2.Effect on invasion ability of cervical cancer cells after silence heparanase gene expression in Hela cells
Qiongying Lü ; Wei ZHANG ; Jing CHENG ; Wenting ZHANG ; Yajuan ZHONG
Chinese Journal of Obstetrics and Gynecology 2013;48(7):532-537
Objective Design and synthesize short hairpin RNA (shRNA) expression vector of RNA for specific silencing of heparanase (HPA) gene,screened plasmid which silence effects is the best.Observe the function of ceil invasion after inhibiting the expression of HPA in cervical carcinoma cell lines (HeLa).Methods The genomic sequence of HPA gene was retrieved from GenBank database.Designed four pairs of specific oligonucleotide sequences and a negative control according to the shRNA design principles.They were inserted into the vector pYr-1.1,vectors,and transfected into HeLa cells via lipofectamine.Reverse transcription (RT)-PCR and immunofluorescence were employed to detect the expression of HPA gene in the transfected cells at the mRNA and protein levels,respectively.The plasmid were screened and transfected into HeLa cells,then transwell small room stromal invasion experiment were employed to observe the cervical carcinoma cell invasion.Results RT-PCR results of transfected HeLa cells shown that the mRNA amplification multiples were 0.54 ±0.05 in the HPA-592 group,0.89 ±0.18 in HPA-995 group,0.82 ±0.22 in the HPA-1351 group,0.91 ±0.47 in HPA-1658 group.While,they were 1.31 ±0.72 and 1.09 ±0.16 in negative control and blank control group,respectively.Green fluorescence was visible in the cytoplasm,which indicated that the HPA protein was expressed in the cytoplasm,of them the weakest green fluorescence in the HPA-592 group.The relative numbers of invasive cells among the HeLa cells were as follows:182 ±6 in the blank control group,258 ± 17 in the negative control group,and 44 ± 4 in the HPA-592-specific interference group(P < 0.01).Conclusion Successfully screened shRNA vector targeting human HPA,efficiently inhibit expression of HPA gene when transfected into HeLa cells,and significantly reduced the invasion capacity of cervical carcinoma cells.
3.Impact of quercetin on the expression of heparanase in cervical cancer cells
Wenting ZHANG ; Wei ZHANG ; Yajuan ZHONG ; Qiongying Lü ; Jing CHENG
Chinese Journal of Obstetrics and Gynecology 2013;(3):198-203
Objective To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA,and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells.Methods The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20,40 and 80 μmol/L) in the culture medium.The control groups included a negative control group,which was cultured with RPMI 1640 only,and a positive control group,in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression.The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24,48 and 72 hours after treatment.Results (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski),and it exists both nucleus and cytoplasm.(2)The real-time PCR shows as follows:as the quercetin concentration increased (20,40 and 80 μmol/L),the mRNA expression level of HPA decreased (P <0.01),in which the inhibition of HPA expression was concentration dependent.In addition,the inhibition of HPA expression was also time dependent.As time growth,the expression level of HPA mRNA (24,48 and 72 hours) in HeLa and Caski cells decreased (P < 0.01).Compared with negative control group,the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P < 0.05) ; Compared with positive control group,the expression level of HPA mRNA expressed no obvious difference in quercetin (80 μmol/L) group (P > 0.05) in HeLa cells,while it was opposite in Caski cells(P <0.01).(3)The result of western blot shown that,as the quercetin concentration increased(20,40 and 80 μmol/L)and time growth (24,48 and 72 hours),the expression level of HPA protein decreased (P < 0.01),and the inhibition of HPA protein expression was concentration and time dependent.Compared with negative control group,the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P < 0.05) ;Conpared with positive control group,the expression level of HPA protein expressed no obvious difference in quercetin (80 μmol/L) group (all P > 0.05) in both HeLa cells and Caski cells (all P>0.05).Conclusion Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines,which inhibition is concentration and time dependent.
4.Influential factors of delayed therapy in dementia praecox patients with various clinical backgrounds
Zhaoxi ZHONG ; Luxian Lü ; Xueqin SONG ; Meizhi JIA
Chinese Journal of Tissue Engineering Research 2007;11(52):10708-10709,10716
BACKGROUND:Delayed therapy widely occurs in patients with dementia praecox;therefore,it brings a series of difficulties for clinical treatment and rehabilitation.OBJECTIVE:To analyze the influential factors of delayed therapy in patients with dementia praecox.DESIGN:Cross-sectional study based on patients with dementia praecox.SETTING:The Second Affiliated Hospital of Xinxiang Medical College.PARTICIPANTS:A total of 96 patients with dementia praecox,including 52 males and 44 females,were selected from the Second Affiliated Hospital of Xinxiang Medical College from January to June 2005.Their ages ranged from 16 to 55 years and the mean age was (32.7±12.3) years.Among them,54 patients had middle-school education and 42 patients had high-school education.METHODS:A domestic inventory was used to carefully record basically clinical data of each patient.and the inventory included sex,marriage status,educational level,attack styles,home address,home environment,economic status,family history,etc.All data were classified and analyzed in details.In addition,condition of delayed therapy,which determined as the duration over 1 year from onset of psychiatric symptoms to accepted treatment,in each classification was surveyed gradually.MAIN OUTCOME MEASURES:Delayed therapeutic rate and occurent rate of related factors.RESULTS:Among 96 patients,60 patients had delayed therapy,and the delayed therapeutic rate was 62.5%.Influential factors of delayed therapy:Logistic regression analysis demonstrated that regression equation was involved in educational level,family history,attack style and economic status.Meanwhile,the standard regression coefficient was 0.332 1,0.210 1,0.190 3 and 0.101 2. CONCLUSION:Educationallevel,family history,attack style and economic status of patients with dementia praecox are risk factors of delayed therapy. It is of importance for strengthening these factors to interfere and reduce delayed therapeutic rate at an early phase.
5.The clinical characteristics of intra-acinar pulmonary artery inflammation and its effect on clinical parameters in smokers with normal lung function and patients with chronic obstructive pulmonary disease
Qifang LAO ; Xiaoning ZHONG ; Zhiyi HE ; Guangnan LIU ; Zili Lü ; Peng WAN
Chinese Journal of Internal Medicine 2011;50(10):839-844
ObjectiveTo study the pathological characteristics of intra-acinar pulmonary artery inflammation and its correlation with smoking index and disease progression in smokers with normal lung function and smokers with chronic obstructive pulmonary disease (COPD).MethodsPatients requiting lung resection for peripheral lung cancer were divided into group A (nonsmokers with normal lung function,n = 10), group B (smokers with normal lung function, n = 13), and group C (smokers with stable COPD,n = 10).The lung tissue far away from rumor were resected to compare the pathological changes of intraacinar pulmonary arteries and infiltration level of inflammatory cell in pulmonary non-muscularized arteries (NMA), pulmonary partially muscularized arteries (PMA) and muscularized arteries (MA) among the three groups.The correlation analysis was made among infiltration level, smoking index, percentage of predicted value of forced expiratory volume in one second (FEV,% Pred), six-minute-walk distance (6MWD) and BODE index.Results (1) Both group B and group C showed the intima and media thickness of MA was significantly higher, the lumen area of MA was narrower and the proportion of MA was higher, and collagenous fiber of MA adventitial proliferated and area increased in group C(P <0.05 or P <0.01).(2) In group B and group C, the percentage of the intra-acinar pulmonary arteries that contained leukocytes, T lymphocytes, CD8+ T lymphocytes and the number of these positive cells infiltrating the intraacinar pulmonary arteries were increased, especially an increased number of CD8+ T lymphocytes infiltrating in the arterial adventitia as compared with group A, moreover there were significant difference between group C and group B (P < 0.05 or P < 0.01).In group B and group C, the degree of these positive cellsinfiltrating NMA, PMA and MA presented a decreasing sequence (P < 0.05 or P < 0.01).Among the intima, media and adventitia of MA, the infiltration of these positive cells was the highest in the adventitia.Among group A, group B and group C, infiltration degree of CD4+ T lymphocyte, B lymphocyte, macrophage and neutrophil demonstrated no significant difference, also among NMA, PMA and MA (P > 0.05).(3)The number of leukocytes, T lymphocytes, CD8+T lymphocytes infiltrating MA showed a positive correlation with the thickness of MA (r =0.563,0.627,0.589 ,P <0.01 ,respectively) and smoking index (r =0.551,0.665, 0.600, P < 0.01, respectively), moreover the degree of these cells infiltrating MA demonstrated negative correlation with FEV1 % Pred (r = - 0.763, - 0.703, - 0.767, P < 0.01, respectively).Also infiltrating degree of T lymphocytes and CD8+ T lymphocytes was positively correlated with BODE(r = 0.390,0.476,P < 0.05, respectively). Furthermore the infiltrating degree of CD8+ T lymphocytes had negative correlation with 6MWD (r = - 0.356, P < 0.05).Conclusions(1) Pulmonary arterial inflammation appears in smokers with normal lung function and smokers with COPD patients.It involves in all types of intra-acinar pulmonary arteries especially NMA and infiltrates whole layer of MA with a characteristic of CDs+T lymphocytes infiltrating in the adventitia of intra-acinar pulmonary arteries. (2)Pulmonary inflammation is closely correlated to cigarette smoking and clinical parameters such as BODE index, FEV1%pred and 6MWD.It is one of the key factors affecting the progression of COPD.
6.The clinical study of chemically-modified chitosan anti-adhesion film in the prevention of postoperative intra-abdominal adhesion
Li REN ; Ye WEI ; Yunshi ZHONG ; Shixu Lü ; Dexiang ZHU ; Jianmin XU
Chinese Journal of General Surgery 2011;26(5):414-416
Objective To evaluate a chemically-modified chitosan anti-adhesion film in the prevention of intestinal adhesion after abdominal surgery.Method In this study 240 patients at the Department of Surgery, Fudan University Zhongshan Hospital undergoing abdominal surgery from Jan 2006 to Dec 2006 were randomly divided into two groups.In the research group, chemically-modified chitosan antiadhesion film was put both at the area of operation and under the incision before closing the abdomen.The recovery procedures were recorded including the recovery of gut movement, the degree and the lasting time of abdominal pain, complication after surgery, the abdominal pain and ileus within 1 year.Result Postoperative incision pain was less significant in research group.The gut function recovered quicker and dietary began earlier.The ratio of early ileus after the surgery decreased significantly.The abdominal adhesion symptom in 1 year after surgery ameliorated significantly.There was no significant difference in other postoperative complications in the two groups.Conclusion The use of chemicallymodified chitosan anti-adhesion film helps to prevent the intestinal adhesion after the abdominal surgery.
7.Application of MR spectroscopy in differential diagnosis between basicranial tumor recurrence and radiation encephalopathy after radiotherapy for nasopharyngeal carcinoma
Yanchun Lü ; Weijun FAN ; Xian LI ; Chuanmiao XIE ; Jingxian SHEN ; Haoqiang HE ; Rui ZHONG
Chinese Journal of Radiology 2008;42(7):714-719
Objective To evaluate the value of MR spectroscopy (MRS) in the differential diagnosis between recurrence and radiation encephalopathy after radiotherapy for nasopharyngeal carcinoma (NPC). Methods Muhi-voxel proton MRS was performed on 50 patients with NPC, who were suspected of intracalvarium tumor recurrence or radiation encephalopathy after radiotherapy by conventional MRI,including 44 males and 6 females. Among the 50 patients, 26 cases were finally diagnosised as basicranial tumor recurrence and 24 cases as radiation encephalopathy by clinical and MRI follow-up. The following metabolites, such as Cho, NAA, Cr, lactate and lipid, were analyzed comparatively between basicranial tumor recurrence and radiation encephalopathy(RE), and between the lesions and the relative normal brain tissue. Wilcoxon's rank sum test was used to analyze the data. Results The median of Cho/Cr, Cho/NAA,LI/Cr in tumor recurrence group were 2. 22, 2. 13, and 1.77, respectively, and 1.40, 1.31, and 0. 57,respectively, in RE group. The difference of Cho/Cr, Cho/NAA, and LL/Cr between the two groups were statistically significant (P < 0. 01). Those in tumor recurrence group were higher than in RE group. The median of Cho, Cr, NAA in tumor recurrence group and in RE group were 3366. 00, 1023.00, 1930. 00 and 2469.50, 1864.50, 1734.00. There were no significant difference of Cho, Cr, and NAA between the two groups (P > 0. 05). In the 14 cases whose normal brain tissue were compared with the recurrent tumor tissue in tumor recurrence group, the median of Cr, NAA, LL, Cho/Cr, Cho/NAA, LL/Cr of recurrent tumor tissue and normal brain tissue were 1023.00, 1930.00, 2090.00, 3.76, 2. 13, 3.39 and 2370.00, 3012.00, 1680.00, 1.64, 1.17, 0.75,The difference of Cr, NAA, LL, Cho/Cr, Cho/NAA, LL/Cr between the normal tissue and recurrent tumor tissue were significant (P <0.05). LL, Cho/Cr, Cho/NAA, LL/Cr of recurrent tumors were higher than those of the normal brain tissue,while NAA and Cr of recurrent tumors were lower than those of the normal brain tissue. In the 12 cases whose normal brain tissue were compared with the RE tissue in RE group, the median of Cho, Cr, NAA, LL, Cho/Cr, IX,/Cr of RE tissue and normal brain tissue were 390.00, 217.50, 427.50, 39.00, 1.30, 0.40 and 680.00, 360.00, 610.00, 30.00, 1.54, 0. 09. The difference of above-mentioned parameters between RE tissue and normal tissue were significant. Cho, Cr, NAA, Cho/Cr of RE were lower than those of normal tissue (P <0. 05) ,while LL and LL/Cr of RE were higher than those of normal tissue (P < 0. 05). Conclusion The changes of the metabolites in recurrent lesions and RE lesions were different on MRS. Parameters such as Cho/Cr, Cho/NAA and LL/Cr, which were higher in recurrent lesions than those of RE, were valuable for the differential diagnosis between basicranial tumor recurrence and radiation encephalopathy after radiotherapy for NPC.
8.Efficacy and safety of GnRH-a combine with laparoscope conservative surgery in the treatment of the moderate or severe endometriosis
Yajuan ZHONG ; Wei ZHANG ; Wenting ZHANG ; Jing CHENG ; Qiongying Lü ; Kangkang ZENG
Chinese Journal of Obstetrics and Gynecology 2013;(3):180-182
Objective To study the efficacy and safety of Gonadotropin-releasing hormone agonists (GnRH-a) combined with laparoscope conservative surgery in treatment of moderate or severe endometriosis.Methods From Jan.2007 to Jan.2010,68 patients with moderate or severe undergoing treatment in Renmin Hospital of Wuhan University were enrolled in this retrospective study.Three groups were classified,which were 25 patients in GnRH-a group,subcutaneous injection Leuprorelin on the second day of menstruation,every 4 weeks for 3 months.Twenty-three patients in Marvelon group,orally one marvelon tablet on the second day of menstruation,continuous 21 days for one period of treatment for 3 courses.Twenty patients in surgery group,without any medicine used preoperatively.All patients were followed by 12 months and compare their surgery time,blood loss,recovery,visual analog scale (VAS),and recurrence and so on.Results The operating time were (68 ± 18) min in GnRH-a group,(80 ± 21) min in Marvelon group and (90± 24) min in surgery group.The amount of bleeding were (118 ± 15) ml in GnRh-a group,(161 ± 18) ml in Marvelon group and (193 ± 13) ml in surgery group.There was significant lower in the operating time and amount of bleeding in GnRH-a group than those in other two groups (P < 0.05).The activity time and the anus exhaust time were shorter in patients in GnRh-a group than those in the other two groups significantly(P < 0.05).When followed up in 12 months after treatment,visual analogue scale had dropped from 3.8 (1.9-6.8) to 1.9 (1.1-2.8) in GnRh-a group,from 2.7 (1.3-5.5) to 1.8 (1.2-3.2) in Marvelon group and from 1.9(1.0-4.9) to 1.6(1.0-3.6) in surgery group.It was showed the most remarkable decreased VAS in GnRHa group when compared with the other two groups(P < 0.05).The recurrence rates were 12% (3/25) in GnRH-a group,22% (5/23) in Marvelon group and 25 % (5/25) in surgery group.It was found that the most significant lower recurrence was in GnRH-a group when compared with the other two groups (P < 0.05).Conclusions It was safe and efficacy that GnRH-a combined with laparoscopic conservative surgery were used in treatment of endometriosis.It could bring shorter operation time,less intraoperative blood loss,quick postoperative recover,the lower recurrence rate.
9.Imaging sectional anatomy study on the optic canal by 64-slice spiral CT
Pengfei JIANG ; Ying SUN ; Yongbin Lü ; Yuanbin LI ; Xiuyu DAI ; Xiaoning ZHONG
Chinese Journal of General Practitioners 2011;10(9):668-669
This study was aimed to evaluate the application of64-slice spiral CT in imaging anatomy of the normal optic canal. The optic canals of 100 healthy subjects were examined by 64-slice spiral CT,and the imaging measurement of the anatomy structures were performed by the multi-planar reconstruction. The longest and the shorzest walls of the optic canal were the medial wall and the inferior wall, respectively.Results indicate that the imaging anatomic measurement of the optic canal can be accurately and easily performed by means of multi-planar reconstruction of 64-slice spiral CT.
10.Treatment of retinal detachment after laser in situ kera-tomileusis
Ling-Yan, CHEN ; Lin, LÜ ; Yong-Hao, LI ; Shi-Yi, LI ; Hui, ZHONG ; Wang, FANG
International Eye Science 2011;11(6):947-950
AIM: To study the characteristics of retinal detachment surgery after laser in situ keratomileusis (LASIK).METHODS: Eleven eyes of ten patients that experienced rhegmatogenous retinal detachment after LASIK procedure participated in the study. The characteristics of retinal detachment, management and complications after surgery were analyzed . RESULTS: Retinal detachment was characterized by the large percentage of multiple peripheral holes (73%) and giant tears (27%). All eyes underwent sclera buckling, and three of them combined with pars plana vitrectomy (PPV) and silicone oil tamponade. Silicone oil was removed after 1 month. Retina was reattached successfully at the first retinal detachment surgery in all eyes except one that succeeded at the fourth time. One eye of LASIK flap dehiscence and one eye of corneal subepithelial opacity occurred after surgery.CONCLUSION: Patients after LASIK should be carefully examined under pupillary dilation during follow-up. Sclera buckling is necessary to most retinal detachment after LASIK, and corneal protection is important in the treatment.