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OBJECTIVE@#To analyze the clinical phenotype and genetic basis for a Chinese pedigree suspected for branchiootic syndrome (BOS).@*METHODS@#The proband was subjected to target-capture high-throughput sequencing to detect potential variant of deafness-associated genes. Candidate variants were verified by Sanger sequencing of the family members.@*RESULTS@#The proband was found to harbor a c.1627C>T (p.Gln543Ter) nonsense variant of the EYA1 gene. Sanger sequencing confirmed that all of the 4 patients with the BOS phenotype from the pedigree have harbored the same heterozygous variant. Based on the guidelines of the American College of Medical Genetics and Genomics, the variant was predicted to be pathogenic (PVS1+PS+PP3+PP4).@*CONCLUSION@#The c.1627C>T (p.Gln543Ter) variant of the EYA1 gene probably underlay the BOS phenotype in this pedigree. Above finding has provided a basis for its clinical diagnosis.
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Humans , Branchio-Oto-Renal Syndrome , China , Intracellular Signaling Peptides and Proteins/genetics , Mutation , Nuclear Proteins/genetics , Pedigree , Protein Tyrosine Phosphatases/geneticsABSTRACT
Objective:To construct the pET30a-EgG1Y162-2 prokaryotic expression plasmid and induce the expression of EgG1Y162-2 protein, so as to provide a research basis for development of Echinococcus granulosus vaccine. Methods:Using Echinococcus granulosus cDNA as a template, the target gene of EgG1Y162-2 was synthesized by PCR, and after digestion with restriction enzymes EcoRⅠ and Hind Ⅲ, it was connected to the prokaryotic expression vector pET30a to construct the recombinant plasmid pET30a-EgG1Y162-2. The recombinant plasmid was transformed into competent cell BL21 (DE3) and induced by isopropyl β-D-thiogalactoside (IPTG) to express a large number of proteins. The recombinant protein was purified by affinity chromatography. The purification level was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the expression product was identified by Western blotting. Results:The recombinant plasmid pET30a-EgG1Y162-2 was successfully constructed. After inducting expression, the bacterial supernatant and the eluate were both at a relative molecular weight of about 15 × 10 3, and the protein antigen component eluted with 200 mmol/L imidazole was relatively pure. Western blotting results showed that the purified recombinant protein EgG1Y162-2 with His tag could be recognized by His monoclonal antibody. Conclusion:The pET30a-EgG1Y162-2 prokaryotic expression plasmid of Echinococcus granulosus is successfully constructed, and the recombinant protein of EgG1Y162-2 is induced to express, laying a foundation for further study on anti- Echinococcus granulosus vaccine.
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Objective:To investigate the changes of sTim-3,HMGB1 and TGF-β in the brucellosis patients and to analyse the relationship between the changes of these molecules and brucella infection. Methods:28 cases of brucellosis patient untreated and 28 healthy control cases in the age and gender matched with brucellosis cases were collected. The serum levels of sTim-3 and HMGB1 were detected by ELISA,and the levels of Spot forming cells secreting TGF-β were measured by ELISPOT in patients and healthy control group. Results: Compared with healthy controls, sTim-3/HMGB1 expression levels and Spot forming cells secreting TGF-β were significantly increased in the brucellosis patients ( P<0. 01 ) . The changes of Spot forming cells secreting TGF-β were positively correlated with the levels of HMGB1 (P<0. 05). Conclusion:The serum levels of sTim-3/HMGB1 and Spot forming cells of secreting TGF-β from peripheral blood mononuclear cell are significantly increased in the brucellosis patients. Those molecules may be involved in the process of brucella infection and may play a significant role in the immune escape of patients infected with brucella.
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To investigate the potential role of transforming growth factor (TGF)-β1 in liver fibrosis during Echinococcus granulosus infection, 96 BALB/c mice were randomly divided into 2 groups, experimental group infected by intraperitoneal injection with a metacestode suspension and control group given sterile physiological saline. The liver and blood samples were collected at days 2, 8, 30, 90, 180, and 270 post infection (PI), and the expression of TGF-β1 mRNA and protein was determined by real-time quantitative RT-PCR and ELISA, respectively. We also evaluated the pathological changes in the liver during the infection using hematoxylin and eosin (H-E) and Masson staining of the liver sections. Pathological analysis of H-E stained infected liver sections revealed liver cell edema, bile duct proliferation, and structural damages of the liver as evidenced by not clearly visible lobular architecture of the infected liver, degeneration of liver cell vacuoles, and infiltration of lymphocytes at late stages of infection. The liver tissue sections from control mice remained normal. Masson staining showed worsening of liver fibrosis at the end stages of the infection. The levels of TGF-β1 did not show significant changes at the early stages of infection, but there were significant increases in the levels of TGF-β1 at the middle and late stages of infection (P<0.05). RT-PCR results showed that, when compared with the control group, TGF-β1 mRNA was low and comparable with that in control mice at the early stages of infection, and that it was significantly increased at day 30 PI and remained at high levels until day 270 PI (P<0.05). The results of this study suggested that increased expression of TGF-β1 during E. granulosus infection may play a significant role in liver fibrosis associated with E. granulosus infection.
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Animals , Mice , Bile Ducts , Echinococcus granulosus , Echinococcus , Edema , Enzyme-Linked Immunosorbent Assay , Eosine Yellowish-(YS) , Hematoxylin , Injections, Intraperitoneal , Liver Cirrhosis , Liver , Lymphocytes , RNA, Messenger , Transforming Growth Factors , VacuolesABSTRACT
Objective To discuss the clinical significant of detecting cytoplasm phospholipase A2 alpha (cPLA2α)in patients with chronic obstructive pulmonary disease (COPD).Methods Selected treated 90 cases of patients with COPD.According to the COPD severity classification standard,they were divided into mild group of 30 cases,moderate group of 25 cases and severe group of 35 cases.In the same period of physical examination,90 cases of normal lung function healthy were control group.Used Enzyme linked immunosorbent test (ELISA)to detect cPLA2αlevel of these groups.At the same time,detected cPLA2αexpression of these groups by RT-PCR.Results cPLA2αlevel in serum of mild group was (0.039 2±0.005 1)pg/ml,gene expression was (0.68±0.01),in moderate group (0.049 8±0.007 4)pg/ml and (0.92±0.02),in severe group (0.055 4±0.008 1)and (1.35±0.02).and in Healthy controls group (0.010 2±0.006 6)pg/ml and (0.11±0.01).There were significant differences among four groups (t = 3.013 ~ 5.817,5.039 ~ 11.667,P < 0.05).Conclusion Detection of blood cPLA2αcan indicate COPD severity,and cPLA2α is the new molecular targets of diagnosis,treatment and classifica-tion COPD.
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Objective To establish the reference ranges of blood reticulocyte (Ret)among healthy Uygur adults in Xinjiang and to compare the difference between genders.Methods The Beckman Coulter LH750 instrument was adopted to determine the pe-ripheral blood Ret multiple parameters in 1024 Uygur healthy adults.The SPSS 17.0 statistical software was adopted to establish the database and perform the statistical analysis.Results The reference range of Ret multiple parameters in Uygur healthy adult men and women aged 20-79 years old in Xinjiang region were as follows:Ret were (1.5 ±0.5 )% and (1.4 ±0.4)%;IRF were (68.4±17.3)and (58.9±15.4);HFR were (0.5±0.3)% and (0.5±0.4)%;MFR were (4.5±2.3)% and (4.6±2.4)%;LFR were (94.7±2.6)% and (95.1 ±3.0)% respectively.Ret among 1024 cases of healthy Uighur population had statistically signifi-cant difference between genders,while IRF,LFR,MFR,HFR had no statistically significant differences.Conclusion The Ret multi-ple parameters reference ranges in Xinjiang Uygur healthy adults are established and the obtained conclusion is that the nationality and gender factors can affect the Ret multiple parameters in healthy adults.
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Recent studies show that the vector of recombinant Bacillus Calmette-Guérin (rBCG) has a series of advantages. With exogenous gene and vaccine in one inoculation, it can obtain strong and persistent immune response at one time so that BCG is considered as a kind of ideal vector for live recombinant vaccine. This review outlines the application of rBCG vaccine and its vector in infectious diseases caused by bacteria, viruses, other microorganisms and parasites.
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AIDS Vaccines , Genetics , Allergy and Immunology , Antigens, Bacterial , Genetics , Allergy and Immunology , BCG Vaccine , Genetics , Allergy and Immunology , Metabolism , Communicable Disease Control , Methods , Genetic Vectors , Genetics , HIV Infections , HIV-1 , Mycobacterium tuberculosis , Recombinant Proteins , Genetics , Allergy and Immunology , Tuberculosis , Vaccines, Synthetic , Allergy and ImmunologyABSTRACT
It is an essential educational goal to strength experimental teaching and improving the practical skill of medical postgraduate students in medical colleges and universities.We made anexploration and practice on how to improve the practical skill and innovative ability of postgraduate students in medical immunology experimental teaching.The article points out that optimizing the design of experimental course,innovating the teaching methods,standard appraised of experiment and improving the quality of teacher are the efficient ways to enhance the practical skill of postgraduate students.
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Objective: To study the expression of PDCD5 mRNA and its significance in esophageal cancer in Xinjiang Kazakh and Han nationality. Methods: RT-PCR was used to detect the mRNA of PDCD5 in 40 cases of esophageal cancer (18 cases of Kazakh, 22 cases of Han). Results: The positive rate of PDCD5 mRNA expression in 40 samples of esopha-geal cancer tissue, adjacent tissue, and normal tissue was 80.0% (32/40), 80.0% (32/40) and 87.5% (35/40), respectively, with no significant difference (P>0.05). The Ods of cancer tissues, adjacent tissues, and normal tissues were 0.7644± 0.1444, 0.9341 ±0.1631 and 1.8703±0.4767, respectively. The expression of PDCD5 was significantly increased in cancer tissues compared with that in normal tissues (P<0.05). The expression level of PDCD5 mRNA was not significantly correlat-ed with the degree of differentiation and lymph node metastasis. Conclusion: No significant difference was found in PDCD5 mRNA expression in esophageal cancer between Xinjiang kazakh and Han nationality (P>0.05). The expression of PDCD5 is not correlated with the degree of differentiation, depth of invasion or lymph node metastasis. Detection of PDCD5 mRNA expression in esophageal cancer tissues may provide valuable information for patient prognosis.
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Kazakh of Xinjiang is the region with a high incidence of esophageal cancer,genetic research is quite active in recent years.Through the research on biological activity of P53,RB gene during esophageal cancer process.We tried to find potential differences in the national heredity susceptibility and so to support treatment and the research.
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So in pharmaceutical microbiology teaching, we made education reform according to pharmaceutical characteristics, that is to revise syllabus, update teaching content, reform of teaching methods, use modern means of teaching to train students in innovative spirit and ability. Besides, we strengthen the practice of teaching, conduct comprehensive and applicative experiment, So that our educational content is more in line with teaching objectives of Chinese pharmacy specialty, and help students to engage in basic knowledge of microbiology, experiment skills, capacity to analysis and solve problems.
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Immunology teaching course includes both basic immunology and clinical im-munology.This paper points out the problems of teaching and knowledge integration between the basic immunology and clinical immunology and suggests that teaching team should include the basic immunology and clinical immunology related content clinical experts to promote teaching quality.
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Objective To observe the change of six cytokines in mice infected with Echinococcus multilocularis as part of the study on immunological mechanism in the infection. Methods Mice were infected by abdominal inoculation of echinococcus protoscoleces. The change of serum level of the cytokines IL-2、IFN-?、TNF-?、IL-4、 IL-5 and IL-10 was determined by ELISA during the infection which lasted for 260 d. Results Compared with uninfected control, the levels of the cytokines all significantly increased in the 260 d. The level of IL-2 reached a peak after 80 d post-infection (p.i.), then decreased quickly after 140 d p.i., High level of TNF-? was detected after 40 d, compared to uninfected control, reached a peak at 100 d p.i., and decreased quickly after 140 d. The level of IFN-? reached a peak after 80 d p.i., and decreased slowly after 140 d p.i., The levels of IL-4, IL-5 and IL-10 remained lower before 80 d, and increased sharply after 100 days. The levels of IL-4 and IL-10 reached peaks at 100 d p.i., and that of IL-5 at 140 d p.i. Conclusion The data suggest that the induction of Th2 antibody-mediated immunity (AMI) with a parallel expansion of Th1 cell-mediated inflammatory (CMI) responses are important mechanism of the host in defending against the metacestodes. Th1 CMI plays an important role at the early stage of infection, and Th2 AMI is important in the later stage of infection.