1.Purification and ATPase activity analysis of human PIF1 helicase participating in DNA repair
Chinese Journal of Endemiology 2009;28(3):276-279
Objective To clone, express and purify human PIF1 protein and analyze its ATPase activity. Methods The PIF1 cDNA was amplified by PCR from HeLa cell cDNA library and inserted to pET24b with histidine tag at its terminus to form pET24b-PIF1 plasmid. The recombinant pET24b-PIF1 plasmid was transformed to RosettaTM 2 (DE3) and the expression of PIF1 protein was monitored by SDS-PAGE analysis. By using fast protein liquid chromatograph (FPLC) system, the PIF1 protein was purified by affinity chromatograph and gel filtration. The ATPase activity of PIF1 was checked by thin layer chromatograph(TLC). Results The PIF1 protein was successfully cloned and expressed in E.coli. Conclusions The purification procedure of PIF1 protein was established using FPLC. The overexpressed and the purified PIF1 helicase has DNA and Mg2+ dependent ATPase activity.
2.Changes and correlation between the levels of plasma lysophosphatidic acid,acid phospholipid,low density lipoprotein and carotid artery atherosclerosis in patients with acute cerebral infarction
Zhiye WANG ; Zuonian ZHANG ; Wei GU
Journal of Clinical Neurology 2001;0(05):-
Objective To investigate the changes and the correlation between the levels of plasma lysophosphalidic acid(LPA),acid phospholipid(AP),low density lipoprotein(LDL) and carotid artery atherosclerosis in patients with acute cerebral infarction(ACI).Methods The levels of plasma LPA,AP and LDL were examined in 67 ACI patients and 30 normal controls,the carotid arteries were investigated with B mode ultrosand.Results The levels of plasma LPA,AP and LDL were higher in ACI group than those in the normal control group(all P
4.Association between IRF6 gene SNPs and non-syndromic cleft lip with or without cleft palate in Xinjiang Uyghur population
Ling WANG ; Li GU ; Wei AN ; Moming ADILI
Journal of Practical Stomatology 2015;(4):502-505
Objective:To study the association between rs2235371,rs2013162,rs2235377 SNPs in interferon regulatory factor 6 (IRF6)gene and non-syndromic cleft lip with or without cleft palate(NSCL/P)in Xinjiang Uyghur population.Methods:100 Uyghur NSCL/P patients from Xinjiang were included in the case group and 60 Uyghur inpatients with upper respiratory tract infection were se-lected in the control group.Next,generation sequencing was used,DNA sequencing results were compared with the information on the genome database and genetic analysis were made.Results:There were no significant differences in the frequency distribution of both genotypes and alles when the cases were campared with the controls at the rs2235371,rs2013162 and rs2235377 loci(P >0.05). Above three loci were located in the same block,rs2235371 and rs2235377 loci presents the strong linkage disequilibrium(r2 =0.949, D'=0.974).Possible haplotypes were:CCT >CAT >TAC,and there was no significant difference between the cases and controls in haplotype distribution(P >0.05).Conclusion:Polymorphisms of rs2235371,rs2013162 and rs2235377 in IRF6 gene may be associ-ated with NSCL/P in Xinjiang Uygur people.
6.Genetic analysis of thalassemia among Tujia and Miao people of reproductive age in Chongqing
Dan HE ; Huayan GU ; Haiyan ZHANG ; Wei WANG ; Xinyue WANG
Chinese Journal of Endemiology 2021;40(2):109-113
Objective:The aim of this study was to investigate the carrier ratio and the genotype of thalassemia among Tujia and Miao people of reproductive age in Chongqing.Methods:According to forward-looking design and multi-stage stratified cluster sampling method, fasting venous blood samples of Tujia and Miao people of reproductive age were collected from 11 survey sites in Chongqing from March to July 2019. Gap-PCR and high-throughput sequencing were used to screen thalassemia genes.Results:A total of 516 Tujia people (258 males, 258 females) and 270 Miao people (139 males, 131 females) were included in this study, and their age were (28.63 ± 5.26) and (28.62 ± 5.35) years, respectively. About 5.04% (26/516) Tujia people carried thalassemia gene, with 1.94% (10/516) and 2.52% (13/516) for α and β thalassemia, respectively. Three kinds of new variants (1 case of each variant), HBA 2: c.46G>A (Gly>Ser), HBB: c.*+129T>A and HBB: c.-39T>G with unclear pathogenicity, were identified in Tujia people. About 7.78% (21/270) Miao people carried thalassemia gene, among these, α and β thalassemia were 3.33% (9/270) and 4.44%(12/270), respectively. The most common mutation type of α-globin gene was -α 3.7/in the two ethnic groups. Three kinds of β-globin gene mutation types, Codons 41/42 (-TTCT) beta 0, Codon 17 (A>T) beta 0 and IVS-Ⅱ-654 (C>T) beta +, were the most common in Tujia people. Meanwhile, the chief β-globin gene mutation type was Codons 41/42 (-TTCT) beta 0 in Miao people. Conclusions:The carrying rate of thalassemia gene is higher in Tujia and Miao people in Chongqing, and the genotypes of thalassemia gene are different between Tujia and Miao people. The clinical significance of three kinds of new variants with unclear pathogenicity should be focused on.
7.Estrogen down-regulates the expression of high mobility group box 1 in human breast cancer cell line MCF-7
Rujun GU ; Dongliang LIU ; Yun MA ; Wei WANG
Basic & Clinical Medicine 2006;0(01):-
Objective To investigate the effects of estrogen and tamoxifen on the expression of high mobility group box 1(HMGB1) in human breast cancer cell line MCF-7.Methods MCF-7 cells were incubated with 17-beta E2 and tamoxifen at different concentrations for 4 days,respectively,with ethanol as control.Real time RT-PCR and Western blotting were used to detect the expression of HMGB1 at mRNA and protein level after different treatment of the cell line,respectively.Results Compared with ethanol control,HMGB1 mRNA and protein level were significantly decreased when 17-beta-E2 was added at concentrations of 10-6 and 10-4 mol/L(P
8.An Analysis on Nosocomial Infection after Surgical Operation
Xuefang WEI ; Yibo SHAO ; Lei ZHANG ; Chunhua WANG ; Youwei GU
Chinese Journal of Nosocomiology 2009;0(21):-
0.05);the main infection site was lower respiratory tract(61.97%),followed by surgical site infection(SSI)(23.94%).CONCLUSIONS In order to reduce postoperation nosocomial infection rate,we should take following measures:Improve pulmonary function,remove tracheal intubation as early as possible,shorten lasting time of surgery and days before operation,keep aseptic technique,strengthen nursing,enhance immunity,and use antimicrobial agents reasonably.
9.The expression of p53 in preventing restenosis after rabbit carotid artery balloon injury
Wei JIANG ; Shumei WANG ; Xinyuan GU ; Liangqiu TANG
Journal of Chinese Physician 2011;13(3):300-301,327
Objective To observe the effect of p53 protein on smooth muscle cell(VSMC)in rabbit artery balloon injury.Methods Restenosis model of carotid artery after balloon injury was established in rabbits.30 rabbits were divided into 2 groups,the sham group(n = 6)and the vascular injury group(n = 24).With H.E.staining and automatic image analysis system,we investigated artery morphology alteration and measured the area of arterial intima and media.The expressions of p53 protein were detected by immunohistochemical analysis.Results With H.E.staining and automatic image analysis and immunohistochemistry,the results showed that the expression of p53 was significantly reduced and the intima area was increased in model group compared with the sham group(P <0.01).But the expression of p53 in media was remarkably reduced compared with intima(P < 0.01).Conclusions The possible mechanism of preventing arterial restenosis in the balloon injury might be related with p53,which may be through inhibiting neointimal proliferation in arterial restenosis .
10.Influence of familial predisposition on the prognosis in young patients with colorectal cancer
Shilin WANG ; Guoli GU ; Yingqin DING ; Juxi OUYANG ; Xueming WEI
Chinese Journal of General Surgery 1997;0(04):-
Objective To investigate if the prognosis of young patients with colorectal cancer (CRC) is influenced by their familial predisposition. Methods The patients with CRC under 40 years old, underwent operation, had pathological diagnosis , family survey, followed up etc were selected into this research. According to familial predisposition they were divided into two groups: group A (with familial predisposition) and group B (without familial predisposition). Results Among the 44 patients twelve were in group A, thirty-two in group B. The 5-year survival rate in group A and B was 83.30%(10/12) and 59.38%(19/32) respectively,the difference was significant between the two groups(? 2=5.64,P