1.Specific Ribozyme Induced Apoptosis on Human Cervical Carcinoma Cell Line CaSKi
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To study the characterization of the cultured cervical cancer cell line transfected with anti- HPV16E6-ribozyme, and to investigate the effect of ribozyme on proliferation and apoptosis of cervical cancer cell. Metliods: Anti-HPV16E6-ribozyme had been designed to cleave the HPV16E6 gene. With the method of lipofectin transfec- tion, the anti-HPVI6E6-ribozyme and empty eucaryotic expressing plasmids were transfected into CaSKi cell, which named as CaSKi-R, CaSKi-P respectively. The amounts of E6 mRNA in the three kinds of cells were detected by northern blot. Cell cycle was detemined by flow cytometry, and cell apoptosis was examined by fluorescent (Hoechst) staining and TUNEL. The expression of some genes, including c-myc, bcl-2, p53, and fas, was also detected by flow cytometry analy- sis. Results: Northern blot showed that E6 mRNA was less in CaSKi-R than in CaSKi. In CaSKi-R cells, cycle was arres- ted in G1 phase, with decreasing in percentage of S phase cells. The apoptosis rate of CaSK1-R cell was much higher than those of CaSKi and CaSK1-P. Anti-HPV16E6-ribozyme could reduce the expression of E6, c-myc, bcl-2 genes on CaSKi- R cells, and increased the expression of p53. While this phenomenon was not found on the CaSK1-P cells. The expression of fas was similar in the three kinds of cells. Conclusion: Anti-HPVE6-rivozyme induces apoptosis of human cervical cancer CaSKi cells. The mechanisms may be the decrease of E6 gene's expression, and the succedent changing of some genes'expression.
2.Treatment and control status of elderly prostatic hyperplasia with metabolism syndrome
Chinese Journal of Geriatrics 2013;(2):175-177
Objective To investigate the clinical features,treatment,medication and abnormal metabolism control status in elderly benign prostatic hyperplasia (BPH) with metabolic syndrome (MS).Methods The 397 patients with BPH aged >60 years (294 with MS) were retrospectively analyzed.The clinical features,treatment and control status were analyzed.Results Body mass index,waist circumference (BMI),2 hPBG,systolic pressure,diastolic pressure,TG and uric acid,urinary albumin/creatinine ratio,urinary albumin were higher and HDL-C was lower in BPH with MS than without MS (P<0.05).Prostate volume,PSA level,IPSS score,quality of life score were higher in BPH patients with MS than without MS (P<0.05).Treatment modality analysis showed that lower urinary tract symptoms was mild,watchful waiting accounted for 22.3%,urinary retention or obstruction related operation treatment for 8.7%,moderate to severe symptoms related oral drugs treatments for 67%,the oral drug treatment was still the main mode of BPH.In the patients with abnormal levels of glucose metabolism,the target arrival rate (TAR) on FBG,2 hPBG and HbAlc were 49.6%,34.9% and 37.5%,TAR on all the three was 19.1%.In the hypertensive population,TAR on systolic and diastolic pressure were 44.1% and 82.5%,TAR on both two was 41.7%.TG,TC,LDL-C and HDL-C were 31.3%,35.4%,48.2% and 64.9%,on all the four was 29.9%.Conclusions The ratio of elderly BPH with MS is high,but the target arrival rate on control of urinary tract symptoms and abnormal metabolism component is not optimistic in spite of a positive drug administration.
3.Integrin alpha v regulates anchorage-independent growth and survival in squamous cell carcinoma cells
Yanfang ZHENG ; Hai LU ;
China Oncology 2001;0(05):-
Purpose:During the malignant transformation, most tuomr cells will become resistant to anoikis (the apoptosis induction upon loss of anchorage), thereby gaining an important growth advantage and providing the basis for tumor spread, growth at distant places, and formation of metastasis.The present study focus on the role of soluble fibronectin, integrin and their signal transduction in the process of tumor cells anoikis apoptosis.Methods:Flow cytometry detected integrin subunits' expression on the human squamous cell carcinoma cell (HSC 3) surface. Establish anoikis induction model. The role of different integrin subunits during anoikis induction were detected by antibody blocking method, p53 and Bcl 2/Bax expression were measured during this anoikis process.Results:There is no or low expression of integrin av on the normal epithelial cells, whilst it expresses relative higher on the surface of HSC 3 cells. The treatment of soluble fibronectin decreased p53 expression, increased the ratio of Bcl 2/Bax and then protected HSC 3 cell from anoikis. Anti integrin ?v blocking antibody resumed the anoikis that was inhibited by soluble fibronectin evidently. Conclusions:We now provide the evidence that the binding of integrin ?v and soluble fibronectin protected HSC 3 cells from anoikis apoptosis, via decreasing the expression of p53, increase the ratio of Bcl 2/Bax.
4.Effects of anti-HPV16E6-ribozyme on telomerase activity in cervical carcinoma cells and the related mechanisms
Zhiguo RAO ; Jiren ZHANG ; Yanfang ZHENG
Chinese Journal of Cancer Biotherapy 1996;0(04):-
Objective: To investigate the effects of HPV16E6-ribozyme on telomerase activity in cervical carcinoma cell line CaSKi and the related mechanisms. Methods: Anti-HPV16E6-ribozyme and blank eucaryotic plasmids were transfected into CaSKi cells via lipofectin, and the resultant cells were named as CaSKi-R and CaSKi-P, respectively. The expression of ribozyme in transfected cells was observed by RNA dot blotting. The expression of E6 mRNA and protein in the 3 kinds of cells were detected by Northern blotting and Western blotting, respectively. Telomerase activity was determined by TRAP-Elisa method; the expression of P53, c-myc, hTERT and hRT mRNA were examined by RT-PCR.Results: RNA dot blotting showed that anti-HPV16E6-ribozyme was stably expressed in transfected CaSKi-R cells. Western blotting showed that the expression of E6 mRNA and protein in CaSKi-R cells was obviously lower than that in CaSKi and CaSKi-P cells. The telomerase activities in CaSKi,CaSKi-P and CaSKi-R cells were (0.89?0.14), (0.90?0.11) and(0.36?0.06),respectively. The inhibitory rate of telomerase activity in CaSKi-R cells was 59.55%, which was significantly lower than those in CaSKi and CaSKi-P cells (P
5.Clinical investigation and drug resistance of fungal hospital-acquired infection in a certain hoapital
Yanfang ZHANG ; Lei LIU ; Xiufeng ZHENG
International Journal of Laboratory Medicine 2008;29(5):407-408,411
Objective To explore the distribution,fundamental diseases,risk factors and drug-resistance of hospital- acquired infection caused by fungi in Department of Respiratory Medicine.Methods The clinical data of 142 patients were retrospectively reviewed,and then with statistical analysis was performed.Results The main pathogen which caused fungal hospital-acquired infection was Candida albicans. The most three possible fundamental diseases were chronic obstructive pulmonary disease,bronchiectasis and lung cancer. The main risk factors were lower resistibility of patients and invasive diagnostic and therapeutic measure. The fungus resistance was gradually rising.Conclusion The hospital infection caused by fungus should be emphasized in respiratory medicine department. In order to effectively prevent the hospital infection in respiratory medicine department,we should improve the basic conditions of patients,rationally diagnose and treat,and rationally use the drug against fungi.
6.The Effect of Anti-HPV16 E6-Ribozyme on Malignant Phenotypes of Cervical Carcinoma Cell Line
Yanfang ZHENG ; Zhiguo RAO ; Jiren ZHANG ;
Chinese Journal of Cancer Biotherapy 1995;0(03):-
Objective: To investigate the characteristics of the cultured cervical cancer cell line transfected with anti HPV16E6 ribozyme, and to investigate the possibility and practicality of ribozyme in treatment of cervical cancer. Methods: The anti HPV16E6 ribozyme and empty eucaryotic expressing plasmids were transfected by lipofectin transfection into CaSKi cell, which named as CaSKi R, CaSKi P respectively. The morphology and the soft agar forming ability were studied. The expression of E6, PCNA and C erbB 2 genes was studied through Flow Cytometry. The tumorgenicity of each cell was detected by injecting cells into the nude mice skin. Three groups of nude mice were injected by CaSKi, CaSKi R and CaSKi P cell separately. Another group of mice was injected by CaSKi cell on right side and CaSKi R cell on left side. Results: There is no distinct difference of the morphology and growth rate between CaSKi and CaSKi P, but the growth rate of CaSKi R decreased. The soft agar forming rate of CaSKi P was similar with that of CaSKi cells, while that of CaSKi R was found decreased. The result of flow cytometric analysis showed that anti HPV16E6 ribozyme could reduce the expression of E6, PCNA and C erbB 2 genes on CaSKi R cells, while this phenomenon was not found on the CaSKi P cells. The tumorgenicity of CaSKi R in nude mice was decreased compared with CaSKi and CaSKi P cells. Conclusion: Anti HPVE6 rivozyme could partly reverse the malignant phenotypes of CaSKi cells. The reason may be the decrease of E6 gene expression, and the succeeding decrease of the PCNA and C erbB 2 genes′ expression.
7.Influence of ligustrazine on neuronal structure in brain of epileptic rats
Xiaoqin ZHU ; Yanfang ZHENG ; Xingsan TANG ; Shuisheng LEI
Chinese Journal of Tissue Engineering Research 2005;9(25):236-237
BACKGROUND: Ligustrazine inhibits discharge of cerebral hippocampal neuron, penetrates blood-brain barrier effectively after absorbed in the body and is distributed extensively in cerebral cortex, brain stem, striate body, hippocampus, cerebellum and midbrain.OBJECTIVE: To probe into the influence of ligustrazine and its different concentrations after abdominal injection on cerebral cortical neural cell structure in epileptic rats induced by penicillin.DESIGN: Randomized control experiment.SETTING: Physiological Department of Xianning Medical College.MATERIALS: The experiment was performed in Anatomy Department of Tongji Medical College of Huazhong University of Science and Technology from September 2004 to March 2005. Forty healthy SD rats of clean grade were employed, of either sex, mass weighted varied from 200 to 250 g.They were randomized into 5 groups, named operation control, penicillininduced epilepsy group and ligustrazine groups of 10 mg/kg, 20 mg/kg and 40 mg/kg, 8 rats in each group.METHODS: After anesthetized, the cranium was opened to expose cerebral cortical record region. BL-410 biofunctional experimental system was used to record brain electricity bilaterally and epileptic discharge of cerebral cortex in penicillin-induced epilepsy group and ligustrazine groups of 10 mg/kg,20 mg/kg and 40 mg/kg. In the control, 1 hour after anesthesia and craniotomy, cerebrum was collected. In penicillin-induced epilepsy group, 1 hour after induction, cerebrum was collected. In ligustrazine groups of 10 mg/kg,20 mg/kg and 40 mg/kg, after penicillin-induced epileptic discharge was stable, ligustrazine of 10 mg/kg, 20 mg/kg and 40 mg/kg was injected abdominally successively, and cerebrum was collected when the most remarkable inhibition was achieved. Brain tissue section was prepared separately, with HE staining, the observation was done under optic microscope.MAIN OUTCOME MEASURES: Structure changes in cerebral cortical neural cells in rats of each group.In the control, the morphological structure of cerebral cortical neural cell alternations on cerebral cortical neural cell structure, karyopykosis, plasmarrhexis and vacuolar structure, but there was no Nissel bodies in cytomarrhexis, vacuolar structure and decreased Nissel bodies in cytoplasm with the control, there were decreased vacuoles in neural cell, increased cytoplasm and few Nissel bodies in cytoplasm and cell structural morpholcontrol, karyon was big, round and light stained; clot-like Nissel bodies were visible and cell structural morphology was in tendency to be normal.CONCLUSION: In penicillin-induced epilepsy, morphological structure of cerebral cortical neural cell in rats is abnormal. Tetramethylpyrazine of various dosages may improve at different degrees morphological structure of cerebral cortical neural cell, especially significantly at high dosage, by which, its inhibition on epileptic discharge in rats is achieved.
8.The expression of CREB in the Glioma
Kebin ZHENG ; Jing XIE ; Yanmeng LI ; Shuai SHI ; Yanfang SHI
Chinese Journal of Nervous and Mental Diseases 2016;42(2):70-73
Objective To investigate the clinical significance and abnormal expression of the CREB in different grade gliomas. Methods The expression of CREB was examined by using immunohistochemistry in brain tissues from the brain injury (5 cases) and different grade gliomas (55 cases).The mRNA and protein levels of CREB were further as?sessed using Western blot and RT-PCR in brain tissues from the patients with brain injury (10 cases) and those with dif?ferent grade gliomas (30 cases). Results The positive rates of CREB immunohistochemistry were 2/5 in control, 10/15 inⅠ-,Ⅱ11/12 in Ⅲ, 28/28 in Ⅳ. The positive rates of CREB were significantly different among different groups (H=28.183,P<0.05).The mRNA levels of CREB were 1.00 ± 0.000 in control, 1.35 ± 0.068 inⅠ-Ⅱ, 2.88 ± 0.111 in Ⅲand 3.75 ± 0.196 in Ⅳ. The expression of CREB was higher in the glioma than in control group, and the mRNA levels of CREB were significantly different among different groups(F=1.208,P<0.05). The protein levels of CREB were 0.311 ± 0.014 in control, 0.469±0.026 inⅠ-Ⅱ, 0.641±0.028 inⅢand 0.896±0.024 inⅣ. The protein levels of CREB were sig?nificantly different among different groups(F=1.123,P<0.05). Conclusion The expression of CREB is elevated in glio?mas with different differentiation degrees. The expression of CREB was positively correlated with the degree of differentia?tion, indicating that CREB may have an important regulatory role in the progress of gliomas.
9.A Comparative Study of Auditory Memory in sentences between the Hearing-Impaired Children and Normal Children of 3~4 Years
Junlan CHEN ; Wei LUO ; Yanfang MA ; Jing ZHANG ; Wenfang ZHENG
Journal of Audiology and Speech Pathology 2017;25(1):24-27
Objective To investigate the differences of the correct memorization of a sentence with auditory between the hearing-impaired children after rehabilitation training and normal children.Methods Twenty-four hearing-impaired children and 14 normal hearing children at age 3~4 were selected.The hearing-impaired chil-dren were divided into two groups according to the length of the hearing device.The closed-set sentence test of e-valuation of auditory response to speech (EARS)was used.Results 1 .In the first test,the hearing-impaired chil-dren with hearing device 2 to 3 years whose correct rate was 57.14%were close to the normal children whose correct rate was 58.93%.The hearing-impaired children with hearing devices 1 to 2 years whose correct rate was 32.5%were worse than the children of other two groups 2 .In the second test,the hearing-impaired children with hearing devices 2 to 3 years whose correct rate was 44.64% were better than the hearing-impaired children with hearing devices 1 to 2 years whose the correct rate was 22.5%.The normal children whose the correct rate was 55.36%were better than the children of other two groups.3.There was no significant difference to correctly memorize a sen-tence between three groups in the first test,but there was significant difference to correctly memorize a sentence between the hearing-impaired with hearing device 1 to 2 years and the hearing-impaired children with hearing de-vice 2 to 3 years (P<0.05),and significant differences also existed between the hearing-impaired with hearing de-vice 1 to 2 years and normal children (P<0.05),but there was no significant difference between the hearing-im-paired children with hearing device 2 to 3 years and normal children (P>0.05).Conclusion The length with hear-ing device strengthens the ability of hearing-impaired children in correctly memorizing a sentence after rehabilitation training.And the content of test affects the result ,it provides reference to the hearing-impaired children's auditory training .
10.RAPD and SCAR Markers Analysis on the Indeterminate Growth Mutant of Tomato Derived by Space Mutagenesis
Yanfang WANG ; Shiheng WANG ; Yanhong ZHAO ; Jirong ZHENG ; Shuijin ZHU
Space Medicine & Medical Engineering 2006;0(05):-
Objective To evaluate the indeterminate growth mutant of tomato derived by space mutagenesis to provide the basis for selecting and cultivating the molecular markers of tomato growth habit. Methods Fifty 10-mer randomly amplified polymorphic DNA(RAPD) primers were used to examine the polymorphism of M1 and 10-3-2. Their polymorphic fragments were cloned, and then were transferred to SCAR markers. Results Of all the 50 10-mer RAPD primers, 44 primers amplified polymerase chain reaction(PCR) products and 2 primers (S165 and S168) amplified stable reproducible polymorphic products. The molecular weight of the specific amplified products were 300 bp and 1 500 bp respectively, therefore they were named as TRS165300 and TRS1681500 temporarily. And TRS1681500 was transferred into stable sequence characterized amplified region(SCAR) marker and this marker could be a specific genetic marker of this indeterminate growth habit mutant. Conclusion Space mutation can produce mutants at DNA level from the loaded materials. The indeterminate growth mutant of tomato is derived by space mutagenesis which can provide a valuable material for studying the regulation and control of tomato growth habit.