Objective To investigate the expression of mRNA of α 1 adrenoceptor in hepatic tissues of cirrhotic patients with portal hypertension Methods Semi quantitative reverse transcription and polymerase chain rea...Objective To investigate the expression of mRNA of α 1 adrenoceptor in hepatic tissues of cirrhotic patients with portal hypertension Methods Semi quantitative reverse transcription and polymerase chain reaction (RT PCR) were used in 12 hepatic virus B related cirrhotic patients with portal hypertension and 15 controls from routine liver cutting biopsy for gallstone patients undergoing cholecystectomy who have no history of hypertension Special α 1 adrenoceptor subtype product and internal standard GAPDH product were amplified in the same RT PCR system simultaneously The ratio of their integral optical density (IOD) was calculated to stand for the relative expression of the α 1 adrenoceptor subtypes Results The relative mRNA expression of α 1a adrenoceptor subtype in hepatic tissues of cirrhotic patients Department of Surgery, People's Hospital, Beijing Medical University, Beijing 100044, China (Zhu JY, Chen L, Leng XS and Du RY) This work was supported by the National Natural Science Foundation and the Scientific Research Foundation of the Ministry of Health.(0 26±0 12) was significantly lower than that of controls (0 86±0 38, P <0 01) The relative mRNA expression of α 1b adrenoceptor subtype in hepatic tissues of cirrhotic patients (0 03±0 01) was significantly lower than that of controls (0 23±0 10, P <0 01) There was no expression of α 1d adrenoceptor subtype in hepatic tissues of either cirrhotic patients or controls The relative mRNA expression of α 1a adrenoceptor subtype is significantly higher than that of α 1b adrenoceptor subtype in hepatic tissues of both cirrhotic patients and controls ( P <0 01) Conclusion α 1a adrenoceptor mRNA expression predominated in both normal and cirrhotic hepatic tissues, came in second the expression of α 1b adrenoceptor and no expression of α 1d adrenoceptor The decreased expression of mRNA of α 1a and α 1b adrenoceptor may underlie the decrease of α 1 adrenoceptor proteins often seen in the hepatic tissues of cirrhotic patients, and may play an important role in the pathogenesis and maintenance of portal hypertension展开更多
AIM:To determine the utility of connective tissue growth factor(CCN2/CTGF) for assessing hepatic fibrosis in hepatitis B virus(HBV)-induced chronic liver diseases(CLD-B).METHODS:Enzyme-linked immunosorbent assay was u...AIM:To determine the utility of connective tissue growth factor(CCN2/CTGF) for assessing hepatic fibrosis in hepatitis B virus(HBV)-induced chronic liver diseases(CLD-B).METHODS:Enzyme-linked immunosorbent assay was used to measure CCN2 in sera from 107 patients with chronic hepatitis B(CHB) and 39 patients with HBVinduced active liver cirrhosis and 30 healthy individuals.Liver samples from 31 patients with CHB,8 patients with HBV-induced liver cirrhosis and 8 HBV carriers with normal liver histology were examined for transforming growth factor β-1(TGF-β1) or CCN2 mRNA levels by in situ hybridization,and computer image analysis was performed to measure integrated optimal density(IOD) of CCN2 mRNA-positive cells in liver tissues.Histological inflammation grading and fibrosis staging were evaluated by H and E staining and Van Gieson's method.RESULTS:Serum CCN2 concentrations were,respectively,4.0-or 4.9-fold higher in patients with CHB or active liver cirrhosis as compared to healthy individuals(P < 0.01).There was good consistency between the levels of CCN2 in sera and CCN2 mRNA expression in liver tissues(r = 0.87,P < 0.01).The levels of CCN2 in sera were increased with the enhancement of histological fibrosis staging in patients with CLD-B(r = 0.85,P < 0.01).Serum CCN2 was a reliable marker for the assessment of liver fibrosis,with areas under the receiver operating characteristic(ROC) curves(AUC) of 0.94 or 0.85 for,respectively,distinguishing normal liver controls from patients with F1 stage liver fibrosis or discriminating between mild and significant fibrosis.CONCLUSION:Detection of serum CCN2 in patients with CLD-B may have clinical significance for assessment of severity of hepatic fibrosis.展开更多
Objective To observe the changes in hepatic histomorphology and the differences in the expression of PD-1 after different moxibustion interventions in the immunosuppressed rabbits caused by cyclophosphamide and analyz...Objective To observe the changes in hepatic histomorphology and the differences in the expression of PD-1 after different moxibustion interventions in the immunosuppressed rabbits caused by cyclophosphamide and analyze the intervention effects of different moxibustion therapies.Methods A total of 40 big-ear white rabbits were randomized into 4 groups,i.e.a blank group,an immunosuppressed model group,a moxibustion group and a herbal-paste isolated moxibustion group,10 rabbits in each one.In the immunosuppressed model group,the moxibustion group and the herbal-paste isolated moxibustion group,the intraperitoneal injection with cyclophosphamide was administered consecutively for 7 days,60 mg/kg,and then,the immunosuppressed models were prepared.After the models were established successfully,in the moxibustion group and the herbal-paste isolated moxibustion group,moxibustion was administer at"Shénquè(神阙CV 8)""Guānyuán(关元CV 4)""Zúsānlĭ(足三里ST 36)""Píshū(脾俞BL20)"and"Shènshū(肾俞BL23)",once every other day,10 times in total.In the blank group and the immunosuppressed model group,the rabbits were restricted on the rabbit platform at the same time periods,without intervention.On the second day after the end of intervention,the animals were weighed and anesthetized,and the livers were extracted.After routine HE staining,the changes of hepatic histomorphology were observed under the microscope and the morphometric analysis was conducted in each group.Besides,PD-1 expression was observed with immunohistochemical method.Result Compared with the blank group,after modeling of each animal in three groups,the body mass decreased obviously(all P<0.01),the hepatocyte count decreased(all P<0.01),the inflammatory cellular infiltration area was increased obviously(all P<0.01),necrosis appeared in hepatic tissue and the positive expression of PD-1 in hepatic tissue was significantly increased(all P<0.01).After moxibustion intervention,compared with the immunosuppressed model group,the body masses of the animals were increased obviously(P<0.05,P<0.01),the hepatocyte count was increased obviously(both P<0.01),the infiltration area of inflammatory cells decreased significantly(both P<0.01)and the expression of the PD-1 decreased in the hepatic tissue(both P<0.05)in both the moxibustion group and the herbal-paste isolated moxibustion group.Compared with the moxibustion group,the animal body mass was higher obviously(P<0.01),the hepatocyte count was raised apparently(P<0.01)and the infiltration area of inflammatory cells were reduced obviously(P<0.05)in the herbal-paste isolated moxibustion group.Conclusion The intervention with different moxibustion therapies improves the condition of body mass reduction and hepatic tissue damage of the immunosuppressed rabbits caused by cyclophosphamide.The effect of herbal-paste isolated moxibustion therapy is better than that of simple moxibustion therapy.展开更多
Objective: To study the relationship between the de- gree of hepatic fibrosis and serum fibrosis markers. Methods: Liver biopsies were performed in 67 pa- tients with hepatitis. The sections were stained with hematoxy...Objective: To study the relationship between the de- gree of hepatic fibrosis and serum fibrosis markers. Methods: Liver biopsies were performed in 67 pa- tients with hepatitis. The sections were stained with hematoxylin eosin and immunohistochemical stain. Staging of hepatic fibrosis was made microscopically. The serum levels of hyaluronic acid (HA), type Ⅲ procollagen (PC-Ⅲ), laminin (LN), and type Ⅳ collagen (Ⅳ-C) were measured by radioimmunoas- say. Results: The serum levels of HA, PC-Ⅲ, LN and Ⅳ-C were elevated from S1 to S4 because of the in- crease of hepatic fibrosis. The serum concentrations of HA, PC-Ⅲ, LN and Ⅳ-C were increased with the progress of disease, with the highest concentration at the stage of cirrhosis. Conclusion: The stages of hepatic fibrosis are corre- lated with the serum levels of HA, PC-Ⅲ, LN and Ⅳ-C, which as markers may play a role in detecting the degree of hepatic fibrosis.展开更多
AIM: DMA mismatch repair (MMR) is an important mechanism for maintaining fidelity of genomic DNA. Abnormalities in one or more MMR genes are implicated in the development of many cancers. We investigated the role of e...AIM: DMA mismatch repair (MMR) is an important mechanism for maintaining fidelity of genomic DNA. Abnormalities in one or more MMR genes are implicated in the development of many cancers. We investigated the role of expression of MMR genes (hMLH1, hPMS1, hPMS2, GTBP/hMSH6, hMSH2) in hepatocellular carcinogenesis. METHODS: We evaluated the expression level of MMR genes in 33 hepatocellular carcinoma (HCC) cases using the multiplex reverse transcription (RT) PCR assays, as well as in 16 cases of normal adjacent hepatic tissues. β-actin gene was used as an internal control and calibrator for quantification of gene expression. RESULTS: Out of the 33 studied cases, 25 were HCV positive and 30 (90.9%) showed reduced expression in one or more of the studied MMR genes. Reduced expression was found in hMSH2(71.9%), hMLH1 (53.3%), GTBP(51.1%), hPMS2 (33.3%) and hPMS1 (6%). A significant correlation was found between reduced expression of hPMS2(P= 0.0069) and GTBP(P= 0.0034), hPMS2 and non-cirrhosis (P= 0.0197), hMLH1 and high grade. On the other hand, 57.1%, 50%, 20%, 18.8%, and 6% of the normal tissues distant to tumors showed reduced expression of hMSH2, hMLH1, GTBP, hPMS2, and hPMS1 respectively. Multivariate analysis revealed a significant correlation between the expression level of hMSH2(P= 0.008), hMLH1 (P= 0.001) and GTBP (P= 0.032) and HCC, between hPMS2, GTBP and HCV-associated HCC (P<0.001, 0.002). CONCLUSION: Reduced expression of MMR genes seems to play an important role in HCV-associated HCC. hPMS2 is likely involved at an early stage of hepatocarcinogenesis since it was detected in normal adjacent tissues. Reduced expression of hPMS2 provides a growth advantage and stimulates proliferation which encourages malignant transformation in non-cirrhotic HCV-infected patients via acquisition of more genetic damages.展开更多
文摘Objective To investigate the expression of mRNA of α 1 adrenoceptor in hepatic tissues of cirrhotic patients with portal hypertension Methods Semi quantitative reverse transcription and polymerase chain reaction (RT PCR) were used in 12 hepatic virus B related cirrhotic patients with portal hypertension and 15 controls from routine liver cutting biopsy for gallstone patients undergoing cholecystectomy who have no history of hypertension Special α 1 adrenoceptor subtype product and internal standard GAPDH product were amplified in the same RT PCR system simultaneously The ratio of their integral optical density (IOD) was calculated to stand for the relative expression of the α 1 adrenoceptor subtypes Results The relative mRNA expression of α 1a adrenoceptor subtype in hepatic tissues of cirrhotic patients Department of Surgery, People's Hospital, Beijing Medical University, Beijing 100044, China (Zhu JY, Chen L, Leng XS and Du RY) This work was supported by the National Natural Science Foundation and the Scientific Research Foundation of the Ministry of Health.(0 26±0 12) was significantly lower than that of controls (0 86±0 38, P <0 01) The relative mRNA expression of α 1b adrenoceptor subtype in hepatic tissues of cirrhotic patients (0 03±0 01) was significantly lower than that of controls (0 23±0 10, P <0 01) There was no expression of α 1d adrenoceptor subtype in hepatic tissues of either cirrhotic patients or controls The relative mRNA expression of α 1a adrenoceptor subtype is significantly higher than that of α 1b adrenoceptor subtype in hepatic tissues of both cirrhotic patients and controls ( P <0 01) Conclusion α 1a adrenoceptor mRNA expression predominated in both normal and cirrhotic hepatic tissues, came in second the expression of α 1b adrenoceptor and no expression of α 1d adrenoceptor The decreased expression of mRNA of α 1a and α 1b adrenoceptor may underlie the decrease of α 1 adrenoceptor proteins often seen in the hepatic tissues of cirrhotic patients, and may play an important role in the pathogenesis and maintenance of portal hypertension
基金Supported by National Natural Scientific Foundation,No. 30872236,81070370(to Gao RP)NIH 5R01AA016003 to (Brigstock D)
文摘AIM:To determine the utility of connective tissue growth factor(CCN2/CTGF) for assessing hepatic fibrosis in hepatitis B virus(HBV)-induced chronic liver diseases(CLD-B).METHODS:Enzyme-linked immunosorbent assay was used to measure CCN2 in sera from 107 patients with chronic hepatitis B(CHB) and 39 patients with HBVinduced active liver cirrhosis and 30 healthy individuals.Liver samples from 31 patients with CHB,8 patients with HBV-induced liver cirrhosis and 8 HBV carriers with normal liver histology were examined for transforming growth factor β-1(TGF-β1) or CCN2 mRNA levels by in situ hybridization,and computer image analysis was performed to measure integrated optimal density(IOD) of CCN2 mRNA-positive cells in liver tissues.Histological inflammation grading and fibrosis staging were evaluated by H and E staining and Van Gieson's method.RESULTS:Serum CCN2 concentrations were,respectively,4.0-or 4.9-fold higher in patients with CHB or active liver cirrhosis as compared to healthy individuals(P < 0.01).There was good consistency between the levels of CCN2 in sera and CCN2 mRNA expression in liver tissues(r = 0.87,P < 0.01).The levels of CCN2 in sera were increased with the enhancement of histological fibrosis staging in patients with CLD-B(r = 0.85,P < 0.01).Serum CCN2 was a reliable marker for the assessment of liver fibrosis,with areas under the receiver operating characteristic(ROC) curves(AUC) of 0.94 or 0.85 for,respectively,distinguishing normal liver controls from patients with F1 stage liver fibrosis or discriminating between mild and significant fibrosis.CONCLUSION:Detection of serum CCN2 in patients with CLD-B may have clinical significance for assessment of severity of hepatic fibrosis.
基金Supported by National Natural Science Foundation of China:81674062。
文摘Objective To observe the changes in hepatic histomorphology and the differences in the expression of PD-1 after different moxibustion interventions in the immunosuppressed rabbits caused by cyclophosphamide and analyze the intervention effects of different moxibustion therapies.Methods A total of 40 big-ear white rabbits were randomized into 4 groups,i.e.a blank group,an immunosuppressed model group,a moxibustion group and a herbal-paste isolated moxibustion group,10 rabbits in each one.In the immunosuppressed model group,the moxibustion group and the herbal-paste isolated moxibustion group,the intraperitoneal injection with cyclophosphamide was administered consecutively for 7 days,60 mg/kg,and then,the immunosuppressed models were prepared.After the models were established successfully,in the moxibustion group and the herbal-paste isolated moxibustion group,moxibustion was administer at"Shénquè(神阙CV 8)""Guānyuán(关元CV 4)""Zúsānlĭ(足三里ST 36)""Píshū(脾俞BL20)"and"Shènshū(肾俞BL23)",once every other day,10 times in total.In the blank group and the immunosuppressed model group,the rabbits were restricted on the rabbit platform at the same time periods,without intervention.On the second day after the end of intervention,the animals were weighed and anesthetized,and the livers were extracted.After routine HE staining,the changes of hepatic histomorphology were observed under the microscope and the morphometric analysis was conducted in each group.Besides,PD-1 expression was observed with immunohistochemical method.Result Compared with the blank group,after modeling of each animal in three groups,the body mass decreased obviously(all P<0.01),the hepatocyte count decreased(all P<0.01),the inflammatory cellular infiltration area was increased obviously(all P<0.01),necrosis appeared in hepatic tissue and the positive expression of PD-1 in hepatic tissue was significantly increased(all P<0.01).After moxibustion intervention,compared with the immunosuppressed model group,the body masses of the animals were increased obviously(P<0.05,P<0.01),the hepatocyte count was increased obviously(both P<0.01),the infiltration area of inflammatory cells decreased significantly(both P<0.01)and the expression of the PD-1 decreased in the hepatic tissue(both P<0.05)in both the moxibustion group and the herbal-paste isolated moxibustion group.Compared with the moxibustion group,the animal body mass was higher obviously(P<0.01),the hepatocyte count was raised apparently(P<0.01)and the infiltration area of inflammatory cells were reduced obviously(P<0.05)in the herbal-paste isolated moxibustion group.Conclusion The intervention with different moxibustion therapies improves the condition of body mass reduction and hepatic tissue damage of the immunosuppressed rabbits caused by cyclophosphamide.The effect of herbal-paste isolated moxibustion therapy is better than that of simple moxibustion therapy.
文摘Objective: To study the relationship between the de- gree of hepatic fibrosis and serum fibrosis markers. Methods: Liver biopsies were performed in 67 pa- tients with hepatitis. The sections were stained with hematoxylin eosin and immunohistochemical stain. Staging of hepatic fibrosis was made microscopically. The serum levels of hyaluronic acid (HA), type Ⅲ procollagen (PC-Ⅲ), laminin (LN), and type Ⅳ collagen (Ⅳ-C) were measured by radioimmunoas- say. Results: The serum levels of HA, PC-Ⅲ, LN and Ⅳ-C were elevated from S1 to S4 because of the in- crease of hepatic fibrosis. The serum concentrations of HA, PC-Ⅲ, LN and Ⅳ-C were increased with the progress of disease, with the highest concentration at the stage of cirrhosis. Conclusion: The stages of hepatic fibrosis are corre- lated with the serum levels of HA, PC-Ⅲ, LN and Ⅳ-C, which as markers may play a role in detecting the degree of hepatic fibrosis.
文摘AIM: DMA mismatch repair (MMR) is an important mechanism for maintaining fidelity of genomic DNA. Abnormalities in one or more MMR genes are implicated in the development of many cancers. We investigated the role of expression of MMR genes (hMLH1, hPMS1, hPMS2, GTBP/hMSH6, hMSH2) in hepatocellular carcinogenesis. METHODS: We evaluated the expression level of MMR genes in 33 hepatocellular carcinoma (HCC) cases using the multiplex reverse transcription (RT) PCR assays, as well as in 16 cases of normal adjacent hepatic tissues. β-actin gene was used as an internal control and calibrator for quantification of gene expression. RESULTS: Out of the 33 studied cases, 25 were HCV positive and 30 (90.9%) showed reduced expression in one or more of the studied MMR genes. Reduced expression was found in hMSH2(71.9%), hMLH1 (53.3%), GTBP(51.1%), hPMS2 (33.3%) and hPMS1 (6%). A significant correlation was found between reduced expression of hPMS2(P= 0.0069) and GTBP(P= 0.0034), hPMS2 and non-cirrhosis (P= 0.0197), hMLH1 and high grade. On the other hand, 57.1%, 50%, 20%, 18.8%, and 6% of the normal tissues distant to tumors showed reduced expression of hMSH2, hMLH1, GTBP, hPMS2, and hPMS1 respectively. Multivariate analysis revealed a significant correlation between the expression level of hMSH2(P= 0.008), hMLH1 (P= 0.001) and GTBP (P= 0.032) and HCC, between hPMS2, GTBP and HCV-associated HCC (P<0.001, 0.002). CONCLUSION: Reduced expression of MMR genes seems to play an important role in HCV-associated HCC. hPMS2 is likely involved at an early stage of hepatocarcinogenesis since it was detected in normal adjacent tissues. Reduced expression of hPMS2 provides a growth advantage and stimulates proliferation which encourages malignant transformation in non-cirrhotic HCV-infected patients via acquisition of more genetic damages.
