1.The condition for error-prone PCR induced mutation
Xiaosui CHEN ; Baoan WANG ; Yan WANG ;
Academic Journal of Second Military Medical University 1981;0(03):-
Objective: To evaluate the effectiveness of random mutagenesis via mutator E.coli strain and error prone PCR. Methods: A ScFv containing phagemid was transformed into mutator strain XL1 Red and subjected to 7 overnight growth phases with 1/100 dilution of each phase. DNA was extracted and sequenced. The ScFv gene was also subjected to PCR mutagenesis. Mutation effects of Mg 2+ concentration, alteration of individual dNTP amounts and addition of dITP were investigated. Results: The mutation rate of 7 growth phases (about 50 cell cycles) in XL1 Red was less than 0.1%. In error prone PCR, higher concentration of Mg 2+ increased the mutation rate. Increased content of dTTP and dCTP had better effect than that of dATP and dGTP. Addition of dITP plus low concentration dATP or dGTP caused lower mutation rate. More than 2% mutation could be reached by 2 rounds PCR containing 5 mmol/L MgCl 2, 0.5 mmol/L MnCl 2, 1 mmol/L dTTP and 1 mmol/L dCTP. But the mutation showed obvious base bias with most substitutions at A/T positions and a prevalence of transition over transversion. Conclusion: Random mutagenesis in mutator strain has too low mutation rate for antibody affinity maturation. A more than 2% mutation rate can be obtained by error prone PCR, which is suitable for constructing mutated antibody libraries, but the base bias of error prone PCR should be considered.
2.EXPERIMENTAL OBSERVATION OF 65Zn METABOLISM ON RATS FED ON CROPS FROM ENDEMIC AND NON- ENDEMIC DISTRICTS OF KESHAN DISEASE
Baoan KANG ; Zhongbai WANG ; Yin SU
Acta Nutrimenta Sinica 1956;0(04):-
Albino rats fed on crops from endemic and non-endemic districts of Ke-shan desease, were used in this experiment, which was studied by 85Zn tracer metabolic method after 8 weeks feeding. Uptake of 85Zn (ZnCl2) by blood, heart, liver, kidneys and brown adipose tissue were determined at 24 and 72 hours after injection of this tracer respectively, and its distribution was thoroughly examined. The results revealed that the uptake of 8aZn was higher in the groups fed on crops from endemic districts, particularly significant in liver, kidneys and brown adipose tissue. This may reflect the lacking or deficiency of zinc in the bodies.The regular differences of 85Zn between two groups were certainly related to the crops produced from endemic or aon-endemic districts. In other words, there should be some differences between crops from endemic and non-endemic areas. Thus more attention should be paid to the factors in endemic crops, which induced lacking or deficiency of zinc in the experimental animal and to its relationship with etiology of Keshan disease.
3.Practice and exploration of competitive intelligence teaching
Lei WANG ; Jianhua ZHANG ; Baoan CHEN
Chinese Journal of Medical Education Research 2013;(8):800-802
Objective To improve the students' awareness of competition and innovation and to explore the teaching methods of competitive intelligence. Methods Totally 130 juniors(46 males and 84 females)in the college of pharmacy of Shihezi University were enrolled as research object. Theory teach-ing+seminar+practice teaching method was employed and knowledge of competitive intelligence was in-tegrated into the teaching of pharmacy literature retrieval course. Investigation and evaluation on stu-dents' competitive intelligence knowledge were conducted before and after teaching. SSPS 17.0 was used to do statistical analysis and chi-square test was performed to analyze data,with P≤0.05 being statistical significant. Results 100.00%students thought that they had mastered the concepts of competitive intelli-gence and anti-competitive intelligence concept;90.77%students already had the ability to collect and sort out information. Information retrieval ability,interests in knowledge of competitive intelligence,competi-tive consciousness and professional information quality of students were improved. Conclusion Increas-ing competitive intelligence teaching can help improve students' sense of competition and innovation consciousness. Proportion of practice teaching should be appropriately increased to cultivate students' collection and analysis abilities of intelligence information.
4.Assessment on the efficacy of intraperitoneal chemotherapy on progressive gastric cancer after radical treatment
Cailian WANG ; Baoan CHEN ; Zao JIANG
China Oncology 2000;0(06):-
Purpose: To observe the efficacy of intraperitoneal chemotherapy on advanced gastric cancer after radical treatment. Methods: 64 cases of patients with stage Ⅱ - Ⅲ treated by radical surgery were selected and randomly divided into systematic plus intraperitoneal chemotherapy group and systemic chemotherapy group. 3-year survival rate and 3-year recurrence rate were recorded. Results: The recurrence rate for the intraperitoneal chemotherapy group was 18.75% while the control was 43. 33% (P
5.Importance and Challenge of Non-Affiliated Scientific Member in Ethics Review Committee
Xiaoru SUN ; Huiping WANG ; Xiuqin WANG ; Baoan CHEN ; Hongli TANG
Chinese Medical Ethics 2015;(5):686-688
After discussing the reason for insufficiency of committee member in current ethical review , and considering the requirement for improving the function of Ethics Review Committee , this paper proposes that Ethics Review Committee should appoint more non -affiliated scientific members to ensure the independence of ethical re-view , thus protecting the rights and beneficence of subjects , and to secure the justice in ethical review of protocols , the paper also discussed the resolutions to address the challenge brought by appointing non -affiliated members .
6.Survival and differentiation of endogenous neural stem cells in the hippocampus of post-stroke depression rats
Shaohua WANG ; Zhijun ZHANG ; Yijing GUO ; Goojun TENG ; Baoan CHEN
Chinese Journal of Neurology 2008;41(9):614-617
Objective To investigate the proliferation and survival of endogenous new born cells in the hippocampus of post-stroke depression (PSD) rats. Methods Male SD rats were divided into 4 groups (n = 18): control, stroke, depression and PSD group. The animals were subjected to left middle cerebral artery occlusion (MCAO) reaulting in consistent focal cerebral infarcts, followed by an 18-day exposure to chronic mild stress (CMS) and single housing to induce PSD animal model The dynamic expression of brodmodeoxyuridine (BrdU), neuron-specific nuclear protein (NeuN) and glial fibrillary acidic protein (GFAP) in the left dentate gyrus of hippocampus were determined by immunohistochemistry or double-lable immunofluorescence staining. BrdU was a marker of new born cell, NeuN and GFAP marked that new born cell differentiated into neuron and astrocyto respectively. Results Compared with ischemia animals (232.2±8.6,123.7±2.6,136.2±2.6), in the left dentate gyrus, PSD samples had significantly less BrdU-positive cells ( 156.2±2.5, t =28.83, P < 0.01 ) on the 21st day after ischemia, and on day 30 and 45 ( 70.2±2.0, 81.2±1.1, t = 52.27, 62.08, both P < 0.01 ). The ratio of BrdU-positive cells to NeuN had decreased on day 30 (79.3%±2.8% vs 69.0%±3.4%, t =5.871, P < 0.01 )and day 45 (87.7% ±4.6% vs 78.3%±2.4%, t =4.403, P < 0.01) in PSD animals. There was a statistically significant increased proportion of BrdU-positive cells co-stained with GFAP in PSD animals compared with the ischemia ones on day 30 and45 (t =4.226, 8.945, both P < 0.01). Conclusion CMS following ischemia exerts an inhibitory effect on hippocampal survival of new born cells as well as their differentiation into neurons, while promotes new born cells differentiating into astrocytes.
7.Observation of the effect of alprostadil combined with valsartan in the treatment of chronic glomerulonephritis proteinuria
Xiaoyan XUAN ; Aicheng YANG ; Zian LIANG ; Jiefeng LI ; Baoan WANG
Chinese Journal of Primary Medicine and Pharmacy 2014;21(11):1676-1677
Objective To observe the clinical efficacy of alprostadil combined with valsartan in treatment of chronic glomerulonephritis proteinuria.Methods 78 patients with chronic glomerulonephritis proteinuria were randomly divided into two groups,and each group had 39 cases.The control group was given conventional treatment,while the observation group was given alprostadil combined with valsartan on the basis of the control group.The clinical outcomes were compared between the two groups.Results The total effective rate of the observation group was 92.31%,which was significantly higher than 74.36% of the control group (x2 =9.825,P < 0.05).After treatment,the 24h Upro,BUN and SCr of the observation group were (1.00 ± 0.39) g/24h,(7.11 ± 0.15) mmol/L and (80.86 ± 0.65) μmol/L,which were significantly lower than those of the control group [(1.30 ± 0.48) g/24h、(9.18 ± 2.21) mmol/L and (98.71 ± 4.34) μmol/L],the differences were statistically significant (t =9.32,7.83,7.12,all P < 0.05).Conclusion Alprostadil combined with valsartan in the treatment of chronic glomerulonephritis proteinuria has significant effect,and it can significantly alleviate clinical symptoms,improve renal function,which should be widely applied in clinical.
8.Research progress on myelodysplastic syndromes:report of the 2011 annual meeting of American Society of Hematology
Tiantian WANG ; Liting GUO ; Baoan CHEN ; Yunyu SUN
Journal of Leukemia & Lymphoma 2012;21(7):393-396,411
Myelodysplastic syndromes (MDS) are a heterogenous group of hematologic malignancies characterized by clonal expansion of BM myeloid cells with impaired differentiation. Of particular interest mutations is the recent recognition that genes involved in the regulation of histone function (EZH2, ASXL1,and UTX) and DNA methylation (DNMT3A,IDH 1/IDH2,TET2) are recurrently mutated in MDS,providing an important link between genetic and epigenetic alterations in this disease. Ongoing analysis of the seminal AZA-001study has taught many important lessons in the use of DNA methyltransferase (DNMT) inhibitors.Improved survival in patients with high-risk MDS treated with azacitidine extends to patients with any International Working Group-defined hematologic response.New information on the impact of DNMT inhibitors on the immune system and on stem cells will likely lead to novel uses of these drugs in MDS and other hematologic and nonhematologic malignancies. The immunomodulating drug thalidomide and its derivative lenalidomide have been used in the treatment of MDS,principally in lower-risk MDS.
9.Live-attenuated Salmonella typhimurium as a Oral Delivery Vehicle for SARS-CoV N DNA Vaccine
Hui HU ; Ling TAO ; Baoan CUI ; Yi HUANG ; Hanzhong WANG
China Biotechnology 2006;0(01):-
Objective:To investigate the immunogenicity of SARS-CoV N DNA vaccine and the feasibility of live-attenuated Salmonella typhimurium as the carrier to deliver the N DNA vaccine.Method:The recombinant attenuated salmonella strain CS022 harboring the pcDNA-N DNA vaccine was constructed.And mouse was immunized with the recombinant strain via intranasal and oral routes.Cellular and humoral immune responses were assessed by ELISA,lymphocyte proliferation assays,ELISPOT and FACS.Result:The oral immunization with the transformed salmonellae elicited strong immune responses mainly including high level of N-specific antibody,a dramatic activation of IFN-?-secreting cells,a high level of lymphocyte proliferation,and a high level of activated CD8+ T cells.Conclusion:Live-attenuated Salmonella typhimurium could effectively deliver the SARS-CoV N DNA vaccine in vivo.These encouraging pre-clinical data provide a rational basis for undertaking a new immune style to investigate SARS vaccine.
10.Effect of exogenous nitric oxide on Schistosoma japonicum in mice
Yanrui WANG ; Xinsheng ZHENG ; Baoan YAO ; Cunzheng DUAN
Chinese Journal of Schistosomiasis Control 1989;0(02):-
Objective To evaluate the effect of exogenous nitric oxide (NO) on Schistosoma japonicum in mice. Methods The emulsion of nitric oxide was performed by the reversal emulsifying method and beeswax was used as an agent absorbing NO. Each mice on the day 22 post-infection with cercariae was orally administracted with exogenous NO emulsion 0.5 ml once a day for five days. The worms of Schistosoma japonicum were collected by perfusion and counted to observe the effect of exogenous NO on Schistosoma japonicum. Results The concentration of exogenous NO was 536.2 ?mol/L. Exogenous NO could reach certain therapy efficacy with the worm reduction rate of 45.0% and the egg reduction rate of 42.7%. Conclusion Exogenous NO may be considered as a novel medicament of schistosomiasis japonica.