.Expression and purification of Mycobacterium tuberculosis MPT83 protein and its application in tuberculosis immunological diagnosis / 中国热带医学

@#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.

Willingness to pay integrated medical and elderly care services among the elderly in Hohhot City / 预防医学

Objective@#To investigate the willingness to pay integrated medical and elderly care services and identify the influencing factors among the elderly in Hohhot City, Inner Mongolia, so as to provide insights into promoting integrated medical and elderly care services.@*Methods@#The elderly at ages of 60 years and older were sampled using a convenient sampling method from public venues in four districts of Huimin, Saihan, Yuquan and Xincheng in Hohhot City from June to December 2021. Basic characteristics, health and disease burdens, social support and willingness to pay integrated medical and elderly care services were collected, and factors affecting the willingness to pay integrated medical and elderly care services were identified among the elderly using a multiple linear regression model. @*Results@#A total of 1 008 valid questionnaires were recovered, with an effective recovery rate of 96.74%. The respondents included 519 men (51.49%) and 489 women (48.51%), and had a mean age of (69.47±6.42) years. The monthly average fees of willingness to pay integrated medical and elderly care services were (2 076.49±36.79) Yuan, and there were 636 participants with 2 000 Yuan and less monthly average fees of willingness to pay integrated medical and elderly care services (63.10%). Multiple linear regression analysis showed that place of residence (β=180.832), satisfaction with housing (satisfied, β=-140.760), physical self-care ability (completely self-care: β=-238.244; mostly self-care: β=-254.557), burdens of disease diagnosis and treatment (able to afford: β=452.488; partly afford: β=228.626), monthly income (β=347.144), expenses of medications (β=0.019) and total score of social support (β=17.116) were factors affecting the willingness to pay integrated medical and elderly care services among the elderly. @*Conclusions@#The willingness to pay integrated medical and elderly care services among the elderly in Hohhot City is associated with place of residence, monthly income, satisfaction with housing, physical self-care ability, burden of disease diagnosis and treatment, expenses of medications and social support.

Anatomy of the original short head of biceps-muscular structure / 解剖学报

Objective To analyze and describe the morphological characteristics of the proximal origin of the biceps brachii, and measure the anatomical data of its tendons, so as to provide references for related clinical applications of the biceps brachii. Methods Six adult shoulder specimens (4 on the left and 2 on the right) were dissected to observe the origin, morphology, and course of the biceps brachii tendon and its adjacent relationship with the coracobrachialis muscle, musculocutaneous nerve and other structures. And then the length, width, and thickness of the biceps tendon were measured. Results The origin of the short head of biceps consisted of muscle fibers attaching directly to the tip of the coracoid process, with a thin, tendinous aponeurosis covering its anterior surface, rather than a true tendon as previously described. The mean of length,widths and thickness of the short head of biceps were (107.7±9.6) mm,(20.0±1.5)mm and (5.8±2.0)mm. Conclusion The short head of the biceps brachii is directly from the coracoid process with muscle fibers.

Genetic Mutation Characteristics of Glucose-6-Phosphate Dehydrogenase Deficiency Patients in Wuhan / 中国实验血液学杂志

OBJECTIVE@#To explore the genotype mutation characteristics of patients with glucose-6-phosphate dehydrogenase(G6PD) deficiency in Wuhan.@*METHODS@#A total of 1 321 neonates with positive screening and outpatients were received G6PD mutation detection, 12 kinds of common G6PD mutation in Chinese people was detected by using multicolor melting curve analysis (MMCA) method, for those with negative results, the enzyme activity and clinical information were analyzed, sequencing was recommended after informed consent when it is necessary.@*RESULTS@#Among 1321 patients, a total of 768 mutations were detected out, with a detection rate of 58.1%. A total of 18 types of G6PD genotypes were identified, including c.1388G>A, c.1376G>T, c.95G>A, c.1024C>T, c.871G>A, c.392G>T, c.487G>A, c.1360C>T, c.1004C>A, c.517T>C, c.592C>T, c.94C>G, c.152C>T, c.320A>G, c.1028A>G, c.1316G>A, c.1327G>C and c.1376G>C, including 683 male hemizygotes, 3 female homozygotes, 80 female heterozygotes and 2 female compound heterozygous.@*CONCLUSION@#A total of 18 types of G6PD mutations are identified in the reaserch, and c.94C>G, c.1028A>G and c.1327G>C are first reported in Chinese population. The most common G6PD mutation types in Wuhan are c.1388G>A, c.1376G>T, c.95G>A.

Progress for targeting adenosine A2A receptors in cancer immunotherapy / 药学学报

Immunotherapy has completely changed the paradigm of clinical tumor treatment, but immune checkpoint inhibitors still have low objective response rates and are prone to drug resistance for most solid tumors. The immune suppression tumor microenvironment and complicated tumor immune escape mechanisms are key factors that affect the clinical outcome and response rates. Therefore, it is critical to reverse the obstacle of the tumor microenvironment to improve immunotherapy efficacy. The immune suppression caused by the increased level of adenosine in the tumor microenvironment raises the attention of people. Targeting adenosine receptors, especially A2AR, will be an effective strategy to improve immunotherapy efficacy. Targeting the adenosine-A2A pathway can increase immune infiltration, enhance immune cell function, and partially reverse immunotherapy-insensitive "cold tumors" to "hot tumors" to enhance treatment response rates and improve the efficacy of current immunotherapy. At present, many adenosine receptor inhibitors have shown good results in clinical trials, especially in combination with immune checkpoint inhibitors, chemotherapy, and adoptive cell transfer therapeutic drugs, which are expected to be used for tumor immunotherapy to bring new breakthroughs. This article reviews the accumulation mode of adenosine in the tumor microenvironment, the role of A2AR and their regulatory mechanism in immune response, the progress of A2AR inhibitors in clinical trials, potential risks to target A2AR, and the prospects for therapeutic targeting A2AR.

Relationship between chronic constipation and brain-gut microflora axis / 中华胃肠外科杂志

The role of the brain-gut microbiota axis in functional gastrointestinal diseases has been gradually recognized. According to the ROME IV diagnostic criteria, functional gastrointestinal diseases are classified as diseases caused by abnormal brain-gut interaction. This concept is of great significance to the change of diagnosis and treatment paradigm of functional gastrointestinal diseases. Chronic constipation is the most common functional gastrointestinal disease. The pathogenesis of chronic constipation is closely related to the imbalance of intestinal flora, the abnormality of enteric nervous system and neurotransmitter in brain. Therefore, in the diagnosis and treatment of chronic constipation, enough attention should be paid to the concept of integration of brain-gut microflora axis, but the clinical application of brain-gut microflora axis is still limited. This may be one of the factors for high incidence but poor treatment efficacy of chronic constipation. Based on the global research progress and our clinical experience, this article expounds the clinical significance of the brain-gut microbiota axis in chronic constipation.

Flourishing Epitranscriptomics Research / 中国生物化学与分子生物学报

So far, researchers have found more than one hundred different kinds of chemical modifications on RNA. Most of these modifications are distributed on high abundant non-coding RNAs and are important for maintaining the functions of these non-coding RNAs. In recent years, thanks to the application of high-resolution mass spectrometry and the development of whole-transcriptome sequencing technologies, more and more modifications on mRNA have been discovered, and accurately mapped and localized, including N

Competency Model of Physical Therapists in China: An Exploratory Research / 中国康复理论与实践

Objective:To construct the competency model of physical therapists in China. Methods:A set of physical therapists' competency elements was established based on literature review and expert interviews. A questionnaire was designed based on the set, and 641 rehabilitation practitioners were investigated (521 valid questionnaires returned) with the questionnaire. Exploration factor analysis was used to construct the competency model of physical therapists, and confirmatory factor analysis was used to confirmed. Results:The set of physical therapists' competency elements included 74 elements in four dimensions. There were 44 competency elements in the competency model of physical therapists, including five dimensions that were Professionalism and Moral Conduct, Knowledge and Skills of Evaluation and Intervention, Abilities of Educational-Research and Advocate, Abilities of Cooperation and Management, and Basic Knowledge of Discipline, covering 74.41% of the total variance. The Cronbach's α coefficient of the whole questionnaire and each dimension was more than 0.8. Model fitness indexes of confirmatory factor analysis were as follow: χ2/df = 2.340, Root Mean Square Error of Approximation = 0.060, Goodness of Fit Index = 0.746, Normed Fit Index = 0.811, Incremental Fit Index = 0.850, Comparative Fit Index = 0.850. Conclusion:The competency model can provide a basis for physical therapists to become independent occupation, as well as for the education, selection, evaluation of physical therapists in China.

Seed oil of Brucea javanica induces apoptosis through the PI3K/Akt signaling pathway in acute lymphocytic leukemia Jurkat cells / 中国天然药物

Brucea javanica oil emulsion (BJOE) has been used to treat tumor in China for more than 40 years. However, its components and effectiveness in the treatment of acute lymphocytic leukemia (ALL) and its mechanism of anti-cancer activity remain unknown. In the current study, high-performance liquid chromatography-evaporative light scattering detector (HPLC-ELSD) was used to analyze the components of BJOE. Then, the anti-leukemia effects of BJOE were examined both in vitro and in vivo using ALL Jurkat cells and the p388 mouse leukemia transplant model, respectively. The primary ALL leukemia cells were also used to confirm the anti-leukemia effects of BJOE. The apoptotic-related results indicated that BJOE induced apoptosis in Jurkat cells and were suggestive of intrinsic apoptotic induction. Moreover, BJOE inhibited Akt (protein kinase B) activation and upregulated its downstream targets p53 and FoxO1 (forkhead box gene, group O-1) to initiate apoptosis. The activation of GSK3β was also involved. Our findings demonstrate that BJOE has anti-leukemia effects on ALL cells and can induce apoptosis in Jurkat cells through the phosphoinositide3-kinase (PI3K) /Akt signaling pathway.

Effect analysis of 71 cases of refractory diarrhea with severe malnutrition treated by fecal microbiota transplantation(FMT) combined with enteral nutrition / 中国实用外科杂志

OBJECTIVE: To investigate the therapeutic effect of fecal microbiota transplantation(FMT) combined with enteral nutrition on patients with refractory diarrhea and severe malnutrition.METHODS: Seventy-one patients with refractory diarrhea complicated with severe malnutrition in Department of Colorectal Disease,the Tenth People ' s Hospital of Tongji University and Department of General Surgery,General Hospital of Eastern Theater Command from June 2014 to October 2018 were prospectively collected for basic information,symptoms and signs,number of bowel movements,Bristol stool scores,and nutritional indicators,which were followed up after bacterial transplantation combined with enteral nutrition.RESULTS: After following up for 6 month,FMT combined with enteral nutrition in the treatment of severe diarrhea with severe malnutrition,the effective rate of treatment of diarrhea was 95.8%,of which the cure rate was 74.7%,and the remission rate was 21.1%.3 cases(4.2%) were ineffective,6 cases(8.5%) recurred.The frequency of bowel movements decreased from(8.3±2.2) before treatment to(5.3±1.3) after 1 month,(3.2±0.8)after 3 months,and(1.9±0.6) after 6 months.The Bristol score was reduced from(6.6±0.3) points before treatment to(4.8±0.7) points 1 month after treatment,(4.6±0.8)points after 3 months,and(4.3±0.8) points after 6 months.The body mass index(BMI) of the patients before treatment was 15.3±1.3,the BMI increased to 17.4±2.4 after 1 month of treatment,increased to 19.0±3.2 after 3 months,and 19.3±3.8 after 6 months.The total protein was increased from(48.6±4.7)g/L before treatment to(55.4±5.7)g/L after1 month of treatment,increased to(60.3±5.4) g/L after 3 months,and(68.4±6.9)g after 6 months.Albumin increased from(22.4±1.7)g/L before treatment to(31.7±2.4)g/L after 1 month of treatment,increased to(35.8±2.7)g/L after 3 months,and(37.5±3.7)g/L after 6 months.Fibrinogen increased from(1.6±0.4)g/L before treatment to(2.0±0.7)g/L after 1 month of treatment,increased to(2.4±0.8) g/L after 3 months,and(2.4±0.7) g/L after 6 months.Prealbumin increased from(82.0±18.6)mg/L before treatment to(178.3±25.3)mg/L after 1 month of treatment,increased to(235.7±28.2) mg/L after 3 months,and(259.6±33.5) mg/L after 6 months.The were significant differences before and after treatment(All P<0.05) CONCLUSION: After 6 months of follow-up,FMT combined with enteral nutrition can effectively reduce the number of diarrhea,improve stool characteristics,and improve nutritional indicators such as BMI,total protein,albumin,fibrinogen and prealbumin.