1.Effect of paclitaxel on telomerase activity, apoptosis and p53/bcl-2 gene protein expression of breast cancer MCF-7 cell line
Rongqing PANG ; Buzhen ZHANG ; Xinghua PAN
Chinese Journal of General Surgery 2001;0(10):-
Objective To investigate effect on telomerase activity, apoptosis and p53/bcl-2 gene expression of MCF-7 cells line induced by paclitaxel.Methods By techniques of cell culture in vitro, telomeric repeat amplification protocol with ELISA(TRAP-ELISA) and flow-cytometry (FCM), MCF-7 cell line was treated by paclitaxel in various concentration for 72h.Results Paclitaxel down-regulated telomerase activity of MCF-7 cell, induced apoptosis of the cell in a concentration-dependent manner, significantly decreased expression of bcl-2 gene protein and increased expression of p53 gene protein. There was a positive correlation between telomerase activity and apoptosis and the expression of p53/bcl-2 gene protein.ConclusionPaclitaxel could down-regulate telomerase activity,induce apoptosis, decrease expression of bcl-2 gene protein and increase expression of p53 gene protein, which may be one of important mechanisms of Paclitaxel′s anticancer action.
2.HLA-peptide tetramers and adoptive immunotherapy in prevention of cytomegalovirus disease
Guangping RUAN ; Xiang YAO ; Xinghua PAN ; Rongqing PANG ; Yongli DENG
Chinese Journal of Tissue Engineering Research 2010;14(31):5878-5882
BACKGROUND: Antiviral drugs can reduce the incidence of early-onset cytomegalovirus(CMV)disease,but are associated with strong toxicity and the development of late-onset CMV disease.In order to prevent CMV disease better,cytotoxic T lymphocytes(CTL)may play a critical role in controlling CMV reactivation.Fluorescent HLA-peptide tetramers are used to monitor the recovery of CMV CTL in recipients of allogeneic transplants.OBJECTIVE: To explore the effect of HLA-peptide tetramers and adoptive immunotherapy in treating CMV disease.METHODS: A computer-based online search of Pubmed and Wanfang databases was performed for articles related to CTL detection,application of antiviral drugs and HLA-peptide tetramers,and adoptive immunotherapy with key words"HLA-peptide tetramers,cytomegalovirus,specific CTL,adoptive immunotherapy"in English and Chinese.Repetitive articles were excluded and 29 articles were included.RESULTS AND CONCLUSION: Adoptive immunotherapy with CMVs cytotoxic T cells as preemptive therapy is a very elegant strategy; however,generation of these cells is costly and time-consuming,and therefore the therapy is not available at every transplantation center.Magnetic selection of CMVs CD8+T cells from peripheral blood of CMV-seropositive donors by using HLA-peptide tetramers may be very hopeful,which simplifies adoptive immunotherapy.
3.Umbilical cord-derived mesenchymal stem cell culture:dyeing and tracer technique
Xia HUANG ; Xinghua PAN ; Rongqing PANG ; Guangping RUAN ; Xuemin CAI
Chinese Journal of Tissue Engineering Research 2014;(23):3751-3755
The culture of umbilical cord-derived mesenchymal stem cells is extremely important for studies on umbilical cord mesenchymal stem cells. Optimization of cellculture technology is crucial for clinical application of mesenchymal stem cells and even celltherapy. Meanwhile, the labeling and tracer technique of umbilical cord-derived mesenchymal stem cells is a hotspot in stem celltransplantation. OBJECTIVE:To review the research and development of the cellmarkers and tracer methods of umbilical cord-derived mesenchymal stem cells. METHODS:A computer-based search of VIP, CNKI, Medline, Highwire and Foreign Journals Integration System databases was performed for articles concerning culture and labeling of umbilical cord-derived mesenchymal stem cells published from January 2001 to October 2013. The keywords were“stem cells, mesenchymal stem cells, umbilical cord-derived mesenchymalstem cells, cellculture, labeling methods”in Chinese and English, respectively. Final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Umbilical cord-derived mesenchymal stem cells have not yet been widely used, mainly because of the immature isolation, culture and staining techniques of umbilical cord-derived mesenchymal stem cells. These techniques are worthy of further optimization studies. Although in recent years, cellmarkers and tracer technology of umbilical cord-derived mesenchymal stem cell s have made great progress, there are stil many problems need to be solved.
4.Clinical value of 99Tcm-MIBI scintigraphy in patients with differentiated thyroid carcinoma after surgery and first 131I therapy
Xiaomei PAN ; Dong DUAN ; Yuquan ZHU ; Hua PANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2015;35(5):383-386
Objective To evaluate the clinical value of 99Tcm-MIBI scintigraphy for diagnosis of residual thyroid tissue and metastasis in patients with DTC after their first 131I therapy.Methods From February 2010 to March 2014,192 DTC patients (38 males,154 females,average age (43.2±8.6) years) who received total or near-total thyroidectomy and were pathologically diagnosed as DTC (171 papillary and 21 follicular carcinomas) underwent 99Tcm-MIBI scan (average dosage:740-925 MBq) 6 months after their first 131 I therapy.131 I scans was performed 4 d after oral administration of 131I of therapeutic dose (average dosage:5 550-8 140 MBq).Pre-and post-therapeutic images and the serum Tg level (detected before the imagings) were compared and analyzed.Any abnormal uptake of agent found inside or outside the thyroid was regarded as positive result.Patient-based and lesion-based data analysis were performed by x2 test and two-sample t test.Results A total of 191 patients were finally included,of which 65 positive cases were found.The sensitivity of 99Tcm-MIBI imaging was significantly lower than that of 131I imaging(56.9% (37/ 65) vs 92.3% (60/65);x2 =14.7,P<0.01).Among 43 thyroid remnants and 22 metastatic lesions,99Tcm-MIBI imaging detected 39.5% (17/43) of thyroid remnants and 90.9% (20/22) of metastases,and those of 131I imaging were 100% (43/43) and 77.3% (17/22) respectively.The sensitivity of 131 I imaging in detecting thyroid remnants was significantly higher than that of 99Tcm-MIBI imaging(x2=24.0,P<0.01).The sensitivities in detecting metastasis were not significantly different (x2=0.57,P>0.05).The serum Tg level of positive groups (99Tcm-MIBI positive + 131I positive or 131I negative) were significantly higher than that of 99Tcm-MIBI negative + 131I negative group (t =-20.7 and-6.0,both P<0.01),and that of 99Tcm-MIBI positive + 131I negative group was higher than that of 131I positive + 99Tcm-MIBI negative group(t=-2.7,P<0.05).Conclusion 99Tcm-MIBI imaging could detect metastasis of DTC patient after first radioiodine therapy,but the value in detecting thyroid remnants is limited.
5.Construction of eIF-4E Adenovirus and Its Effect on Metastasis Ability of Human Breast Cancer Cell Line MCF-7
Guo-Qiang ZHANG ; Wei PAN ; Liang ZHAO ; Da PANG ;
China Biotechnology 2006;0(03):-
Objective: To reconstruct adenovirus vector of breast eukaryotic initiation factor 4E and to observe its effect on the metastasis ability of breast cancer cell line MCF-7. Methods: eIF-4E gene was constructed into adenovirus vector pAD-X by gene recombination technique, which was transformed into 293 packaged cell for high titer adenovirus. Real-time PCR was applied to detect eIF-4E gene expression. eIF-4E siRNA was applied and then transwell cabin assay was used to observe changes of invasion and motor ability of MCF-7 cells transfected with reconstruction adenovirus. Result:The finding of digestion was coincided with expected. eIF-4E gene over-expression was detected in transfected MCF-7 cells with real-time PCR. And the invasion and motor abilities of transfected MCF-7 cells were more significantly inhibited in transwell cabin assay (respectively p
6.Cultivating High-quality Medical Talents
Feng PAN ; Houjie LIANG ; Xueli PANG ; Zhihua RUAN
Chinese Journal of Medical Education Research 2003;0(04):-
This paper discussed that how to cultivate high-quality medical talents by the use of basic education,medical moral education and psychologic education etc.
7.Effect of different anticancer drugs on the hyperplasia and telomerase activity of breast cancer cell MCF-7
Rongqing PANG ; Buzhen ZHANG ; Jian CHEN ; Xinhua PAN ; Zhilong CHEN ;
Chinese Journal of General Surgery 1997;0(06):-
Objective To study the alteration of telomerase activity of MCF 7 cell line of breast cancer in the presence of different anticancer drugs. Methods The hyperplasia and viability of MCF 7 cell were detected by cell counting and trypan blue exclusion, and the telomerase activity was measured by TRAP.The alteration of MCF 7 cell and its related factors of telomerase activity were observed on cell growing in different condition. Results In abscence of drug, there was a positive correlation between hyperplasia and telomerase activity of the cell(r= 0.901 ). Adriamycin, paclitaxel and cisplatin could obviously inhibit the growth and reduce the telomerase activity of the cell in a dose-dependent and time-dependent fashion, and this reduction in telomerase activity closely correlated with the reduction in the number of viable cells. Conclusions Adriamycin, paclitaxel and cisplatin can inhibit the growth of MCF 7 cell, which may be correlated with the reduction in telomerase activity and cell viability.
8.Efficient purification and label of anti-human beta 2-microglobulin light chain monoclonal antibody
Guangping RUAN ; Xiang YAO ; Rongqing PANG ; Xingming WANG ; Ying DAI ; Xinghua PAN
Chinese Journal of Tissue Engineering Research 2011;15(24):4557-4560
BACKGROUND: The inoculation of hybridoma cell strain onto mouse abdominal cavity may obtain ascites containing mass antibody. Previous method to purify monoclonal antibody in ascites is complex and difficult to operate.OBJECTIVE: To prepare, purify and label anti-human leukocyte antigen (HLA)-I class molecule light chain monoclonal antibody, and to detect the expression of tumor cell surface HLA-I class molecules. METHODS: Hybridoma cells were inoculated onto the mouse abdominal cavity. Ascites containing anti-human light chain beta2-microglobulin antibody were obtained and purified with the modified caprylic acid-ammonium sulfate method. The purified monoclonal antibody was labeled with fluorescein isothiocyanate to detect peripheral blood mononuclear cells, T2 cells expressing blank HLA-A2 molecule and K562 cells surface HLA-I class molecules. The expression of HLA-I class molecules was determined by using flow cytometry and fluorescent microscopy. RESULTS AND CONCLUSION: The purified anti-human light chain beta2-microglobulin-fluorescein isothiocyanate monoclonal antibodies accounted for 96% purity. Flow cytometry results showed that, the HLA-I class molecules were highly expressed in peripheral blood mononuclear cells surface, lowly expressed in T2 cells, and not expressed in K562 cells surface. It is a simple and convenient method to purify ascites with the modified caprylic acid-ammonium sulfate method, and according prepare anti-human light chain beta2-microglobulin-fluorescein isothiocyanate. This method is effective to distinguish the levels of HLA-I class molecules expressed in various cells.
9.Intervention of nicotine on MNU-induced bladder cancer in rats.
Di, LIU ; Feng, PAN ; Bing, LI ; Xiaomin, HAN ; Wencheng, LI ; Ying, SHI ; Zili, PANG ; Qijun, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(1):103-6
This study examined the effect of nicotine on the expression of mutant p53 (mt-p53) in bladder cancer rats. The rat models of bladder cancer were established by infusing N-methyl-nitroso-urea (MNU, 10 mg/kg every 2 weeks for 8 weeks) into the bladder. Pathological examination on the bladder was conducted to confirm the establishment of the model. All the bladder cancer rats were randomly divided into an MNU group and 3 nicotine groups. In the nicotine groups, the rats were intragastrically administered nicotine at different concentrations (25, 15, 5 mg/kg respectively) 3 times per week for 8 weeks. The mt-p53 expression was detected by the immunohistochemical method. The results showed that rat bladder cancer models developed histopathological changes of bladder transitional cell carcinoma. The positive rate of mt-p53 expression in the 3 nicotine groups (25, 15, 5 mg/kg) was 75.00%, 58.33% and 41.67% by the 14th week, respectively, significantly higher than that in the MNU group (33.33%) (all P<0.05). The mt-p53 expression rate was positively correlated with the medication dose and time (P<0.05). It is concluded that nicotine may play an important role in the development of bladder cancer partially by increasing the expression of mt-p53.
10.Emergency treatment of testicular torsion and postoperative follow-up: a 71 case report.
Feng, PAN ; Zhaohui, ZHU ; Bing, LI ; Zili, PANG ; Yajun, XIAO ; Fuqing, ZENG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2012;32(5):704-6
This study presented our experience in the treatment of testicular torsion, which may help achieve early diagnosis and improve therapeutic effects. A retrospective analysis was conducted in 71 patients with testicular torsion who were treated in our hospital from October 2007 to April 2011. The age of the patients ranged from 16 days to 34 years. All the patients had unilateral testicular torsion, which took place on the left side in 43 cases and on the right side in 28 cases. The course of the disease varied between three hours to 30 days. Post-operative follow-up was conducted until October 2011. Items examined included signs and symptoms at their first clinical visit, ultrasound findings, treatment in emergency surgery, and post-operative follow-up. In this study, the 71 patients were diagnosed with testicular torsion by color Doppler sonography, 7 had testicular fixation, 63 patients received orchiectomy, while 1 patient did not undergo surgery due to pressure from family members. Post-operative follow-up showed that the one patient's testicle, which had been reserved, atrophied, while all the other survived. No recurrence was found during the follow-up visits. It is concluded that an early diagnosis and surgery is important in improving the survival rate of testicular torsion, and the diagnosis and treatment by the first attending clinician is of critical importance.