The development of metagenomics revealed a novel role of microorganism in lots of diseases.Emerging researches at home and abroad illustrated that microbiome changes from nasopharynx, oropharynx and/or lung in quality and/or quantity exist in many respiratory diseases including chronic obstructive pulmonary disease, asthma, cystic fibrosis, pneumonia as well as upper respiratory infection, which play an important role in immune system, metabolism and neuroregulation.These research results may provide us new strategy for the diagnosis, therapy, surveillance and prognosis of respiratory diseases.

AIM: To investigate the differentiation of human bone marrow mesenchymal stem cells (MSC) into chondrocytes in vitro and determine factors involving in the differentiation process. METHODS: MSC were separated from iliac bone marrow with lymphocyte separating medium using density centrifugation. Cells were cultured and expanded in medium until reaching required number. MSC was induced to differentiate into chondrocytes by adopting high cell density, supplying growth factor and using micromass culture. Cells were observed by HE staining. Matrix of cartilage was detected by alcian blue and toludine blue and cartilage specific collagen II was detected by immunohistochemistry. RESULTS: The structure of the micromass assumed that of cellular cartilage, alcian blue staining were uniformly positive and toludine blue detected diffuse metachromasia substance, cells uniformly expressed collagen Ⅱ. CONCLUSION: High cell density, growth factor and appropriate culture conditions are critical to induce differentiation of MSC into chondrocytes. [

An optimized two-dimensional polyacrylamide gel electrophoresis (2-DE) system for analyzing plant proteins was developed by evaluating different reagents and concentrations used in the sample extraction solutions and lysis buffers. Two main sample preparation methods, referred to as trichloroacetic acid (TCA)-acetone method and phenol extraction-ammonium acetate/methanol (phenol-NH4Ac/methanol) precipitation method, were compared. Four ecotypes of reed plants (Phragmites communis Trin.) from the desert region of north-western China were used as experimental materials: (1) swamp reed (SR) which grows in water about 1 m deep; (2) dune reed (DR) which grows on 5～10 m high sand dunes; (3) heavy salt meadow reed (HSMR) which grows on low-lying salt flats; and (4) light salt meadow reed (LSMR) which grows in the transition area between DR and HSMR growing areas. The optimized phenol-NH4Ac/methanol precipitation method consisted of extracting leaf proteins of different ecotypes of reed with water-saturated phenol and then precipitating with a 5-fold volume of 0.1 mol/L NH4Ac in methanol, followed by dissolving in the lysis buffer. The optimized protein lysis buffer consisted of 7 mol/L urea, 2 mol/L thiourea, 4% CHAPS, 2% Ampholine(pH 3.5～10∶pH 5～8 = 1∶4) and 65 mmol/L DTT. The prepared protein sample (80 ?g) was then separated by 2-DE gel and detected by silver staining method. This improved 2-DE system resulted in a 2-D protein profile of higher resolution and higher protein yields as analyzed by PDQuest software. Good results were also obtained when this 2-DE system was used in 2-D analysis of proteins from other plant materials, such as rice leaves, indicating that it is a suitable 2-DE system for analyzing leaf proteins of different plant species.

Objective To study the effect of genetic polymorphism of leukotriene C4 synthase (LTC4S) and 5-lipoxy-genase (ALOX5) on efifcacy of leukotriene receptor antagonist (LTRA) in children with moderate persistent asthma. Methods Seventy-two children with moderate persistent asthma who visited the out-patient clinic of Shanghai Children’s Medical. Center from June 2011 to June 2013 were divided into two groups, each of which ifrst had ICS or LTRA+ICS for twelve weeks and then had the other for another twelve weeks. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to assess the genetic polymorphism of LTC4S RS730012 and ALOX5 RS2115819. Pulmonary function, clinical symptoms and C-ACT score were evaluated before and after treatment. Results After the treatment with LTRA, 75%forced expiratory lfow (FEF75) was improved more signiifcantly in patients with A/C or C/C genotype at LTC4S (RS730012) locus than in patients with A/A genotype. After the treatment with LTRA+ICS, there was no difference of pulmonary function among patients with different genotypes at ALOX5 (RS2115819). Conclusions The SNP of LTC4S (RS730012) is associated with the efifcacy of montelukast in asthmatic patients because of the improvement of small airway function.

Objective To investigate the association of nitrotyrosine with coronary heart disease (CHD) in type 2 diabetes mellitus.Methods The nitrotyrosine levels were determined in 109 patients of type 2 diabetes mellitus without CHD (T2DM).One hundred and fifty-two patients of type 2 diabetes mellitus with CHD (T2DM-CHD) and 103 healthy control subjects by ELISA.Results T2DM-CHD patients had significantly increased nitrotyrosine compared with T2DM group and the control group [ ( 78.17±10.68 )nmol/L,(70.50 ± 9.13) nmol/L vs ( 63.23 ± 11.55 ) nmol/L,Ps ＜ 0.01 ].Nitrotyrosine was correlated with total cholesterol,triglyceride,fasting glucose and Gensini Score (r=0.361,P=0.009;r =0.206,P=0.001 ;r=0.347,P=0.026; r=0.466,P ＜ 0.001 ).Multivariable logistic regression showed nitrotyrosine was independently associated with CHD combined with type 2 diabetes mellitus ( OR=1.094,95％ CI:1.053-1.137 ; P ＜ 0.01 ).Conclusion Nitrotyrosine plays an important role in the formation and development of cardiovascular disease in tvoe 2 diabetes.

AIM: To observe the effect of antibiotic treatment on the inflammatory mediator expression in peritoneum and the peritoneal transport function in rats with acute peritonitis, and explore its mechanisms. METHODS: Eighty-six SD rats were randomly divided into three groups. Control group (n=28) were treated with PBS (ip), peritonitis group (n=28) and treatment group (n=28) were challenged with the E.coli (ip), but at 3 h and 9 h gentamicin was given (ip) in treatment group. Seven rats of every group were randomly sacrificed at 24 h, 48 h, 72 h and 7 d. Peritoneal equilibration test (PET) was did before they were killed. Leukocyte count, pathological changes and the expression of CD45, NF-?B, IL-1?, TNF-? in peritoneum were examined. RESULTS: (1)The blood leukocytes in peritonitis group decreased strikingly, but did not change obviously in other two groups. The peritoneal fluid leukocytes in peritonitis group increased significantly from 24 h to 72 h, while in treatment group, it enhanced more strikingly than peritonitis group at 24 h, and recovered earlier. (2) Both in peritonitis group and treatment group, the expression of activated NF-?B, IL-1?, TNF-? and CD45 increased significantly, but the treatment group was lower than model group at 48 h and 72 h. The mRNA level of IL-1? and TNF-? had the same trend as their protein expression. (3) Compared with the control group, UF and D/D_0 Glu decreased significantly in model group and treatment group, and D/PTP increased dramatically. The D/P TP in treatment group lowered obviously compared with peritonitis group, while the net UF and D/D_0 Glu had not significant difference between treatment group and model group. CONCLUSION: Antibiotic treatment can partly decrease the expression of inflammatory mediators in peritoneum of rats with acute peritonitis and also can improve the protein transport ability to some extent, but can not improve the peritoneal ultrafiltration and the glucose transport function.

Rosmarinic acid (RA), a phenolic acid, is one of the important secondary metabolites produced in Salvia miltiorrhiza. To observe the influence of salicylic acid (SA), an elicitor, on the synthesis of RA and related enzymes, we treated the cell suspension cultures of S. miltiorrhiza with SA and L-a-aminooxy-beta-phenylpropionic acid (AOPP), a competitive inhibitor of tyrosine aminotransferase (TAT). Under this condition, the activities of related enzymes, such as phenylalanine ammonia-lyase and TAT were traced and assayed; the accumulative amount of RA was measured. The results showed that the PAL activity reached the peak at 4 h, 124% higher than that of the control, and the content of RA reached its maximum ((5.914 +/- 0.296) mg/g dry weight) at 8 h, after treated by 6.25 mg/L SA on day 6 of the suspension culture. The results of treatment with 0.1 micromol/L AOPP showed that AOPP affected little on the TAT activity, while the PAL activity was significantly influenced, with 44% lower than that of the control at 6 h. Meanwhile, the reduced accumulation of RA ((4.709 +/- 0.204) mg/g dry weight) paralleled with the decrease in PAL activity. The co-treatment by 0.1 micromol/L AOPP and 6.25 mg/L SA relieved the restriction imposed by AOPP on PAL, and made the cell cultures accumulate more RA than sole treatment with AOPP, indicated that SA induced the accumulation of RA in suspension cell culture of S. miltiorrhiza, and the rate-limiting effect of PAL was stronger than TAT.
Cell Culture Techniques ; methods ; Cinnamates ; metabolism ; Depsides ; metabolism ; Phenylalanine Ammonia-Lyase ; metabolism ; Plant Cells ; metabolism ; Salicylic Acid ; pharmacology ; Salvia miltiorrhiza ; cytology ; growth & development ; metabolism ; Suspensions ; Tyrosine Transaminase ; metabolism

Objective:To analyze the clinical features and ultrasonographic features of Williams syndrome(WS) in children, and to explore the diagnostic value of ultrasonography in this disease.Methods:A retrospective analysis was performed on 19 children with abnormal echocardiography at Hunan Children′s Hospital from January 2016 to December 2020, and the abnormal echocardiography was evaluated by WS phenotypic scoring method of American Academy of Pediatrics.Cardiological CT and genetic tests were performed, and the clinical data and ultrasound image characteristics were analyzed.Results:The WS score of 19 cases was more than six, which was highly suspicious clinically.In 19 cases, supra-aortic stenosis and pulmonary stenosis were the most common findings on echocardiography, accounting for 63.2%(12/19), among which three cases were complicated with aortic arch constriction and one case with mitral lobe deformity.There were 15.8%(3/19)children with supra-aortic stenosis associated with pulmonary artery velocity increase, and 10.5%(2/19)children with left and right pulmonary artery branch stenosis associated with aortic inner diameter narrowing.Other cardiac malformations were found in 42.1%(8/19) of the children.Chromosome 7 microdeletions accounted for 68.4%(13/19) in genetic examination, and no abnormalities were found in three cases(15.8%, 3/19). The genetic results of three cases were still under follow-up.Conclusion:Echocardiography can accurately and sensitively detect cardiac lesions in children with WS.Combined with WS score of American Academy of Pediatrics, the diagnosis rate of WS can be improved.

【Objective】 To investigate the expression level of liver kinase B1 (LKB1) gene in bone marrow of patients with acute non-lymphoblastic leukemia (AML) and its correlation with prognosis. 【Methods】 A total of 90 AML patients from May 2015 to January 2017 were selected as study subjects, and 30 cases of bone marrow specimens from non-malignant hematologic diseases were selected as control group. The expression of LKB1 mRNA in bone marrow was detected by real-time fluorescent quantitative PCR (qRT-PCR). The expression of LKB1 protein was detected by Western blot. The correlation between LKB1 mRNA and prognosis of AML was analyzed by Kaplan-Meier survival analysis. 【Results】 The mutation rate of LKB1 gene, the mRNA and LKB1 protein expression in bone marrow of AML patients was lower than those of control group (χ²=13.274, t=34.134, t=45.235, P<0.05). The mutation rate of LKB1 gene and the mRNA expression from high to low order is M1(81%, 17/21)>M5(78.6%, 11/14)>M6(75%, 3/4)>M2(42.4%, 14/33)>M4(41.7%, 5/12)>M3(35.3%, 6/17). Thefollow-up survival rate of patients with AML in the LKB1 high amplification group was higher than that of patients with LKB1 low amplification(χ²=8.039, P<0.05) The median survival time of the LKB1 high amplification group was higher than that of the LKB1low amplification group (27.3 months vs 19.8 months) (χ²=5.552, P<0.05). The incidence of post-chemotherapy infection, post-chemotherapy recurrence and extramedullary infiltration in the LKB1 high amplification group was lower than that in patients with LKB1 low amplification (P>0.05). 【Conclusion】 The expression level of LKB1 gene in patients with AML is low, moreover the more low expression level of LKB1 gene were, the more severe ill condition and more poor prognosis.