Objective To improve the purifying method of Jo-1 antigen from rabbit thymus used for detection of anti-Jo-1 antibody by dot-blotting immunoassay(DB).Methods The rabbit thymus glands were cut into pieces,homogenized and extracted by PBS.Total protein was precipitated by acetone to get acetone powder(RTAP).The RTAP was solved in PBS and separated by an by anti-Jo-1 IgG affinity column.Results 5～7 g RTAP was obtained from 100g rabbit thymus glands.There was 19%～24% of protein in RTAP.Jo-1 antigen was enriched around 1900 folds through affinity chromatography,with 2.5% recovery of antigenic activity.In this preparation,there were several bands on SDS-PAGE,but only one band about 50 ku,reacted with anti-Jo-1 antisera on immunoblotting.Dot-blotting also showed that the antigen only reacted with Jo-1 antisera.The purified Jo-1 antigen was not stable for long time,but the antigenic activity could maintain for a long time when there was MgCl2 in the solution.Conclusion Affinity chromatography was a simple and easy method for purifying Jo-1 antigen from rabbit thymus.The antigen purified by affinity chromatography could meet the requirement for detecting Jo-1 antibody bydot-blotting.

Purpose To observe the effect of Rongban Tongmai Granules on thrombosis and blood viscosity of in vitro rat model of blood stasis, and to study the activating blood circulation effect of the drug.Methods To observe the effect of the Rongban Tongmai Granules on thrombosis and blood viscosity of in vitro rat model of stasis, subcutaneous inject rat with epinephrine hydrochloride, and then copy "blood stasis" model by ice water stimulation in rats.Results According to a continuous 7 days′intragastric administration of Rongban Tongmai Granules, thrombus length of blood stasis model rats in vitro reduced significantly (P<0.05-0.01),wet and dry weight of thrombus reduced significantly (P<0.05), the shear rate of the whole blood viscosity under 100 S~(-1), 30 S~(-1), 5 S~(-1) decreased significantly as well (P<0.05-0.01), and the shear rate of whole blood viscosity had decreasing tendency under 200 S~(-1).Conclusion Rongban Tongmai Granules can inhibit thrombosis and lower blood viscosity.

OBJECTIVE: To study the clinical efficacy of Nalmefene in the treatment of chronic type Ⅱrespiratory failure. METHODS: A total of 40 patients with chronic type Ⅱ respiratory failure were randomly divided into treatment group and the control group: the treatment group received Nalmefene (1.0 mg) plus 5% GS (250 mL) q.d by iv gtt. in addition to the routine treatment, and the control group received 1.875 g Nikethamide plus 5% GS (250 mL) q.d by iv gtt. After 5-day treatment, the clinical symptom, physical signs, adverse drug reactions, lung function testing and blood gas analysis in two groups were evaluated. RESULTS: The overall response rate was 95.0% in the treatment group vs. 60.0% in the control group(P

Objective To investigate the effects of silencing Smo gene on proliferation and apoptosis of rat prima-ry chondrocyte in vitro.Methods The primary chondrocyte was obtained by mechanical-enzyme digestion and identified by Immunohistochemical cells ( ColⅡ) .The animals were divided into control group , control siRNA group and Smo siRNA 1 ~3 group.The siRNA was transfected into chondrocytes by lentivirus vector .After 72 h, the cell viability was detected by MTT, Smo expression was detected by RT-PCR and Western blot, and the apoptosis of chondrocyte was assessed by flow cytometry .Results All types of siRNA were transfected into primary chondrocyte by vectors, the Smo siRNA 1 ~3 may inhibit the expression of Smo mRNA and protein in chondrocytes, and Smo siRNA2 had the highest silencing rate ( the expressions of Smo mRNA and protein were 0.19 ±0.03 and 0.39 ±0.07 ) .The cell viability in Smo siRNA2 group was lowest ( 77.38% ±7.19%) , while the apoptosis rate of Smo siRNA2 was highest ( 21.43%±2.97%) .Conclusions Silencing Smo gene in primary chondrocytes may inhibit proliferation and promote apoptosis , Smo may have a protecting role from apop-tosis of the chondrocyte.

Objective To study the proliferation and apoptosis and investigate the expression of Indian hedgehog (Ihh),parathyroid hormone-related peptide (PTHrp),smoothened (Smo) protein and mRNA in the cultured rat primary chondrocytes exposed to different doses of NaF.Methods The third generation articular chondrocytes of neonate rat were cultured in vitro and treated with 0 (control),5,10,20 and 40 mg/L of fluoride.The proliferation activities of cells at different times (24,48 and 72 h) were tested by Thiazolyl Blue Tetrazolium Bromide (MTT).The apoptosis rate was determined by flow cytometry.The expressions of protein and mRNA of Ihh,Smo and PTHrp at 48 h were determined by Western blotting and semi-quantitative RT-PCR,respectively.Results After exposed to 5 mg/L of fluoride for 24,48 and 72 h,the proliferation rates were significantly increased [(1.17 ± 0.07)％,(1.20 ±0.06)％,(1.16 ± 0.08)％] compared with those of control group [(1.10 ± 0.08)％,(1.13 ± 0.08)％,(1.15 ± 0.08)％],but the proliferation activity at 48 and 72 h in 40 mg/L group [(0.72 ± 0.11)％,(0.68 ± 0.04)％] was significantly lower than those in control group (all P ＜ 0.05).Compared with the control group,apoptosis rate of cartilage cell in fluoride treatment group increased gradually [(1.47 ± 0.05)％,(19.87 ± 3.03)％,(25.30 ± 1.28)％,(45.73 ± 4.63)％,F =123.328,P ＜ 0.01].Western blot analysis and RT-PCR results showed that the Ihh,PTHrp,Smo mRNA and protein expression increased in the fluoride groups at 48 h (Ihh protein:0.77 ± 0.08 vs.0.98 ±-0.07,1.23 ± 0.06,1.37 ±0.07,1.34 ± 0.07;PTHrp protein:0.68 ± 0.04 vs.0.89 ± 0.05,0.83 ± 0.05,1.29 ± 0.05,1.16 ± 0.08;Smo protein:0.37 ± 0.01 vs.0.64 ± 0.06,0.67 ± 0.03,0.96 ± 0.06,0.69 ± 0.06;Ihh mRNA:0.77 ± 0.05 vs.0.98 ± 0.05,1.09 ±0.05,1.27 ± 0.03,1.46 ± 0.06;PTHrp mRNA:0.67 ± 0.07 vs.0.97 ± 0.05,1.07 ± 0.08,1.37 ± 0.05,1.45 ± 0.05;Smo mRNA:0.45 ± 0.03 vs.0.63 ±-0.04,0.71 ± 0.05,0.81 ± 0.01,1.00 ± 0.02,all P ＜ 0.05).Conclusions Low doses of fluoride can promote the proliferation of chondrocytes cultured in vitro,and high doses of fluoride can promote the apoptosis of chondrocytes cultured in vitro.The expression of Ihh signaling pathway RNAs and proteins of the cartilage cells are increased following increased levels of fluoride.The results suggest that fluorine has activated the Ihh signaling pathway in chondrocytes and promoted the proliferation and apoptosis processes which might be involved in chondrocytes injury.

OBJECTIVE: To observe the clinical efficacy of Jinhua xiaocuo pill and 0.05% taiarotene cream in the treatment of acne vulgaris. METHODS: 64 patients with acne vulgaris were randomly divided into experimental group and control group according to visiting order, the cases of 2 groups were 32. Experimental group were treated with 4 g Jinhua xiaocuo pill p.o. and 0.05% taiarotene cream for external use once every evening. Control group were treated with chloramphenicol/metronidazole liniments three times a day, the treatment course of 2 groups were 6 weeks. Efficacy before and after treatment were evaluated and analyzed with SPSS 13.0 software. RESULTS: The effective rate was 93.8% for experimental group and 65.6% for control group. Significant different was noted between two groups(?2=7.169,P

Objective To evaluate antimicrobial resistance and antimicrobial resistance mechanisms of Enterococcus faecalis (E.faecalis)to linezolid (LNZ),and provide basis for clinical rational drug use.Methods Twelve E.faecalis strains isolated from sputum of patients who received LNZ therapy in a hospital between January 2012 and January 2013 were collected.The minimum inhibitory concentrations (MICs)of antimicrobial agents were de-termined by agar dilution method,23S rRNA V region gene of E.faecalis was amplified by polymerase chain reac-tion,the amplified products were sequenced.Results Of 1 2 isolates,2 were intermediate strains and 1 0 sensitive strains.The G2576U mutation was detected in 2 intermediate strains,1 of which was also detected G2424U muta-tion;the variations were not detected in 10 sensitive strains.C2424U and G2576U mutation existed in R1 and R4 region respectively.Conclusion 23S rRNA V region gene mutations are found in the intermediate strains of E.faecalis.Change in MIC values of linezolid should be paid close attention in clinical use.

Objective To study the clinical features and possible pathogenesis of acute ischemic stroke in renal cell cancer patients. Methods The clinical data from in-hospital patients with renal cell cancer who developed acute ischemic stroke were collected, including the patients with renal cell cancer who developed acute ischemic stroke during anti-cancer therapies and those patients with acute ischemic stroke who were firstly diagnosed to have renal cell cancer during anti-stroke therapies between January 2003 and December 2015. Results A total of 2516 patients with renal cell cancer were screened, and there were 36 patients (1.43%) with acute ischemic stroke. Out of the 36 patients, there were 29 men (80.56%) and 7 women (19.44%). Their age ranged from 45 to 68 years, with a average age of (65.11 ± 14.77) years. Eight patients (22.22%) had some conventional cardiovascular risk factors, while the other 28 patients (77.78%) had no such risk factors. Magnetic resonance imaging (MRI) scans at the acute stage of ischemic stroke were carried out for all these patients. Based on the diffusion weighted imaging (DWI) of MRI, 8 patients (22.22%) had single lesion and 28 patients (77.78%) had multiple lesions in different arterial territories in their brains. The pathological types of renal cell cancer were:suprarenal epithelioma (18 patients, 50.00%), papillary cell carcinoma (12 patients, 33.33%) and chromophobe renal cell carcinoma (6 patients, 16.67%). Metastases were found 10 patients (27.78%) out of the 36 patients. Blood biochemical examination showed that 28 patients had elevated plasma D-dimer level, 22 patients had elevated plasma cancer antigen (CA)125 level, and 17 patients had elevated plasma carcinoembryonic antigen (CEA) levels. Conclusions It is suggested that the renal cell cancer associated stroke is characterized by lacking of traditional risk factors and having multiple lesions in brain;and that the elevated plasma D-dimer, CA125 and CEA levels may lead to hypercoagulable state and lead to ischemic stroke eventually .

Objective To study the relationship between single nucleotide polymorphisms(SNPs)of thrombospondin-1 gene(TSP-1)and coronary artery disease(CAD).Methods Fragment 8831A→G of Exon thirteen in TSP-1 gene from 437 cases of CAD and 423 subjects without coronary heart disease from November 2003 to May 2007 in The First Affiliated Hospital of Nanjing Medical University and Affiliated Yueyang Hospital of Shanghai Traditional Chinese Medicine University were analysed by polymerase chain reaction and restriction fragment length polymorphism(PCR-RFLP),and sequence analysis for confirmation.Results The prevalence of the 8831G in the 860 subjects was rare.No association of the N700S polymorphism with an altered risk of ACS was found in our study (GA VS AA:OR=1.699;95%CI 0.309-9.348,P＞0.05).Conclusion The TSP-1 8831A→G polymorphism is rare and unrelated to CAD in the Chinese Han population.

Abstract: Objective　To investigate the antimicrobial activity of omadacycline (OMC) against clinical Streptococcus agalactiae (GBS) isolates, as well as its relationship with biofilm formation, resistance genes and virulence genes. Methods　A total of 136 strains of Streptococcus agalactiae isolated from Shenzhen Nanshan People's Hospital between 2015 to 2020. The minimum inhibitory concentration (MIC) of OMC against Streptococcus agalactiae was determined by broth microdilution. Crystal violet staining was used to detect the biofilm formation ability of GBS. Resistance genes (tetM, tetO, tetK, ermB, OptrA) and virulence genes (cpsⅢ, bca, fbsA, cpsA, scpB) were investigated by polymerase chain reaction (PCR). Results　Among the 136 clinical isolates of GBS, 20 strains (14.7%) were resistant to OMC, 64 (47.1%) were intermediate, and 52 (38.2%) were sensitive. Fifty-seven strains (41.9%) were biofilm-positive, 20 of which (35.1%) were sensitive to OMC. Seventy-nine strains (58.1%) were biofilm-negative, 32 of which (40.5%) were susceptible to OMC. There was a statistically significant difference in the sensitivity rates between the two groups of strains (χ2=63.062, P<0.001), but there was no significant difference in the sensitivity of OMC among the biofilm-positive strains (Fisher's exact test, P=0.824). The resistance rates of tetM, tetO, ermB and OptrA positive strains were higher than those of negative strains, while tetK was opposite. The presence of tetM (Z=0.815, P=0.415), tetO (Z=0.151, P=0.88), tetK (Z=0.567, P=0.571), ermB (Z=1.198, P=0.231) resistance genes in Streptococcus agalactiae had no significant impact on the sensitivity of OMC. However, the presence of the OptrA resistance gene showed a statistically significant effect on the sensitivity of OMC (Z=2.913, P=0.004). The virulence factors cpsⅢ, bca, fbsA, cpsA and scpB were all detected at a rate higher than 50%. The presence of the virulence genes cpsⅢ (Z=0.222, P=0.824), bca (Z=0.141, P=0.888), fbsA (Z=0.813, P=0.416), and cpsA (Z=1.615, P=0.106) in Streptococcus agalactiae had no significant impact on the sensitivity of OMC. However, there was a significant inter-group difference in the scpB virulence gene (Z=2.844, P=0.004), but the rank mean values and resistance rates of scpB-positive strains were lower than those of the negative strains. Conclusions　The formation of biofilm in Streptococcus agalactiae reduces its sensitivity to OMC, but there was no significant difference in the sensitivity to OMC among the biofilm-positive strains. The presence of resistance genes tetM, tetO, tetK, ermB, and virulence genes cpsⅢ, bca, fbsA, cpsA, scpB in Streptococcus agalactiae is not associated with OMC resistance, but the presence of the resistance gene OptrA is correlated with OMC resistance..