1.Effects of atorvastatin on apoptosis and cytochrome c expression in perihematoma tissue after intracerebral hemorrhage in rats
Yuehua WANG ; Guijun LIN ; Zhiguo RAO
International Journal of Cerebrovascular Diseases 2013;(3):191-196
Objective To investigate the effect of atorvastatin on cytochrome c (CytC) expression and neuronal apoptosis after intracerebral hemorrhage in rots.Methods A total of 108 male Sprague-Dawley rats were randomly allocated into 3 groups:sham operation group,saline control group,and atorvastatin group (n =36 each group).All the groups were redivided into 6 h,12 h,day 1,3,5 and 7 time points (n =6 at each time point).An intracerebral hemorrhage model was induced by using a modified two-step injection method.After modeling,atorvastatin was used for gavages (20 rng/kg,once a day) in the atorvastatin group.The saline control group was given the same volume of saline.Behavior evaluation was used for neurological score.TUNEL staining was used to detect apoptosis in perihematoma tissue.Immunohistochemical method was used to detect the CytC expression in perihematoma tissue.Results Behavior evaluation showed that the neurological scores decreased gradually with the passage of time in the atorvastatin group and the saline control group.There were no significant differences at 6 h,12 h,day 1 and day 3,but the neurological scores in the atorvastatin group were significantly lower than those in thc saline control group at day 5 (0.50 ± 0.55 vs.1.50 ± 0.55; t =3.162,P =0.010) and day 7 (1.00 ±0.63; t =2.712,P =0.022).TUNEL staining showed that the numbers of apoptotic cells increased first and then decreased in the saline control group and the atorvastatin group.They reached the peak at 1 hour after modeling.There were significant differences in the number of apoptotic cells in each group in perihematoma tissue at the same time point (all P =0.000),and the significance in the saline control group was more than that in the sham operation group and the atorvastatin group (all P <0.05),but at day 7,there was no significant difference in the number of apoptotic cells between the atorvastatin group and the sham operation group (12.69 ± 3.35 vs.9.33 ± 2.07; P =0.148).Immunohistochemical method showed that the numbers of CytC positive cells increased first and then decreased in the saline control group and the atorvastatin group,reached the peak at 12 h after modeling in te saline control group (68.19 ± 11.93) and at 1 d in the atorvastatin group (35.64 ± 9.12).There were significant differences in the numbers of CytC positive cells in perihematoma tissue at the same time point in each group (P =0.000).The numbers of CytC positive cells in the saline control group was significantly more than that in the sham operation group and the atorvastatin group (all P <0.05),but there was no significant difference in the numbers of CytC positive cells between the atorvastatin statin group and the sham group at day 7 (16.08 ± 3.80 vs.13.67 ± 2.94; P =0.349).Conelusions Atorvastatin may inhibit the release of CytC of nerve cells in perihematoma tissue after intracerebral hemorrhage,and thus reduce CytC-mediated apoptosis and neurological deficit after intracerebral hemorrhage.
2.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
3.A meta-analysis of compound kushen injection in the treatment for advanced hepatocellular carcinomas
Yuwei JIANG ; Bo YANG ; Zhiguo RAO
Journal of Chinese Physician 2015;17(6):803-806
Objective To investigate the efficacy and safety of compound kushen injection (CKI)in the treatment for patients with advanced liver cancers.Methods Relevantly randomized controlled trials from China National Knowledge Infrastructure (CNKI),Chinese Science and Technology Journal Database (VIP),Wanfang Data,and Pubmed were searched until November,2014.Randomized controlled trials (RCTs) of supportive care compared with combined therapy of CKI in the treatment for patients with advanced liver cancers were included.The methodological quality of RCTs was assessed independently with bias risk according to the Cochrane collaboration.All data were analyzed with the Review Manager 5.3.Results Eight RCTs involving 472 patients were included.The meta-analysis results suggested that the shortterm efficacy of the treatment group be higher (P =0.008) and that the pain relief be a significant benefit in the treatment group (P <0.01).Adverse reactions were not observed.Conclusions CKI is an effective and safe adjuvant drug for advanced liver cancers.
4.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.
5.Mechanism of anti-tumor effect of dihydroartemisinin
Wenmiao PENG ; Hongxing FU ; Lifang YU ; Zhiguo RAO
Journal of International Oncology 2017;44(6):448-451
Dihydroartemisinin (DHA),the major active metabolite of artemisinin,participates in tumor progression through the following ways:forming free radicals to induce cancer cells death dependent on iron,inducing apoptosis,inhibiting angiogenesis,tumor cells invasion and metastasis,modulating muhidrug resistance,controlling intracellular Ca2+ concentration,regulating cell cycles,cell autophagy and the immune system and so on.Generally,it is considered to be a potential anti-tumor drug.
6.Risk factors for nosocomial pneumonia in elderly stroke patients
Zhiguo RAO ; Zongming LIU ; Guijun LIN ; Hui GUO
Chinese Journal of Geriatrics 2008;27(7):518-520
Objective To investigate the risk factors for nosoeomial pneumonia in elderly stroke patients(aged 60 years and over). Methods The clinical data of 259 patients with nosoeomial pneumonia from Jan 2002 to June 2007 were collected and the risk factors were retrospectively analyzed. Results The morbility rate of nosocomial pneumonia in elderly stroke patients was 41.3%,and the risk factors were aging,Iong hospitalization,unconsciousness,type of stroke,and underlying diseases,smoking,tracheal intubation,tracheotomy,application of respiratory machine (all P>0.05). Conclusions The morbility rate of nosoeomial pneumonia in elderly stroke patients was high,and the risk factors are aging,long hospitalization,unconsciousness,type of stroke,and underlying diseases,smoking,tracheal intubation,tracheotomy,application of respiratory machine.
7.Research of ribomomal protein L23 in tumor progression
Wenmiao PENG ; Chuanrong QIN ; Zhimin ZHANG ; Meng HU ; Zhiguo RAO
Journal of International Oncology 2016;43(11):835-837
Ribosomal protein L23 is a new target for gene therapy of cancer.It participates in tumor progression by activating p53,inactivating murine double minute 2,regulating the carcinogenic activity of c-Myc,inducing the multi-drug resistance,and affecting the biologic behaviour of tumors.Generally,it′s con-sidered to be a potential prognostic factor in human cancers.
8.Bone marrow mesenchymal stem cell transplantation for the treatment of systemic lupus erythematosus
Zhiguo WANG ; Xueming LIU ; Shengquan TONG ; Zhequn SHI ; Li RAO
Chinese Journal of Tissue Engineering Research 2016;20(10):1461-1467
BACKGROUND:The non-specific immune suppression method is generaly used for treatment of systemic lupus erythematosus, but poor prognosis, such as infection and high recurrence rate, exists.
OBJECTIVE:To evaluate the therapeutic effect of bone marrow mesenchymal stem cel transplantation on systemic lupus erythematosus in mice.
METHODS:Sixteen mice with systemic lupus erythematosus were equivalently randomized into control and experimental groups, or then subjected to passage 3 bone marrow mesenchymal stem cel transplantation or the equal volume of normal saline via the tail vein, respectively. Mouse urine samples were colected to detect urine protein levels by Bradford method. Blood samples from the tip of the mouse tail were extracted to detect serum anti-ds-DNS antibody concentration by radioimmunoassay. Mouse kidney tissues were taken and observed pathohistologicaly through hematoxylin-eosin staining and immunohistochemistry staining under microscope. Flow cytometry was used to detect the expression of CD4+CD25+T cels in the inner canthus blood, fresh spleen and thymus.
RESULTS AND CONCLUSION:Within 10 weeks after cel transplantation, the urine protein levels in the two groups were gradualy increased, and the rising velocity was higher in the control group than in the experimental group. From the 4th to 10th week, the urine protein levels in the experimental group were significantly lower than those in the control group (P < 0.05). In the control group, lymphocyte infiltration was visible in the kidney tissues with a few of plasmocytes, and pathological findings showed the mice presented with interstitial nephritis; in the experimental group, the mice had no pathological changes in the kidney. In the two groups, immune complexes were found in the mesangial area, which showed a patch-like distribution in the control group and a punctate distribution in the experimental group; the relative proportion of the occupied area in the experimental group was significantly lower than that in the control group. The expression level of CD4+CD25+T cels in the blood and thymus were significantly higher in the experimental group than the control group (P < 0.05), and the expression level of CD4+CD25+T cels in the spleen was slightly higher in the experimental group than the control group with no significant difference (P > 0.05). The serum anti-ds-DNA antibody concentration in the experimental group was significantly lower than that in the control group (P < 0.05). Taken together, bone marrow mesenchymal stem cel transplantation can improve the pathological damage in systemic lupus erythematosus mice, and has a certain therapeutic effect on systemic lupus erythematosus.
9.Therapeutic Effects of Pregabalin Combined with Hydrochloric Oxycodone on 33 Casesof Malignant Neuropathic Pain
Bicheng ZHANG ; Zhihuai ZHANG ; Jun WANG ; Zhigang WANG ; Tingting WU ; Zhiguo RAO ; Jianfei GAO
Herald of Medicine 2015;(3):325-328
Objective To evaluate the effects of pregabalin combined with hydrochloric oxycodone on patients with ma-lignant neuropathic pain (MNP). Methods A total of 66 patients with MNP was divided into control group or treatment group randomly. The patients in control group received only hydrochloric oxycodone, and treatment group were treated with the combina-tion of pregabalin and hydrochloric oxycodone. Numeric rating scale (NRS) score was used to evaluate the analgesic effects. Med-ical outcomes study sleep scale (MOS-SS,Chinese version) was used to evaluate the improvement of sleep disorder. The changes of depression or anxiety were investigated by 17-item Hamilton Depression Rating Scale (HAMD-17) or Hamilton Anxiety Scale (HAMA), respectively. Side effects were accessed by Acute and Subacute Toxicity Grading Criteria of Anticancer Drugs (WHO). Results The pain control rate of treatment group was 87. 1% , which was superior to that of control group (58. 6% ) (P<0. 05). The improvement of sleep interference, and the quality and quantity of sleep in treatment group were also superior to that in control group (P<0. 05). After the treatment, depression and anxiety was attenuated in both groups, and the improvement degree in treatment group was higher than that in control group (P<0. 05). No obvious side effects were found in either groups. Conclusion The combination therapy of pregabalin and hydrochloric oxycodone is the better way to treat MNP.
10.Construction and screening of RPL23-siRNA interference fragments
Wenmiao PENG ; Zhimin ZHANG ; Meng HU ; Lifang YU ; Bicheng ZHANG ; Zhiguo RAO ; Chuanrong QIN
Practical Oncology Journal 2016;30(6):487-491
Ob jective To construct and screen out the RPL 23-siRNA interference fragments ,providing the basis for the following experiments about the correlation with RPL 23 and gastric cancer .Methods The RPL23-siRNA,synthesized chemically through lipofection ,were selected from three target sequences by RNA in-terference and detected by real -time PCR and Western blot .Results Compared with normal cell group and RPL23 control group ,the mRNA and protein expression of RPL 23 in the other 3 interference groups were signifi-cantly decreased(P<0.01).Multiple comparisons showed that the interference efficiency of RPL 23 -siRNA1 group was significantly higher than that of RPL 23-siRNA2 group and RPL23-siRNA3 group(P<0.01).Con-clusion The RPL23-siRNA interference fragment can be successfully constructed and screened out ,which pro-vides the basis for the following experiments .