Objective To investigate the effects of heat shock protein Gp96 on alcoholic liver fibrosis in mice. Methods A total of 220 male healthy C57BL/6 J mice were randomly divided into four groups; normal control group (n = 10), saline+alcohol induced liver fibrosis group (n = 70), the injection of CRISPR expression Gp96-sgRNA3 by tail vein+ alcohol induced liver fibrosis group (n = 70), the intraperitoneal injection of nuclear factor kappa B(NF-κB) inhibitors PDTC+alcohol induced liver fibrosis group (n = 70). The blood was got from eyeballs and the mice were killed after 8 weeks of ethanol induction. We detected the activity of serum aspartate aminotransferase (AST) in mice of different groups. The pathological changes were detected by HE staining, sirius red staining and periodic acid-Schiff (PAS) staining in the liver of mice. The expression of Gp96 and transforming growth factor βl ( TGF-βl ) were detected by Western blotting. Results Compared with the normal control group, the AST enzyme activity and liver fibrosis increased significantly, glycogen decreased significantly in other three groups (P<0.01). Compared with the saline+alcohol group, the AST enzyme activity and liver fibrosis increased more significantly, glycogen decreased more significantly, Gp96 expression decreased significantly and TGF-βl expression increased significantly in Gp96-sgRNA3+ alcohol group and NF-κB inhibitors PDTC+ alcohol group (P<0.01 or P<0.05). Conclusion The injection of CRISPR expression plasmid Gp96-sgRNA3 by tail vein significantly inhibited the Gp96 expression, promoted the degree of alcoholic liver fibrosis in mice, and NF-κB signaling pathway played a certain role in regulating the expression of Gp96.

Objective To explore the damage mechanism of dopamine cells induced by amphetamine (AMPH). Methods The damage model of dopaminergic cells in mice was established by intraperitoneal injection of AMPH. The mice were randomly grouped into control, saline, amphetamine treatment for 1 day, 7 days, 14 days and 28 days. Each group contained 10 mice. The model of cell injury was established by use of AMPH in PC12 cells. The dopaminergic fibers of corpus striatum and PC12 cells were observed by the immunohistochemistry and immunofluorescence method, and changes of proteins in the protein kinase B (Akt) / glycogen synthase kinase 3β(GSK-3β) / collapsin response mediator protein 2 (CRMP-2) signal pathway were detected by Western blotting. Results AMPH caused the damage of dopaminergic fibers in the mouse corpus striatum and PC12 cells. Meanwhile, AMPH inhibited Akt and GSK-3β phosphorylation levels, and increased phosphorylated CRMP-2 level. Nerve growth factor(NGF), an agonist of Akt, or SB216763, an inhibitor of GSK-3β protected PC12 cells against AMPH-induced toxicity through upregulation of Aat and GSK-3β phosphorylation and downregulated of phosphorylation CRMP-2. Conclusion AMPH causes damage of dopamine cells via inhibition of Akt/ GSK-3β/ CRMP-2 signal pathway.

Purpose: The aim of this study was to compare the efficacy of liver transplantation (LT) and liver resection (LR) for hepatocellular carcinoma (HCC) patients with portal vein tumor thrombus (PVTT) and to investigate risk factors affecting prognosis. Materials and Methods: A total of 94 HCC patients with PVTT type I (segmental PVTT) and PVTT type II (lobar PVTT) were involved and divided into LR (n=47) and LT groups (n=47). Recurrence-free survival (RFS) and overall survival (OS) were compared before and after inverse probability of treatment weighting (IPTW). Prognostic factors for RFS and OS were explored. Results: Two treatment groups were well-balanced using IPTW. In the entire cohort, LT provided a better prognosis than LR. Among patients with PVTT type I, RFS was better with LT (p=0.039); OS was not different significantly between LT and LR (p=0.093). In subgroup analysis of PVTT type I patients with α-fetoprotein (AFP) levels >200 ng/mL, LT elicited significantly longer median RFS (18.0 months vs. 2.1 months, p=0.022) and relatively longer median OS time (23.6 months vs. 9.8 months, p=0.065). Among patients with PVTT type II, no significant differences in RFS and OS were found between LT and LR (p=0.115 and 0.335, respectively). Multivariate analyses showed treatment allocation (LR), tumor size (>5 cm), AFP and aspartate aminotransferase (AST) levels to be risk factors of RFS and treatment allocation (LR), AFP and AST as risk factors for OS. Conclusion LT appeared to afford a better prognosis for HCC with PVTT type I than LR, especially in patients with AFP levels >200 ng/mL.

The mammalian epididymis not only plays a fundamental role in the maturation of spermatozoa, but also provides protection against various stressors. The foremost among these is the threat posed by oxidative stress, which arises from an imbalance in reactive oxygen species and can elicit damage to cellular lipids, proteins, and nucleic acids. In mice, the risk of oxidative damage to spermatozoa is mitigated through the expression and secretion of glutathione peroxidase 5 (GPX5) as a major luminal scavenger in the proximal caput epididymidal segment. Accordingly, the loss of GPX5-mediated protection leads to impaired DNA integrity in the spermatozoa of aged Gpx5

Objective To observe the clinical efficacy and safety of benazepril tablets combined with metoprolol tablets in the treatment of elderly hypertension complicated with heart failure.Methods One hundred elderly hypertension complicated with heart failure were randomly divided into A group (n =33 cases),B group (n =33 cases) and C group (n =34 cases).A group received benazepril 10 mg each time,qd,orally.B group received metoprolol 100 mg each time,bid,orally.C group received benazepril 10 mg each time,qd,orally + metoprolol 100 mg each time,bid,orally.Three groups were treated for 6 months.The clinical efficacy,blood pressure,cardiac ftmction indexes,and adverse drug reactions were compared between the three groups.Results After treatment,the total effective rates of A,B and C groups were 75.76％ (25 cases/33 cases),72.73％ (24 cases/33 cases) and 91.18％ (31 cases/34 cases),respectively.The differences between C group and A,B groups were statistically significant (all P ＜ 0.05).After treatment,the main indexes in A,B,C groups were compared:systolic blood pressure were (141.61 ± 17.22),(142.23 ± 15.34) and (121.73 ± 13.23) mmHg,diastolic blood pressure were (88.04 ± 9.38),(86.96 ± 9.54) and (79.81 ± 9.19) mmHg,left ventricular ejection fraction were (41.03±4.31)％,(41.10 ± 4.32)％ and (51.54 ± 5.38)％,left ventricular end systolic diameter were (53.41 ±5.04),(52.83 ±5.13) and (43.41 ±4.17)mm,left ventricular end diastolic diameter were (45.93 ±4.86),(45.80 ±4.93) and (38.92 ±3.81)mm,distance of 6 min walking test were (328.95 ±68.50),(319.61 ±67.83) and (360.80 ± 69.89) m.The differences between C group and A,B groups were statistically significant (all P ＜ 0.05).The adverse drug reactions in three groups were gastrointestinal reactions and dry cough.The incidences of adverse drug reactions in A,B,C groups were 12.12％,15.15％ and 14.70％ without significant differences (all P ＞ 0.05).Conclusion Benazepril tablets combined with metoprotol tablets have a definitive clinical efficacy in the treatment of elderly hypertension complicated with heart failure,which can significantly reduce the patient's blood pressure and improve the patient's heart function,without increasing the incidence of adverse drug reactions.

AIM:To investigate the protective effect of curcumin analogue L 6H4 on diaphragm of type 2 dia-betic rats.METHODS: SPF male Sprague-Dawley rats ( n=40) were randomly divided into 5 groups: normal control (NC) group, high fat (HF) group, high fat+L6H4 treatment (FT) group, diabetes mellitus (DM) group and DM +L6H4 treatment (DT) group.The rats in the later 4 groups were fed with high-fat diet.After 4 weeks of high-fat diet fee-ding, the rats in DM and DT groups were intraperitoneally injected with streptozotocin to induce type 2 diabetes melliutus. The rats in FT and DT groups were given L6H4 by gavage for 8 weeks.Blood glucose and blood lipid levels were detected biochemically .Fasting serum insulin ( FINS ) level was measured by radioimmunoassay and insulin resistance index (HOMA-IR) was calculated.Serum adiponectin (APN) level was measured by ELISA.The morphological changes of the diaphragm were observed under light and transmission electron microscopes .Lipid deposition and the activity of succinate dehydrogenase (SDH) and NADH-tetrazolium reductase (NADH-TR) were observed by enzyme histochemical staining . The content of malondialdehyde ( MDA) and the activity of superoxide dismutase ( SOD) in the diaphragm were measured by thiobarbituric acid method and hydroxylamine method , respectively .The protein expression of adiponectin receptor 1 ( AdipoR1) in the diaphragm was determined by immunohistochemistry and Western blot .RESULTS:The levels of blood lipids , blood glucose , FINS and HOMA-IR in HF and DM groups were higher than those in NC group , but decreased after L6H4 treatment.The serum APN level in HF and DM groups was lower than that in NC group , but increased after treat-ment with L6H4.The muscle fibers of the diaphragm were shrunk , fat particles accumulated in the muscle fibers , and the mitochondria were slightly swollen in HF and DM groups .The diaphragmatic fibrosis was obvious in DM group .These le-sions were relieved after L6H4 treatment.Compared with NC group, the level of MDA and the activity of SDH and NADH-TR in the diaphragm were increased in HF and DM groups , but decreased after treatment with L 6H4.The activity of SOD and the expression of AdipoR1 in the diaphragm were lower than those in NC group , but increased after L6H4 treatment. CONCLUSION: The curcumin analogue L6H4 exerts a protective effect on diaphragm in type 2 diabetic rats.The strengthened protein expression of AdipoR 1, the increased serum level of APN , and anti-lipid peroxidation may be involved in the process .

OBJECTIVETo investigate the effects and mechanisms of diethylhexylphthalate (DEHP) on morphology and function of progenitor Leydig cells (PLC) in rats.

METHODSTwenty pregnant SD rats were randomly divided into 4 groups ( n = 5): normal control group, DEHP low dose group , middle dose group, and high dose group, which were treated from postnatal day (PND) 1 to PND 21 of the pubs with DEHP at the doses of 0, 10, 100, 750 mg/(kg · d) in 0.5 ml of corn oil by gavage respectively. At the end of the treatment, the male pups were killed and blood samples were collected for determination of serum testosterone concentration by chemiluminescence method. The body weight, testis weight and anogenital distance (AGD) were measured. The morphology of PLC was observed by light and transmission electron microscopy. The protein expression of steroidogenic acute regulatory protein(StAR) in PLC was determined by immunohistochemistry. The mRNA expression of insulin-like growth factor-I (IGF-I) in the testis was assayed by real-time PCR.

RESULTSCompared with normal control group, the serum testosterone and AGD of male pubs from the middle and high dose groups were declined significantly (P < 0.01), the testis weight and body weight from high dose group were decreased significantly (P < 0.01), while the testis weight increased in the low dose group (P < 0.05). Under light microscope, PLC showed hyperplasia and cluster aggregation in the low dose group and focal hyperplasia in the middle and high dose group. The spermatogenic cells in seminiferous tubules showed decrease, apoptosis and unfix in the high dose group. Under transmission electron microscope, the PLC showed decreased lipid droplets, smooth endoplasmic reticulum and mitochondriae in the treated group. The mRNA expression of IGF-I increased in the low dose group, and the protein expression of StAR decreased in the middle and high dose group.

CONCLUSIONLactating exposure to DEHP may interfere with the synthesis of testosterone of PLC in male pubs, the decrease of StAR and the damage of PLC may be involved in it.

Animals ; Body Weight ; Diethylhexyl Phthalate ; adverse effects ; Female ; Germ Cells ; drug effects ; Insulin-Like Growth Factor I ; metabolism ; Lactation ; Leydig Cells ; cytology ; drug effects ; Male ; Organ Size ; Phosphoproteins ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; drug effects ; Testis ; Testosterone ; blood

OBJECTIVETo explore the relationship between CpG island methylator phenotype(CIMP) and genetic instability in sporadic colorectal cancer(SCRC).

METHODSSeventy-one SCRC patients were enrolled in this study. Promotor methylation status of five genes including P14(ARF ), hMLH1, P16(INK4a), MGMT and MINT1 was detected with methylation specific PCR to confirm CIMP. Microsatellite instability (MSI) status was evaluated with two microsatellite loci of BAT25 and BAT26, and the ploidy was detected with flow cytometry. The association between CIMP and MSI as well as chromosomal instability(CIN) was examined.

RESULTSThe positive rates of CIMP, MSI and aneuploidy were 21.1% (15/71), 9.9% (7/71) and 73.5% (50/68) respectively. The positive rate of MSI in positive CIMP patients was higher than that in negative CIMP ones, but the difference was not significant (20.0% vs 7.1%,P=0.158). The positive rate of MSI was 57.1% in patients with hMLH1 gene promotor hypermethylation, which was significantly higher than that (4.7%) in patients without hMLH1 gene promotor hypermethylation (P=0.001). SCRCs with positive CIMP displayed significant inclination of diploidy (P=0.003). The positive rate of diploidy among SCRCs with CIMP was 61.5% while only 18.2% of cases without CIMP demonstrated diploid.

CONCLUSIONSSCRCs with positive CIMP are significantly more likely to be diploid. Simultaneous multiple genes hypermethylation represented by CIMP may be an epigenetic mechanism competing with the genetic mechanism of CIN.

Chromosomal Instability ; Colorectal Neoplasms ; genetics ; CpG Islands ; DNA Methylation ; Genome, Human ; Humans ; Microsatellite Instability ; Phenotype

OBJECTIVETo summarize our clinical experience in adult-to-adult living donor liver transplantation (ALDLT).

METHODSClinical data of 12 patients with ALDLT performed in our center from September 2000 to June 2005 were analyzed, retrospectively.

RESULTSLeft lobe (segments II, III, IV, including the middle hepatic veins) transplantation was performed in 3 patients and right lobe (segments V, VI, VII, VIII, with or without the middle hepatic veins) transplantation was performed in 9 patients. Donors: There were no operative deaths. The median operative time was 6.20+/-1.40 hours and their blood loss ranged from 300 ml to 1200 ml. Postoperative complications included biliary fistula (1 donor) and wound fat liquefaction (1 donor). During a 6-12 months follow-up, no long-term complications were found. Recipients: The operating time ranged from 5 to 11 hours and their blood loss ranged from 800 to 7000 ml. Modified outflow reconstruction, microvascular reconstruction of the hepatic artery and duct-to-duct biliary reconstruction were done during the recipient operations. The median cold ischemia time was 1.90+/-0.50 hours. The median anhepatic phase of recipients was 1.63+/-0.43 hours. Graft/recipient weight ratio (GRWR) was (1.20+/-0.26)%. One recipient presented a postoperative complication of biliary fistula and another recipient died 1 month after the operation from serious infection. The other 11 recipients had long-term survivals.

CONCLUSIONALDLT is an effective treatment for decompensated end-stage liver disease patients and is relatively safe for the donors.

Adult ; Female ; Hepatolenticular Degeneration ; surgery ; Humans ; Liver Cirrhosis ; surgery ; Liver Transplantation ; Living Donors ; Male

ObjectiveTo explore the effect of the Breviscapine (Bre) on rabbit's cardiac muscles after ischemic preconditioning (IP).MethodsThe myocardial ischemic reperfusion model was made with 32 New Zealand white rabbits by silk thread passed around the left circumflex coronary artery and the apex. Model animals were randomly divided into four groups: myocardial ischemia-reperfusion (I/R) group, Bre+I/R group, IP group and Bre+IP group. The changes of the endothelin (ET), nitrous oxide (NO) and the enzymes of the cardiac muscle were measured, and the areas of myocardium infarction were analyzed.ResultsBre and IP could decrease the content of ET, the enzymes of the cardiac muscle and myocardial infarction area; increase the content of the NO. Bre+IP could strengthen the role of protecting the ischemic myocardial cells.ConclusionThe Bre can protect the ischemic cardiac muscle. The Bre+IP can strengthen the protective effect of the IP.