1.Effects and mechanisms of hepatitis B virus X protein on invasion and migration of hepatocellular carcinoma cells
Kuan CAO ; Zhongming BAO ; Xinyu ZHOU ; Guangyu JIA ; Bin ZHANG ; Quan WEN ; Renhao WANG
Chinese Journal of Digestive Surgery 2017;16(2):177-182
Objective To explore the effects and mechanisms of hepatitis B virus-X protein (HBx) on invasion and migration of hepatocellular carcinoma (HCC) cells.Methods The retrospective cohort study was conducted.The clinicopathological data of 30 patients with liver tumor (20 with HCC and 10 with benign tumor of liver) who were admitted to the Affiliated Hospital of Xuzhou Medical College between July 2014 and July 2015 were collected.HCC tissues of 20 patients with HCC (with history of HBV infection) were collected by surgical resection and peritumoral normal tissues (outside of tumor capsule) of 10 patients with benign tumor of liver (without history of HBV infection) were collected.The expressions of epidermal growth factor receptor 3 (ErbB3)in HCC tissues and peritumoral normal tissues were detected by immunohistochemistry (IHC).The relative expressions of ErbB3 and HBx in HCC tissues and peritumoral normal tissues were detected by Western blot,and relative expressions of ErbB3 in HepG2 of which green fluorescent protein (GFP) and GFP-HBx were respectively transfected were detected.The relative expressions of ErbB3 mRNA in HepG2 transfected by GFP and GFP-HBx were detected by real-time polymerase chain reaction (RT-PCR).The migration and invasion of HepG2 were respectively detected by Transwell assay with and without matrix.The measurement data with normal distribution were represented as $± s.The comparisons between groups were evaluated with the independent-sample t test.Correlation analysis was done by the Pearson test.Results (1) The expressions of ErbB3 were detected by IHC:relative value of mean optical density (MOD) of ErbB3 in HCC tissues of 20 patients with HCC and peritumoral normal tissues of 10 patients with benign tumor of liver were 2.54± 1.33 and O.99±0.29,respectively,with a statistically significant difference (t =6.542,P < 0.05).(2) The relative expressions of ErbB3 and HBx were detected by Western blot:relative expressions of ErbB3 and HBx were respectively 0.79±0.13,1.10±0.28 in HCC tissues of 10 patients with HCC and 1.07±0.17,0 in peritumoral normal tissues of 10 patients with benign tumor of liver,with statistically significant differences (t =3.229,19.486,P<0.05).The results of Pearson test showed that there was a positive correlation of expression between ErbB3 and HBx in HCC tissues (r=O.637,P< 0.05).(3) The relative expressions and transcriptional levels of ErbB3 were detected by Western blot and RT-PCR:relative expressions of ErbB3 in HepG2 of which GFP and GFP-HBx were respectively transfected were O.75±0.11 and 1.10±0.10,respectively,with a statistically significant difference (t=4.291,P<0.05).The relative expressions of ErbB3 mRNA in HepG2 of which GFP and GFP-HBx were respectively transfected were O.38±0.03 and O.94±0.07,respectively,with a statistically significant difference (t=11.703,P<O.05).(4) The effects of ErbB3 on migration and invasion of HepG2:numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay with matrix were respectively 271± 18 and 463± 31,respectively,with a statistically significant difference (t =8.202,P<0.05).Numbers of transmenbrane cell in HepG2 of which His and His-ErbB3 were respectively transfected by Transwell assay without matrix were respectively 315±38 and 549±34,respectively,with a statistically significant difference (t =8.310,P<0.05).Conclusion HBx protein can promote the invasion and migration of hepatocellular carcinoma cells through up-regulating expressions of ErbB3 protein.
2.Effect of Lotensin on inflammatory factors, vascular endothelial function and heart function in patients with acute myocardial infarction
Xiaogang JIA ; Sheng HU ; Zhongnan CAO ; Guoxing ZUO ; Kuan WANG ; Xinping DU
Chinese Journal of Biochemical Pharmaceutics 2017;37(8):228-230
Objective To investigate the effects of Lotensin on inflammatory factors, vascular endothelial function and heart function in patients with acute myocardial infarction. Methods 100 cases with acute myocardial infarction from March 2015 to January 2016 in the fifth central hospital of Tianjin were selected as the research object, which were randomly divided into the control group and the observation group. The control group were given routine treatment, at this basis, the observation group were given Lotensin. After treatment, the cardiac function, the levels of inflammatory factors, the blood vessel endothelial function, the serum NO and endothelin 1 and the therapeutic effect in the two groups were compared. Results LVESV, LVEDV (156.28±3.29、213.45±6.12) mL in the observation group were better than (162.98±4.16、202.83±7.16) mL in the control group (P<0.05). LVEF was (48.72± 2.13)% in the observation, which was higher than (40.62±3.29)% in the control group(P<0.05). Hs-CRP, IL-6 were (2.66±0.68) mg/L、(4.90±0.92) ng /L in the observation group , which were less than (6.35±1.50) mg/L、(9.38±2.01) ng/L in the control group (P<0.05). FMD(10.37±0.62)% in the observation group was bet er than (6.16±0.92)% in the control group (P<0.05)、 NO, ET-1 level (71.52±13.21) μmol/L、(56.27±7.10) ng/L in the observation group were bet er than (60.63 ±10.57) μmol/L、(69.72±9.50) ng/L in the control group (P<0.05). The total effective rate in the observation group was 94.00% (47/50), which was better than 62.00% (31/50) in the control group (P<0.05). Conclusion The effect is significant which Lotensin is used in the treatment of acute cerebral infarction, which can reduce inflammatory factors, improve endothelial function and cardiovascular function.
3.Therapeutic effect of human bone marrow mesenchymal stem cell lysates on rat arthritis induced by collagen.
Xiao-Fang CAO ; Yong-Qi WANG ; Zhi-Yong LI ; Zi-Kuan GUO ; Heng-Xiang WNAG
Journal of Experimental Hematology 2013;21(3):765-769
Our previous work has shown that mesenchymal stem cells (MSC) have little therapeutic effect on rat arthritis induced by collagen. This study was aimed to further investigate whether the MSC lysates exhibit beneficial effects on rheumatoid arthritis. Aliquots of cell lysates from 1×10(7) human bone marrow MSC were intraperitoneally injected into collagen-induced arthritis (CIA) Wistar rats weekly for 4 consecutive weeks. Methotrexate at a dose of 1 mg/kg or normal saline was served as positive and negative controls respectively. On week 4 the symptom scores were recorded and the hind joints of the rats were pathologically examined and X-ray examination was performed. The results showed that on week 4, the symptom scores of the rats that received MSC lysates (6.87 ± 0.83) and MTX (6.44 ± 1.13) were significantly lower than that of control rats (7.33 ± 0.77, P < 0.01). Meanwhile, pathological examination on the involved ankle showed that the synovitis and arthritis scores of MSC lysates and control groups were 2.28 ± 0.48 and 2.28 ± 0.55 respectively, significantly higher than that of MTX treatment rats (0.71 ± 0.48, P < 0.05). However, X-ray examination on the ankle joints showed that the injury score of control rats was 4 ± 0.57, greatly higher than those from MSC lysates (2.71 ± 0.75) and MTX treatment groups (2.57 ± 0.78, P < 0.05 for both groups). It is concluded that MSC lysate infusion has beneficial effects on CIA rat, but the effectiveness seems inferior to MTX.
Animals
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Arthritis, Experimental
;
chemically induced
;
therapy
;
Bone Marrow Cells
;
cytology
;
Cells, Cultured
;
Collagen
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Humans
;
Male
;
Mesenchymal Stromal Cells
;
cytology
;
Methotrexate
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pharmacology
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Rats
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Rats, Wistar
4.Study on the seropositive prevalence of human immunodeficiency virus in a village residents living in rural region of central China.
Hua CHENG ; Xu QIAN ; Guang-hua CAO ; Chang-kuan CHEN ; Yan-ning GAO ; Qing-wu JIANG
Chinese Journal of Epidemiology 2004;25(4):317-321
OBJECTIVETo study the human immunodeficiency virus (HIV) prevalence of residents living in a village located in rural region of central China where the paid blood donors used to be popular.
METHODSAll residents of one village "ZY" were asked to participate in HIV serologic screening test. Those over the age of 16 and participated in HIV serologic test was followed to undergo an indoor interview a week later, to finish a questionnaire, which including information on demography, marital status, smoking and drinking habits, history of blood donation, sexual behavior and history of drug use. Names on those either died from AIDS or having HIV serologic testing before were collected from local hospitals and the Center of Disease Control and Prevention.
RESULTSThere were in total 2364 residents in 527 households in "ZY" village including 1551 residents participated in the HIV serologic screening test, and 926 completed questionnaires. A total of 115 HIV seropositive residents were found in the HIV screening. 5 had been undergone HIV screening tests before, and 24 died of AIDS. The HIV prevalence of the whole population was 9.1% (CI: 7.7% - 10.6%). There were 19.6% households had at least one member living with HIV, out of which 25% had over two HIV seropositive members. No significant difference was found between genders with respect to HIV prevalence. 40 - 50 years old group had the highest HIV prevalence, significantly differed from other age groups. The HIV prevalence among former paid blood donors was 22.4%. The proportions of getting HIV infection through routes of blood transfusion, sex and mother-to-child, were 0.9%, 9.6% and 5.2% respectively among 115 HIV seropositive residents.
CONCLUSIONSome villages in the rural regions of central China showed high HIV prevalence. Former paid blood donation was responsible for the main transmission route in these villages. However, sexual transmission and mother-to-child transmission were becoming dominant routes in general population which called for effective action be taken to contain the spread of HIV.
Adolescent ; Adult ; Aged ; Blood Donors ; Child ; Child, Preschool ; China ; epidemiology ; Enzyme-Linked Immunosorbent Assay ; Female ; HIV Seropositivity ; diagnosis ; epidemiology ; HIV Seroprevalence ; Humans ; Infant ; Male ; Mass Screening ; Middle Aged ; Rural Population
5.Effects of tumor necrosis factor-α on immunoregulatory activities of mesenchymal stem cells in vitro and in vivo.
Yong-Qi WANG ; Zhi-Yong LI ; Xiao-Fang CAO ; Heng-Xiang WANG ; Zi-Kuan GUO
Journal of Experimental Hematology 2012;20(4):981-984
Mesenchymal stem cells (MSC) are characterized by their potent immuno-regulatory activity, however our previous data have shown that MSC have no therapeutic effects on collagen-induced arthritis (CIA). To further clarify the complexity, the effects of tumor necrosis factor-alpha (TNF-α) on the in vitro and in vivo immunoregulatory activity of MSC were investigated in this study, as TNF-α is recognized as the key factor in the development of rheumatoid arthritis. The nuclear translocation of the inflammation-associated factor NF-κB was observed after human umbilical cord MSC were treated with TNF-α and the cell proliferation status was assessed by MTT test. The inhibitory effects of MSC or TNF-α-treated MSC on the mixed lymphocyte reaction, in which Wistar rat spleen mononuclear cells were served as the responders and the splenocytes from SD rat spleens as the stimulators, were also determined by the MTT test. Further, the therapeutic potentials of MSC or TNF-α-treated MSC were observed in a Wistar rat CIA model. The results showed that NF-κB translocated into the nuclei promptly after TNF-α treatment, though TNF-α had little effect on the MSC proliferation. MSC, whether pre-stimulated by TNF-α or not and when different doses were tested, exhibited obviously inhibitory effects on the proliferation of the lymphocytes (P < 0.001 for all groups tested), while MSC-treated by TNF-α displayed more potent suppression especially when low-density were used. Unexpectedly, the infiltration of inflammatory cells into the involved knees was aggravated by cell treatment and the pathological scores were significantly higher than those of controls (P < 0.05). It is concluded that the TNF-α exhibits different effects on immune regulation activity of MSC, and its underlying mechanism needs to further investigate.
Animals
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Arthritis, Experimental
;
metabolism
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pathology
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Cell Proliferation
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drug effects
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Cells, Cultured
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Humans
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Male
;
Mesenchymal Stromal Cells
;
cytology
;
drug effects
;
immunology
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NF-kappa B
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metabolism
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Rats
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Rats, Sprague-Dawley
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Rats, Wistar
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Tumor Necrosis Factor-alpha
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pharmacology
6.Human bone marrow mesenchymal stem cells have little preventive or therapeutic effect on rat arthritis induced by collagen.
Yong-Qi WANG ; Xiao-Fang CAO ; Zi-Ming HE ; Feng-Jun XIAO ; Heng-Xiang WANG ; Zi-Kuan GUO
Journal of Experimental Hematology 2012;20(2):433-437
The aim of this study was to investigate if transfusion of mesenchymal stem cells (MSC) could exhibit beneficial effects on rheumatoid arthritis. Human bone marrow MSC were intraperitoneally injected into Wistar rats with collagen-induced arthritis at a dose of 10(7) on the next day (preventive group) or 2 weeks (treatment group) after collagen II induction, once a week for 2 weeks (preventive group) or 4 weeks (treatment group). The control group was given normal saline (NS) at corresponding time. The symptom scorings were documented weekly from the second week of the induction. On week 6, the hind joints of the rats were pathologically examined and the activation status of splenocytes was analyzed by flow cytometry. The results showed that all the rats developed arthritis and subsequent joint abnormality. On the sixth week, symptom scores of the rats that received MSC preventive (9.5 ± 0.5) or therapeutic (9.4 ± 0.6) infusions had no significant difference between each other, but were significantly greater than those of the NS controls (7.6 ± 0.6, P < 0.05). Consistently, pathological examination on the involved knees showed that the synovitis and arthritis scorings of MSC treated rats were greatly elevated compared with NS controls. Furthermore, the ratios of CD86(+) cells in the spleens of MSC prevention, MSC treatment and NS control groups were (4.16 ± 1.48), (4.06 ± 1.97) and (4.15 ± 2.04) respectively, while those of CD11b/c(+)CD86(+) cells were (1.04 ± 0.68), (0.95 ± 0.56) and (0.98 ± 0.44), all of which were significantly higher than those of healthy controls [(0.97 ± 0.18) and (0.30 ± 0.17), P < 0.05 for both parameters]. It is concluded that MSC infusion has little beneficial effects on collagen-induced arthritis in rats, conversely, MSC therapy aggravated the damage of the involved joints, its underlying mechanisms need to be further investigated.
Animals
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Arthritis, Experimental
;
pathology
;
prevention & control
;
therapy
;
Bone Marrow Cells
;
Cells, Cultured
;
Humans
;
Male
;
Mesenchymal Stem Cell Transplantation
;
Mesenchymal Stromal Cells
;
cytology
;
Rats
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Rats, Wistar
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Transplantation, Heterologous
;
Treatment Outcome
7.Expression of BATF, a member of the activator protein-1 family, in renal tissues of MRL/lpr mice.
Yusheng PENG ; Qian LI ; Menglei WANG ; Can CAO ; Kuan LAI ; Kang ZENG
Journal of Southern Medical University 2015;35(6):816-819
OBJECTIVETo investigate the expression of BATF, a member of the activator protein-1 family, in the renal tissues of mice with lupus nephritis.
METHODSThe renal tissues from 24-week-old female MRL/lpr mice and age- and sex-matched C57BL/6 mice were examined for BATF protein expressions using Western blotting and for expressions of BATF, RORγt and IL-17A mRNA using quantitative real-time PCR. The results of laboratory examinations were analyzed in relation with the histopathology of the mice.
RESULTSThe urinary protein and ds-DNA levels were significantly higher in MRL/lpr mice than in the control mice (P<0.05). Compared to normal control mice, MRL/lpr mice showed obvious glomerular fibrosis and inflammatory cell infiltrating with significantly increased BATF protein and mRNA expressions (P<0.05) and RORγt and IL-17 mRNA expressions in the renal tissues (P<0.05). In MRL/lpr mice, the expression of BATF mRNA was positively correlated with the RORγt and IL-17A mRNA expressions in the renal tissues.
CONCLUSIONThe renal expressions of BATF protein and mRNA is increased in MRL/lpr mice. BATF may participate in the immunopathogenesis of lupus nephritis by enhancing Thl7 cell response.
Animals ; Basic-Leucine Zipper Transcription Factors ; metabolism ; Blotting, Western ; Female ; Interleukin-17 ; metabolism ; Kidney ; metabolism ; Kidney Glomerulus ; pathology ; Lupus Nephritis ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred MRL lpr ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; metabolism ; RNA, Messenger ; Real-Time Polymerase Chain Reaction
8.Treatment of secondary intra-and extrahepatic cholangiolithiasis with electrohydraulic shock wave lithotripsy combined with laparoscope and choledochoscope
Bin ZHANG ; Kuan CAO ; Quan WEN ; Guang-Yu JIA ; Ren-Hao WANG
Journal of Regional Anatomy and Operative Surgery 2018;27(3):187-191
Objective To explore the clinical application value of electrohydraulic shock wave lithotripsy combined with laparoscope and choledochoscope in treatment of intra-and extrahepatic cholangiolithiasis.Methods The clinical data of 42 patients with intra-and extrahepatic cholangiolithiasis treated by laparoscope and choledochoscope(control group) in hepatobiliary surgery department of affiliated hospital of Xuzhou medical university from May 2012 to December 2015,and patients with intra-and extrahepatic cholangiolithiasis treated by electrohydraulic shock wave lithotripsy combined with laparoscope and choledochoscope (combined group) from January 2016 to February 2017,were retrospectively analyzed.The rate of transferring to laparotomy (transfer to laparotomy due to inflammatory adhesion or difficulty to remove),residual rate of choledocholithiasis,incidence of bile leakage and incidence of common bile duct stricture between two groups were compared.Results All patients in both groups were recovered.In the control group,there were 6 cases(6/42) transferred to laparotomy,among which 2 cases were transferred to laparotomy due to severe abdominal inflammatory adhesion and 4 cases were due to the difficulty to remove the calculus,without bile leakage or common bile duct stricture.There were 3 cases with residual choledocholithiasis,but without bile leakage or common bile duct stricture.In the combined group,there were 2 cases(2/42) transferred to laparotomy due to severe abdominal inflammatory adhesion and 5 cases used the technique of electrohydraulic shock wave lithotripsy without post-operation residual choledocholithiasis,bile leakage or bile duct stricture.In the control group,the residual choledocholithiasis was removed with T tube fistula choledochoscope.Conclusion In the process of minimally invasive surgery to treat intra-and extrahepatic cholangiolithiasis,electrohydraulic shock wave lithotripsy under laparoscope and choledochoscope can decrease the incidence of transferring to laparotomy due to the difficulty to remove calculus and the residual rate of calculus.It can be applied safely and efficiently to patients with intra-and extrahepatic cholangiolithiasis but with laparoscopic surgery indications.
9.Expression of BATF, a member of the activator protein-1 family, in renal tissues of MRL/lpr mice
Yusheng PENG ; Qian LI ; Menglei WANG ; Can CAO ; Kuan LAI ; Kang ZENG
Journal of Southern Medical University 2015;(6):816-819
Objective To investigate the expression of BATF, a member of the activator protein-1 family, in the renal tissues of mice with lupus nephritis. Methods The renal tissues from 24-week-old female MRL/lpr mice and age- and sex-matched C57BL/6 mice were examined for BATF protein expressions using Western blotting and for expressions of BATF, RORγt and IL-17A mRNA using quantitative real-time PCR. The results of laboratory examinations were analyzed in relation with the histopathology of the mice. Results The urinary protein and ds-DNA levels were significantly higher in MRL/lpr mice than in the control mice (P<0.05). Compared to normal control mice, MRL/lpr mice showed obvious glomerular fibrosis and inflammatory cell infiltrating with significantly increased BATF protein and mRNA expressions (P<0.05) and RORγt and IL-17 mRNA expressions in the renal tissues (P<0.05). In MRL/lpr mice, the expression of BATF mRNA was positively correlated with the RORγt and IL-17A mRNA expressions in the renal tissues. Conclusion The renal expressions of BATF protein and mRNA is increased in MRL/lpr mice. BATF may participate in the immunopathogenesis of lupus nephritis by enhancing Thl7 cell response.
10.Expression of BATF, a member of the activator protein-1 family, in renal tissues of MRL/lpr mice
Yusheng PENG ; Qian LI ; Menglei WANG ; Can CAO ; Kuan LAI ; Kang ZENG
Journal of Southern Medical University 2015;(6):816-819
Objective To investigate the expression of BATF, a member of the activator protein-1 family, in the renal tissues of mice with lupus nephritis. Methods The renal tissues from 24-week-old female MRL/lpr mice and age- and sex-matched C57BL/6 mice were examined for BATF protein expressions using Western blotting and for expressions of BATF, RORγt and IL-17A mRNA using quantitative real-time PCR. The results of laboratory examinations were analyzed in relation with the histopathology of the mice. Results The urinary protein and ds-DNA levels were significantly higher in MRL/lpr mice than in the control mice (P<0.05). Compared to normal control mice, MRL/lpr mice showed obvious glomerular fibrosis and inflammatory cell infiltrating with significantly increased BATF protein and mRNA expressions (P<0.05) and RORγt and IL-17 mRNA expressions in the renal tissues (P<0.05). In MRL/lpr mice, the expression of BATF mRNA was positively correlated with the RORγt and IL-17A mRNA expressions in the renal tissues. Conclusion The renal expressions of BATF protein and mRNA is increased in MRL/lpr mice. BATF may participate in the immunopathogenesis of lupus nephritis by enhancing Thl7 cell response.