The deubiquitinating enzyme 13 retards non-alcoholic steatohepatitis via blocking inactive rhomboid protein 2-dependent pathway

Nowadays potential preclinical drugs for the treatment of nonalcoholic steatohepatitis (NASH) have failed to achieve expected therapeutic efficacy because the pathogenic mechanisms are underestimated. Inactive rhomboid protein 2 (IRHOM2), a promising target for treatment of inflammation-related diseases, contributes to deregulated hepatocyte metabolism-associated nonalcoholic steatohepatitis (NASH) progression. However, the molecular mechanism underlying Irhom2 regulation is still not completely understood. In this work, we identify the ubiquitin-specific protease 13 (USP13) as a critical and novel endogenous blocker of IRHOM2, and we also indicate that USP13 is an IRHOM2-interacting protein that catalyzes deubiquitination of Irhom2 in hepatocytes. Hepatocyte-specific loss of the Usp13 disrupts liver metabolic homeostasis, followed by glycometabolic disorder, lipid deposition, increased inflammation, and markedly promotes NASH development. Conversely, transgenic mice with Usp13 overexpression, lentivirus (LV)- or adeno-associated virus (AAV)-driven Usp13 gene therapeutics mitigates NASH in 3 models of rodent. Mechanistically, in response to metabolic stresses, USP13 directly interacts with IRHOM2 and removes its K63-linked ubiquitination induced by ubiquitin-conjugating enzyme E2N (UBC13), a ubiquitin E2 conjugating enzyme, and thus prevents its activation of downstream cascade pathway. USP13 is a potential treatment target for NASH therapy by targeting the Irhom2 signaling pathway.

Expression of miR-4687-5P and STIM1 in esophageal squamous cell carcinoma and their regulation mechanism / 临床与实验病理学杂志

Purpose To investigate the expression and the methylation status of miR-4687-5P and STIM1 gene in esophageal squamous cell cancer (ESCC) cell lines and ESCC tissue samples,in order to explore the correlation between miR-4687-5P and STIM1 expression,as well as whether they have a common expression regulation mechanism. Methods The qRTPCR and methylation specific PCR (MSP) methods were applied respectively to examine the expression and methylation of miR-4687-5P and STIM1 genes in ESCC cell lines (TE13, KYSE150,T. Tn) and ESCC samples,and further to analyze their correlation. Results The expression of miR-4687-5P and STIM1 genes in ESCC was significantly decreased,and consistent. The weak expression of miR-4687-5P and STIM1 genes was detected in three ESCC cell lines. After treated with 5-Aza-2'-deoxycytidine (5-Aza-Dc,a demethylation agent) ,the expression levels of these two genes were obviously increased. Meanwhile, the methylation bands were obviously weakened or disappeared. The promoter region of STIM1 gene was hypermethylated in ESCC tissues,and its methylation frequency was correlated with the expression of STIM1 and miR-4687-5P (P < 0. 01) . Conclusion miR-4687-5P and STIM1 genes are down-regulated in esophageal carcinoma,and the expression of miR-4687-5P may be regulated by the promoter of its host gene STIM1,and the hypermethylation may be one of the common mechanisms leading to down-regulatory expression of miR-4687-5P and STIM1 genes in ESCC.

Effect of DAPPER1 on malignant biological behavior of esophageal carcinoma cells and its epigenetic regulatory mechanism / 临床与实验病理学杂志

Purpose To investigate the effect of DAPPER1 on the malignant biological behavior in esophageal carcinoma cells,and further to analyze the expression and the possible regulated mechanism of DAPPER1 in esophageal squamous cell cancer (ESCC) samples.Methods MTT assay,colony formation assay and wound healing were used to examine the effect of DAPPER1 on malignant biological behavior in esophageal carcinoma cells,RT-PCR and MSP methods were applied respectively to examine the expression and the methylation status of DAP-PER1 in three ESCC cell lines (TE13,T.Tn,Eca109).Immunohistochemistry was used to examine the DAPPER1 protein expression.Results The negative or weak expression of DAP-PER1 was detected in ESCC cell lines.After treated with 5-Aza2'-deoxycytidine (5-Aza-dC,a demethylation agent),the expression level of DAPPER1 was obviously increased.Meanwhile,over expression of DAPPER1 or treated with 5-Aza-Dc could obviously inhibit proliferation and immigration abilities of TEl3 cell line.The level of DAPPER1 expression had no obviously change after treated with trichostatin A (TSA).Decreased protein expression of DAPPER1 was observed in ESCC tumor tissues compared with non-cancerous tissues (P ＜ 0.01),and associated with the methylation status of this gene (P ＜ 0.01).Conclusion DAPPER1 plays an important role of tumor suppressor gene in esophageal carcinoma,and the aberrant hypermethylation may be one of the main mechanisms in abnormal expression of this gene.

Abnormal expression and its regulation mechanism of Cav-1 gene in gastric cancer / 临床与实验病理学杂志

Purpose To investigate the expression of Cav-1 in gastric cancer ( GC) cell lines and GC samples, and to analyze the pos-sible effect of gene methylation in expression of Cav-1. Methods Methylation specific PCR ( MSP) method was applied to examine the CpG methylation of the Cav-1 promoter in GC cell lines (AGS, MKN45, BGC-823) and 104 samples of GC and corresponding ad-jacent tissues. RT-PCR method was applied to examine the mRNA expression in GC cell lines. IHC were applied to examine the pro-tein expression of Cav-1 in the GC tissues. Results The expression level of Cav-1 mRNA was obviously increased after treated with 5-aza-2’-deoxycytidine (5-Aza-Dc, a demethylation agent) in AGS cell line. We detected the positive expression of Cav-1 gene in MKN45 and BGC-823 cell lines before and after treated with 5-Aza-Dc. The level of Cav-1 mRNA expression was no any change in AGS, MKN45 and BGC-823 cell lines treated with trichostatin A (TSA). MSP results showed that it can be amplified methylated bands in AGS cell line, and the methylated bands disappeared after treated with 5-Aza-Dc. MKN45 and BGC-823 cell lines were no any methylated bands amplified before and after treatment. The methylation frequency of Cav-1 gene was 29. 8% (31/104), which was significantly higher than that in adjacent tissues (P=0. 000). Furthermore, Cav-1 gene hypermethylation status was correlated with lymph node metastasis and family history of upper gastrointestinal cancers ( UGIC) , but not with pathological grade and clinical stage (P>0. 05). The positive frequency of Cav-1 expression was 51. 9%(54/104) in GC, which was significantly lower than that in adja-cent tissues (P=0. 000). The expression of Cav-1 was correlated with the frequency of gene methylation in GC tissues (P=0. 000). Conclusion The expression of Cav-1 was reduced in GC tissues and the gene hyermethylation may be one of the mechanisms causing Cav-1 gene silencing.

Effect of ulinastatin on intestinal mucosal barrier function of rats with obstructive jaundice / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of ulinastatin on intestinal mucosal barrier function of rats with obstructive jaundice.</p><p><b>METHODS</b>Seventy-two male SD rats were randomly divided into sham operation, obstructive jaundice, and ulinastatin treatment groups (groups A, B, and C, respectively). In groups B and C, the common bile duct was ligated to induce obstructive jaundice. The rats in group C were given intraperitoneal injection of ulinastatin at the daily dose of 40,000 IU/kg after the operation, while those in groups A and group B received equal amount of normal saline. At 3, 5, 7 and 10 days after the operation, the liver function and plasma endotoxin level were evaluated and measured, and bacterial culture of the mesenteric lymph nodes, liver and spleen was performed. The terminal ileum mucosa was observed under light microscope, and the intestinal villi and mucosal thinckness was examined with image analysis system.</p><p><b>RESULTS</b>The indices relative to the liver function and plasma endotoxin level were higher at different time points of observation in group B than in group A (P<0.01), and were lower in group C than in group B (P<0.01). Plasma endotoxin level was similar between groups A and C 3 days after the operation (P>0.05). The rate of bacterial translocation was higher in group B than in group A and C (P<0.01, P<0.05), but comparable between groups A and C (P>0.05). Intestinal mucosal injury was observed in group B 3 days after operation, and aggravated with the passage of time. The injury was milder in group C. The intestinal villus length and mucosal thickness were greater in groups A and C than in group B (P<0.01 or P<0.05), but comparable between the former two groups 3 days after operation (P>0.05).</p><p><b>CONCLUSION</b>In early stage of obstructive jaundice, the intestinal mucosal barrier may sustain injuries which aggravate with time; ulinastatin has significant effect in protecting the mucosal barrier function especially against early pathological changes.</p>

Effect of shenqi fuzheng injection on pre/post-operational change of argyrophilic-nucleolar organizer regions in peripheral T-lymphocyte in patients with gastric carcinoma / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the pre-post operational change of argyrophilic-nucleolar organizer regions (Ag-NORs) in peripheral T-lymphocyte of patients with gastric carcinoma (GC), and to explore the effect of shenqi fuzheng injection(SFI) on it.</p><p><b>METHODS</b>Eighty five patients were divided into two groups according to the operation performed was radical or non-radical, and the two groups were subdivided into two by additional intravenous dripping of SFI was given to them or not. The content of Ag-NORs in peripheral T-lymphocyte in all patients before and after operation as well as in 12 healthy subjects was determined.</p><p><b>RESULTS</b>Content of Ag-NORs in GC patients was significantly lower than that in the healthy subject (P < 0.01), which significantly increased after patients underwent radical operation (P < 0. 01 or P < 0.05), especially in those treated with SFI (P < 0.01). While in patients underwent non-radical operation but not treated with SFI, it showed insignificant change after operation, however it did significantly increase in those treated with SFI (P < 0.05).</p><p><b>CONCLUSION</b>The immune function of T lymphocyte was low in patients with gastric carcinoma, post-operational adjuvant treatment of SFI can significantly improve the cellular immunity of patients.</p>

The NAD(P)H: quinone oxidoreductase 1 C609T polymorphism and susceptibility to esophageal cancer / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the association of the NAD(P)H: quinone oxidoreductase 1 (NQO1) C609T polymorphism with susceptibility to esophageal squamous cell carcinoma (ESCC) in a northern Chinese population.</p><p><b>METHODS</b>The NQO1 C609T genotypes were determined by polymerase chain reaction-restriction fragment length polymorphisms (PCR-RFLP) analysis in 193 patients with ESCC and 141 unrelated healthy controls.</p><p><b>RESULTS</b>The frequency of the T allele (null) among ESCC patients was significantly higher than that among healthy controls (Chi-square=4.86, P=0.028). The NQO1 C/C and C/T genotype distribution among ESCC patients was not significantly different from that among healthy controls (Chi-square= 2.27 and 0.127; P=0.132 and 0.721, respectively). However, the T/T genotype frequency among ESCC patients was significantly higher than that among healthy controls (Chi-square=4.39, P=0.036). The NQO1 T/T genotype significantly increased the risk for developing ESCC, compared to the combination of C/C and C/T genotypes, with the adjusted odds ratio (OR) of 1.81 (95%CI: 1.04-3.15). This increased susceptibility exhibited pronouncedly in patients with family history of upper gastrointestinal cancers (adjusted OR=2.22, 95%CI 1.18-4.17).</p><p><b>CONCLUSION</b>Determination of the NQO1 C609T genotype may be used as a stratification marker to predicate high-risk individuals for ESCC.</p>

The protective mechanism of insulin-like growth factor-1 from hypoxic-ischemic brain damage in neonatal rats / 中华围产医学杂志

Objective To evaluate the protective effect of insulin-like growth factor-1 (IGF-1) on hypoxic-ischemic brain damage (HIBD) in newborn rats. Methods One hundred and twenty 7-day-old Wistar rats were randomly divided into sham control,HIBD,HIBD treated with 0.2 mg/kg RH-IGF-1 ,HIBD treated with 0.066 mg/kg RH-IGF-1,and normal saline control groups. Those groups were further divided into 24 h、 48 h and 72 h groups respectively after damage with 8 rats in each group. The histological feature,levels of glutamate (Glu) and apoptosis,and expression of Bcl-2 protein in brain were observed in each group. Results (1) Level of Glu in HIBD 48 h group (1162.2?108.1) mg/kg and levels of apoptosis in every HIBD groups[24 h: (7.6?1.9),48 h: (12.6?1.2),72 h: (13.8?0.9) %] are significantly higher than those in sham groups of the same time,[Glu: (750.9? 53.4) mg/kg and apoptosis: 24 h: (2.0?0.2)%,48 h: (2.0?0.3)%,72 h: (2.0? 0.2)%] respectively ( P