autoimmune diseases [SLE, rheumatoid arthritis (RA), IBD, and experimental autoimmune encephalomyelitis (EAE)]. More importantly, the therapeutic potentials of small molecular autophagy modulators (including autophagy inducers and inhibitors) on autoimmune diseases are comprehensively analyzed.
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@#AIM: To investigate the curative effect of coaxial microincisional phacoemulsification combined with AcrySof ReSTOR intraocular lens implantation on senile cataract. <p>METHODS: Totally 66 patients(66 eyes)with age-related cataract who were admitted to the hospital from April 2015 to August 2017 were enrolled in the study. All of them were treated with coaxial microincisional phacoemulsification combined with intraocular lens implantation. All 32 cases underwent AcrySof ReSTOR intraocular lens implantation were included in observation group, while 34 cases underwent monofocal intraocular lens implantation were included in control group. The visual acuity, contrast sensitivity, stereoacuity, pseudo-accommodation power and satisfaction with visual function were compared between the two groups. <p>RESULTS: The near and middle vision, near stereoacuity, near and distant pseudo-accommodation power of naked eyes in observation group were significantly better than those in control group at 3mo after surgery(<i>P</i><0.05). When the spatial frequencies were 3, 6, 12 and 18 c/d, contrast sensitivities of observation group were significantly lower than those of control group under glare and no glare conditions(<i>P</i><0.05). The scores of visual function-related quality of life scales, satisfaction with near vision and glass-wearing at near vision of observation group were significantly better than those of control group(<i>P</i><0.05). <p>CONCLUSION: The curative effect of coaxial microincisional phacoemulsification combined with AcrySof ReSTOR intraocular lens implantation is significant on cataract. Compared with monofocal lens, it can improve the visual acuity, pseudo-accommodation power and visual related quality of life. However, the contrast sensitivity is reduced.
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Parthenogenetic embryonic stem (pES) cells isolated from parthenogenetic activation of oocytes and embryos, also called parthenogenetically induced pluripotent stem cells, exhibit pluripotency evidenced by both in vitro and in vivo differentiation potential. Differential proteomic analysis was performed using differential in-gel electrophoresis and isotope-coded affinity tag-based quantitative proteomics to investigate the molecular mechanisms underlying the developmental pluripotency of pES cells and to compare the protein expression of pES cells generated from either the in vivo-matured ovulated (IVO) oocytes or from the in vitro-matured (IVM) oocytes with that of fertilized embryonic stem (fES) cells derived from fertilized embryos. A total of 76 proteins were upregulated and 16 proteins were downregulated in the IVM pES cells, whereas 91 proteins were upregulated and 9 were downregulated in the IVO pES cells based on a minimal 1.5-fold change as the cutoff value. No distinct pathways were found in the differentially expressed proteins except for those involved in metabolism and physiological processes. Notably, no differences were found in the protein expression of imprinted genes between the pES and fES cells, suggesting that genomic imprinting can be corrected in the pES cells at least at the early passages. The germline competent IVM pES cells may be applicable for germ cell renewal in aging ovaries if oocytes are retrieved at a younger age.