Research progress on the relationship between liver cirrhosis and thyroid hormones / 中华肝脏病杂志

There exists a complex relationship between liver and thyroid hormones. Liver plays an important role in the activation, inactivation, transportation, and metabolism of thyroid hormones. At the same time, thyroid hormones also affect hepatocytes activity and liver metabolism, such as lipid and bilirubin metabolism. Importantly, thyroid hormone levels often change abnormally in patients with liver cirrhosis. Therefore, studying the change of thyroid hormone levels in patients with liver cirrhosis has a certain clinical value for assessing the severity, prognosis, diagnosis and treatment. This paper reviews the research progress on the relationship between liver cirrhosis and thyroid hormone.

Effects of nerve growth factor on proliferation of hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To determine the effects of nerve growth factor (NGF) on proliferation of hepatic stellate cells (HSCs) and investigate the related molecular mechanism.</p><p><b>METHODS</b>After incubating cultured HSCs for 24 h with different concentrations of NGF (100, 200 or 400 ng/mL), the cell proliferation was observed by XTT colorimetric assay and cell cycle was detected by flow cytometry. Morphological changes in response to a 24 h exposure to 100 ng/mL NGF were observed by transmission electron microscopy.</p><p><b>RESULTS</b>NGF significantly inhibited HSC proliferation (P less than 0.05) in a dose-independent manner. The optical densities of the XTT colorimetric assay were 0.66+/-0.03 for 100 ng/mL NGF, 0.69+/-0.03 for 200 ng/mL NGF, and 0.66+/-0.03 for 400 ng/mL NGF, all of which were significantly lower than that of the control group (0.73+/-0.01; P less than 0.05). All concentrations of NGF led to significantly higher numbers of HSCs in the G2 phase (100 ng/mL: 14.83+/-5.41%, 200 ng/mL: 14.73+/-2.50%, and 400 ng/mL: 14.87+/-2.06%), compared to that detected in the control group (7.47+/-4.39%; P less than 0.05). Twenty-four hours of exposure to 100 ng/mL NGF caused morphological changes indicative of apoptosis.</p><p><b>CONCLUSION</b>NGF inhibits the proliferation of HSCs, possibly by arresting the cells in the G2 phase of the cell cycle. NGF-inhibited cells may also undergo apoptosis.</p>

Percutaneous endoscopic gastrostomy/jejunostomy combined with percutaneous transhepatic biliary drainage in treating malignant biliary obstruction

To investigate the safety and efficacy of percutaneous endoscopic gastrostomy/jejunostomy [PEG/PEJ] combined with percutaneous transhepatic biliary drainage [PTCD] in treating malignant biliary obstruction. Nine patients [6 males and 3 females, average age 71.3 +/- 5.5 years] with complete obstruction of the biliary tract were treated with PEG/PEJ after PTCD. The PEG/PEJ and PTCD tubes were linked outside of the abdominal wall to direct the externally drained bile back to the jejunum through the PEG/PEJ intestinal tube. Clinical symptoms and liver function were assessed following the treatment. The operations were successfully completed in the 9 patients within 40 min [average 35 +/- 2.9 min]. Clinical symptoms such as jaundice, abdominal distension, stomachache and diarrhea appeared but improved within 7 days of the operation. Serum levels of bilirubin, aspartate aminotransferase and alanine aminotransferase were reduced [p < 0.01] 4 weeks following the treatment. There were no procedural complications. Combined PEG/PEJ and PTCD appeared to be safe and effective in the management of malignant biliary obstruction. Further, larger-scale studies will be needed to verify findings of this report

Preliminary study on mechanism of therapeutic effect of Huganjiexian decoction on hepatic fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the effects of Huganjiexian decoction on rat hepatic fibrosis and the creation of cytokines.</p><p><b>METHODS</b>Rat hepatic fibrosis was induced by intraperitoneally injection of carbon tetrachloride. At the same time, these rats were treated with different dosages of Huganjiexian decoction. Sho-saiko-to compound treating group and Fufangbiejiarangan Tablets treating group were used as positive controls. After twelve weeks, all rats were executed. Histopathologic changes were observed after H.E and Masson stainings. The expression of collagen type I, collagen type III, TGF-beta 1 and PDGF-BB in liver were detected by immunohistochemical staining.</p><p><b>RESULTS</b>Compared with fibrotic group, hepatic fibrosis in decoction groups was significantly improved. In decoction groups, levels of collagen type I, collagen type III, TGFbeta1 and PDGF-BB were decreased, especially in the low-dose curcumin group. The TGF-beta 1 positive percentage were 7.56%+/-2.18%, 29.25%+/-7.84%, 13.54%+/-4.15%, 21.82%+/-6.64%, 20.06%+/-7.14%, 13.78%+/-4.35%, 12.75%+/-3.98% in liver tissues from normal group, model group, low, middle, high curcumin, Sho-saiko-to compound and Fufangbiejiarangan Tablets treating groups respectively (P less than 0.05); while the PDGF-BB positive percentage were 1.68%+/-0.41%, 11.70%+/-2.28%, 3.65%+/-0.76%, 5.24%+/-1.04%, 6.37%+/-1.12%, 4.16%+/-0.61%, 3.38%+/-0.56% in liver tissues from those groups respectively (P less than 0.05).</p><p><b>CONCLUSION</b>Huganjiexian decoction can improve rat hepatic fibrosis, possibly via inhibiting the expression of collagen type I, collagen type III, TGFbeta1 and PDGF-BB.</p>

The role and mechanisms of cyclooxygenase-2 inhibitors on the proliferation of hepatic stellate cell / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To observe the effects of NS-398 on proliferation of hepatic stellate cells (HSCs) in vitro, and to investigate the possible molecule mechanism.</p><p><b>METHODS</b>HSCs were incubated with different concentrations of NS-398. The effects of NS-398 on cell proliferation was detected by MTT colormetric assay. The cell cycle of HSCs was analyzed by Flow Cytometry (FCM), cyclooxygenase-2 (COX-2) and proliferating cell nuclear antigen (PCNA) proteins in HSCs were detected by immunocytochemistry.</p><p><b>RESULTS</b>Administration of 20-160 micromol/L NS-398 significantly inhibited HSCs proliferation in dose-dependent manner compared with the control group (P less than 0.01). After treated with NS-398 at concentrations of 90, 120, and 150 micromol/L for 48 h, the number of HSCs in G(2)/M phase increased and the number of HSCs in G(0)/G(1) phase decreased (P less than 0.05); Incubated with 120 micromol/L NS-398 for 48 h, percentage of masculine cell of PCNA was 28.91%+/-0.11%, which was significantly lower than that of the control group (85.99%+/-0.13%) (P less than 0.01). Percentage of masculine cell of COX-2 was 13.80%+/-0.43%, which was not significantly different from that of the control group (14.07%+/-0.59%) (P more than 0.05).</p><p><b>CONCLUSIONS</b>NS-398 could inhibit the proliferation of HSC-T6 and arrest HSCs in G2/M phase. Down-regulation of PCNA protein may partially accounted for the proliferation inhibition effect on HSCs induced by NS-398.</p>

Therapeutic effects of curcumin treatment on hepatic fibrosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic effects and mechanisms of curcumin treatment on hepatic fibrosis.</p><p><b>METHODS</b>A model of hepatic fibrosis was established using carbon tetrachloride intraperitoneal injections in rats. Curcumin was administered to one group of the model rats (curcumin group) and the other rats were used as controls (control group). Serum levels of ALT, AST, HA, LN, PCIII, and NO were measured, and Hyp, MDA, and SOD in liver tissues were measured. Liver tissue slides were stained with HE and Masson staining to study the pathological changes in the livers. Grades of hepatic fibrosis were evaluated according to a semiquantitative scoring system.</p><p><b>RESULTS</b>In the curcumin group, serum levels of ALT, AST, NO, HA, LN, PCIII, MDA, and Hyp, were (218.50+/-48.89) U/L, (376.60+/-79.13) U/L, (47.96+/-6.53) micromol/L, (289.96+/-60.43) mg/L, (107.35+/-27.24) mg/L, (148.95+/-28.63) microg/L, (236.10+/-30.54) nmol/g, (478.40+/-75.74) microg/g and all were lower than those of the control group (693.75+/-117.57) U/L, (892.50+/-105.69) U/L, (70.95+/-10.23) micromol/L, (468.22+/-93.45) mg/L, (346.44+/-75.08) mg/L, (279.82+/-54.00) microg/L, (402.25+/-39.16) nmol/g, and (752.50+/-77.62) microg/g. The differences were significant. In the curcumin group, the level of SOD (90.39+/-21.23) in the liver tissues was significantly higher than that of the control group (46.52+/-20.01). The hepatic fibrosis scores in the curcumin group were significantly lower than those of the control group. These effects were dose-dependent.</p><p><b>CONCLUSIONS</b>Curcumin reduces rat hepatic fibrosis. Anti-peroxidation and regulation of collagen metabolism in liver tissues may be involved in the therapeutic effectiveness of curcumin on hepatic fibrosis.</p>

Ultrastructural changes of duodenal mucosas and their significance in patients with liver cirrhosis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the ultrastructural changes of duodenal mucosas and their significance in patients with liver cirrhosis (PLC).</p><p><b>METHODS</b>Endoscopic biopsy duodenal mucosa specimens of 60 PLC and 18 healthy volunteers as controls were obtained. Ultrastructural changes of them were studied with transmission electron microscopy. These PLC were divided into groups A, B and C according to the Child-Pugh classification. The ultrastructural changes in the duodenal mucosas of each group were rated and compared with those of the other groups. PLC with and without ultrastructural changes of duodenal mucosas were divided into a positive group and a negative group. Levels of plasma Alb, TBil, PT, plasma endotoxin, and blood ammonia of the PLC were detected and compared.</p><p><b>RESULTS</b>There were 20 PLC each in groups A, B, and C. Ultrastructural changes of duodenal mucosas were found in 5 PLC of group A, 9 in group B and 17 in group C. Among the 60 PLC, 52% had some changes in their duodenal mucosas. The changes included decrease and rupture of the microvilli; also karyopyknosis, karyorrhexis, widening of the gaps of the tight junction and tumefactions of mitochodrion of duodenal mucosa epithelial cells. No ultrastructural changes of duodenal mucosas were found in the control group. The rate of changes in the three Child-Pugh class groups and in the control group were 25%, 45%, 85%, 0% respectively (P < 0.01). The level of Alb of the positive group was significantly lower than that of the negative group (P < 0.01). Levels of plasma TBil, PT, endotoxin and blood ammonia of the positive group were significantly higher or longer than those of the negative group (P < 0.01). Levels of plasma Alb of the positive and negative groups were significantly lower than those of the control group (P < 0.01). Levels of TBil, PT, plasma endotoxin and blood ammonia of the positive and negative groups were significantly higher or longer than those of the control group (P < 0.01).</p><p><b>CONCLUSION</b>There were ultrastructural changes of duodenal mucosas in PLC, especially in end-stage PLC. Ultrastructural changes of intestinal mucosas in the PLC may have important pathophysiological and clinical significance.</p>

Prophylactic effect of curcumin on hepatic fibrosis and its relationship with activated hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To observe the prophylactic effect of curcumin on hepatic fibrosis and the number, location, apoptosis of activated hepatic stellate cells (HSCs) in the livers and to discuss the relationship between the prophylactic effects and activated HSC.</p><p><b>METHODS</b>A rat model of hepatic fibrosis was established by intraperitoneal injection of carbon tetrachloride. Curcumin doses of 5 mg, 10 mg, 20 mg per 100 gram per 100g of body weight were given to three groups of the model rats. No curcumin was given to one group of the model rats and it served as the control. After eight weeks, all rats were sacrificed and their left liver lobes were examined histopathologically with H.E and Masson stainings. Grades of hepatic fibrosis were evaluated according to the SSS system. Activated HSC was detected by the alpha-SMA immunohistochemistry staining. HSC apoptosis was detected by double-stainings of terminal deoxynucleotidyl transferase (TdT) dUTP nick-end labeling (TUNEL) and desmin immunohistochemistry staining.</p><p><b>RESULTS</b>Degrees (SSS system scores) of hepatic fibrosis in the curcumin groups were all less severe in comparison with those of the control group. Activated HSCs in the livers of the rats of the control group increased significantly compared with that of the treatment groups, and also fewer apoptotic HSCs were detected in the control group. On the contrary, fewer activated HSCs and more apoptotic HSCs were detected in the curcumin groups compared with those of the control group. The degrees of the effects were curcumin dose-dependent.</p><p><b>CONCLUSIONS</b>Curcumin can prevent hepatic fibrosis. It can inhibit activation and proliferation of HSCs and induce HSCs apoptosis, which may be the mechanism(s) contributing to the prophylactic effects of curcumin on hepatic fibrosis.</p>

Relationship between alterations of p16INK4a and p14ARF genes of CDKN2A locus and gastric carcinogenesis / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between alterations of p16INK4a and p14ARF genes and gastric carcinogenesis.</p><p><b>METHODS</b>Tumors and gastric tissues neighboring carcinoma from 48 patients with gastric cancer were studied. Homozygous deletion, mutation, methylation of the CpG islands, mRNA expression of p16INK4a and p14ARF genes were assessed by polymerase chain reaction (PCR), PCR-single strand conformation polymorphism (SSCP), PCR based methylation assay, and reverse transcription (RT)-PCR.</p><p><b>RESULTS</b>(1) The overall homozygous deletion rate of p16INK4a and p14ARF was 35.4% (17/48), and no homozygous deletion was examined in all the gastric tissues neighboring tumor. (2) There was no point mutation of p16INK4a and p14ARF in 31 gastric cancers without homozygous deletion and in the matched gastric tissues adjacent to tumor. (3) Methylation of the CpG islands of p16INK4a and p14ARF was detected in 47.9% (23/48) of gastric cancers, while methylation was observed only in 2 of 48 gastric tissues neighboring cancers with a significant difference (P <0.01). (4) The rate of p16INK4a mRNA loss was 47.9% (23/48) in gastric cancer, and the cases of combined methylation of exons 1alpha and 2 had a higher loss rate (100%, 6/6) of p16INK4a mRNA than those of methylation form the other exons (11.8%, 2/17) (P <0.01). The loss rate of p14ARF mRNA was 45.8% (22/48) in gastric cancer, and patients with combined methylation of exons 1beta and 2 had a higher loss rate (100%, 3/3) of p14ARF mRNA than those of the methylation from the other exons (15%, 3/20) (P < 0.05). (5) The combined loss of p16INK4a and p14ARF mRNAs was examined in 1 (5.6%) of 18 cases of well and moderately-differentiated carcinomas, and 11 (36.7%) of 30 cases of poorly and not-differentiated carcinomas with significant difference (P <0.05).</p><p><b>CONCLUSION</b>p16INK4a and p14ARF genes were frequently inactivated by homozygous deletion and methylation of the 5' CpG islands in gastric cancer, which might have played an important role in the development of gastric cancer.</p>

An improved method for the isolation of rat hepatic stellate cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To present an improved method to obtain pure, viable, freshly isolated hepatic stellate cells.</p><p><b>METHODS</b>Adult male SD rats were used. All procedures were performed with the animals under sodium pentobarbital anesthesia. Three days after the single intravenous administration of 1 ml liposome-encapsulated CL2MDP, which has selective cytotoxicity of Kupffer cells, livers were perfused with D-Hank's solution containing 100 U/ml heparin for 10 to 15 minutes, and then with 0.05% collagenase dissolved in D-Hank's solution for 25 to 30 minutes. The liver was then gently homogenized and further incubated in 0.025% collagenase, and 0.005% DNAase I for 30 minutes at 37 degrees C under constant stirring. This suspension was filtered through stainless steel gauze and centrifuged for 2 minutes at 50 x g to remove parenchymal cells. Sinusoidal cells in the supernatant were recovered by centrifugation for 10 minutes at 300 x g. The cells were resuspended in the presence of 28.7% Nycodenz stock solution. The final concentration of Nycodenz at this stage was 11.5%. Following centrifugation for 17 minutes at 1400 x g, The cells at the top of this Nycodenz solution were collected. Cells were resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum, The cells were seeded in 50 ml culture flask at a density of 500,000 cells/ml, The cell viability was determined by trypan blue exclusion staining, the purity of hepatic stellate cells was identified by the expression of Desmin using immunocytochemistry method. Endogenous peroxidase staining was used to detect Kupffer cells.</p><p><b>RESULTS</b>The yield rate of hepatic stellate cells was 3 x 10(7) per rat, the cell viability was more than 95%, the desmin positive cell rate was 90%, no endogenous peroxidase positive cells were detected.</p><p><b>CONCLUSION</b>The method for the isolation of hepatic stellate cells was developed without Kupffer cells confusion. The availability of highly purified stellate cells will facilitate the investigation of their functions in primary culture.</p>

Cox regression analysis of predictive factors of hepatorenal syndrome / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the predictive factors and the best prognostic parameter of hepatorenal syndrome (HRS).</p><p><b>METHODS</b>71 patients with cirrhosis who underwent HRS were included for multivariate Cox regression analysis. 35 possible predictive factors that included clinical and biochemical features contributing to the survival of these patients were analyzed.</p><p><b>RESULTS</b>Only Child-Pugh's score at the time of diagnosis was the only dependent risk factor of HRS, RR=1.333, 95% CI (1.026, 1.731).</p><p><b>CONCLUSION</b>Child-Pugh's score at the time of diagnosis of HRS is the best parameter for predicting the clinical outcome of HRS.</p>