Lipid nanoparticle delivery of siRNA targeting Cyp2e1 gene attenuates subacute alcoholic liver injury in mice / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effect and mechanism of lipid nanoparticle (LNP) delivery of small interfering RNA (siRNA) targeting Cyp2e1 gene on subacute alcoholic liver injury in mice.@*METHODS@#siRNA targeting Cyp2e1 gene was encapsulated in LNP (si-Cyp2e1 LNP) by microfluidic technique and the resulting LNPs were characterized. The optimal dose of si-Cyp2e1 LNP administration was screened. Forty female C57BL/6N mice were randomly divided into blank control group, model control group, si-Cyp2e1 LNP group, LNP control group and metadoxine group. The subacute alcoholic liver injury mouse model was induced by ethanol feeding for 10 d plus ethanol gavage for the last 3 d. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and the superoxide dismutase (SOD) activity as well as malondialdehyde, reactive oxygen species, glutathione, triacylglycerol, total cholesterol contents in liver tissue were measured in each group, and liver index was calculated. The expression of genes related to oxidative stress, lipid synthesis and inflammation in each group of mice were measured by realtime RT-PCR.@*RESULTS@#Compared with the model control group, the levels of liver index, serum ALT, AST activities, malondialdehyde, reactive oxygen species, triacylglycerol, total cholesterol contents in liver tissue decreased, but the SOD activity as well as glutathione increased in the si-Cyp2e1 LNP group (all P<0.01). Hematoxylin-eosin staining result showed disorganized hepatocytes with sparse cytoplasm and a large number of fat vacuoles and necrosis in the model control group, while the si-Cyp2e1 LNP group had uniformly sized and arranged hepatocytes with normal liver tissue morphology and structure. Oil red O staining result showed si-Cyp2e1 LNP group had lower fat content of the liver compared to the model control group (P<0.01), and no fat droplets accumulated. Anti-F4/80 monoclonal antibody fluorescence immunohistochemistry showed that the si-Cyp2e1 LNP group had lower cumulative optical density values compared to the model control group (P<0.01) and no significant inflammatory reaction. Compared with the model control group, the expression of catalytic genes P47phox, P67phox and Gp91phox were reduced (all P<0.01), while the expression of the antioxidant enzyme genes Sod1, Gsh-rd and Gsh-px were increased (all P<0.01). The mRNA expression of the lipid metabolism genes Pgc-1α and Cpt1 were increased (all P<0.01) and the lipid synthesis-related genes Srebp1c, Acc and Fasn were decreased (all P<0.01); the expression of liver inflammation-related genes Tgf-β, Tnf-α and Il-6 were decreased (all P<0.01).@*CONCLUSIONS@#The si-Cyp2e1 LNP may attenuate subacute alcoholic liver injury in mice mainly by reducing reactive oxygen levels, increasing antioxidant activity, blocking oxidative stress pathways and reducing ethanol-induced steatosis and inflammation.

Amelioration of ethanol-induced oxidative stress and alcoholic liver disease by in vivo RNAi targeting Cyp2e1

Alcoholic liver disease (ALD) results from continuous and heavy alcohol consumption. The current treatment strategy for ALD is based on alcohol withdrawal coupled with antioxidant drug intervention, which is a long process with poor efficacy and low patient compliance. Alcohol-induced CYP2E1 upregulation has been demonstrated as a key regulator of ALD, but CYP2E1 knockdown in humans was impractical, and pharmacological inhibition of CYP2E1 by a clinically relevant approach for treating ALD was not shown. In this study, we developed a RNAi therapeutics delivered by lipid nanoparticle, and treated mice fed on Lieber-DeCarli ethanol liquid diet weekly for up to 12 weeks. This RNAi-based inhibition of Cyp2e1 expression reduced reactive oxygen species and oxidative stress in mouse livers, and contributed to improved ALD symptoms in mice. The liver fat accumulation, hepatocyte inflammation, and fibrosis were reduced in ALD models. Therefore, this study suggested the feasibility of RNAi targeting to CYP2E1 as a potential therapeutic tool to the development of ALD.

Exploration of the development of Industry-University-Institute cooperation for promoting scientific and technological achievements transformation in medical universities: Taking "Shanghai Pharmaceuticals-SJTUSM Innovative Achievements" incubation platform as an example / 中华医学科研管理杂志

Objective:The article aims to explore the positive promoting effect of the construction of a medical university’s incubation platform on the Industry-University-Institute cooperation and biomedical scientific and technological achievements transformation.Methods:Through literature review, the article studied the existing mode of Industry-University-Institute cooperation in universities, analyzed the main bottlenecks, and summarized the practical exploration experience of ″Shanghai Pharmaceuticals-SJTUSM Innovative Achievements″ incubation platform.Results:The incubation platform effectively promoted the process and system construction of the scientific and technological achievements transformation in the university, and improved the project mining level and platform support function.Conclusions:The practical exploration of the incubation platform lays a foundation for the construction of the biomedical industrial park. Through the in-depth construction of the " Government-Industry-University-Institute-Investment-Inventor" development chain, the platform can help to promote strategic innovation and industrialization of achievements, making the " first-in-class" medicine in China.

Preliminary exploration on the formative assessment model in the integrated curriculum under the network environment / 中华医学教育探索杂志

Curriculum integration has become the core content of the current medical education reform.With the rapid development of information technology,how to reflect the teaching and learning effects and to further improve them,is one of the difficulties in the reform of the integrated curriculum.In view of the problems of single form of assessment model,separate classroom teaching and extracurricular teaching evaluation,low utilization of network and electronic teaching resources,we make full use of network methods to carry out reform on the current formative teaching evaluation system.With the purpose of formative assessment and evaluation,the "Blackboard network teaching system" of the curriculum has been built,and the evaluation indexes of classroom teaching and extracurricular teaching have been established.And at the same time,the formative assessment index has been enriched and perfected in a diversified form,and the formative assessment model adapted to the integrated curriculum is preliminarily explored.

E3 ligase UHRF2 stabilizes the acetyltransferase TIP60 and regulates H3K9ac and H3K14ac via RING finger domain

UHRF2 is a ubiquitin-protein ligase E3 that regulates cell cycle, genomic stability and epigenetics. We conducted a co-immunoprecipitation assay and found that TIP60 and HDAC1 interact with UHRF2. We previously demonstrated that UHRF2 regulated H3K9ac and H3K14ac differentially in normal and cancer cells. However, the accurate signal transduction mechanisms were not clear. In this study, we found that TIP60 acted downstream of UHRF2 to regulate H3K9ac and H3K14ac expression. TIP60 is stabilized in normal cells by UHRF2 ubiquitination. However, TIP60 is destabilized in cancer cells. Depletion or inhibition of TIP60 disrupts the regulatory relationship between UHRF2, H3K9ac and H3K14ac. In summary, the findings suggest that UHRF2 mediated the post-translational modification of histones and the initiation and progression of cancer.

Discussion on Reconciliation of Medical Insurance Payment in Public Hospitals of Tianjin Based on Total Medical Prepayment / 中国卫生经济

To deal with the current large workload and difficulty of reconciliation for medical insurance expenses in public hospitals,it proposed a institutionalized and normalized reconciliation management process.Tianjin was selected as the object of research.Based on hospital and medical insurance institutions,it analyzed the existing problems and reasons of medical insurance on reconciliation.It proposed a medical insurance debt accounting based,medical settlement information establishment supported,medical insurance reconciliation process centered medical insurance reconciliation management work system to solve the problem of medical insurance reconciliation and promote the safety and integrity of the obligatory right for medical insurance.

Recombinant Kluyveromyces lactis expressing highly pathogenic porcine reproductive and respiratory syndrome virus GP5 elicits mucosal and cell-mediated immune responses in mice

Currently, killed-virus and modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccines are used to control porcine reproductive and respiratory syndrome. However, both types of vaccines have inherent drawbacks; accordingly, the development of novel PRRSV vaccines is urgently needed. Previous studies have suggested that yeast possesses adjuvant activities, and it has been used as an expression vehicle to elicit immune responses to foreign antigens. In this report, recombinant Kluyveromyces lactis expressing GP5 of HP-PRRSV (Yeast-GP5) was generated and immune responses to this construct were analyzed in mice. Intestinal mucosal PRRSV-specific sIgA antibody and higher levels of IFN-gamma in spleen CD4+ and CD8+ T cells were induced by oral administration of Yeast-GP5. Additionally, Yeast-GP5 administered subcutaneously evoked vigorous cell-mediated immunity, and PRRSV-specific lymphocyte proliferation and IFN-gamma secretion were detected in the splenocytes of mice. These results suggest that Yeast-GP5 has the potential for use as a vaccine for PRRSV in the future.

Practice and exploration of the reform of web-based course examination of practice pathology / 中华医学教育探索杂志

Test is an important way for getting feedback from students about teaching. The traditional examination is not suitable for the requirements of modernization teaching.Web-Based Course Examination of practice pathology simplifies the process of traditional examination greatly. It is one of the important means in modernization teaching.

Lentivirus PARG-shRNA transfection decreases matrix adhesion,migration and invasion potencies of colon carcinoma lovo cells / 基础医学与临床

Objective To explore the effects of poly(ADP-ribose) glycohydrolase (PARG) gene silencing on colon carcinoma lovo cells matrix adhesion,migration and invasion.Methods Lentivirus PARG-shRNA was transfected into colon carcinoma lovo cells and the lovo cell strain with PARG gene silencing perpetually was selected.Western blot was used to test the expression of PARG,poly(ADP-ribose) polymerase(PARP) and NF-κB.The lovo cell matrix adhesion,migration and invasion potencies were observed by cell matrix adhesion,migration and invasion assay.Results The expression of PARP and NF-κB in experiment groups was weaker than that in the control groups (P＜0.05).PARG gene silencing of lovo cells decreased the cell matrix adhesion,migration and invasion potencies.The inhibitory rates of lovo cell matrix adhesion,migration and invasion were 25.22%,38.71%,35.29% respectively.Conclusion PARG gene silencing of lovo cells could inhibite cell matrix adhesion,migration and invasion potencies,which may relate to that PARG gene silencing and decreasing the activity of PARP and NF-κB.PARG probably plays an important role in the infiltration and metastasis of tumor.

Correlation of PARP expression with P-selectin and ICAM-1 expression in colorectal carcinoma / 基础医学与临床

Objective To investigate the correlation of PARP with the metastasis of colorectal carcinoma.Methods Poly(ADP-ribose) polymerase activity was tested by detecting poly-ADP ribose.Immunohistochemical SP was used to detect the expression of PAR,P-selectin and ICAM-1 in colorectal carcinoma and control colorectal mucosa.The co-expression of PAR with P-selectin and ICAM-1 was investigated by immunofluorescence staining in colorectal carcinoma and control colorectal mucosa.Results The expression of PAR,P-selectin and ICAM-1 was significantly higher in colorectal carcinoma than that in the control colorectal mucosa((P

Practicing and thinking of Web -based supplementary teaching for medical morphology courses / 中华医学教育探索杂志

A well Web-based teaching supplementary system is beneficial for saving teaching resources,arousing students’enthusiasm for study.but there is no the emotion exchange between teachers and students which exists in traditional class.Web-based supplementary teaching is an important supplementary method for medical morphology courses,but it can not replace the traditional teaching method.Combining them each other is needed for raising teaching quality.

Effect of PARP inhibitor 5-AIQ on PARP/NF-?B complex and NF-?B activity in murine colon carcinoma CT26 cells / 基础医学与临床

Objective To investigate the effect of PARP inhibitor 5-AIQ on PARP/NF-?B P65 complex and the NF-?B activity in murine colon carcinoma CT26 cells.Methods The murine colon carcinoma CT26 cells were treated with 5-AIQ in vitro.The expressions of PARP and NF-?B P65 were analyzed by Western Blot.The PARP/NF-?B P65 complex was analyzed by co-immunoprecipitation.The NF-?B binding activity was detected by electrophoretic mobility shift assay(EMSA).Results Compared with 5-AIQ-untreated group,the expressions of PARP and NF-?B P65 were markedly decreased in 5-AIQ-treated colon carcinoma CT26 cell groups(P

The Preparation for English-exclusive Teaching for Young Teachers in Medical Colleges and Universities / 中华医学教育探索杂志

With the development of internationalization of medical education,English-exclusive medical education has gradually been carried out to cultivate international medical students in many general medical colleges and universities.Analyzing the learning character of international medical students,paying attention to accumulation of practical public English and special English,using for reference all kinds of advanced experiences and forming own teaching style are the way to make good preparation for English-exclusive teaching for young teachers in general medical colleges and universities.

Effect of poly (ADP-ribose) polymerase inhibitor on liver metastasis of colorectal carcinoma CT26 cell line in mice / 第三军医大学学报

Objective To investigate the effect of poly (ADP-ribose)polymerase (PARP) expression inhibited by 5-AIQ on the liver metastasis in colorectal carcinoma CT26 cell lines. Methods Liver metastasis model of colorectal carcinoma was established by intrasplenic inoculation of colorectal carcinoma CT26 cell in BALB/c mice. Treated by 5-AIQ (100 or 500 ?mol/L),CT26 cells were inoculated to the spleen of the mice,or the rats were first inoculated of CT26 cells,then treated by 5-AIQ (3 or 10 mg/kg) ip. The changes spleen primary carcinoma and liver metastasis carcinoma were observed. The PARP expression in spleen primary carcinoma was detected by Western blotting. Results Significant difference was found in the splenic carcinoma size,the number of liver metastatic nodules,the grading of liver metastatic nodules in the 5-AIQ treated group and control group(P0.05). The PARP expression of spleen primary carcinoma in 5-AIQ treated groups was lower significantly than that in control group (P

Exploration of molecular pathology teaching reform for postgraduated students / 中华医学教育探索杂志

The aim of medical postgraduate education is to train innovative person with high quality. It is a good way ,that teachers with special research experiences who comes from different department teach postgraduated students,which can not only teach students theoretical knowledge but also train them to have innovative ability. Traditional teaching way should be changed,which is benefitial to bring active study of postgraduated students into full play.

Regulatory effects of PARG on PARP in colorectal carcinoma / 第三军医大学学报

Objective To study the relationship between Poly(ADP-ribose) glycohydrolase(PARG) and Poly(ADP-ribose) polymerase(PARP) in colorectal carcinoma.MethodsImmunohistochemical S-P staining was used to examine the expressions of PARG and PARP in 44 specimens human colorectal carcinoma.The positive rate of PARG and PARP in murine colon carcinoma CT26 cells was detected by flow cytometry before and after the treatment of PARG inhibitor Gallotannin.ResultsPARG and PARP expressed in colorectal carcinoma specimens was of positive correlation(r=0.300 01,P

Effect of poly(ADP-ribose)polymerase inhibitor 5-AIQ on invasion and metastasis of CT26 cells / 第三军医大学学报

Objective To study the effect of poly(ADP-ribose)polymerase(PARP)inhibitor 5-AIQ on the adhesion,migration and invasion of mouse colon adenocarcinoma cell line(CT26).Methods The expression of PARP after 5-AIQ treatment was detected Western blot.The cell adhesion,migration and invasion of CT26 after 5-AIQ treatment were observed by cell-matrix adhesion assay,cell migration assay and matrigel invasion assay.Results The expression of PARP in 5-AIQ-treated CT26 was weaker than that in 5-AIQ-untreated cells(P