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Objective To investigate the effect of MicroRNA-9 (miR-9) on cell proliferation and collagen synthesis in hepatic stellate cells (HSCs),and to explore the potential mechanism.Methods HSC-T6 cells were cultured and transfected with miR-9 mimics with lipofectamine 2000.After incubation 48 h,the cells were collected and total proteins and RNAs were extracted.The expression of miR-9 was detected by quantitative real time polymerase chain reaction (qRT-PCR).The protein expression of type Ⅰ collagen and type Ⅲ collagen were measured by Western blot.The methyl thiazolyl tetrazolium (MTT) method was used to asses the proliferation of HSC-T6 cells.The expression of transforming growth factor-β1 receptor 2 (TGFBR2) was detected by qRT-PCR and Western blot.Results Compared to the control group,miR-9 expression in HSCs was increased in the miR-9 mimics group (P ＜ 0.05),type Ⅰ and type Ⅲ collagen protein expression was reduced by (44 ± 2) ％ and (50 ± 3) ％ (P ＜ 0.01),respectively.The proliferation activity of HSCs was decreased by (48 ± 4)％ (P ＜ 0.05).The expression of TGFBR2 was inhibited in the miR-9 mimics group.Conclusions Upregulation of miR-9 plays a role on suppressing cell proliferation and collagen synthesis in HSCs.This process might be mediated by downregulation of TGFBR2.
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Objective To investigate the changes of vitamin D level and its relationship with bone alkaline phosphatase(BAP)and parathyroid hormone(PTH)in infants fed with breast milk and vitamin D supplementation for 0-8 months in People`s Hospital of Longhua New District in Shenzhen city.MethodsTotally 67 newborns in the Department of Obstetrics and Gynecology of People`s Hospital of Longhua New District were selected.These newborns were breastfed and began to supplement vitamin D 400 U/d in the 15th day after birth.The serum 25-(OH)D3 and BAP and PTH levels were measured in newborns and 4-/8-month-old infants,and the correlations of serum 25-(OH)D3 with BAP and PTH were analyzed by Pearson correlation analysis.ResultsCompared with the newborns,the serum 25-(OH)D3 [(78.47±36.41)nmol/L,(75.26±34.73)nmol/L],BAP[(231.43±78.81)μg/L,(205.97±71.64)μg/L],PTH[(31.76±11.54)pg/ml,and(32.94±13.82)pg/ml] levels of 4-and 8-month-old infants were significantly higher,and the difference were statistically significant(t4 months=7.823,12.568,11.852,t8 months=7.364,9.473,11.947;all P=0.000).There was a negative correlation between serum 25-(OH)D3 and BAP in newborns and 4-/8-monthold infants(r=-0.372,-0.463,-0.309;P=0.008,0.016,0.021),the serum BAP was positively correlated with PTH in infants aged 4 months and 8 months(r=0.487,0.338;P=0.005,0.032),and the serum 25-(OH)D3 was negatively correlated with PTH in infants aged 4 months(r=-0.297,P=0.041).Conclusions Infants at birth are prone to vitamin D deficiency in Shenzhen City.After 4 months of breastfeeding and vitamin D400U/d supplementation,the vitamin D levels in most infants become normal.25-(OH)D3 is correlated with BAP and PTH in different months.
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Objective To evaluate the correlation between transient elastrography (TE) and liver function Child-Pugh grade in patients with liver cirrhosis. Methods The clinical data of 120 patients with liver cirrhosis were retrospectively analyzed. Among the patients, hepatitis-B virus-related cirrhosis was in 103 cases, and 57 patients had ascites. The liver stiffness measurement (LSM) was measured by FibroScan. The liver function Child-Pugh grade was evaluated by liver function Child-Pugh system score. The LSM was compared in patients with different liver function Child-Pugh grade. Results Among the 120 patients with liver cirrhosis, Child-Pugh A grade was in 39 cases, Child-Pugh B grade in 28 cases, and Child-Pugh C grade in 53 cases. The LSM in Child-Pugh B grade patients and C grade patients were significantly higher than that in Child-Pugh A grade patients: (20.2 ± 1.1) and (30.8 ± 1.2) kPa vs. (15.7 ± 1.4) kPa, the LSM in Child-Pugh C grade patients was significantly higher than that in Child-Pugh B grade patients, and there were statistical differences (P<0.05). Among the 103 patients with hepatitis-B virus-related cirrhosis, Child-Pugh A grade was in 33 cases, Child-Pugh B grade in 24 cases, and Child-Pugh C grade in 46 cases. The LSM in Child-Pugh B grade patients and C grade patients were significantly higher than that in Child-Pugh A grade patients: (18.7 ± 0.9) and (26.9 ± 0.6) kPa vs. (12.6 ± 1.7) kPa, the LSM in Child-Pugh C grade patients was significantly higher than that in Child-Pugh B grade patients, and there were statistical difference (P<0.05). Among the 57 patients associated ascites, Child-Pugh B grade was in 11 cases, and Child-Pugh C grade in 46 cases. The LSM in Child-Pugh C grade patients was significantly higher than that in Child-Pugh B grade patients:(42.3 ± 1.4) kPa vs. (35.1 ± 1.0) kPa, and there was statistical difference (P<0.05). Among the 103 patients with hepatitis-B virus-related cirrhosis, associated ascites was in 49 cases, Child-Pugh B grade was in 10 cases, and Child-Pugh C grade in 39 cases. The LSM in Child-Pugh C grade patients was significantly higher than that in Child-Pugh B grade patients: (40.6 ± 0.9) kPa vs. (33.2 ± 1.5) kPa, and there was statistical difference (P<0.05). Conclusions The LSM values of patients with liver cirrhosis are higher with the elevation of liver function Child-Pugh grade. There is a correlation between LSM values and Child-Pugh scores. The LSM can partly evaluate the severity of liver disease in patients with liver fibrosis.
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Objective To estimate the clinical effect of fuzhenghuayu capsule in treatment of early hepatocirrhosis by liver instantaneous elastic imaging. Methods Eighty patients with early hepatocirrhosis were divided into experiment group and control group according to the treatment method with 40 cases each. All the patients in 2 groups were given the same antiviral treatment with entecavir dispersible tablets, and the patients in experiment group combined with fuzhenghuayu capsule. All treatment lasted for 6 months. The liver stiffness measurement (LSM) was measured before treatment and 1, 3 and 6 months after treatment by liver instantaneous elasticity imaging. The serum hyaluronic acid was measured, and the Child-Pugh score was evaluated at the same time. Results The LSM, hyaluronic acid and Child-Pugh scores 6 months after treatment in experiment group were significantly lower than those in control group:(19.3 ± 0.9) kPa vs. (29.6 ± 1.3) kPa, (215.6 ± 59.3)μg/L vs. (344.4 ± 39.6)μg/L and (2.1 ± 1.3) scores vs. (3.9 ± 0.9) scores, and there were statistical differences (P<0.05). No obvious adverse reactions related to the use of fuzhenghuayu capsule were found during the course of treatment. Conclusions Liver instantaneous elasticity imaging can be used to evaluate and monitor early hepatocirrhosis. Fuzhenghuayu capsule on patients with early hepatocirrhosis has a certain degree of curative effect.
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This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.
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This study was aimed to examine the correlation of the cytotoxic effects induced by two types of TNF-α to cell cycle. Hoechst 33342 and PI were used to detect the morphological changes in the cell death induced by the two types of TNF-α. TdT and PI co-staining was performed to determine the phase of cell cycle of apoptotic cells. L929 cells in different phases of cell cycle were further synchronized and their sensitivity to the two types of TNF-α was observed. Our results showed that the apoptosis of HepG2 cells triggered by tm-TNF-α mainly occurred in G(1) phase while in HL-60, Raji and K562 cell lines it mainly took place in S phase. The apoptosis of L929 cells induced by tm-TNF-α mainly occurred in S phase while the apoptosis induced by s-TNF-α mainly appeared in G(1) phase. L929 cells were sensitive to s-TNF-α when synchronized in G(1) phase (cytotoxicity 49.8%) while their sensitivity to tm-TNF-α was highest in S phase (45.7%) and G(1)/S phase (cytotoxicity 40.6%). It was concluded that tm-TNF-α-induced apoptosis of different target cells took place in different phases of cell cycle. The apoptosis of the specific cell line induced by the two types of TNF-α occurred in different phases of cell cycle. The sensitivity of the specific cell line to the two types of TNF-α was correlated with the phase of cell cycle.