Due to the serious shortage of corneal donor, the development of penetrating keratoplasty and corneal endothelial transplantation is severely restricted in clinical practice.The root cause is the limited proliferation capacity of healthy corneal endothelial cells.With the continuous development of tissue engineering technology and cell engineering technology, the research of tissue-engineered cornea has made some progress. In vitro culture of corneal endothelial cells with high density and healthy endothelial function for transplantation is a hot topic in current tissue engineering research.The keys of tissue-engineered corneal endothelial technology include seed cells, vector materials and the strategy of cell transplantation.At present, many research teams domestic and abroad have reported that the source of seed cells includes human corneal endothelial cells, stem cells, vascular endothelial cells and human amniotic epithelial cells.Vector materials include amniotic membrane, acellular corneal stroma, posterior elastic layer, anterior capsular membrane and various biomaterials.The cultured cells are transplanted by penetrating keratoplasty, corneal endothelial transplantation or anterior chamber injection.This review summarized the latest progress in the research on the source of corneal endothelial seed cells, the selection of vectors and the methods of corneal endothelial transplantation, and summed up the problems faced in the current research and looked forward to its prospects.

Objective To analyse the experience of out-of-hospital self-management in patients with temporary enterostomy after colorectal cancer surgery,aiming to provide a guidance for developing an effectively continuous nursing plan. Methods In this descriptive qualitative study,the purpose sampling method was applied to include 11 patients with temporary enterostomy discharged after colorectal cancer surgery as the study participants between January and October 2023. NVivo12 software and content analysis method were used to analyse the data acquired from interview and to extract the themes. Results Totally,4 themes and 8 sub-themes were extracted,including the lack of stoma management skills and low willingness to learn the skills ( lack of stoma management skills and unwillingness to actively learn skills for stoma management),difficulty in living with stoma (difficulty in living with changes of daily life brought by stoma,and expectations for the life advices of care providers),insufficient psychological and social support (negative emotions towards the life with stoma and lack of psychological support from family,friends and medical staff),and a gap between patients' expectations and actual effects on continuous nursing management (poor effectiveness of continuity of care and the high expectations for continuity of care). Conclusion Further improvement are required for the out-of-hospital self-management skills for the patients with temporary colostomy after colorectal cancer surgery. The skills can be improved by taking multiple measures such as education of nursing skills,advice on daily life,psychological support,and improvement of nursing effectiveness,therefore to enhance the out-of-hospital self-management behaviour of patients with colostomy.

Objective To analyse the experience of out-of-hospital self-management in patients with temporary enterostomy after colorectal cancer surgery,aiming to provide a guidance for developing an effectively continuous nursing plan. Methods In this descriptive qualitative study,the purpose sampling method was applied to include 11 patients with temporary enterostomy discharged after colorectal cancer surgery as the study participants between January and October 2023. NVivo12 software and content analysis method were used to analyse the data acquired from interview and to extract the themes. Results Totally,4 themes and 8 sub-themes were extracted,including the lack of stoma management skills and low willingness to learn the skills ( lack of stoma management skills and unwillingness to actively learn skills for stoma management),difficulty in living with stoma (difficulty in living with changes of daily life brought by stoma,and expectations for the life advices of care providers),insufficient psychological and social support (negative emotions towards the life with stoma and lack of psychological support from family,friends and medical staff),and a gap between patients' expectations and actual effects on continuous nursing management (poor effectiveness of continuity of care and the high expectations for continuity of care). Conclusion Further improvement are required for the out-of-hospital self-management skills for the patients with temporary colostomy after colorectal cancer surgery. The skills can be improved by taking multiple measures such as education of nursing skills,advice on daily life,psychological support,and improvement of nursing effectiveness,therefore to enhance the out-of-hospital self-management behaviour of patients with colostomy.

OBJECTIVE: To explore the laboratory phenotype and molecular pathogenesis in a Chinese pedigree affected with Hereditary coagulation factor Ⅻ (FⅫ) deficiency. METHODS: A male proband admitted to Ningbo No.2 Hospital on July 17, 2021 due to chronic gastritis and members of his pedigree (7 individuals from three generations) were selected as the study subjects. Prothrombin time (PT), activated partial thromboplastin time (APTT), FⅧ activity (FⅧ: C), FⅨ activity (FⅨ: C), FⅪ activity (FⅪ: C), FⅫ activity (FⅫ: C), and FⅫ antigen (FⅫ: Ag) were determined. All of the exons, exon-intronic boundaries, as well as the 5'- and 3'-untranslated regions of the F12 gene were subjected to Sanger sequencing. Candidate variants were verified by cloning sequencing. The effect of candidate variants on the protein function was analyzed by bioinformatics software. RESULTS: The proband, a 47-year-old male, had significantly prolonged APTT (180.0 s) and decreased FⅫ:C and FⅫ:Ag levels (< 1%). His father, mother, brother and two sons also showed certain degrees of reduction. Genetic testing revealed that the proband has harbored compound heterozygous variants of the F12 gene, namely c.1092_1093insC (p.Lys365Glnfs*69) in exon 10 and c.1792_1796delGTCTA (p.Val579Hisfs*32) in exon 14. His mother and elder son were heterozygous for the c.1092_1093ins variant, whilst his father, brother, and younger son were heterozygous for the c.1792_1796delGTCTA variant. Analysis of the promoter region of exon 1 also showed that the proband and both sons had harbored a 46T/T polymorphism, whilst other family members were 46C/T. Bioinformatic analysis suggested that the p.Val579 is a highly conserved site. Protein model analysis showed that, with the p.Val579Hisfs*32 variant, a benzene ring was added and the hydrogen bond of surrounding amino acids was changed. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1792_1796delGTCTA was classified as a pathogenic variant (PVS1+PM2_Supporting+PM4). CONCLUSION The c.1092_1093insC (p.Lys365Glnfs*69) and c.1792_1796delGTCTA (p.Val579Hisfs*32) compound heterozygous variants of the F12 gene probably underlay the decreased FXII levels in this pedigree. Above finding has also enriched the mutational spectrum for FⅫ deficiency.
Male ; Humans ; Aged ; Middle Aged ; Pedigree ; East Asian People ; Exons ; Introns ; Family ; Factor XII Deficiency/genetics* ; 3' Untranslated Regions ; Factor XII/genetics*

ObjectiveTo investigate the prevalence of overweight or obesity in community patients with schizophrenia in Shanghai and to explore the related factors. MethodsStratified cluster sampling method was used and the general condition， physical examination and laboratory examination data of patients with schizophrenia who voluntarily participated in 2020 free health examination of National Basic Public Health Service were analyzed. ResultsA total of 3 200 patients were included into the study ，and the prevalence of overweight and obesity was 36.75% and 17.19%， respectively. Multivariate logistic regression analysis indicated that age between 40 and 60 （OR=1.333， 95%CI： 1.030‒1.724）， intake of first-generation antipsychotics （OR=1.413， 95%CI： 1.112‒1.796）， intake of second-generation antipsychotics （OR=1.573， 95%CI： 1.288‒1.921）， high-normal blood pressure （OR=1.549， 95%CI： 1.245‒1.927）， high-abnormal blood pressure （OR=2.824， 95%CI： 2.204‒3.619）， elevated ALT （OR=1.874， 95%CI： 1.386‒2.535）， elevated FBG （OR=1.270， 95%CI： 1.066‒1.513）， and elevated TG （OR=1.652， 95%CI： 1.335‒2.044） were the related factors that associated overweight or obesity in patients with schizophrenia. ConclusionOverweight and obesity are highly prevalent among community patients with schizophrenia in Shanghai. Age between 40 and 60， taking first-generation and second-generation antipsychotics， blood pressure higher than 120/80 mmHg， elevated ALT， elevated FBG， and elevated TG are associated with overweight or obesity in patients with schizophrenia. To provide personalized health guidance， medical staff in primary health care institutions should pay more attention to high-risk groups of overweight and obesity in schizophrenia patients at annual physical examination.

Objective:To systematically review the effects of limited fluid intake on the prognosis of patients with heart failure.Methods:RCTs about the effects of limited fluid intake on the prognosis of patients with heart failure published up to March 31, 2021 were retrieved from PubMed, The Cochrane Library, CINAHL, Embase, Web of Science, CNKI, Wanfang, VIP, and SinoMed. Two independent researchers were employed to extract data and evaluate the quality of included literature. Rev Man 5.3 was used for Meta-analysis, sequential analysis to evaluate the reliability and authenticity of the research results, and the Egger's test for publication bias.Results:A total of 7 articles were included, with a sample size of 867 cases. Meta-analysis showed that the body weight [ MD=-3.04, 95% CI (-4.70, -1.38) , P<0.001], B-type natriuretic peptide [ MD=-249.32, 95% CI (-305.00, -193.63) , P<0.001], blood creatinine [ MD=-22.03, 95% CI (-24.98, -19.09) , P<0.001], readmission rate [ OR=0.30, 95% CI (0.20, 0.45) , P<0.001] of the limited fluid intake group in the chronic phase of heart failure were lower than those in the control group; the body weight [ MD=1.41, 95% CI (-3.73, 6.55) , P=0.59], B-type natriuretic peptide [ MD=64.52, 95% CI (-50.01, 179.06) , P=0.27], serum creatinine [ MD=3.83, 95% CI (-9.69, 17.36) , P=0.58], readmission rate [ OR=1.21, 95% CI (0.65, 2.27) , P=0.55] of the acute fluid intake group were not statistically different from those in the control group. Conclusions:Limiting fluid intake in the chronic phase of heart failure can effectively improve the patients' heart and kidney function and the prognosis of the disease. In the acute phase of heart failure, it may be necessary to combine multiple treatments to keep the patients at the best volume state.

MicroRNA (miRNA) is a non-coding single strand endogenous RNA, with a length of 20-25 bp. It regulates the transcription and translation of target mRNA by binding to its 3' untranslated regions (UTR). Abnormal expression of miRNA plays an important role in the occurrence and development of digestive system diseases. Clarifying the regulatory mechanism of miRNA will be of great help for early diagnosis and treatment of diseases. This article reviewed the advances in research on regulatory mechanism of abnormal miRNA expression in digestive system diseases.

Objective: To construct the mmu-miR-155 eukaryotic overexpression vector pmR-155 and to investigate its effect on HBV replication and expression of PTEN in vivo. Methods: The mmu-mir-146a precursor gene fragment pre-mmu-mir-146a was amplified by PCR, then connected to the pmR-mCherry plasmid vector after double enzyme digestion, the accuracy of recombinant vector was verified by colony PCR、double enzyme digestion and sequencing; then the recombinant vector was transfected HBV transgene mice(Experimental Group)with hydrodynamics-based injection via vena caudalis, and pmR-mCherry plasmid、PBS were respectively transfected into the mice as Empty plasmid Group、Blank Group. The concentration of IFN-γ in the serum was detected by ELISA. The expression of SOCS1、PTEN mRNA in the liver was detected by qPCR at 30d post-transfectioned. The Western blot was performed to detect the changes in SOCS1、PTEN、HBX in the liver tissue at 30 d post-transfectioned. The results were analyzed with Student’s t-test, or one-way analysis of variance and the least significant difference test. Results: the colony PCR、double enzyme digestion and sequencing verified that the gene was inserted into the pmR-mCherry vector. Compared with Blank Group, the expression of miR-155 in the Experimental Group was significantly increased(t = 8.90, P < 0.01); the concentration of IFN-γ in the Experimental Group was significantly increased(F = 26.58, P < 0.01); the mRNA(F_{SOCS1 mRNA} = 19.72, P < 0.01; F_{PTEN mRNA} = 7.38, P < 0.05) and protein(F_SOCS1 = 50.30, P < 0.01; F_PTEN = 129.00, P < 0.01) expression of COCS1、PTEN was significantly decreased in the Experimental group and the protein of HBX was also significantly(F_HBX = 77.97, P < 0.01). Conclusion The pmR-155 eukaryotic overexpression vector is successfully constructed, this recombinant vector can express miR-155 stably; miR-155 can down-regulate cocs1、PTEN gene expression and up-regulate the expression of IFN-γ, it can inhibit the replication of HBV and a potential targets to treating hepatocellular carcinoma.

Background Recent researches show that oxidative stress is involved in the progress of keratoconus.Nuclear factor-E2-related factor 2-antioxidant response element (Nrf2-ARE) pathway plays a critical role in the defense against oxidative stress,but its function in keratoconus is unclear.Objective To investigate the differences of Nrf2-ARE signaling activation and matrix degenerating enzymes between keratoconus and normal corneal stromal cells.Methods Corneal stromal cells were isolated from keratoconus and normal cornea by using dispase and collagenase digestion.The cells were treated with hydrogen peroxide (H2O2) to mimic in vivo oxidative stress condition.Reactive oxygen species (ROS) production was measured by fluorescence substrate DCHF-DA incubation.Nrf2 level and the expression of Nrf2-ARE downstream antioxidant genes were analyzed by Western blot and real-time quantitative-PCR(RT-qPCR).The activity of matrix degenerating enzymes,including urokinase-type plasminogen activator (uPA)-uPA receptor (uPAR) system and matrix metalloproteinase-2 (MMP-2) were assessed by Western blot and gelatin zymography respectively.Results In normal culture,keratoconus corneal stromal cells assumed increased basal ROS and Nrf2 level when compared with normal cells(t =18.155,P＜0.01).However,after H2O2 treatment,the keratoconus corneal stromal cells showed increased ROS production,while decreased Nrf2 translocation and no significant difference in expression levels of Nrf2-ARE downstream antioxidant genes (Nrf2:t =62.123,P＜ 0.01 ; (nicotinamide adenine dinucleotide phosphate quinine oxidoreductase-1 [NQO-1]:t =2.209,P =0.092 ; hemo oxygenase-1 [HO-1]:t =0.293,P =0.784 ; superoxide dismutase [SOD2]:t =0.749,P =0.495).The contents of uPA-uPAR and the activity of MMP-2 also showed a higher level in keratoconus corneal stromal cells than normal cells,with significant differences between them (t =19.164,15.458,4.818,all at P＜0.01).Conclusions The defect of Nrf2-ARE signaling activation exists in the keratoconus corneal stromal cells,and correlats with the abnormal expression level of stromal degeneration enzymes,which suggests that the defect of Nrf2-ARE signaling activation may be involved in the progression of keratoconus.

Background and purpose: MicroRNA(miRNA) is a class of small non-coding RNA playing an important regulatory role in many diseases. In this study, we explore the levels of miR-192 and zinc ifnger E-box binding homeobox 2 (ZEB2) in CRC, the clinical signiifcance of miR-192 and the correlation between the expression of miR-192 and ZEB2 protein. Methods: The expression levels of miR-192 and ZEB2 mRNA in 30 colorectal carcinoma samples, 30 corresponding cancer-adjacent tissue samples (from the edge of tumor≥5 cm), 25 colorectal adenoma samples, and 15 normal tissue samples were quantiifed using quantitative real-time polymerase chain reaction (qRT-PCR). ZEB2 protein expression was determined using Western blot. The relationship between the miR-192 and clinicopathological factors, miR-192 and ZEB2 protein expression was analyzed. Results:Signiifcant upregulation of miR-192 expression and reduction of ZEB2 mRNA and protein expression were identiifed in CRC tissues, compared to cancer-adjacent tissues, colorectal adenoma samples, and normal tissues (P<0.05). Low miR-192 levels were signiifcantly associated with lymph node (P=0.021) and distant metastasis (P=0.023). An inverse relationship between miR-192 and ZEB2 protein expression was identified in CRC group (r=-0.365, P<0.05). Conclusion: MiR-192 downexpression was correlated with CRC metastasis. MiR-192 may play a role in the development and progression of CRC through ZEB2.