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The in vitro passage number and proliferation of non-immortalized cells are limited, which restrictions cell therapy or in vitro studies. Cells transfected with temperature sensitive simian virus 40 large T antigen (ts-SV40LT) gene could show the greatest proliferation. The cells can be amplified with compensating the lack of limited number of cells under the permissive temperature. Non-permissive temperature can be used in studying the cell therapy or its other physiological characteristics. This research field involves peritoneal stromal cells, satellite cells of urinary tract, oral epithelial cells, adre?nal medullary cells, bone marrow-derived endothelial cells, retinal progenitor cells, mesenchymal stem cells, hematopoietic stem cells, mast cells, podocytes and Kupffer cells. In this study, the current research on Ts-SV40-mediated temperature-sensitive cells was reviewed.
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Objective To investigate the therapeutic effect of Xuebijing injection on stroke associated pneu monia the changes of plasma C-reactive protein level .Methods 80 cases of post-stroke patients with pneumonia were randomly divided into Xuebijing injection treatment group 40 cases and control group of 40 cases, two groups were given conventional antibiotics and anti-inflammatory treatment ,treated with Xuebijing injection group was dealed with Xuebijing injection 50ml plus 0.9% sodium chloride solution 100ml intravenous drip on the basis of conventional therapy,2/d,for 7d,changes before and after treatment in the two groups were evaluated the body temperature ,periph-eral white blood cell count ,neutrophil percentage ,C-reactive protein index .Results The treatment group after treat-ment for 7d body temperature,blood routine,neutrophils and C-reactive protein were compared before treatment were significantly improved(t=9.99,24.09,12.44,43.98;all P<0.05),all of the indexes in the control group compared before treatment were significantly improved,the differences were significant(t=15.95,20.12,4.14,16.53;all P<0.05),after treatment the observation index except temperatrue decreased significantly ,with statistically significant differences compared with control group (t=4.83,6.15,7.93,all P<0.05).Conclusion Xuebijing injection syner-gistic effect of stroke-associated pneumonia antibiotic treatment significantly , more effective than antibiotic therapy alone,has the very good application and promotion of clinical value .
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<p><b>OBJECTIVE</b>To compare the safety and effectiveness of DPPHR with PPPD/PD for treating chronic pancreatitis with an inflammatory mass in the head of pancreas.</p><p><b>METHODS</b>The relative data bases such as Medline, EMBase, Biosis, COCHRANE Library, Science Citation Index, SinoMed, Chinese Journal Full-text Database, Wangfang, CNKI were searched systematically, researchers selected randomized controlled trials (RCT) and prospective clinical controlled trials (CCT) . The assessment of the bias risk of the included trials was according to the assessing tools suggested by Cochrane Handbook 5.1. The Review Manage 5.2 was used to perform the statistical analysis.</p><p><b>RESULTS</b>In total, 5 RCTs and 2 CCTs were included, 381 patients involved. Comparing with PPPD/PD procedure, DPPHR has no significant difference in terms of the mortality of perioperative period (RD = 0.01, P = 0.51), the incidence of bleeding (RD = -0.01, P = 0.72), pancreatic fistula(RD = -0.01, P = 0.59) and delayed gastric emptying (RD = -0.15, P = 0.10), the ration of complete pain relief after operation (RR = 1.06, P = 0.32) and the score of global quality of life (WMD = 10.31, P = 0.19).While DPPHR had significant superiorities in terms of the total morbidity of perioperative period (RR = 0.60, P = 0.008), the duration of the operations(WMD = -71.60, P = 0.03), the postoperative hospitalization duration(WMD = -3.95, P < 0.01), weight gain(WMD = 3.68, P < 0.01), occupational rehabilitation after the operations (RR = 1.38, P = 0.008).</p><p><b>CONCLUSIONS</b>In terms of reducing the morbidity of perioperative period, shortening the duration of the operations and the postoperative hospitalization duration, weight gain, occupational rehabilitation after the operations, the DPPHR is more favorable for improving patients' life qualities comparing with PPPD/PD.</p>
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Humans , Duodenum , General Surgery , Pancreas , General Surgery , Pancreatectomy , Methods , Pancreaticoduodenectomy , Methods , Pancreatitis, Chronic , General Surgery , Prospective Studies , Quality of LifeABSTRACT
OBJECTIVE@#To over-express cyclin-dependent kinase 2-associated protein 1 (CDK2-AP1) gene, and investigate its effect on the proliferation and cell cycle regulation in breast cancer cell line MCF-7.@*METHODS@#CDK2-AP1 gene coding region was cloned into lentivirus vector. Lentivirus particles were infected into MCF-7 cells to upregulate the expression of CDK2-AP1 gene. The expression level of CDK2-AP1 was detected at both mRNA and protein levels by real-time PCR and Western blot. MTT assay, colony formatting assay, and flow cytometry were performed to detect the change of proliferation and cell cycle in MCF-7 cells. We examined the expression of cell cycle associated genes (CDK2, CDK4, P16Ink4A, and P21Cip1/Waf1) followed by CDK2-AP1 over-expression by Western blot.@*RESULTS@#CDK2-AP1 gene was up-regulated significantly at both mRNA (6.94 folds) and protein level. MTT based growth curve, colony formatting assay and flow cytometry showed that CDK2-AP1 over-expression lentivirus inhibited the proliferation of MCF-7 cells with statistical difference (P<0.05). In addition, with CDK2-AP1 over-expression, MCF-7 cells were arrested in G1 phase accompanied by apoptosis. Western blot showed that the expression level of P21Cip1/Waf1 and P16 Ink4A was upregulated, while the expression level of CDK2 and CDK4, members of the CDK family, was downregulated.@*CONCLUSION@#CDK2-AP1 gene plays a cancer suppressor role in breast cancer. Its function includes inhibiting the proliferation of MCF-7 cells and arresting the cell cycle in G1 phase.
Subject(s)
Humans , Breast Neoplasms , Cell Cycle , Cell Division , Cell Proliferation , Cyclin-Dependent Kinases , Down-Regulation , MCF-7 Cells , Protein Kinases , Genetics , Metabolism , Tumor Suppressor Proteins , Genetics , MetabolismABSTRACT
OBJECTIVE@#To investigate dosimetric characteristics and to evaluate the clinical efficacy of intensity modulated radiotherapy (IMRT) as compared with conventional radiotherapy in nasopharyngeal carcinoma (NPC).@*METHODS@#Forty-seven NPC patients who accepted IMRT served as the IMRT group, and conventional radiotherapy plan was also made for each patient in this group using the treatment planning system. Dosiological evaluation of the 2 radiotherapy plans was made through dose volume histogram, 95% target volume dose (V(95)) and normal tissue complication probability. Another 47 patients who underwent conventional radiotherapy (CRT) at the same period formed the control group. The therapeutic effect as well as the acutes and late toxicities of normal tissues in the 2 groups were observed.@*RESULTS@#V(95) of the IMRT was more than 96% (96.83%-99.99%) for each target area, obviously superior to CRT in the sub-clinical target area. The radiation dose of normal tissues such as the brainstem and the spinal cord in the IMRT was much less than that in the CRT. Consistant with this, the part and complete remission rate, the 3-year loco-regional progress free survival rate, and overall survival rate in the IMRT group were all higher than those in the CRT group. For most patients in the IMRT group, the grade of acute toxicities was much lower than that in the CRT group. Patients in the IMRT group showed no more than grade 3 xerostomia, while in the CRT group still 21% of the patients suffered grade 3 or higher xerostomia a year later.@*CONCLUSION@#Compared with CRT, IMRT can improve the target volume dose and decrease the dose of surrounding tissues, resulting in higher control rate and fewer side effects.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Young Adult , Carcinoma, Squamous Cell , Radiotherapy , Nasopharyngeal Neoplasms , Radiotherapy , Radiotherapy Dosage , Radiotherapy Planning, Computer-Assisted , Methods , Radiotherapy, Intensity-Modulated , MethodsABSTRACT
Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the racliosensitivity of CNE2-implanted tumors, and to investigate the related mechanism. Methods: E1A gene was transfected into CNE2 cells using adenovirus system, and sta-ble E1A positive clones were established. The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was ob-served in nude mice. The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated. The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR. Results: CNE2 cells stably transfected with E1A gene (CNE2-Ad-E1A) were successfully established. The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell- and CNE2-Ad-β-gal cell-implanted mice (CNE2 cells stably transfected with Ad-β-gal). Radiotherapy, E1A gene therapy and E1A gene + radio-therapy all suppressed the growth of implanted tumors, with the tumor suppression rates being (60.32±5.34) %, (70.53±6.12) %, and (97.15±4.87) % , respectively. E1A gene therapy significantly increased the expression of P53 gene in tumor tissues. Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells, and enhance its sensitivity to radiotherapy, which may be related to the increased expression of P53 gene in tumor tissues.
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Objective To study the effect of E1A gene on the radiosensitivity of nasopharyngeal carcinoma (NPC) cells and its mechanism. Methods Ad-E1A gene was transfected into human NPC cells (CNE2), then the positive clones (CNE2-Ad-E1A) were identified by RT-PCR. CNE2 cells, CNE2 cells transfected with Ad-β-gal (CNE2-Ad-β-gal) and CNE2-Ad-E1A cells were irradiated with 0 Gy,2 Gy,4 Gy,6 Gy and 8 Gy respectively using 6 MV X-ray. Clone forming assays were carried out, cell survival curves were drawn and the sensitivity enhancing ratio (SER) was calculated. The redistributions of cell cy-cle were analyzed by flow cytometry. RT-PCR was used to detect the expression of wtp53. Results RT-PCR confirmed that E1A gene had been integTated into positively transfected cells and stably expressed. Cell survival curves showed that the SER of D0,Dq and SF_2 value was 1.37, 1.95 and 1.46 in CNE2-Ad-E1A cells. The D_0,D_q and SF_2 value was 1.57 Gy,1.82 Gy, 0.89 in CNE-2 cells and 1.53 Gy,1.78 Gy,0.82 in CNE2-Ad-β-gal cells, respectively. The G_2/M arrest was shown in CNE2-Ad-E1A cells. Moreover, the expression of wtp53 gene was markedly enhanced in Ad-E1A-CNE2 cells. Conclusions E1A gene can ef-fectively enhance the radiosensitivity of human NPC cells, which may be associated the enhancement of wt-p53 expression and G_2/M arrest.
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OBJECTIVE@#To determine the effect of Ad-E1A gene therapy on the radiosensitivity of nasopharyngeal carcinoma cell by downregulating the expression of VEGF in vitro.@*METHOD@#The human nasopharyngeal carcinoma CNE-2Z cell lines were investigated. The recombinant adenovirus vector containing E1A gene was used for this study. After CNE-2Z cells was treated with PBS, Ad-beta-gal and Ad-E1A for 48 h, the three groups were irradiated in different doses at 0, 2.4, 6, 8 and 10 Gy, the cytotoxicity was determined by MTT assay and cell cycle was analysis by flow cytometry. The VEGF expression were evaluated by RT-PCR assay and immunocytochemical analysis.@*RESULT@#Significant cell deaths by IR were observed in a dose dependent manner in the three group CNE-2Z cells. After transduction of the E1A gene into CNE-2Z cells, the sensitivity of these cells to radiation was enhanced than the PBS treated group and Ad-beta-gal treated group. Cell growth inhibition in Ad-E1A group by IR was strongly enhanced than Ad-beta-gal treated group and PBS treated group. RT-PCR assay and immunocytochemical analysis showed VEGF expression was downregulated in Ad-E1A treated group.@*CONCLUSION@#E1A gene therapy can effectively enhance the nasopharyngeal carcinoma cell sensitivity to the radiotherapy by down-regulating VEGF expression. These findings may pave the way for efficient radiation-gene therapy to NPC in future.
Subject(s)
Humans , Adenoviridae , Genetics , Cell Line, Tumor , Gene Expression Regulation, Neoplastic , Genetic Therapy , Genetic Vectors , Nasopharyngeal Neoplasms , Genetics , Metabolism , Radiotherapy , Radiation Tolerance , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
Objective To investigate the proteomics differences between the high-sensitivity(HS) and the low-sensitivity(LS)groups of cervical carcinoma treated by concurrent chemoradiotherapy,and to confirm the sensitivity associated proteins in intermediate stage and advanced cervical carcinoma.Methods Fresh carcinoma tissues were collected from 10 untreated cervical carcinoma patients.According to the response to concurrent chemoradiotherapy,the tissues were classified into HS group and LS group.In the first part of our experiment,protein separation was performed using two-dimensional gel electrophoresis(2-DE)with Amersham 18 am linear pH 3-10 immobilized pH gradient(IPG)strips.The images of the gels were analyzed by PD-quest 7.0 software to find the differentially expressed protein-spots in each group.Then the differentially expressed protein-spots were incised from the gels and digested by trypsin.The peptide mass flngerprintings(PMF)was acquired by matrix assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF-MS).The proteins were identified by data searched in the Mascot-database.Two differentially expressed proteins were assayed by western blot and immunohistochemical methods.Results Most of the gels were clear and successfully analyzed by PD-quest 7.0 software.Most of the protein-spots concentrated on the area of 20-100 KDa(Mw)and pH4-8.The average number of the protein-spots was 781±74 in HS group and 766±52 in LS group.The match rate was 87.6%between the two groups.Eight proteins highly in HS group but lowly expressed in LS group included hemoglobin subunit beta,caspase-14 precursor,calmodulin-like,S100-A9 protein(MRP-14),galectin-7,HSKERC4,keratin 19 and actin.Ten proteins highly in LS group but lowly expression in HS group included anti HBs antibody light-chain Fab,laminB1,WARS protein,flavin reductase,glutamate dehydrogenase 1,nuclear matrix protein 238,retinal dehydrogenase 1,AFl 65172,subunit of replicative DNA polymerase and HSP70.The higher expression of HSP70 in LS group and galectin7 in HS groups were further confirmed by western blot and immunohistochemical method.Conclusions The 2-DE gels images are successfully acquired from high-sensitivity group and low-sensitivity group of intermediate stage and advanced cervical carcinoma tissues treated by concurrent chemoradiotherapy.Some differentially expressed proteins between the two groups can be further confirmed by western blot and immunohistochemical method.
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Objective To investigate the effect of El A gene on the radiosensitivity of human laryngeal carcinoma cells and its correlated mechanisms. Methods The Ad-E1A and Ad-β-gal were amplifieated in Hek293 cells, extracted by freezing (-80℃) and thawing(37℃) repeatedly (3 times) , purificated by the method of density gradient of CsC1 and titrated by plaque assay method. Then they were transfected into human laryngeal carcinoma cells (Hep-2) and authenticated by RT-PCR. The radiosensitivity of Hep-2 cells transfeeted with or without El A were studied by cell surviral curve. Finally we investigated the correlated mechanisms including cell apoptosis studied by flow cytometry and VEGF content studied by RT-PCR. Resuits The radiosensitivity of Hep-2 cells transfected with El A was intensified, Do and Dq were lowered and α was increased. Flow cytometry showed that the apoptosis rate of cells with E1A or with El A and radiotherapy was increased. The VEGF content of the cells transfeeted with E1 A or treated by radiotherapy was decreased, which reached the lowest level when the cells were treated with the both mathods. Conclusions E1 A gene can intensify the radiosensitivity and contribute to the apoptosis of human laryngeal carcinoma cells. El A gene and radiotherapy can markedly decrease the VEGF content.
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Objective:To study the inhibitory effect of E1A gene on the growth of tumors in nude mice implanted with nasopharygeal carcinoma CNE2 cells and its promotion effect on the radiosensitivity of CNE2-implanted tumors,and to investigate the related mechanism.Methods: E1A gene was transfected into CNE2 cells using adenovirus system,and stable E1A positive clones were established.The inhibitory effect of E1A on tumor formation-ability of CNE2 cells was observed in nude mice.The efficacy of E1A gene therapy with or without radiotherapy against CNE2 cell-implanted tumors was evaluated.The effect of E1A gene therapy on the expression of P53 was detected by RT-PCR.Results: CNE2 cells stably transfected with E1A gene(CNE2-Ad-E1A)were successfully established.The tumor formation time was later and tumor size was smaller in CNE2-Ad-E1A cell-implanted mice compared with those in CNE2 cell-and CNE2-Ad-?-gal cell-implanted mice(CNE2 cells stably transfected with Ad-?-gal).Radiotherapy,E1A gene therapy and E1A gene+radiotherapy all suppressed the growth of implanted tumors,with the tumor suppression rates being(60.32?5.34)%,(70.53?6.12)%,and(97.15?4.87)%,respectively.E1A gene therapy significantly increased the expression of P53 gene in tumor tissues.Conclusion: E1A can inhibit the growth of tumors in mice implanted with nasopharygeal carcinoma cells,and enhance its sensitivity to radiotherapy,which may be related to the increased expression of P53 gene in tumor tissues.