Obstract: Objective To study the effect of Guyanding Decoction(GYD) on the metabolism of oxygen-derived free radicals in the chicken model of knee osteoarthritis and to explore its therapeutic mechanism.Method Forty chickens were randomized into 5 groups: normal group,model group,low-dose GYD group,high-dose GYD group and Fenbid group.Low-dose group was administered 5 g/kg GYD and high-dose group with 10 g/kg GYD by gastric gavage.Fenbid group was given Fenbid 0.15 g/kg.The SOD activity and the MAD content in serum were detected in each group.Result The activity of SOD in model group was significantly lower than that in normal group(P 0.05).For the content of MDA,model group had a higher level than normal group(P 0.05).Conclusion The decoction of GYD exerts curative effect on experimental osteoarthritis by improving the activity of SOD,eliminating excessive oxygen-derived free radicals,decreasing the content of MDA,and preventing osteoarthritis patients from the injury of oxygen-derived free radicals.

Objective To evaluate the expression of CyclinD1 in gallbladder carcinoma and its relationship with clinlicopathological parameters.Method SABC immunohistochemistry was used to detect the expression of CyclinD1in 41carcinoma,15 chronic cholecystitis,10 normal tissue of gallbladder .Gallbladder carcinoma cell proliferation condition was detected by PCNA counting score .Results There were no positive signal of CyclinD1 deteted in chronic cholecystitis and in normal tissue,the positive rate of CyclinD1 in gallbladder carcinoma was 58.5%(24/41).The expression of CyclinD1 was related to tumor TNM stage, lymph node metastasis and cell differentiation (P

Obgective To analyze the relevant risk factors of recurrent wheezing(≥3 attacks) in the first 3 years of life.Methods Wheezing,respiratory sounds,risk factor were used as key words to retrieve papers in Chinese literature databases including Sinomed,Wanfang and Weipu databases.The same strategy was used to retrieve English papers in English literature databases including PubMed,Cochrane library and Embase.Time range was from 31th May 2004 to 1 rd June 2014.The execution of quality evaluation of the included documents was in compliance with Newcastle-Ottawa Scale and cross-sectional study standard recommended by Agency for Healthcare Research and Quality.The evidence quality evaluation was conducted with GRADEpro and followed by the Meta analysis with RevMan 5.2.R~ults A total of 13 studies were included in this Meta-analysis.Several factors were related to recurrent wheezing episodes,including risk factors such as maternal smoking during pregnancy (OR =1.47,95％ CI:1.30-1.66),asthma in parents (OR =1.94,95 ％ CI:1.72-2.19),family history of atopy (OR =1.94,95％ CI:1.72-2.19),male (OR =1.42,95 ％ CI:1.19-1.69),history of eczema (OR =2.36,95 ％ CI:1.69-3.30),colds (＞ 6 times) (OR =2.02,95 ％ CI:1.54-2.64),history of bronchopneumonia (OR =1.85,95 ％ CI:1.46-2.34),exposure to cigarette smoking(OR =2.30,95％ CI:1.68-3.14),daycare attendance(OR =2.27,95％ CI:1.97-2.60);Education received by the mother ＞ 12 years (OR =0.80,95％ CI:0.70-0.92) was the protective factor.Conclusions The risk factors of recurrent wheezing(≥3 attacks) in the first 3 years of life are maternal smoking during pregnancy,asthma in parents,family history of atopy,male,history of eczema,colds (＞ 6 times),history of bronchopneumonia,exposure to cigarette smoking and daycare attendance.The protective factor is education received by the mother ≥ 12 years.The prerequisite in precaution of infants recurrent wheezing is to ensure the utmost avoidance of hazardous factors and reinforcement of protective factors.

【Objective】To observe the effect of Guyanding(GYD)decoction,a prescription with the actions of tonifying kidney,strengthening Qi and activating blood on proliferation of vascular endothelial cells(VEC).【Methods】New Zealand rabbits were randomized into blank control groups and GYD decoction groups in the dosages of 2.5,5 and 10g/kg respectively.GYD in the dosage of 10g/kg was given to rats for 7 successive days,and blood sampling was collected 0.5,1,1.5,2,2.5,3,3.5 and 4 hours after the last administration to prepare serum containing GYD.Then human umbilical vein endothelial cells were cultured with serum containing GYD,and the effects of GYD-containing serum at different time points on the proliferation of VEC were observed by methylthiazolyltetrazolium(MTT)assay.GYDcontaining serum collected 2h after the last administration was partially inactivated and its effect on VEC proliferation was compared with that of non-inactivated GYD-containing serum.Meanwhile,the effect of non-inactivated GYD-containing serum on VEC proliferation was observed 12,24 and 36 hours after the culturing.GYD in three different dosages was given to rats for 7 successive days,and serum containing GYD collected 2h after the last administration was prepared to observe the effect of GYD in different doses on VEC proliferation.【Results】Serum containing high-dose GYD collected 0.5～4h after the administration had obvious effect in promoting VEC proliferation(P

Objective To analyze the relevant risk factors of recurrent wheezing(≥3 attacks) in the first 3 years of life in Shanghai Pujiang.Methods A case-control study was conducted.Two hundred and sixty-two research children were chosen for clinical visits (＜ 3 years old) with wheezing at the Pediatric Department of Shanghai Renji Hospital (South Campus),School of Medicine,Shanghai Jiaotong University,from January to December 2014.According to the frequency of wheezing,the subjects were divided into 75 cases of recurrent wheezing group (≥ 3 attacks),110 cases of occasional wheezing group(1-2 attacks) and 77 cases of no wheezing group.Probable risk factors were inquired by using face-to-face questionnaire.The passive agglutination method was used to detect the Mycoplasma pneumoniae antibody immunoglobulin M (IgM).The indirect immunofluorescence was used to detect the respiratory pathogens.The Western blot was used to detect 20 items of serum allergen.Chi-square test was firstly used for univariate analysis,and then the multivariate stepwise Logistic regression was used to analyze the independent risk factors associated with infant recurrent wheezing.Results A total of 20 factors were found relevant to infant recurrent wheezing by univariate analysis,which included boys (OR =4.030,95％ CI:1.937-8.388),personal atopy (OR =13.125,95％ CI:5.951-28.946),allergic dermatitis (OR =9.833,95％ CI:4.663-20.737),allergic rhinitis (OR =40.327,95％ CI:5.300-306.842),like rubbing eyes or nose(OR =6.487,95％ CI:3.190-13.191),food allergy (OR =6.689,95 ％ CI:1.860-24.051),premature birth (OR =3.795,95 ％ CI:1.001-14.385),low birth weight (OR =9.075,95％ CI:1.106-74.450),parental atopy (OR =10.667,95％ CI:4.824-23.587),parental allergic dermatitis (OR =8.072,95 ％ CI:2.634-24.734),parental allergic rhinitis (OR =6.524,95 ％ CI:2.920-14.577),parental allergic conjunctivitis (OR =1.087,95％ CI:1.017-1.162),parental asthma history (OR =1.119,95％ CI:1.035-1.210),colds ＞ 6 times (OR =9.111,95％ CI:3.970-20.909),history of bronchopneumonia(OR =7.554,95％ CI:3.588-15.903),age at first time use of antibiotics less than 6 months (OR =2.388,95％ CI:1.129-5.052),exposure to cigarette smoking (OR =1.922,95 ％ CI:1.004-3.681),maternal passive smoking during pregnancy (OR =2.508,95 ％ CI:1.298-4.848),living close to wood stove (OR =3.342,95 ％ CI:1.427-7.827) and positive results of inhaled allergens (OR =1.821,95 ％ CI:1.420-2.336).Keeping cats was the protective factor(OR =0.922,95％ CI:0.864-0.984).The forward Logistic regression analysis showed that personal atopy (OR =10.278,95 ％ CI:2.503-42.202),like rubbing eyes or nose (OR =1 0.316,95 ％ CI:2.722-39.101),food allergy (OR =10.370,95％ CI:1.248-86.145),parental atopy (OR =5.402,95％ CI:1.340-21.778),colds ＞ 6 times (OR =7.048,95 ％ CI:1.688-29.423),history of bronchopneumonia (OR =7.876,95 ％ CI:2.040-30.407) and maternaal passive smoking (OR =3.696,95 ％ CI:1.013-13.494) during pregnancy were the independent risk factors of infants recurrent wheezing.Conclusion Personal atopy,like rubbing eyes or nose,food allergy,parental atopy,colds ＞ 6 times,history of bronchopneumonia,maternal passive smoking are the independent risk factors of recurrent wheezing in infants less than 3 years old.

Objective:To analyze the mistakes in the application of tuina therapy in the rehabilitation of stroke,in order to elevate the clinical effects in the treatment of stroke by tuina.Methods:The issues are found out by searching and analyzing the relevant articles on tuina treatment of stroke.Results:In the tuina treatment of stroke,mistakes exist in the intervening time of the treatment,in the emphasis of tuina with negligence of rehabilitation,in the assessment of therapeutic effects,in the selection of the manual techniques and acupoints,and in the negligence of health education.Conclusion:In the tuina treatment of stroke,it is advisable to start treatment in the early stages,use the manual techniques in different phases,to cooperate with rehabilitation at the proper time and to emphasize health education,in order to avoid or reduce the occurrence of misuse syndrome,and eliminate the appearance of disuse syndrome,for displaying the unique advantage of tuina therapy fully and promoting the innovation and development of TCM rehabilitative techniques.

The ompA gene of Chlamyia psittaci in cows was amplified by PCR with primers designed based on those reported in GenBank.The amplified ompA gene was inserted into the bacterial plasmid vector pGEX-4T-1 and then transformed into E.coli BL21(DE3) with IPTG induction. The gene was derived from plasmid pMD18-T vector and then sequenced.It was demonstrated that this recombinant fusion protein of approximately 68kD in molecular mass was highly expressed in inclusion body and more pure proteins would be produced after purification.The fusion protein specifically reacted with positive sera of bovine Chlamydia as demonstrated by Western blotting. These results indicate that this recombinant fusion protein shows good reactivity and could be used to develop the diagnostic kit for bovine Chlamydia and genetic engineering vaccine.

Objective: To analyze the clinical features and factors associated with osteonecrosis in children with systemic lupus erythematosus (SLE). Methods: A retrospective analysis of 15 SLE patients with osteonecrosis in Department of Pediatrics, Renji Hospital Affiliated to School of Medicine of Shanghai Jiaotong University from January 2013 to May 2017 was carried out.Forty-two SLE patients without osteonecrosis were selected as control group.The clinical, laboratory variables and the treatment were compared among SLE patients who were with and without osteonecrosis. Results: (1) Fifteen patients developed osteonecrosis that constituted 8.6% of all the 175 hospitalized SLE patients during the same period.(2) Of 15 patients, 2 patients were male, 13 patients were female, who developed osteonecrosis with an average age of (13.9±2.7) years (range: 10-18 years old). The duration of SLE before the diagnosis of osteonecrosis ranged from 6 days to 141 months, the median was 10 months, and 80.0% (12/15 cases) was diagnosed with osteonecrosis within 2 years of SLE diagnosis.There were 36 joints involved in 15 patients, all of which were detected by magnetic resonance imaging(MRI). The knees were the most commonly involved joints(14/15 cases, 93.3%), followed by hip and ankle joints.(3) Univariate analysis revealed that the level of Triglyceride [(2.080±1.500) mmol/L vs.(1.350±0.945) mmol/L], maximum daily dose of glucocorticoid[(1.25±0.33) mg/kg vs.(1.07±0.22) mg/kg], positive rate of gene associated with glucocorticoid-induced osteonecrosis of femoral head(100.0% vs.54.8%)were significantly higher in SLE with osteonecrosis than those in controls(all P<0.05). While the level of 25(OH)D₃[(21.37±11.29) μg/L vs.(31.45±17.73) μg/L] was significantly lower than that of controls(P<0.05). Multiple factor Logistic regression analysis showed that hypertriglyceridemia and daily maximum dose of glucocorticoid were the risk factors for osteonecrosis. Conclusions Osteonecrosis mostly occurred in children over 10 years old, knee joint involvement is the most common.The high-risk time of osteonecrosis is within 2 years of SLE diagnosis.Hypertriglyceridemia and daily ma-ximum dose of glucocorticoid are risk factors associated with osteonecrosis in children with SLE.

Human beta defensin 4 is a small cationic peptide with a broad range of antimicrobial activity. It plays an important role in innate immunity of human body, especially in mucosal and epithelial defense. In this study, the full-length encoding gene of HBD4 was synthesized by overlap extension polymerase chain reaction and inserted into cloning vector pMD18-T. The gene encoding mature peptide of HBD4 was amplified by PCR and cloned into prokaryotic expression vector pGEX-4T-2. Then pGEX-4T-2/mHBD4 was transformed into E. coli DH5 alpha, which was induced by isopropy-beta-D-thiogalactoside (IPTG). The identification was made by means of endonuclease digestion, DNA sequencing, sodium dodecyl sulphate-polyacrylamine gel electrophoresis (SDS-PAGE). The results showed that the synthesized gene and cloned gene were identical to the HBD4 gene sequence registered in GenBank and were successfully cloned into cloning vector pMD18-T and prokaryotic expression vector pGEX-4T-2. After IPTG induction, the GST-HBD4 fusion protein was successfully expressed in E. coli.
Anti-Infective Agents ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Humans ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; beta-Defensins ; biosynthesis ; genetics

Abstract:Objective To study the structure and function of a novel serine protease gene associated with human colorectal cancer SNC19.Methods The cDNA sequence was determined by both manual and automatic sequencing techniques. The full length cDNA sequence was obtained by the 5'-Rapid Amplification of cDNA Ends technique and web-based analysis. Open reading frame analysis and protein function prediction were also performed. Northern blot was used to detect the expression of SNC19 in various human normal tissues and tumor cell lines. Fluorescent in situ hybridization combined with fluorescent R-banding technique was employed to map the SNC19 gene on human chromosome.Results Full length SNC19 cDNA, size 3152 bp, encodes a protein highly homologous to a mouse serine protease epithin. In normal human tissues, high SNC19 expression levels were observed in the kidney, pancreas, prostate, small intestine and colon; moderate SNC19 expression levels were observed in the placenta, lung, liver, spleen thymus, testis and peripheral blood lymphocytes; and extremely low expression levels were observed in the heart, brain, skeletal muscle and ovary. In tumor cell lines, colorectal cancer cells SW480, SW620, SW1116 and Colo205, breast cancer cell Bcap37 and gastric cancer cells MKN28 and SGC7901 showed high levels of SNC19 expression; cervical cancer cell HeLa-S3, lung cancer PAA, oral epithelial cancer cell KB and lymphoma cell Raji showed moderate levels of SNC19 expression; and tongue squamous cancer cell Tca8113, leukemia cells HL-60, K562, MOLT-4, lung cancer cell A549 and melanoma cell G361 showed very low levels of SNC19 expression. SNC19 was mapped to human chromosome 11q24-25. Conclusion SNC19 encodes a novel human serine protease with 855 amino acid residues. As a novel serine protease associated with human colorectal cancer, the expression of SNC19 in various tissues and cell lines may have very important impact on their phenotypes and biological behaviors.