1.The effects of DNA repair enzyme1 hMTH1 on HBx-induced malignant transformation of L02 cells
Bin CHENG ; Yaochu ZHENG ; Xiaorong GUO ; Songting LIN ; Peiyuan LI
Chinese Journal of Internal Medicine 2008;47(10):830-833
Objective To study the effects of the HBV x gene (HBx) on the biological characteristics and the expression of DNA repair enzyme hMTH1 mRNA of the L02/HBx transgene cell model. Methods Light microscopy was used to observe the morphologic characteristics of gene-transfected cell strain Lff2/HBx that stably expressed the HBx protein and the control groups of L02 and L02/PcDNA3.1. The changes of L02/HBx on the proliferation, cell cycle and apoptosis were observed by MTT assays and flow cytometry analysis respectively. Moreover, the malignant transformation of L02/HBxwas assayed by colony formation in soft agar and the expression of DNA repair enzyme hMTH1 mRNA was assayed in each group by real-time qPCR. Results Inversion phase contrast microscope showed that the morphologic characteristics of L02/HBx had changed obviously compared with control groups. The MTT showed that L02/HBx proliferated more quickly and flow cytometry analysis indicated that HBx could accelerate the progression of cell cycle and inhibit apoptosis. Colony formation in soft agar demonstrated that the rate of colony formation of L02/HBx was remarkably higher than the L02 and the L02/peDNA3. 1 cells (P<0. 05). The real-time qPCR detection showed that the expression of hMTH1 mRNA in L02/HBx was significantly higher than that in the control groups ( P < 0. 05 ). Conclusion HBx could play an important role in the malignant transformation of L02/HBx and the over expression of hMTH1 mRNA.
2.The effects of HBx gene on the expression of DNA repair enzymes hOGG1 and hMYHalpha mRNA in HepG2 cells.
Bin, CHENG ; Xiaorong, GUO ; Yaochu, ZHENG ; Ying, WANG ; Chunyan, LIU ; Peiyuan, LI
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(2):187-92
To observe the alteration in the expression of DNA repair enzymes hOGG1 and hMYHalpha and the change in 8-OHdG levels in the HBx gene-transfected cells HepG2/HBx and to explore the mechanisms of the HBV-associated hepatocellular carcinoma, the gene-transfected cells HepG2/HBx which stably expressed HBx was established, and the effect of HBx on the cell cycle and proliferation of HepG2 was examined. By using the beta-actin as the interior control, real-time polymerase chain reaction (Real-time qPCR) was employed to quantitatively detect the expression of DNA repair enzymes hOGG1 and hMYHalpha in the HepG2/HBx, the control cells HepG2 and HepG2 transfected with pcDNA3.1 vector (HepG2/pDNA3.1). The 8-OHdG levels were determined by HPLC/ECD in the established gene-transfected cells HepG2/HBx and the control cells HepG2 and HepG2/pcDNA3.1. Our results showed that the expression of DNA repair enzyme hMYHalpha in the HepG2/HBx (0.021+/-0.007) was significantly lower than that of HepG2 (0.099+/-0.041) (P<0.05) and HepG2/pDNA3.1 (0.121+/-0.005) (P<0.05). However, the no significant differences existed in the expression of DNA repair enzyme hOGG1 among the three cell strains (P>0.05). The 8-OHdG level in the HepG2/HBx was significantly higher than that in HepG2 and HepG2/pcDNA3.1 (P<0.05). It is concluded that HBx gene may inhibit the expression of DNA repair enzyme hMYHalpha mRNA to impair the ability to repair the intracellular DNA oxidative damage, to increase the oxidative DNA-adduct 8-OHdG and to affect the nucleotide excision repair function, thus participate in the occurrence and development of hepatocellular carcinoma.
DNA Glycosylases/genetics
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DNA Glycosylases/*metabolism
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Deoxyguanosine/analogs & derivatives
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Deoxyguanosine/metabolism
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Hep G2 Cells
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RNA, Messenger/genetics
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RNA, Messenger/metabolism
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Trans-Activators/*genetics
3.The therapeutic efficacy of Shixiaosan and Siwu decoction combined with rehabilitation intervention used for pelvic floor dysfunction
China Modern Doctor 2014;(27):51-53
Objective To explore the therapeutic efficacy of Shixiaosan and Siwu decoction combined with rehabilita-tion intervention used for pelvic floor dysfunction. Methods The 126 cases were randomly divided into test and control groups. There were 63 patients in every group. The control group accepted only two courses of pelvic floor rehabilitation therapy. However treatment of traditional Chinese medicine combined with two courses of pelvic floor rehabilitation therapy were used in the test group. Then the therapeutic effect of all cases was estimated. The incidence of stress in-continence was investigated after one year. Results The amount of pelvic floor muscle strength up to Ⅲ in the test group was 58 cases, however only 41 cases in the control group. There were 5 cases suffered stress incontinence, and the incidence was 7.9% in the test group. But there were 14 cases suffered stress incontinence, and the incidence was 22.2% in the control group. 3 cases occurred anorectum dysfunction, and the incidence was 4.8% in the test group,however there were 11 cases occurred anorectum dysfunction, and the incidence was 17.5% in the control group. 4 cases suffered pelvic organ prolapse,and the incidence was 6.3% in the test group,while there were 13 cases suffered pelvic organ prolapse, and the incidence was 20.6% in the control group. The difference between the two groups was statistically significant (P<0.05). Conclusion Our study indicated that the therapeutic efficacy of Shixiao san and Siwu decoction combined with rehabilitation intervention used for pelvic floor dysfunction was significantly better than simple rehabilitation intervention.
4.The Effects of HBx Gene on the Expression of DNA Repair Enzymes hOGG1 and hMYHα mRNA in HepG2 Cells
CHENG BIN ; GUO XIAORONG ; ZHENG YAOCHU ; WANG YING ; LIU CHUNYAN ; LI PEIYUAN
Journal of Huazhong University of Science and Technology (Medical Sciences) 2009;29(2):187-192
To observe the alteration in the expression of DNA repair enzymes hOGG1 and hMYHα and the change in 8-OHdG levels in the HBx gene-transfected cells HepG2/HBx and to explore the mechanisms of the HBV-associated hepatocellular carcinoma,the gene-transfected cells HepG2/HBx which stably expressed HBx was established,and the effect of HBx on the cell cycle and proliferation of HepG2 was examined.By using the β-actin as the interior control,real-time polymerase chain reaction (Real-time qPCR) was employed to quantitatively detect the expression of DNA repair enzymes hOGG1 and hMYHα in the HepG2/HBx,the control cells HepG2 and HepG2 transfected with pcDNA3.1 vector (HepG2/pDNA3.1).The 8-OHdG levels were determined by HPLC/ECD in the established gene-transfected cells HepG2/HBx and the control cells HepG2 and HepG2/pcDNA3.1.Our results showed that the expression of DNA repair enzyme hMYHα in the HepG2/HBx (0.021±0.007) was significantly lower than that of HepG2 (0.099±0.041) (P<0.05) and HepG2/pDNA3.1 (0.121±0.005) (P<0.05).However,the no significant differences existed in the expression of DNA repair enzyme hOGG1 among the three cell strains (P>0.05).The 8-OHdG level in the HepG2/HBx was significantly higher than that in HepG2 and HepG2/pcDNA3.1 (P<0.05).It is concluded that HBx gene may inhibit the expression of DNA repair enzyme hMYHα mRNA to impair the ability to repair the intracellular DNA oxidative damage,to increase the oxidative DNA-adduct 8-OHdG and to affect the nucleotide excision repair function,thus participate in the occurrence and development of hepatocellular carcinoma.
5.The effect of acupuncture combined with clean intermittent catheteriza-tion used for neurogenic bladder
Yuehong XU ; Yueyuan XU ; Yuemei XU ; Yaochu ZHENG ; Aimei HU ; Cuiping YU ; Wenjuan HUANG ; Aixiu HONG ; Weifang QIU
China Modern Doctor 2014;(29):149-151
Objective To study the effect of acupuncture combined with clean intermittent catheterization in the treat-ment of neurogenic bladder. Methods A total of 46 patients were randomly divided into test and control groups. There were 23 patients in the each group. The clean intermittent catheterization only used in the control group. However inte-grated acupuncture and clean intermittent catheterization were used in the test group. Final the residual urine volume and the incidence of bacteriuria were researched after 5 months. Results There were 17 cases restored the micturating function in the test group, however only 7 cases in the control group. There were 3 cases suffered bacteriuria in the test group, but there were 11 cases suffered bacteriuria in the control group. The difference between the two groups was sta-tistical ly significantly (P<0.05). Conclusion Combined acupuncture with clean intermittent catheterization can effi-ciently maintain the normal function of bladder, decrease the residual urine volume, and reduce the incidence of bac-teriuria.