Danqi Tablet () Regulates Energy Metabolism in Ischemic Heart Rat Model through AMPK/SIRT1-PGC-1α Pathway.

OBJECTIVE: To investigate the cardioprotective effect of Danqi Tablet (DQT, ) on ischemic heart model rats and the regulative effect on energy metabolism through peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α). METHODS: Rat ischemic heart model was induced by ligation of left anterior descending coronary artery. Totally 40 Sprague-Dawley rats were randomly divided into sham group, model group, DQT group (1.5 mg/kg daily) and trimetazidine (TMZ) group (6.3 mg/kg daily) according to a random number table, 10 rats in each group. Twenty-eight days after continuous administration, cardiac function was assessed by echocardiography and the structures of myocardial cells were observed by hematoxylin-eosin staining. The level of adenosine triphosphate (ATP) in myocardial cells was measured by ATP assay kit. Expressions level of key transcriptional regulators, including PGC-1α, Sirtuin 1 (SIRT1), AMP-activated protein kinase (AMPK), and downstream targets of PGC-1α, such as mitofusin 1 (MFN1), mitofusin 2 (MFN2) and superoxide dismutase 2 (SOD2) were measured by Western blot. Expression level of PGC-1α was examined by immunohistochemical staining. RESULTS: The rat ischemic heart model was successfully induced and the heart function in model group was compromised. Compared with the model group, DQT exerted cardioprotective effects, up-regulated the ATP production in myocardial cells and inhibited the infiltration of inflammatory cells in the margin area of infarction of the myocardial tissues (P<0.01). The expressions of PGC-1α, SIRT1 and AMPK were increased in the DQT group (all P<0.05). Furthermore, the downstream targets, including MFN1, MFN2 and SOD2 were up-regulated (P<0.05 or P<0.01). Compared with the TMZ group, the expression levels of PGC-1α, MFN1 and SOD2 were increased by DQT treatment (P<0.05 or P<0.01). CONCLUSION: DQT regulated energy metabolism in rats with ischemic heart model through AMPK/SIRT1 -PGC-1α pathway. PGC-1α might serve as a promising target in the treatment of ischemic heart disease.

[Mechanisms of Fufang Danshen tablets on treating heart failure via regulating AT1-mediated PLA2-COX2/5-LOX metabolic pathway].

Myocardial fibrosis (MF) is an important pathological change involved in the progress from myocardial infarction (MI) to heart failure(HF). Metabolic disorder of arachidonic acid (AA) in cardiomyocytes plays an important role in process of MF. Fufang Danshen tablets is a traditional Chinese medicine (TCM), which showed significant effect on coronary heart diseases and anti-MF. However, the underlying mechanism of anti-MF remains unclear. In this study, HF animal model of myocardial infarction was established by ligation of left anterior descending coronary artery. The heart function of rats in each group was evaluated by echocardiography and hemodynamic measurement. Histological examination, TUNEL and Western blot were used to detect the levels of MF and proteins related to AA metabolism. As a result, MI significantly decreased the levels of ejection fraction (EF), ejection fraction (FS) and left ventricular systolic pressure (LVSP), and these decreases were significantly improved by the treatment of Fufang Danshen tablets. Besides, Fufang Danshen tablets treatment down-regulated the levels of creatine kinase-MB (CK-MB) and lactate dehydrogenase (LDH) in serum. HE, Masson and TUNEL staining results showed that Fufang Danshen tablets treatment could inhibit the inflammatory cells infiltration and attenuate the fibrosis and apoptosis to exert cardioprotective effect. Western blot indicated that Fufang Danshen tablets treatment down-regulated the expressions of AT1, MMP2, MM9, while up-regulated the expression of AT2 to inhibit MF. Further mechanism study indicated that Fufang Danshen tablets inhibited MF by down-regulated the expressions of AA metabolism, such as PLA2, P450, COX2 and 5-LOX. In summary, Fufang Danshen tablets can effectively inhibit MF in the ischemic area after MI in rats. The mechanism is related to the regulation of AT1-mediated PLA2-COX2 metabolic pathway.

Effects of rosuvastatin on expression of angiotensin-converting enzyme 2 after vascular balloon injury in rats.

OBJECTIVE: To investigate the effects and mechanisms of rosuvastatin on angiotensin -converting enzyme 2 (ACE2) in the process of neointimal formation after vascular balloon injury in rats, and to explore the effects of ACE2 and rosuvastatin in restenosis. METHODS: Thirty-six Wistar rats were randomly allocated into three groups: control group (n = 12), surgery group (n = 12), and statin group (n = 12). Aortic endothelial denudation of rats was performed using 2F balloon catheters. At days 14 and 28 after injury, aortic arteries were harvested to examine the following. Intimal thickening was examined by hematoxylin and eosin staining. We measured angiotensin II (Ang II) and angiotensin 1-7 (Ang-[1-7]) levels by a radioimmunological method or enzyme-linked immunosorbent assay. Protein and mRNA expression of ACE2 and Ang II type 1 receptor (AT1) were investigated by immunohistochemistry, Western blots, and Reverse transcriptase-polymerase chain reaction (RT-PCR). We measured changes in proliferating cell nuclear antigen (PCNA) by immunohistochemistry. The level of phosphorylated extracellular signal regulated kinase 1/2 (P-ERK1/2) was evaluated by Western blotting. RESULTS: Proliferation of vascular smooth muscle cells (VSMC) and intimal thickening were higher at day 14 after vascular balloon injury in the surgery group compared with the control group. Proliferation of VSMC was decreased by day 28 after injury, while intimal thickening continued. With rosuvastatin treatment, the extent of VSMC proliferation and intimal thickening was reduced at day 14 and 28 after injury. Ang II and P-ERK levels were significantly increased, Ang-(1-7) levels were significantly decreased, mRNA and protein expressions of ACE2 were significantly decreased, and AT1 expression was significantly increased at days 14 and 28 after vascular balloon injury in the surgery group compared with the control group. PCNA expression was higher in the surgery group than in the control group, and it was significantly decreased after being given rosuvastatin. Expression of ACE2 mRNA and protein, and Ang-(1-7) levels were significantly increased, while AT1 expression and levels of Ang II and P-ERK were significantly decreased in the statin group compared with the surgery group. CONCLUSIONS: Expression of ACE2 mRNA and protein is decreased in the process of intimal thickening after balloon injury. The inhibitory effect of rosuvastatin on intimal thickening is related to upregulation of ACE2, an increase in Ang-(1-7), downregulation of AT1, and activation of the P-ERK pathway.

[Effects of valsartan on aortic angiotensin converting enzyme 2 expression after aortic balloon injury in rats].

OBJECTIVE: To investigate the process and mechanism of neointimal formation, the level of angiotensin II and angiotensin (1-7), the expression of angiotensin converting enzyme 2(ACE2), angiotensin II type 1 receptor (AT(1)R), extracellular signal regulated kinase (ERK) and the effects of valsartan on them after aortic balloon injury in rats. METHODS: Aortic endothelial denudation of rats was induced by 2F balloon catheter. Thirty-six rats were randomly allocated into three groups: Group 1 (n = 12): controls; Group 2 (n = 12): aortic balloon injury; Group 3 (n = 12): valsartan (20 mg×kg(-1)×d(-1)) given from 1 day before injury to 14 and 28 days after aortic injury. The expression of ACE2 and AT1, the level of P-ERK, AngII, Ang(1-7) and intimal thickening were investigated by RT-PCR technique, immunohistochemistry, Western blot, radioimmunological method, enzyme linked immunosorbent assay (ELISA) and HE stain, respectively. RESULTS: (1) The proliferation of vascular smooth muscle cells (VSMC) and the intimal thickening were evidenced at day 14 and 28 after aortic balloon injury. (2) The mRNA and protein expressions of ACE2 decreased significantly, but AT(1)R mRNA and protein expression increased significantly at day 14 and 28 after balloon injury. (3) The level of AngII and p-ERK increased and Ang(1-7) reduced after balloon injury. (4) Valsartan not only attenuated the proliferation of VSMC and the intimal thickening but also upregulated the expression of ACE2 and the level of Ang(1-7) and downregulated the expression of AT(1)R and the level of AngII, p-ERK in this model. CONCLUSION: Intimal thickening after balloon injury is linked with reduced expression of ACE2.Valsartan can inhibit the intimal thickening possibly by upregulating ACE2 and Ang(1-7) and downregulating AT(1) in this model.

Epidemiologic survey on hospitalized neonates in China.

OBJECTIVE: To carry out a nationwide epidemiologic survey on the neonates in urban hospitals with an attempt to understand the disease spectrum and treatment outcomes of hospitalized neonates in China. METHODS: The clinical data of 43,289 hospitalized neonates from 86 hospitals in 47 Chinese cities (22 provinces) between January 1, 2005 and December 31, 2005 were retrospectively analyzed. RESULTS: The male:female ratio was 1.73:1. Premature infants accounted for 26.2% of the hospitalized neonates, which was higher than that reported in 2002 (19.7%). The top three diseases during the neonatal period were jaundice, pneumonia, and hypoxic-ischemic encephalopathy. The incidences of pneumonia, meconium aspiration syndrome, and bilirubin encephalopathy in term infants were higher than those in premature infants, while the incidences of asphyxia, respiratory distress syndrome, and pulmonary hemorrhage in term infants were lower than those in premature infants. The incidences of asphyxia, small for gestational age infant, and wet lung were higher in neonates whose mother had pregnancy induced hypertension. The outcomes of these hospitalized neonates included: recovered, 63.9%; improved, 27.3%; discharged due to the family's own decisions, 7.6%, and died, 1.2%. Nearly half (46.4%) of the neonatal death occurred within 24 hrs after admission. CONCLUSION: The incidence of premature birth shows an increasing trend among hospitalized neonates. Since the neonatal deaths mainly occur within 24 hrs after admission, monitoring during this period should be enhanced.

[Clinical outcomes of women with transfusion-associated hepatitis C after 10-15 years follow-up].

BACKGROUND: To investigate natural history of hepatitis C virus (HCV) infection and related factors in China. METHODS: Totally 41 women infected with HCV 10-15 years ago were evaluated in this study. Their clinical manifestations were recorded and sera tested for alamine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyltransferase (gamma-GT), alkaline phosphatase (ALP), total bilirubin (TBIL), direct bilirubin (DBIL), total protein (TP), albumin (ALB), and alpha-fetoprotein (alpha-FP), as well as virus markers. The B-type ultrasound examination was performed on the liver, gall bladder, spleen and pancreas. For virus markers, HCV RNA was detected with fluorogenic-quantitative polymerase-chain-reaction, HBsAg and anti-HCV antibody, with enzyme-linked immunosorbent assay (ELISA). RESULTS: Ten to 15 years after infection, the infection was cleared spontaneously in 8 cases (19.51%); 73%(30/41) cases had signs at present, 32% (13/41) cases had mild ALT and/or AST elevation. On ultrasound examination, 83%(34/41) had mild degree 17%(7/41) moderate degree changes of the viral hepatitis. CONCLUSION: Women with HCV infection for 13 +/- 1 (10-15) years after transfusion had slight hepatic inflammation. A part of them had cleared the virus spontaneously.

[Clinical outcomes of 283 patients of transfusion-related hepatitis C in the northern areas of China].

OBJECTIVE: To investigate the natural history and clinical outcomes in a cohort of transfusion-related hepatitis C in northern areas of China. Methods The authors studied 283 patients (137 males, 146 females, mean age 45.79+/-9.92 age) who became infected with HCV while donating plasma 12.25 years ago. These cases were subjected to ultrasonography and liver biochemical tests and serologic anti-HCV assays. Statistical analyses were performed using the SPSS software. RESULTS: Ultrasonographic findings suggestive of liver cirrhosis were 8.3% of cases. No decompensated cirrhosis or HCC was detected. The value of ALT was higher in severe chronic hepatitis (mean 62.07+/-50.87 IU/L) and cirrhosis (mean 115.50+/-108.41 IU/L) patients than in the other groups (mean 32.30+/-29.10 IU/L). The abnormal rate of ALT was 53.3% in severe group, 100% in cirrhosis group. CONCLUSION: The natural history of transfusion-related hepatitis C in the areas seemed to be relatively mild when compared with previous data. Our cases showed relatively low rate of positive findings in ultrasonography. None of this cohort had the decompensated cirrhosis or HCC. The factor of sex but not the age at time of infection was found being related to the outcomes. In the absence of liver biopsy, ultrasonography was a suitable and sensitive method for the diagnosis of the progressive hepatitis and cirrhosis.

Association of interleukin-12 p40 gene 3'-untranslated region polymorphism and outcome of HCV infection.

AIM: To investigate the effect of interleukin-12 p40 gene (IL12B) 3'-untranslated region polymorphism on the outcome of HCV infection. METHODS: A total of 133 patients who had been infected with HCV for 12-25 (18.2+/-3.8) years, were enrolled in this study. Liver biochemical tests were performed with an automated analyzer and HCV RNA was detected by fluorogenic quantitative polymerase chain reaction. B-mode ultrasound was used for liver examination. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for the detection of IL12B (1188A/C) polymorphism. RESULTS: Self-limited infection was associated with AC genotype (OR = 3.48; P = 0.001) and persistent infection was associated with AA genotype (OR = 0.34; P = 0.014) at site 1188 of IL12B. In patients with persistent HCV infection, no significant differences were found regarding the age, gender, duration of infection and biochemical characteristics (P>0.05). According to B-mode ultrasound imaging and clinical diagnosis, patients with persistent infection were divided into groups based on the severity of infection. No significant differences were found in the frequency of IL-12 genotype (1188A/C) between different groups (P>0.05). CONCLUSION: The polymorphism of IL12B (1188A/C) appears to have some influence on the outcome of HCV infection.

[Evaluation of health-related quality of life in chronic hepatitis C women patients].

OBJECTIVE: To evaluate the health-related quality of life (HRQOL) in chronic hepatitis C (CHC) female patients, and to identify factors associated with impairment in HRQOL. METHODS: We enrolled 41 women cases with hepatitis C for 10 to 15 years and 51 health women in Guan area of Hebei province, China. HRQOL was assessed by the SF-36 by an interview. RESULTS: CHC female patients had significantly lower SF-36 scors in all subscales when compared to those of the non-HCV infected female population. CONCLUSION: CHC female patients have a reduced HRQOL. CHC alone is associated with significant impairment in HRQOL.

[Nine-year follow-up of hepatitis C virus infection in a rural area of Hebei province, China].

OBJECTIVE: To investigate natural history of hepatitis C virus infection and related factors among plasma donors in China. METHODS: 172 plasma donors in a rural area of Hebei province had been diagnosed as HCV infection in 1993. No antiviral treatment was applied to them during the period of infection. In the present study, we investigated the outcome of HCV infection nine years later and related factors affecting the outcome. In fact, only 142 cases were followed up in the investigation. The mean age of 142 cases of blood donors was 46 +/- 9 and the mean age of infection was 37 +/- 9 years old. RESULTS: After nine-year follow-up, 1.2% died of end-stage liver disease. 130 (91.6%) of 142 cases under investigation were still positive for HCV RNA or anti-HCV in their blood and 12 cases (8.4%) were negative for both HCV RNA and anti-HCV. 3.1% developed liver cirrhosis among the patients with persistent infection. The mean level of ALT, AST, GGT among HCV RNA positive cases were significantly higher than that of HCV RNA negative cases (P < 0.001). The abnormal rates of ALT and/or AST in male patients were significantly higher than those of female patients (P = 0.005). The rate of progression to liver cirrhosis from chronic hepatitis C virus was significantly higher in patients co-infected with HCV/HBV than that of the cases of single HCV infection. CONCLUSION: Higher chronic rate was observed in this research. Superinfection of HBV/HCV may have worse clinical outcomes.

[Relative stasis of hepatitis C virus quasispecies in natural duration].

OBJECTIVE: To investigate the evolution of HCV quasispecies in persistent and self-limited infection patients. METHODS: Peripheral blood HCV preserved for 10 years from 8 naive patients were analyzed. HCV core and E2 genes were cloned and sequenced. Genetic distances of different isolates at different time were analyzed by phylip software. RESULTS: The genetic distances of HCV quasispecies were higher in persistent infection patients than that of self-limited patients. In each patient the genetic distance of HCV E2 region isolates were higher than that of HCV C region which has some relationship between each other. In the 4 persistent patients there exist no differences between within-group and between-group genetic distance in HCV C region and also exist no differences in HCV E2 region in 2 of the 4 persistent patients. CONCLUSIONS: The diversity degree of HCV quasispecies may affect the outcome of hepatitis C. Research on HCV C and E2 region to study HCV quasispecies may obtain different results. The distribution of HCV quasispecies may be stable in hepatitis C natural history.

[Relationship between the genotypes of hepatitis B virus and the severity of liver diseases].

OBJECTIVE: To investigate the relationship between different genotypes of hepatitis B virus (HBV) and the severity of liver diseases. METHODS: The S nucleotide sequences of HBV strains isolated from plasma samples of 284 patients were detected and compared. Among them, 87 patients were HBV asymptomatic carriers (ASC), 157 chronic hepatitis B (CHB), 22 liver cirrhosis (LC), and 18 hepatocellular carcinoma (HCC). RESULTS: Genotypes B and C were predominant, with a 26.1% proportion and a 63.2% proportion respectively. The percentage of genotypes B and C in patients with ASC, CHB, LC, and HCC were significantly different (x(2)=15.09, P<0.001). Compared with genotype B, genotype C was more common in patients with CHB and HCC (59.6% vs 43.2%, chi(2)=10.87, P<0.001; 7.7% vs 1.4%, x(2)=7.41, P<0.001), but in patients with LC there was no different (7.7% vs 8.1%, chi(2)=1.29, P>0.05). CONCLUSION: This study suggests that genotype B and C are predominant. And genotype C may induce more severe the liver inflammation than genotype B may do.

[Comparison of hepatitis B virus serotype and genotype among HBsAg positive hepatitis B patients in a northern and a southern city of China].

OBJECTIVE: To understand HBV serotypes and genotypes epidemiology in a northern city and a southern city in China. METHODS: Using polymerase chain reaction (PCR) and direct sequencing of HBV DNA PCR products, the serotypes and genotypes of HBV in 530 from HBsAg positive samples. The enrolled patients were from Harbin, a northern city and Lianjiang, a southern city in China. RESULTS: Comparison of the serotypes and genotypes of HBV between Harbin and Lianjiang showed that adrq+ was the most predominant hepatitis B virus serotype in both Harbin and Lianjiang (87.2% and 73.5%,respectively), adw2 was the next (12.0% and 25.7%, respectively); genotype C was the most frequent in Harbin and Lianjiang (87.8% and 73.2%, respectively), and genotype B was the next (12.2% and 26.1%, respectively) only 1 patient was infected by genotype D, and 1 patient was found to be co-infected by genotype B and C in Lianjiang. CONCLUSION: The results suggest that the percentage of HBV serotypes and genotypes between Harbin and Lianjiang was significantly different (P less than 0.001), but the main HBV serotype and genotype of the two cities were similar.