1.Relationship between HLA-DRB1 alleles and chronic urticaria with positive autologous serum skin test
Yi WU ; Jinqiu MENG ; Yulin YAN ; Cunwei CAO ; Donghua LIU ; Ling LIANG
Chinese Journal of Dermatology 2012;45(2):78-80
ObjectiveTo investigate the relationship between HLA-DRB1 alleles and chronic urticaria with positive autologous serum skin test (ASST) in Guangxi Zhuang Autonomous Region.MethodsASST was conducted in 144 patients with chronic urticaria,who were subsequently divided into two groups according to the test result:positive group (n =62) and negative group (n =82).PCR amplification with sequence-specific primers was used to determine the genotypes of HLA-DRB1 alleles in the patients and 199 normal human controls.Chi-square test was performed to analyse the difference in the frequency of HLA-DRB1 alleles between the 3 groups by using the SPSS 13.0 statistical software package.ResultsThere were significant differences in the frequency of HLA-DRB1*01,*1401 and *16 alleles among the patients with positive and negative ASST and the controls (x2 =10.92,Pc =0.032;x2 =35.34,Pc < 0.01 ;x2 =12.69,Pc =0.032).Paired comparison revealed significant differences in the frequency of HLA-DRB1*1401 allele between the patients with positive ASST and controls(RR =17.09,Pc < 0.01 ) as well as between the patients with positive and negative ASST (RR =7.20,Pc < 0.01).ConclusionHLA-DRB1*1401 allele may be,or be linked to,the predisposing gene of chronic urticaria with positive ASST in Guangxi Zhuang Autonomous Region.
2.Down-regulation of Twist1 increases the sensitivity of nasopharyngeal carcinoma cell lines HNE1 to taxol.
Da-wei MENG ; Ji-min BAO ; Yun-peng MA ; Zhe LI ; Su-jie LI ; Dan LI
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2009;44(6):504-508
OBJECTIVETo investigate whether down-regulation of Twist1 could change sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol.
METHODSHNE1 cells were transfected with the small interfering RNA (siRNA) expression vector pSuppressor-Retro-Si-Twist, containing the short hairpin RNA (shRNA) sequence targeting the Twist gene-coding region by Fugene 6. Positive clones were then selected in Neomycin (400 microg/ml) for 21 days. The low expressions of Twist1 were examined by real-time reverse-transcription polymerase chain reaction (RT-PCR) and Western blot. Drug sensitivity of si-Twist1 HNE1 to taxol was determined by Annexin V-fluorescein isothiocyanate( FITC)/propidium lodide (PI) double-labeled flow cytometry and detection of DNA ladder. The Effect of Twist1 inactivation on HNE1 cell proliferation was observed by MTT assay and flow cytometry.
RESULTSAnnexin V- FITC-PI assay showed that apoptosis ratio was 40.2% in si-Twist HNE1 after treated with 10 ng/ml taxol, significantly higher than that in the control siRNA group 24.3%. The deference had statistic meaning. After the re-expression of HNE1, apoptosis ratio was 44.80% +/- 4.80% (x +/- s) in low Twist1 protein expression group and that was 27.00% +/- 2.91% in high expression group. The deference had statistic meaning (t = 4.374, P = 0.049). Real time PCR test revealed apoptosis protein bcl-2 expression in si-Twist HNE1 was 0.28 +/- 0.05, significantly lower than that in the control siRNA HNE1 (0.57 +/- 0.08, t = 6.710, P = 0.021), nevertheless, significant bax and bcl-XL changes were not observed (t = 2.000, P = 0.184 and t = 1.502, P = 0.272). MTT and FCM showed that down-regulation of Twist1 did not alter cell proliferation rate (P>0.05).
CONCLUSIONSDown-regulation of Twist1 could increase drug sensitivity of nasopharyngeal carcinoma cell line HNE1 to taxol by inducing apoptosis. These results suggested that Twist1 may be a promising treatment target for nasopharyngeal carcinoma therapy.
Apoptosis ; drug effects ; Cell Line, Tumor ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Nasopharyngeal Neoplasms ; drug therapy ; Nuclear Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA Interference ; RNA, Small Interfering ; Twist-Related Protein 1 ; genetics
3.Dynamic observation and antibiotic susceptibility analysis of biofilm of Staphylococcus aureus isolated from clinical specimens
Jinqiu QIN ; Meng LI ; Yuanyuan QIN ; Faquan LIN
Chinese Journal of Laboratory Medicine 2019;42(2):140-145
Objective To analyze the correlation between antibiotic susceptibility andbiofilm formation of Staphylococcus aureus. Methods According to Standardized Operational Guidance for Clinical Microbiological Testing, fifty-eight non-repetitive pathogenic Staphylococcus aureus isolates were collected from blood, urine, sputum and purulent secretions of inpatients in the Laboratory Department of the First Affiliated Hospital of Guangxi Medical University in January 2018. The antibiotics susceptibility of strains was characterized by disk diffusion method and VITEK-2.96-hole culture. The biofilm formation ability was detected bythe crystal violet assay and Congo redplate methods.The in vitro dynamic forming process of S.aureus′sbiofilm was analyzed by crystal violet staining combined with bacterial culture in 96 wells.Besides, the rate differences of drug resistance between biofilm producers and non-producers was performed by Chi-square test. The diameter of bacteriostasis zone of biofilm producers and non-producers was compared by t test. The drug resistance among strains with different biofilm formation ability was analyzed withnonparametric rank sum test. Results The positive ratio of biofilm producing S. aureus was higher in MRSA (68.42%) than in MSSA(20.00%)(χ2=12.304,P=0.001. Antibiotic resistance rates of biofilm producers were higher than non-producers.The resistance rates of biofilm positive strains to oxacillin and clindamycin were 73.33% and 53.33%, respectively. The antibiotic resistance of the strain was higher along with the biofilm forming ability was increasing. (χ2=9.099, P=0.008). Depths of the S. aureus′s biofilm on the 96-well plates increased significantly over time and reached biofilm maturation after 72 hours′ incubation. Light microscopic observation revealed that the mature biofilm was compact and growing with many layers. Conclusions For the clinical isolates of S. aureus, the antibiotic resistance of biofilm producers is significantly higher than non-producers. Strains with stronger biofilm forming ability had higher antibiotics resistance. Depths of the S. aureus′s biofilm on the 96-well plates increased significantly over time and reached biofilm maturation after 72 hours′incubation.
4.Metamorphopsia change and influence factors before and after idiopathic epiretinal membrane surgery
Xida LIANG ; Yi WANG ; Limei LIU ; Meng GAO ; Yanping YU ; Zengyi WANG ; Jinqiu CHEN ; Xinxin LIU ; Wu LIU
Chinese Journal of Experimental Ophthalmology 2019;37(1):21-28
Objective To investigate the changes of metamorphopsia between before and after surgery in the patients with idiopathic epiretinal membrane and its influence factors.Methods A series cases observitional study included 39 eyes of 39 patients with idiopathic epiretinal membrane.Follow-up was carried out at 1 week before surgery and 3,6 months after surgery respectively.M-chart was used to quantify the severity of metamorphopsia (M-score).EDTRS visual chart was used to quantify best corrected visual acuity (BCVA) (converted to LogMAR).Central subfield thickness (CST),central foveal volumn (CV),cube average thickness (CAT),central foveal thichness (CFT),ganglion cell layer (GCL) thickness,inner nuclear layer (INL) thickness,outer nuclear layer (ONL) and outer plexiform layer (OPL) thickness,the integrity of external limiting membrane,ellipsoid zone and interdigitation zone were analyzed by using spectral domain-optical coherence tomography (OCT).This study protocol was approved by Ethic Committee of Beijing Tongren Hospital (No.TRECKY-012).Written informed consent was obtained from each subject before surgery.Results Mean M-score was significantly decreased from 0.8 (0.3,1.1) before surgery to 0.5 (0.2,0.8) at 3 months after surgery,with a significant difference between the two time points (Z=-2.013,P=0.044).Mean M-score was 0.6(0.2,0.8) at 6 months after surgery,which was not significantly different in comparison with before surgery and 3 months after surgery (Z =-1.873,P =0.061;Z =-0.288,P =0.773).Compared with before surgery,the horizontal M-score was significantly decreased 3 months and 6 months after surgery (Z =-2.329,P =0.020;Z =-2.858,P =0.004).No significant difference was found in vertical M-score among before surgery and 3,6 months after surgery (all at P>0.05).The BCVA was improved from 0.40 (0.30,0.66) before surgery to 0.20 (0.06,0.42) 3 months after surgery and declined to 0.30 (0.10,0.52) at 6 months after surgery,and significant differences were obtained between 3 months after surgery and before surgery or 6 months after surgery (Z =-4.087,P<0.001;Z =-2.235,P =0.025).Compared with before surgery,the BCVA in cataract combined with vitrectomy operative group was significantly improved in 3 months and 6 months after surgery (Z=-2.613,P=0.009;Z=-2.466,P=0.014) and the BCVA in only vitrectomy group was significantly improved at 3 months after surgery but decreased 6 months after surgery,showing significant differences between 3 months after surgery and before surgery or 6 months after surgery (Z =-3.104,P =0.002;Z =-3.464,P =0.001).Preoperative M-score was positively correlated with preoperative BCVA,preoperative CST or preoperative CFT (rs =0.384,P =0.016;rs =0.585,P<0.001;rs =0.601,P<0.001).No correlation was found between BCVA with GCL,INL or ONL + OPL thickness.Horizontal M-score was positively correlated with CST,postoperative CV and postoperative CAT (rs=0.322,P=0.045;rs=0.340,P=0.034;rs =0.336,P=0.036),and no correlation was found between horizontal M-score and BCVA,CFT,GCL thickness,INL thickness,ONL+OPL thickness in 6 months after surgery.The vertical M-score and mean M-score were not correlated with OCT parameters in 6 months after surgery.The mean M-score was positively correlated with preoperative mean M-score,preoperative CST,preoperative CV,preoperative CAT in 6 months after surgery (rs =0.589,P<0.001;rs =0.330,P =0.040;rs =0.404,P =0.011;rs =0.410,P =0.009).In addition,and no significant correlation between mean M-score and preoperative BCVA,CFT,GCL thickness,INL thickness,ONL+OPL thickness.Multivariate stepwise linear regression showed that preoperative M-score was a predictor of postoperative M-values (adjusted R2 =0.211,P =0.002).Conclusions Most metamorphopsia can be alleviated after idiopathic epiretinal membrane surgery.The residue metamorphopsia after surgery probably is correlated with preoperative metamorphopsia and CFT.