2.Study on the inhibition effect of siRNA on herpes simplex virus type 2 ICP4 gene.
Ji-feng LIU ; Cui-ping GUAN ; Xu TANG ; Ai-e XU
Chinese Journal of Experimental and Clinical Virology 2010;24(3):199-201
OBJECTIVETo explore the inhibition effect of RNA interference on the ICP4 expression and DNA replication of herpes simplex virus type 2 (HSV2).
METHODSFour pairs of siRNA targeted to HSV2 ICP4 gene and negative control siRNA were synthetized by chemical method, named as siRNA-1, siRNA-2, siRNA-3, siRNA-4 and siRNA-N respecticely. HSV2 HG52 was used to attack Vero cell after transfection overnight. Vero cell and supernatant were collected at 1d, 2d, 3d, 4d and 5d after virus attacking. Flurogenic quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR)was used to detect the expression of HSV2 ICP4 mRNA, flurogenic quantitative polymerase chain reaction(FG-PCR) was used to detect the expression of HSV2 DNA and Western-Blot was used to detect the expression of HSV2 ICP4 protein.
RESULTSAll the four pairs of siRNA could significantly inhibit the expression of HSV2 ICP4 mRNA and protein, especially siRNA-2. The above siRNAs could significantly decrease HSV2 DNA copy number,too.
CONCLUSIONsiRNAs targeted to HSV2 ICP4 gene could significantly inhibit expression of HSV2 ICP4 mRNA and protein, and decrease HSV2 DNA copy number, suggesting that siRNA can inhibit HSV2 DNA replication through silencing ICP4 gene.
Gene Expression Regulation, Viral ; drug effects ; genetics ; Gene Silencing ; drug effects ; physiology ; Herpesvirus 2, Human ; drug effects ; genetics ; RNA Interference ; drug effects ; immunology ; RNA, Small Interfering ; pharmacology ; RNA, Viral ; drug effects ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction
3.Dynamic contrast-enhanced MRI in breast cancer treated with neoadjuvant chemotherapy:correlation with pathological response
Jie LI ; Xiao-Peng ZHANG ; Ai-Ping LU ; Tao OUYANG ; Kan CAO ; Ying-Shi SUN ; Lei TANG ;
Chinese Journal of Radiology 2000;0(11):-
Objective To investigate the relationship of pathological response of breast cancer after neoadjuvant chemotherapy with the imaging findings in dynamic contrast-enhanced MRI.Methods Forty- five patients with pathologically confirmed breast carcinoma who finished courses of neoadjuvant chemotherapy had breast MRI prior to operation.Dynamic contrast-enhanced MRI scans were performed on a 1.5 T scanner using 3D SPGR sequence before and repeated 6 times after administration of Gd-DTPA. Pathological response was assessed by a pathologist according to Miller & Payne five points classification blinded to breast MRI results.Grade 5 was defined as pCR(pathological complete response).Grade 4 and 5 were defined as major histopathological response(MHR).The type of time signal intensity curve(TIC) (three types),pattern of residual enhancement of each breast cancer were recorded and correlated with pathological findings.Fisher exact test was used for statistical analysis.Results Grade 5 responses were achieved in seven patients;grade 4 in sixteen patients;grade 3 in sixteen patients and grade 1—2 in six patients.70.0%(14/20)of type Ⅰ time signal intensity curve correlated with MHR,while all 6 type Ⅲ curves showed non-MHR response.The type of time signal intensity curve and pathological response grades had statistically significant correlation(P=0.001).18 of the 23 cases with MHR exhibited residual enhancement,while the remaining 5 cases showed no enhancement.Of the 18 MHR cases with residual enhancement,11 showed non-mass-like enhancement and 7 showed mass-like enhancement.The mass(non- mass)morphological pattern in dynamic contrast enhanced-MRI had statistically significant differences in pathological response(P=0.012).Conclusions Pathological response of breast carcinoma after neoadjuvant chemotherapy could be characterized using dynamic contrast-enhanced MRI by identifying patterns of residual contrast enhancement and kinetic curve.Favorable pathological responses correlated with Type Ⅰ TIC,non-enhancement,and non-mass-like residual enhancement.
4.Two methods of hepatic vascular exclusion for hepatectomy
Wei-Ping ZHOU ; Ai-Jun LI ; Si-Yuan FU ; Ze-Ya PAN ; Yuan YANG ; Liang TANG ; Meng-Ehao WU
Chinese Journal of General Surgery 2000;0(12):-
Objective To compare the effects of hepatic vein occlusion with tourniquet and Satinsky clamp in reseeting liver tumor involving the second hepatic portal.Methods From Jan 2003 to Jun 2006,180 patients underwent major liver resection with the selective hepatic vascular exclusion (SHVE).According to methods of hepatic vein occlusion,they were divided into two groups:Occlusion with tourniquet(tourniquet group,n=95)and occlusion with Satinsky clamp(Satinsky clamp group,n= 85).In tourniquet group,the hepatic veins were encircled and occluded with tourniquet,and in Satinsky clamp group,the hepatic veins were not encircled and clamped directly with Satinsky clamp.Data regarding the intraoperative and postoperative courses of the patients were analyzed.Results There was no difference between the two groups regarding the operating time,ischemia time,intraoperative blood loss and postoperative complications rate.The dissecting time of hepatic veins was significantly shorter in Satinsky group(6.2?2.4 min vs 18.3?6.2 min).lu the tourniquet group,five hepatic veins(one fight hepatic vein and four common trunk of left-middle hepatic veins)could not be dissected and encircled because of the tumors involving the cava hepatic junction.Another patient's common trunk of left-middle hepatic vein was inadvertently lacerated during the dissection.Hepatic veins in these 6 patients were occluded with Satinsky clamp successfully.Conclusion Occlusion with Satinsky clamping is safer and easier procedure than tourniquets in the resection of liver tumor involving the second porta hepatis.
5.Clinical observation of gefitinib in treatment of female patients with adenocarcinoma of lung WU
Xiao-Ping WU ; Ying-Zhi ZHUANG ; Hao JIANG ; You-Hua WU ; Wen-Xiang DAI ; Xiao-Hong AI ; San-Yuan TANG ;
Cancer Research and Clinic 2006;0(12):-
Objective To evaluate the efficacy and adverse effects of gefitinib in the treatment of fe- male patients with advanced adenocarcinoma of lung who had failed to previous chemotherapy.Methods These patients received 250mg of gefitinib orally,once daily until disease progression or development of intol- erable toxic reaction.They were evaluated one month after treatment and every other month thereafter.Results Among the 27 evaluable patients,there were 1 CR(3.7%),11 PR(40.8%),10 SD(37.0%)and 5 PD(18.5%). The overall response rate was 44.5%(95% CI 29%~68%);and 22 patients(81.5%)gained profit(CR+PR+ SD)from the clinical therapy(95% CI 62%~94%);the mean TTP was 7.2 months.Symptomatic improvement rate was 80.0%.The main adverse effects were mild rash and diarrhea.Conclusion gefitinib has significant efficacy in the treatment of female patients with advanced tung cancer who had failed to previous chemother- apy.Adverse effects are mild.gefitinib is a suitable therapy for these patients.
6.The treatment of relapsing primary nephrotic syndrome in children.
Ya-ping WANG ; Ai-min LIU ; Yu-wen DAI ; Cheng YANG ; Hong-feng TANG
Journal of Zhejiang University. Science. B 2005;6(7):682-685
OBJECTIVETo explore better therapy and reduce the rate of re-relapse of primary nephritic syndrome in children who had been treated with corticosteroids but relapsed.
METHODSEighty relapsers were enrolled from Jan. 1994 to Apr. 2000, who were randomly divided into two groups. The treatment group (n=39) had been treated with tripterysium glucosides for three months, with the control group (n=41) members were treated with cyclophosphmide (CTX) by intermission intravenous pulse, with total dose of CTX not being more than 150 mg/kg. Prednisone, meanwhile, was given to both groups. The total treatment period of prednisone was prolonged by 12-18 months.
RESULTSAfter following up for 3-7 years, the re-relapse rates of both groups were observed. The re-relapse rate of the treatment group was 28.2% to 29.3% in the CTX-controlled group. The re-relapse rates between two groups were almost similar, and with no observed significant difference (P>0.05). The side effect of tripterysium glucosides was less than that of CTX.
CONCLUSIONFor the treatment of relapsing nephritic syndrome in children, the combination of tripterysium glucosides and prolonged corticosteroid therapy is as effective as the regimen of CTX plus prolonged use of prednisone.
Anti-Inflammatory Agents ; administration & dosage ; Child, Preschool ; Cyclophosphamide ; administration & dosage ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glucosides ; administration & dosage ; Humans ; Immunosuppressive Agents ; administration & dosage ; Longitudinal Studies ; Male ; Nephrotic Syndrome ; diagnosis ; prevention & control ; Prednisone ; administration & dosage ; Secondary Prevention ; Treatment Outcome
7.Expression of vascular endothelial growth factors (VEGF)-A, -C and -D and their prognostic significance and relationship with angio- and lymphangiogenesis in gastric cancer.
Xiao-lei WANG ; Zi-sheng AI ; Jian-ping FANG ; Ru-yong TANG ; Xi-mei CHEN
Chinese Journal of Oncology 2008;30(11):837-843
OBJECTIVETo investigate the expressions of vascular endothelial growth factors (VEGF)-A, -C and -D and their prognostic significance and relation to angio- and lymphangiogenesis in gastric cancer.
METHODSThe expression of VEGF-A, -C and -D in 123 primary gastric cancers was detected by immunohistochemical staining. The lymphatic vessel density (LVD) and microvessel density (MVD) were assessed after immunohistochemical double-staining with D2-40 and CD34, respectively. The correlation between the expression of those VEGF factors and clinicopathological parameters were analyzed by univariate method. The overall survival was evaluated by Kaplan-Meier method and log-rank test. Multivariate analysis was carried out using Cox proportion hazard model.
RESULTSThe positive expression rate of VEGF-A, -C and -D in primary gastric cancer samples were 64.2%, 65.9% and 41.5%, respectively. High expression of VEGF-A, or -C or -D, or any two of them was correlated with high LVD (P < 0.05). High expression of both VEGF-A and -C was associated with high MVD, lymph node metastasis, LVI and MVI (P < 0.05). Both VEGF-C and -D high expression was correlated with LVI and lymph node metastasis (P < 0.05). The patients with high expression of these factors had a statistically shorter overall survival (P < 0.05). The patients with both VEGF-A and -C expression had the shortest survival (56 months). Multivariate analysis showed that VEGF-A high expression, MVD, lymph node metastasis and depth of tumor invasion were independent survival predictors (P = 0.033, 0.002, 0.019 and P < 0.001, respectively).
CONCLUSIONHigh expression of both VEGF-A and -C imply high potential of lymphangiogenesis, metastasis and poorer survival in gastric cancer patients. High expression of VEGF-C and -D may induce lymphangiogenesis and promote lymph node metastasis, but only VEGF-A is an independent predictor of survival.
Adult ; Aged ; Aged, 80 and over ; Female ; Follow-Up Studies ; Humans ; Lymphangiogenesis ; Lymphatic Metastasis ; Lymphatic Vessels ; pathology ; Male ; Microvessels ; pathology ; Middle Aged ; Neovascularization, Pathologic ; Proportional Hazards Models ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism
8.Study on the concordance of objective multi-parameters analysis and perceptual evaluation.
Ping YU ; Gang WANG ; Gui-juan ZHANG ; Ai-rong TANG ; Rong-guang WANG
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2012;47(10):817-822
OBJECTIVEThrough exploring the concordance of objective multi-parameters analysis and perceptual evaluation, to establish an objective multi-parameters evaluation protocol of voice disorder and to make the evaluation of voice objectification and quantification.
METHODSVoice samples from 271 patients (124 female and 147 male)with dysphonia and 69 control subjects with normal voice (37 female and 32 male)were recorded and assessed by a jury composed of 5 experts in phoniatrics from different hospitals. The jury was instructed to classify voice samples according to the G (grade) component of the GRBAS scale on a visual analogue scale secondarily transformed in a 4-point scale ranging from 0 for normal to 3 for severe dysphonia. The voice samples were unified sentences and ordered randomly 3 times, the mean of 3 evaluation scores were the final results. The objective parameters, including fundamental frequency (F0), jitter, shimmer, fundamental frequency standard deviation (F0SD), normalized noise energy (NNE), harmonic-to-noise ratio (HNR) and maximal phonatory time (MPT), were measured on a 2-second sustained vowel/a/including its initial segment, using the software Dr. Speech for Windows. The data were analyzed using SPSS11.0.
RESULTSAll objective parameters except for F0 had high correlation with G and the variance tendency of these parameters values was coherent with the extent of voice disorder. And there were statistical differences between adjacent voice disorder groups. Male and female objective multi-parameters protocols were established respectively consisting of jitter, shimmer, F0SD, NNE, HNR and MPT using discriminant analysis (P < 0.05). The concordance between perceptual evaluation and objective multi-parameters evaluation was 81.6% in male and 83.2% in female. The concordance of evaluation of normal voice and severe voice disorder groups were better than that of mild and moderate voice disorder groups. All mis-grading voices were judged in the adjacent voice group.
CONCLUSIONSThe objective parameters of voice are able to reflect the characteristic of its perceptual evaluation and the concordance between perceptual evaluation and objective multi-parameters evaluation is good. The objective multi-parameters evaluation protocol we established could provide an objective and quantitative evaluation method for voice disorders.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Discriminant Analysis ; Female ; Humans ; Male ; Middle Aged ; Voice Disorders ; Voice Quality ; Young Adult
9.Epidemiological investigation of human papillomavirus infection in men attending a sexually transmitted disease clinic in Hangzhou area.
Xu TANG ; Ai-E XU ; Xiao-Ping DONG ; Xiu-Kun SUN ; Hong SHEN ; Ji-Feng LIU
Biomedical and Environmental Sciences 2006;19(2):153-157
OBJECTIVETo investigate the epidemiological characteristics of human papillomavirus (HPV) infection in men attending a sexually transmitted diseases (STD) clinic in Hangzhou area.
METHODSMale subjects (n=375) aged 18-70 years, attending the STD clinic were recruited. Urethral swabs were assessed for HPV DNA using polymerase chain reaction (PCR) with the consensus primers MY09/11. HPV genotypes of positive PCR products were determined by restriction fragment length polymorphisms and direct sequence analysis.
RESULTSOf the 375 swabs collected, 305 (81.3%) yielded sufficient DNA for the subsequent HPV analysis. Among the 305 subjects, the prevalence of HPV was 13.8%. Nononcogenic HPV types were found in 8.5% (26/305) of subjects, oncogenic types in 4.3% (13/305), and multiple types in 1.0% (3/305). The prevalence of HPV infection was higher in subjects from urban area than in those from rural area (P < 0.05). The prevalence was also higher in those who received fewer years of education (P < 0.05) and those who had more sex partners (P < 0.05).
CONCLUSIONSHPV infection among men at high risk is not uncommon. The detection rate of HPV DNA is significantly related to some sociodemographic factors, such as residence, educational level and the number of sex partners.
Adolescent ; Adult ; Aged ; Ambulatory Care Facilities ; China ; epidemiology ; Humans ; Male ; Middle Aged ; Papillomaviridae ; classification ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; epidemiology ; virology ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Sexually Transmitted Diseases ; prevention & control
10.Stability analysis of reference gene based on real-time PCR in Artemisia annua under cadmium treatment.
Liang-Yun ZHOU ; Ge MO ; Sheng WANG ; Jin-Fu TANG ; Hong YUE ; Lu-Qi HUANG ; Ai-Juan SHAO ; Lan-Ping GUO
China Journal of Chinese Materia Medica 2014;39(5):777-784
In this study, Actin, 18S rRNA, PAL, GAPDH and CPR of Artemisia annua were selected as candidate reference genes, and their gene-specific primers for real-time PCR were designed, then geNorm, NormFinder, BestKeeper, Delta CT and RefFinder were used to evaluate their expression stability in the leaves of A. annua under treatment of different concentrations of Cd, with the purpose of finding a reliable reference gene to ensure the reliability of gene-expression analysis. The results showed that there were some significant differences among the candidate reference genes under different treatments and the order of expression stability of candidate reference gene was Actin > 18S rRNA > PAL > GAPDH > CPR. These results suggested that Actin, 18S rRNA and PAL could be used as ideal reference genes of gene expression analysis in A. annua and multiple internal control genes were adopted for results calibration. In addition, differences in expression stability of candidate reference genes in the leaves of A. annua under the same concentrations of Cd were observed, which suggested that the screening of candidate reference genes was needed even under the same treatment. To our best knowledge, this study for the first time provided the ideal reference genes under Cd treatment in the leaves of A. annua and offered reference for the gene expression analysis of A. annua under other conditions.
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