Aspartate-ornithine granules in the treatment of nonalcoholic steatohepatitis: a multiple-dose parallel controlled clinical trial / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic efficacy and safety of aspartate-ornithine granules in patients with nonalcoholic steatohepatitis (NASH).</p><p><b>METHODS</b>Seventy-two patients with NASH were included in this multiple-dose parallel controlled clinical trial and received a 12-week course of aspartate-ornithine granule treatment at either high-dose (6 g bid po; n = 38) or low-dose (3 g bid po; n = 34). Clinical efficacy was assessed by monitoring data from urinalysis, serologic tests (alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transferase (GGT), and triglyceride (TG)), and abdominal computed tomography (CT) scan. Safety was assessed by occurrence of adverse events (fatigue, anorexia, abdominal distension, nausea, and vomiting). Statistical analyses were conducted to determine the significance of differences between parameters before (baseline) and after treatment.</p><p><b>RESULTS</b>After 12 weeks of treatment, the liver and spleen CT ratios in both the high-dose group (0.89 +/- 0.19) and the low-dose group (0.80 +/- 0.15) were significantly higher than at baseline (S = 329, P less than 0.0001 and S = 246, P less than 0.0001); the overall improvement was more robust in the high-dose group (52.63%) than in the low-dose group (38.23%) (Z = -2.1042, P less than 0.05). After 6 and 12 weeks of treatment, the serum ALT levels in both the high-dose group and the low-dose group were significantly lower than at baseline (6 weeks: S = 324.5, P less than 0.0001 and S = 223, P less than 0.0001; 12 weeks: S = 370.5, P less than 0.0001 and S = 297.5, P less than 0.0001); the overall improvement was more robust in the high-dose group (79.0%) than in the low-dose group (53.0%) (Z = -2.0533, P less than 0.05). Similar trends were seen for the serum levels of AST and GGT after 6 and 12 weeks of treatment (all P less than 0.01) and serum levels of TG after 12 weeks of treatment. The rate of adverse reactions was low and similar between the two groups (high-dose: 4.8% and low-dose: 4.4%; all gastrointestinal).</p><p><b>CONCLUSION</b>Aspartate-ornithine granule therapy was an effective and safe treatment of nonalcoholic steatohepatitis, with the higher dose of 6 g bid po providing more robust clinical benefit without affecting the safety profile.</p>

Gastro-protecting effect of gefarnate on chronic erosive gastritis with dyspeptic symptoms / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The role of gastro-protecting agents on symptomatic chronic gastritis is unclear. This multicenter, open, randomized trial was designed to compare the comprehensive effects of gefarnate with sucralfate on erosive gastritis with dyspeptic symptoms.</p><p><b>METHODS</b>Totally 253 dyspepsia patients confirmed with erosive gastritis were enrolled from six centers in China. They randomly received either daily 300 mg gefarnate or 3 g sucralfate for six weeks. The primary endpoint was the effective rate of both treatments on endoscopic erosion at week six.</p><p><b>RESULTS</b>Gefarnate showed an effective rate of 72% and 67% on endoscopic score and dyspeptic symptom release, which is statistically higher than sucralfate (40.1% and 39.3%, P < 0.001, intension-to-treat). For histological improvement, gefarnate showed both effective in decreasing mucosal chronic inflammation (57.7% vs. 24.8%, P < 0.001, intension-to-treat) and active inflammation (36.4% vs. 23.1%, P < 0.05, intension-to-treat) than the control. A significant increase of prostaglandins and decrease of myeloperoxidase in mucosa were observed in gefarnate group. Severity of erosion is non-relevant to symptoms but Helicobacter pylori (H. pylori) status does affect the outcome of therapy.</p><p><b>CONCLUSIONS</b>Gefarnate demonstrates an effective outcome on the mucosal inflammation in patients with chronic erosive gastritis. Endoscopic and inflammation score should be the major indexes used in gastritis-related trials.</p>

Prevention of variceal rebleeding by TIPS combined with embolization of gastric coronary veins: a clinical controlled study / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To prospectively compare the rates of gastroesophageal variceal rebleeding in patients underwent TIPS alone and TIPS combined with embolization of gastric coronary veins.</p><p><b>METHODS</b>According to the bleeding state within one week before the shunt placement, 122 patients with hepatic cirrhosis indicated for the secondary prevention of gastroesophageal variceal rebleeding were allocated to the shunt group (n = 44, treated with TIPS alone) and the shunt plus embolization group (n = 78, treated with TIPS combined with embolization of gastric coronary veins). All the patients were followed up for 1 year, and the 1-year cumulative rates of rebleeding, shunt patency and mortality were compared.</p><p><b>RESULTS</b>The basic characteristics of patients in the two groups were comparable (P is more than 0.05). The 1-year cumulative re-bleeding rates were 41.5% in the shunt group and 19.5% in the shunt combined with embolization group (x2 = 6.320, P = 0.012). The differences of 1-year cumulative rates of shunt patency and mortality between the two groups were not significant (P is more than 0.05).</p><p><b>CONCLUSIONS</b>TIPS combined with embolization of gastric coronary veins could reduce significantly the rate of rebleeding in 1 year after the shunt placement as compared with TIPS alone.</p>

Induction of necrosis in the hepatocellular carcinoma HepG2 xenografts treated with SOM230 / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of SOM230, a new somatostatin analogue, on the proliferation of hepatocellular carcinoma (HCC) cell line HepG2 in vitro and in vivo, and explore the mechanism underline the necrosis of tumors.</p><p><b>METHODS</b>MTT, TdT-mediated dUTP nick end labeling assay (TUNEL) and flow cytometric assay were used to measure the effects of SOM230 on the proliferation and apoptosis of HCC HepG2 cells. Nude mice bearing HCC xenografts of the HepG2 cell line were treated with SOM230 (100 microg/kg/d subcutaneously injection) and saline as a control for eight weeks. The mass and percentage of necrotic volume of the HCC xenografts in nude mice were determined. Western blot was used to detect SSTR2 in HCC xenografts. Immunohistochemical method was used to detect the expression sites of SSTR2 and VEGF in HCC xenografts. ELISA was used to detect the levels of TNFalpha.</p><p><b>RESULTS</b>No proliferation and apoptosis of HepG2 cells were induced by SOM230 in vitro (F = 0.16, P more than 0.05). The percentage of necrotic volume in SOM230 were significantly higher than that of control group (73.4%+/-7.0% vs 30.2%+/-14.0%, t = -8.02, P more than 0.01). SSTR2 was expressed in blood sinus of HCC xenografts in nude mice. There was no significance difference in the level of SSTR2 expression between SOM230 group and saline treated group. VEGF expression in xenografts was down-regulated by SOM230 treatment. SOM230 treatment did not affect the level of TNFalpha in HCC xenografts (t = -0.24, P more than 0.05).</p><p><b>CONCLUSIONS</b>SOM230 can induce massive necrosis of HCC xenografts only after the blockage of blood flow through down-regulation of VEGF mediated by SSTR2.</p>

Celecoxib induces apoptosis and inhibits angiogenesis in gastric cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to explore the effect of celecoxib, a cyclooxygenase-2 inhibitor, on induction of apoptosis and inhibition of angiogenesis in gastric cancer.</p><p><b>METHODS</b>Fifty nine gastric cancer patients were randomly divided into 2 groups: celecoxib group (n = 37) and control group (n = 22). The patients in the celecoxib group were treated orally with celecoxib 200 mg twice daily for 7 days before resection. The patients in the control group received surgical resection alone. Another group of 20 healthy subjects were recruited as normal control. The number of apoptotic tumor cells was measured by terminal deoxynucleotidyl transferse-mediated dUTP nick end labeling (TUNEL). The expression of COX-2, VEGF and the microvessel density (MVD) were evaluated by immunohistochemistry.</p><p><b>RESULTS</b>The TUNEL results showed an increase of apoptosis in the tumor cells after celecoxib treatment in comparison with that in the control group (7.1% +/- 1.0% vs. 6.2% +/- 0.9%, P < 0.05). The expression level of COX-2 and VEGF in the gastric cancer tissues was significantly decreased in the celecoxib group compared with those in the control group (P < 0.05). Furthermore, MVD was also significantly lower in the celecoxib group when compared with that in the control group (30.48 +/- 5.02 vs. 38.98 +/- 4.58, P < 0.05).</p><p><b>CONCLUSION</b>Oral intake of celecoxib can induce apoptosis and suppress angiogenesis in gastric cancer. It may become an effective agent in the treatment of gastric cancer.</p>

Inhibitory effect of quercetin, rutin and puerarin on LDL oxidation induced by Cu2+ / 中国中药杂志

<p><b>OBJECTIVE</b>To evaluate the inhibitory effect of quercetin, rutin and puerarin on the LDL oxidation induced by Cu2+ and to investigate their action on the prevention and treatment of atherosclerosis.</p><p><b>METHOD</b>The serum LDL was isolated by the one step density gradient ultracentrifugation. The LDL oxidation was induced by Cu2+ in vitro for different time periods. Quercetin, rutin and puerarin at 5 micromol x L(-1) were added respectively, as the experimental groups, 3 hours before oxidation. The oxidation of LDL in experimental and control groups was identified by measuring A234, REM, TBARS and protein carbonyls content, and the values were compared between the two groups.</p><p><b>RESULT</b>(1) The values of A234, REM, TBARS and protein carbonyls formation increased gradually during LDL oxidation induced by Cu2+ in vitro. (2) During LDL oxidation induced by Cu2+ in vitro and incubation with each of quercetin, rutin and puerarin, the kinetic changes of A234, REM, TBARS and protein carbonyls formation showed lag phases of 2-6 h, 2 h and 2 h respectively, and the corresponding values for each of the agents treated group were reduced by 27.7%-49.6%, 24.1%-38.6%, 19.8%-34.3% and 36.4%-56.8%; 12.8%-39.3%, 15.7%-32.0%, 19.0%-28.1% and 12.8%-50.3%; and 3.3%-19.2%, 7.0%-22.5%, 19.5%-22.8% and 8.6%-47.0%, respectively.</p><p><b>CONCLUSION</b>These results suggest that quercetin, rutin and puerarin can substantially inhibit LDL oxidation, and quercetin has antioxidation ability stronger than rutin and puerarin.</p>

Changes of vasoactive intestinal polypeptide and vasoactive intestinal polypeptide receptor 1 in small intestine and liver during macaque development / 生理学报

The present study was aimed to investigate the changes of vasoactive intestinal polypeptide (VIP) and VIP receptor 1 (VIPR1) in small intestinal and hepatic tissues during macaque development. The tissue samples of small intestine, liver and blood samples from peripheral and portal vein of 4 macaques of 6-month fetus, 2-day neonate, 45-day neonate and adult were obtained after anesthetization. The concentration of VIP in blood or tissues of macaques was measured by radioimmunoassay. The distribution of VIP in small intestinal or hepatic tissues was visualized by immunohistochemical staining. The expression of VIPR1 was detected by in situ hybridization. The results showed that: (1) VIP concentration in intestinal tissue of 6-month fetus was (20.7+/-14.3) ng/mg protein, and a few VIP-positive nerve fibers first appeared in intestinal villus root and submucosal layer but not in muscle layer. The intestinal concentration of VIP increased gradually with macaque development and reached (514.8+/- 49.2) ng/mg protein in adult, significantly higher than that in 6-month fetus (P<0.01). (2) In adult animal, VIP-positive nerve fibers became thicker and gradually extended into the mucosal crypt, submucosal layer nerve, myenteric nerve plexus of annular muscle and indulge muscle, and annular muscle. Correspondingly, the expression of VIPR1 in intestine was up-regulated during development. (3) On the contrary, the levels of VIP and VIPR1 in liver were gradually decreased during development. (4) VIP concentration in small intestinal tissue was higher than that in hepatic tissue during development. The VIP level in portal vein was also significantly higher than that in peripheral blood during development. In conclusion, the levels of VIP and VIPR1 in mucosal crypt, submucosal layer nerve, myenteric nerve plexus of annular muscle and indulge muscle increase rapidly after birth. Most of VIP from intestinal tract is degraded in portal vein before entering liver, suggesting that VIP does not metabolize and decompose in liver, and that VIPR1 is only present in embryo hepatic blood vessels.

Inhibition of transcription and expression of X gene by lamivudine / 中华肝脏病杂志

<p><b>OBJECTIVE</b>The X protein of the hepatitis virus B plays an important role in the development of hepatocellular carcinoma (HCC). In this experiment we studied the effects of lamivudine on replication, transcription and expression of the X gene.</p><p><b>METHODS</b>The replication of transfected X gene was measured by polymerase chain reaction. The mRNA of transfected X gene was analyzed by reverse transcriptase polymerase chain reaction. The expression of X protein was detected by biosensor. Moreover, we studied if these effects changed in a dose and time dependent manner.</p><p><b>RESULTS</b>The OD data of purpose band reflected the replication of X gene for control and lamivudine groups were 151.4+/-3.5 and 144.0+/-11.4, respectively. The transcription of mRNA for X gene could be inhibited by lamivudine at a concentration of 4.36 x 10(-4) mol/L for 24 h. The OD data of purpose band for control and treatment groups (16 h) were 243.9+/-9.0 and 133.2+/-7.8. The inhibition was enhanced with the increase of lamivudine concentration and reacting time. The expression of the X protein in HepG2x cells was suppressed by lamivudine at a concentration of 4.36 x 10(-4) mol/L for 16 hours. The resonance unit for expression of the X protein decreased from 353.3+/-15.9 to 252.3+/-18.8.</p><p><b>CONCLUSION</b>Although the replication of transfected X gene of HepG2x cells is not influenced by lamivudine, it may significantly inhibit the transcription and expression of the X protein.</p>

Changes of somatostatin and expression of somatostatin receptor in small intestine and liver tissues during macaque development / 生理学报

Intestinal tract, which produces more than fifty kinds of gut peptides, is regarded as the largest endocrine organ. With regard to the gut peptides, a number of studies were focused on their structure, function and the roles in some diseases. The changes in output or distribution of gut peptides in the intestinal tract during development have been largely unknown. This study was aimed to investigate the changes of somatostatin (SST) and somatostatin receptor 2 (SSTR2) in small intestinal and hepatic tissues during the development of macaque. The tissue samples of small intestine, liver or blood samples from peripheral and portal vein of 4 macaques in 6-month fetus, 2-day neonate, 45-day neonate and adult were obtained after anesthetization. The concentrations of SST in blood or tissues of macaques were measured by radioimmunoassay. The distributions of SST in small intestinal or hepatic tissues were visualized by immunohistochemical staining. The expression of SSTR2 was detected by in situ hybridization. SST concentration of intestinal tissue in 6-month-old macaque was (27.3+/-16.6) ng /mg protein and light positive staining of SST was localized in mucosal crypts but negative in muscle layer. The intestinal concentration of SST increased gradually with macaque development and reached to the peak [(120.1+/-35.3) ng /mg protein] in adult. It was significantly higher than that in fetus (P<0.01). Strong positive staining of SST was found in both mucosal crypts and myenteric nerve plexus of adult animal. SSTR2 was obviously expressed in intestinal epithelium of fetus but its expression was greatly reduced in epithelium and was shifted to mucosal crypts when grown to adult. Negative staining of SSTR2 in muscle layer of fetal or neonatal macaque turned to be positive in myenteric nerve plexus of adult. The levels of SST or SSTR2 in liver decreased gradually during development. SST concentrations of small intestinal tissue kept significantly higher than those of hepatic tissues in the macaque developing stages. SST levels of portal vein were also maintained significantly higher than those of peripheral blood in the macaque developing stages. In conclusion, the level of SST and expression of SSTR2 in mucosal crypt increased gradually with macaque development. SST from intestinal tract was quickly degraded in portal vein before entering into liver. SST positive myenteric nerve plexus was visualized only in mature macaque.

Quantitative variation of vasoactive intestinal polypeptide and its receptor in rat liver during development / 生理学报

Accumulated data have suggested that vasoactive intestinal polypeptide (VIP) and corresponding receptor (VIPR) are involved in the development of hematopoietic stem cells and liver growth. In the present study, radioimmunoassay, biomolecular interaction analysis and reverse transcriptation polymerase chain reaction were used to quantify VIP, VIPR and detect the subtype of VIPR in rat liver during development. VIP concentration of liver in fetal or neonatal rats was significantly lower than that of teens or adult rats (P<0.05). The binding capacities of VIPR in liver of immature rats were much greater than that of the adult rats (P<0.05). The tendency of change in VIP concentration was contrary to that of the binding capacity of VIPR in the liver of rats during development. VIPR-1 was expressed in rat liver in all phases of development. These results may be of benefit to the understanding of the mechanisms of liver growth and fetal liver hemopoiesis shift.

Inhibiting effects of aspirin on the growth of human hepatocellular carcinoma / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To assess the effects of aspirin on the proliferation and apoptosis of human HCC cells.</p><p><b>METHODS</b>The effects of aspirin on the synthesis of DNA in SMMC-7721 HCC cells were determined by using (3)H-thymidine incorporation. Apoptosis of SMMC-7721 was studied by observation of morphologic changes, Tunnel method and flow cytometry after treatment with aspirin. We also assessed the effects of aspirin on the growth of HCC xenografts in nude mice in vivo.</p><p><b>RESULTS</b>A dose-dependent suppression (r=-0.918, P<0.01) of (3)H-TdR incorporation in HCC cell line treated with aspirin was observed in the concentration range of 1 10(-1)~10(-7)mol/L. The mean tumor volume and weight in nude mice treated with aspirin were significantly lower than those of the control group. The inhibiting rate for HCC xenografts was 71.62% in the aspirin group. After exposure to aspirin (31 10(-3)mol/L) for 48 hours, HCC cells presented some morphologic features of apoptosis. The apoptosis index was markedly higher in the aspirin group (8.90% 1.32%) than in the control group (0.50% 0.35%, P<0.01). A typical subdiploid peak before G0/G1 phase with an apoptosis rate as 12.79% was also observed.</p><p><b>CONCLUSIONS</b>Aspirin inhibits the proliferation and increases the apoptosis of human HCC cells not only in vitro but also in vivo.</p>

Establishment of a liver cancer model induced by DENA in Sprague-Dawley rats / 中华放射学杂志

Objective To explore the feasible approach for establishment of a liver cancer model induced by N-nitrosodiethylamine(DENA)with Sprague-Dawley rat,and to provide ideal animal model for imaging diagnosis and interventional therapy.Methods One-hundred and forty male SD rats were administrated with 0.95 g/L DENA for 10 weeks,and MRI was performed for inspecting pathological changes of rat livers on the following week.When the liver tumor was proved then the rat will be the candidate for sequential procedures,otherwise the animal continued under observation until next MRI examination after 4 weeks.DSA was done in 64 rats'for detecting blood supply of liver tumors.Animals were sacrificed with an overdose of chloral hydrate,The representative tumor tissues were fixed in 10% formalin and 2.5% glutaraldehyde for light and electron microscopy analysis respectively.Alpha fetoprotein(AFP) and hepatocyte were assaied by immunohistochemistry technique in order to identify intrinsic trait of the harvested tumors.Results The earliest induced tumor was detected on 11th week and the latest was on 20th week by MRI,and the median period was 13.9 weeks.Tumor size ranged from 2 mm to 40 mm in diameter. The rate of single and multi-induced tumor was 9.7%(7/72)and 90.3%(65/72),respectively.87.7% (57/65)" of the induced multiple tumors was with hepatocirrhosis and 18.1% of these tumors combined with extrahepatic neoplasm or metastasis.Plenty blood supply was proved by DSA in most of those tumors. Tumors not only derived from hepatocyte but also manifested positive expression of AFP.Histological types of these tumors include hepatocellular carcinoma(HCC)(92.0%,66/72),intrahepatic cholangiocarcinoma (ICC)(4.0%,3/72),and combined HCC and ICC(4.0%,3/72),respectively.Electro-microscope analysis indicated that cytoplasm and organelle of induced tumors were abnormal distinctly compared with those of non-carcinomatous cells.Conclusion DENA can induce ideal rat liver cancer as a feasible animal model,MRI is the best approach for scrutinizing pathological changes of rat livers during induced period.