AIM:To explore the role and molecular mechanism of Men1 gene in regulating mouse renal fibro-sis.METHODS:A unilateral ureteral obstruction(UUO)-induced renal fibrosis model was established using C57BL/6 mice,and the mice were randomly divided into 4 groups:sham,UUO-3 d,UUO-7 d,and UUO-14 d,with 15 mice in each group.The C57BL/6 mice with Men1 knockout were randomly divided into 4 groups:sham-Men1-WT,sham-Men1-CKO,UUO-Men1-WT,and UUO-Men1-CKO,with 8 mice in each group.HE staining,Masson staining,and Sirius red staining were used to detect UUO-induced renal injury and renal fibrosis.Human renal tubular epithelial HK-2 cells with MEN1 knockout were constructed.RT-qPCR,Western blot,immunohistochemistry and immunoflurorescnence were per-formed to detect the mRNA and protein expression of MEN1,fibrosis markers(α-smooth muscle actin,collagen type Ⅲ and fibronectin 1)and m6A-related proteins[methyltransferase-like 3(METTL3),METTL14,YTH domain family pro-tein 2(YTHDF2),AlkB homolog 5(ALKBH5),and fat mass and obesity-associated protein(FTO)]in UUO mouse kid-ney tissues and transforming growth factor-β(TGF-β;10 μg/L)-treated HK-2 cells.Dot blot analysis was conducted to measure m6A methylation levels in both mouse kidney tissuess and HK-2 cells.RESULTS:The expression of Men1 de-creased with the aggravation of renal fibrosis(P＜0.01).Men1 inhibited the expression of fibrosis markers in renal tis-sues,and MEN1 knockout increased the accumulation of collagen induced by UUO and TGF-β(P＜0.01).The expres-sion of FTO and ALKBH5 in mouse kidney tissues and HK-2 cells was down-regulated by MEN1 knockout(P＜0.01),and the methylation level of m6A was increased(P＜0.01).Overexpression of FTO significantly reduced the accumulation of m6A modifications and renal fibrosis caused by MEN1 loss,and the methylation level of m6A was increased(P＜0.01).CONCLUSION:Loss of Men1 gene promotes renal fibrosis in mice,and Men1 suppresses renal fibrosis in mice by pro-moting the expression of FTO/ALKBH5 to reduce m6A modifications.

AIM:To explore the role and molecular mechanism of Men1 gene in regulating mouse renal fibro-sis.METHODS:A unilateral ureteral obstruction(UUO)-induced renal fibrosis model was established using C57BL/6 mice,and the mice were randomly divided into 4 groups:sham,UUO-3 d,UUO-7 d,and UUO-14 d,with 15 mice in each group.The C57BL/6 mice with Men1 knockout were randomly divided into 4 groups:sham-Men1-WT,sham-Men1-CKO,UUO-Men1-WT,and UUO-Men1-CKO,with 8 mice in each group.HE staining,Masson staining,and Sirius red staining were used to detect UUO-induced renal injury and renal fibrosis.Human renal tubular epithelial HK-2 cells with MEN1 knockout were constructed.RT-qPCR,Western blot,immunohistochemistry and immunoflurorescnence were per-formed to detect the mRNA and protein expression of MEN1,fibrosis markers(α-smooth muscle actin,collagen type Ⅲ and fibronectin 1)and m6A-related proteins[methyltransferase-like 3(METTL3),METTL14,YTH domain family pro-tein 2(YTHDF2),AlkB homolog 5(ALKBH5),and fat mass and obesity-associated protein(FTO)]in UUO mouse kid-ney tissues and transforming growth factor-β(TGF-β;10 μg/L)-treated HK-2 cells.Dot blot analysis was conducted to measure m6A methylation levels in both mouse kidney tissuess and HK-2 cells.RESULTS:The expression of Men1 de-creased with the aggravation of renal fibrosis(P＜0.01).Men1 inhibited the expression of fibrosis markers in renal tis-sues,and MEN1 knockout increased the accumulation of collagen induced by UUO and TGF-β(P＜0.01).The expres-sion of FTO and ALKBH5 in mouse kidney tissues and HK-2 cells was down-regulated by MEN1 knockout(P＜0.01),and the methylation level of m6A was increased(P＜0.01).Overexpression of FTO significantly reduced the accumulation of m6A modifications and renal fibrosis caused by MEN1 loss,and the methylation level of m6A was increased(P＜0.01).CONCLUSION:Loss of Men1 gene promotes renal fibrosis in mice,and Men1 suppresses renal fibrosis in mice by pro-moting the expression of FTO/ALKBH5 to reduce m6A modifications.

ObjectiveTo provide a new idea for exploring the molecular genetic approach to the pathogenesis of schizophrenia via construction of microRNA-messenger RNA （miRNA-mRNA） regulatory network in schizophrenia. MethodsThe microarray datasets of GSE54578 miRNA expression profiles in peripheral blood and GSE145554 mRNA expression in the anterior cingulate in postmortem brain of schizophrenic subjects were downloaded from Gene Expression Omnibus （GEO） database since July 2021. The GEO2R was used to identify the differentially expressed miRNAs and mRNAs， screen the miRNA with target differentially expressed mRNA， and predict their potential upstream transcription factors. The overlapping genes from the mRNA targeted by the differentially expressed miRNA and the mRNA differentially expressed in GSE145554 dataset were collected. Then the biological features of hub genes were analyzed via Gene Ontology （GO） analysis and Kyoto Encyclopedia of Genes and Genomes （KEGG） pathway enrichment analysis， and the protein-protein interaction （PPI） network and miRNA-mRNA regulatory network of hub genes were constructed. ResultsA total of 8 up-regulated differentially expressed miRNAs with targeted mRNA were screened out in GSE54578 datasets regarding schizophrenia， which involved in the regulation of 10 transcription factors， 247 down-regulated differentially expressed mRNAs were screened out in GSE145554 datasets， and 17 overlapping mRNAs were obtained. GO analysis showed that the target mRNAs were mainly involved in astrocyte differentiation and development. KEGG pathway enrichment analysis showed that the target mRNAs were mainly involved in Rap1 and Ras signaling pathways. PPI network analysis showed that the mRNAs （KRAS and CD28） might be key genes in schizophrenia. ConclusionThe integrated bioinformatics analysis based on GEO database can identify potential susceptibility genes in schizophrenia， and it also contributes to the construction of miRNA-mRNA regulatory network in schizophrenia.

This paper introduced a liver normothermic machine perfusion repair and assessment system. This system consists of a liver normothermic machine perfusion device, a fluorescence imaging system and a tissue oxygen detector. The normothermic machine perfusion device can continuously perfuse the donor liver and monitor and control the perfusion parameters in real time. The fluorescence imaging system can detect the indocyanine green metabolized by the liver to evaluate the microcirculation and the metabolism function of hepatocytes. The tissue oxygen detector can monitor the change of oxygen partial pressure of liver tissue in real time to evaluate the state of cell oxygen consumption.
Humans ; Liver ; Liver Transplantation ; Living Donors ; Organ Preservation ; Perfusion

Objective: To understand the epidemiologic features of the rabies in Xishuang banna prefecture of Yunnan province, China in 2008-2017 and the viral molecular-evolution characteristics. Methods: The data of rabies case questionnaire were collected. The brain tissue samples from mad dogs, suspicious sick dogs and human brain tissue, saliva and cerebrospinal fluid samples from rabies patients were collected in Xishuangbanna. Coding region of nucleoprotein and glycoprotein genes were amplified by RT-PCR and sequenced. Homology and phylogenetic analysis were performed using the relevant bioinformatics software. Results: A total of 62 cases of human rabies were occurred in 28 districts of the 3 counties, Xishuangbanna prefecture in 2008-2017. Of them, 37 cases in Jinghong county, 15 in Menghai county and 10 in Mengla county. In which 48 cases were bitten by domestic dogs (77.42%), 11 cases were bitten by wild dogs (17.74%). Rabies case was occurred every year in the past decade. The seasonal incidence was not obvious. The majority of patients were aged from 30 to 59 years-old, with the youngest 1 year-old and the eldest 91 year-old. The male to female ratio was 1.70∶1, most cases were farmers. The nucleotide sequences of nucleoprotein gene of 9 virus strains (7 from Jinghong, 1 from Menghai and 1 from Mengla) were obtained from the samples of dogs and patients. Homology and phylogenetic analyses indicated that the 5 strains belonged to clade China-Ⅰ, 3 clade China-Ⅱ and 1 clade China-Ⅵ. The nucleotide sequences of glycoprotein gene of 5 virus strains (3 from Jinghong, 1 from Menghai and 1 from Mengla) were obtained from these positive samples, and all were clade China-Ⅰ, it is same with nucleoprotein genes analysis result from these 5 virus strains. These China-Ⅰ and China-Ⅱ strains from Xishuangbanna have a closer genetic relationship with same clade strains isolated from Pu’er and other prefectures of Yunnan province as well as Sichuan, Guizhou and Guangxi. The China-Ⅵ strain from Xishuangbanna share high homology and genetic relationship with China-Ⅵ strains isolated from southwestern Yunnan and neighbouring countries such as Myanmar, Laos and Vietnam in recent years. Conclusions In Xishuangbanna, rabies mainly occurred in rural area and domestic dog was the main source of transmission. These RABV clades China-Ⅰ, China-Ⅱ and China-Ⅵ were found in this region and the China-Ⅰ was principal clade. The transmission source of China-Ⅰ and China-Ⅱ were from adjacent areas in the province and China-Ⅵ was from Myanmar and Laos.

Objective To study the effect of Angong-niuhuang pill on expression of eNOSmRNA in spontaneously hypertensive rats after intracerebral hemorrhage and to explore it's protective mechanism.Methods 120 spontaneously hypertensive rats were randomly divided into four groups: model group, western medicine treated group. Angong-niuhuang pill treated group, combining traditional Chinese medicine western medicine group, expression of eNOSmRNA in rat brain tissue was detected by RT-PCR. Results Expression of BCL-2mRNA in combining traditional Chinese medicine western medicine group was significantly higer than other goups(P＜0.01), Compared with model group, the expression of BCL-2mRNA in western medicine treated group and Angong-niuhuang pill treated group was markedly increased (P＜0.01), the differences between Angong-niuhuang pill treated group and Western medicine treated group were not obvious (P＞0.05).Conclusion Protective effect of Angong-niuhuang pill on spontaneously hypertensive rats after intracerebral hemorrhage is related to the the inducement of eNOS expression.