Aim To establish the fatty liver animal model induced by high-fat diet.Observe the dynamic variety of the hepatic steatosis in the different time.Finding out the ideal animal model-making time of hyperlipidemia and nonalcoholic fatty liver.Methods 50 wistar rats were fed with high-fat diet.The serum TG,TC,HDL,LDL,AST,and ALT were detected from 2 to 6 weeks.At the same time,we analyzed the hepatic steatosis situation. Results The rats developed hyperlipidemia and slight fatty liver after two weeks.In the following weeks,the serum lipid level and liver index kept rising.So did the amount of steatosis cells in liver tissue.The 3～4 weeks animal model had developed moderate fatty liver and the 5～6 weeks animal model had developed serious fatty liver.Liver fibrosis was detected in the 6 weeks animal model.Conclusions Feeding with high-fat diet,different phases of fatty liver had been developed in six weeks,and could been used for correlative pharmacological test.

Objective To study the protection of taurine(TAU) against apoptosis of neurons induced by manganese(Mn) in vitro. Method Cortex neurons were separated from Wistar neonatal rats and cultured in vitro.The assays began when neurons grew under the best conditions. Cells were randomly divided into 7 groups: control group,Mn-added groups (Mn 0.2,0.6 and 1.0 mmol/L respectively),TAU-intervened groups (1.5mmol/L TAU with Mn). All treatments lasted 24h. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) was used to observe the morphology of apoptosic neurons. Flow cytometry (FCM) was used to quantitate neuron apoptosic rates. Results (1) Typical morphologic charateristic was found in Mn-added groups. TAU intervention could protect against the effect of 1.5mmol/l Mn on neurons. (2) FCM indicated that TAU can protect against neurons apoptosis induced by Mn. Conclusion Taurine can protect neurons from apoptosis induced by Mn in vitro.

ATM: To observe the anti-inflammatory, and immunomodulation of Jingchufengshi Ointment (JCFSO) (Semen Strychni, Myrrha Preparata, etc.) on the pathological model. METHODS: Whittle's method, the swelling of rat sole and mice carbon clearance test were used to determine the immunomodulation and anti-inflammatory effect of JCFSO. RESULTS: JCFSO external application could inhibite the swelling of rat sole induced by ameliorates and inhibite the higher permeability of abdominal capillary of rat which was induced by acetic acid and also significantly inhibite the skin delay allergic reaction induced by 2,4-dinitroflurbenzene in rat and reduce immune indices (P

Objective:To explore the relationships between the endothelial microparticles (EMPs)in peripheral blood and CD4+ CD25 + Foxp3+ regulatory T cells and their cytokine levels in the patients with acute coronary syndrome (ACS),and to clarify the mechanism of EMPs in the pathogenesis of ACS by affecting the Treg cell differentiation and function.Methods:Twenty-three patients with stable angina (SAP)were allocated to SAP group,and 52 patients with ACS were allocated to ACS group.Twenty individuals with normal conventional coronary angiography results were recruited as control group.The levels of EMPs and the percents of CD4+ CD25 +Foxp3+ regulatory T cells in peripheral blood of the patients in various groups were measured by flow cytometry. The expressions of Foxp3 mRNA and the plasma levels of TGF-β1 were detected with Real-Time PCR and ELISA, respectively.Correlation analysis was performed between EMPs and regulatory T cells,Foxp3 mRNA expression level and TGF-β1 level.Results:Compared with control group and SAP group,the level of EMPs in peripheral blood of the patients in ACS group was increased significantly,but the percentage of regulatory T cells,the Foxp3 mRNA expression level and the plasma TGF-β1 level were decreased markedly (P <0.01).The correlation analysis showed that there were significantly negative correlation between EMPs and regulatory T cells,Foxp3 mRNA expression level and TGF-β1 level (r = - 0.452,P = 0.001;r = - 0.466,P = 0.001;r = - 0.555,P = 0.000 ). Conclusion:EMPs may involve in the process of occurrence and development of ACS and plaque instability by regulating the differentiation and function of CD4+ CD25 + Foxp3 + regulatory T cells.

Objective To study the defects of bone marrow-derived endothelial progenitor cells (EPCs) in number ratio and biological abilities (proliferation, adhesion and migration) in diabetic rats. Methods (1) Establishment of diabetic rat model:1%STZ solution was quickly injected into the abdominal cavity of the male SD rats with the dose of 60 mg/kg. (2). Isolation, culture and identification of bone marrow-derived EPCs in diabetic and normal rats. Bone marrow mononuclear cells were isolated from diabetic and normal rats by density gradient centrifugation methods and cultured by EGM-2 MV medium. The cells were identified by morphological observation, FITC-UEA-1 binding and Dil-Ac-LDL uptake assay, and fluorescent immunocytochemistry was used for detection of CD34 , CD133 and VEGFR-2 expression. CCK-8 method and Transwell kit method were used to determine biological activities of EPCs. Results (1) When cultured in vitro, both bone marrow-derived EPCs in diabetic and normal rats were fusiform in shape, the cells snuggled up to the wall. The expression of CD34, CDl33, VEGFR-2 could be detected in these cells, and the cells could uptake Dil-Ac-LDL and bind FITC-UEA-1, which proved that these cells were EPCs. (2) No significant difference in the number of EPCs derived from bone marrow existed between diabetic rats and normal rats, but the proliferation ability, migration ability and adhesion ability of bone marrow-derived EPCs in diabetic rats were obviously lower than those in normal rats. Conclusion The number of bone marrow-derived EPCs in diabetic rats is not obviously different from that in normal rats, but the biologic activity of EPCs derived from bone marrow in diabetic rats is degraded, which is manifested as weakened abilities of the proliferation, adhesion and migration.

OBJECTIVETo analyze the clinical phenotype of a Chinese pedigree affected with hereditary dentinogenesis imperfecta and mutation of dentin sialophosphoprotein (DSPP) gene.

METHODSAffected members underwent intraoral photography, dental film and panoramic radiography. Genomic DNA was extracted from peripheral venous blood samples. Coding regions of the DSPP gene were subjected to PCR amplification and Sanger sequencing. Functional effect of the mutation was predicted with SIFT and PolyPhen-2. The tertiary structure of wild type and mutant proteins were predicted by Swiss-Port.

RESULTSA heterozygous c.50C to T (p.P17L) mutation was identified in exon 2 of the DSPP gene in the proband and her father. The same mutation was not found among 200 unrelated healthy controls. The Pro-17 residues and its surrounding positions in DSPP are highly conserved across various species. The mutation was predicted to be damaging to the structure of DSPP protein.

CONCLUSIONThe c.50C to T (p.P17L) mutation of the DSPP gene probably underlies the disease in this pedigree. Above finding has expanded the spectrum of DSPP gene mutations and provided a basis for genetic counseling and prenatal diagnosis for this family.

Objective:To investigate the effects of compound fermented milk on intestinal microbiota, short chain fatty acid (SCFA), intestinal motility and mucosal barrier in mice with constipation.Methods:Twenty-seven C57BL/6JNifdc mice were randomly divided into three groups: control group, model group and intervention group. The model group and the intervention group were given loperamide intragastrically for two weeks. Starting from the second week, the intervention group was treated with compound fermented milk for 7 d. The control group was given normal saline. Food intake, water intake, weight change, fecal moisture content, time of first-time black stool and small intestine propulsion rate were detected. Expression of serotonin C receptor (5-HTR2C), zona occludins-1 (ZO-1) and mucin-2 (MUC-2) at mRNA level in colon was analyzed. Western blot was used to measure the expression of Raf/ERK/MAPK-related proteins. SCFA level in intestinal tract was detected by gas chromatography. Intestinal microbiota diversity was analyzed by high-throughput sequencing.Results:Compared with the control group, the first black stool excretion time was significantly prolonged in the model group ( P<0.01). Moreover, fecal moisture content, small intestine propulsion rate and the expression of 5-HTR2C and ZO-1 at mRNA level in colon were significantly decreased ( P<0.01). Compared with the model group, the first black stool excretion time was significantly shortened ( P<0.05); fecal moisture content, small intestine propelling rate ( P<0.05), the expression of colon 5-HTR2C and ZO-1 at mRNA level ( P<0.05), phosphorylation of Raf/ERK/MAPK pathway in the colon, intestinal SCFA-producing bacteria and intestinal SCFA content were increased in the intervention group. Conclusions:Compound fermented milk had a therapeutic effect on constipation in a mouse model by increasing the abundance of SCFA-producing bacteria and SCFA content, enhancing the phosphorylation of the Raf/ERK/MAPK pathway to up-regulate the expression of 5-HTR2C at mRNA level in the colon, and increasing the expression of ZO-1 at mRNA level in the colon. Intestinal peristalsis and intestinal mucosal barrier function were enhanced, thus improving the symptom of constipation.

Objective:To study the clinical effect of different application times of antibiotics in patients with hypertriglyceridemic severe acute pancreatitis(HTG-SAP).Methods:The clinical data of 92 patients with HTG-SAP who were treated at the Department of Critical Care Medicine of the Second Affiliated Hospital of Anhui Medical University from January 2016 to February 2022 were analyzed retrospectively. There were 65 males and 27 females, aged (40.82±10.55) years old. These patients were divided into three groups based on the time of prophylactic use of antibiotics: the early prevention group (used within 72 h after admission, n=39), the delayed prevention group (used after 72 h after admission, n=30) and the nonprevention group ( n=23). The clinical data such as the general data, length of stay and all cause mortality rate of the three groups of patients were analyzed. Results:Compared with patients in the nonprevention group, the incidence rate of pancreatic/peripancreatic infection [10.0%(3/30) vs. 39.1%(9/23)] and the incidence rate of bloodstream infection [6.7%(2/30) vs. 34.8%(8/23)] in the delayed prevention group were significantly decreased (pancreas/peripancreatic infection: χ 2=6.31, P=0.012; bloodstream infection: χ 2=6.72, P=0.010). The infection rate of multiple/pan resistant bacteria in the early prevention group [23.1%(9/39)] was significantly higher than the nonprevention group [4.3%(1/23)] (χ 2=4.49, P=0.034). There were no significant differences in the all cause mortality rate, length of stay, duration of intensive care, hospitalization cost, incidence of intestinal fistula and retroperitoneal hemorrhage, and the proportion of patients requiring surgery among the 3 groups (all P>0.05). Conclusions:Pophylactic use of antibiotics within 72 h of hospital admission significantly reduced the incidence of pancreatic/peripancreatic infection and bloodstream infection in patients with HTG-SAP. There were no significant differences in the final prognosis among these patients with HTG-SAP treated with antibiotics at different times.

Objective:To study the clinical characteristics and risk factors for hypertriglyceridemia-induced severe acute pancreatitis (HTG-SAP) complicated by acute kidney injury.Methods:The clinical data of HTG-SAP patients admitted to the Department of Critical Care Medicine of the Second Affiliated Hospital of Anhui Medical University from January 2014 to July 2022 were retrospectively collected, and divided into an AKI group and a non-AKI group according to whether AKI occurred. The general condition, laboratory data and clinical characteristics of the two groups were compared. Factors with statistically significant differences were screened for logistic regression analysis and a predictive model was constructed. We plotted the receiver operating characteristic curve and used the area under the curve (AUC) to evaluate the predictive ability of each predictor for HTG-SAP complicated with AKI.Results:A total of 91 patients were included, including 37 (40.7%) with AKI and 54 (59.3%) without AKI. The proportions of extrapancreatic infection, sepsis and multiple organ dysfunction syndrome, requiring mechanical ventilation, continuous renal replacement therapy (CRRT), retroperitoneal puncture, and surgical intervention were higher in the AKI group than in the non-AKI group (all P<0.05). The length of ICU stay, total length of hospitalization, and mortality rate in the AKI group were higher than those in the non-AKI group (all P<0.05). Multivariate logistic regression analysis showed that acute physiology and chronic health status scoreⅡ (APACHEⅡ) ( OR=2.069, 95% CI: 1.286-3.417, P=0.003), abdominal pressure ( OR=1.469, 95% CI: 1.108-1.958, P=0.007), and lactic acid ( OR=3.156, 95% CI: 1.013-9.831, P=0.047) were independent risk factors for AKI in HTG-SAP patients. The AUCs of the APACHEⅡ score, abdominal pressure, lactic acid level and combined prediction model were 0.951, 0.918, 0.837 and 0.986, respectively. Conclusions:Patients with HTG-SAP complicated with AKI were more likely to have complications related to extrapancreatic infection, sepsis and multiple organ dysfunction, and the proportion of mechanical ventilation, CRRT, retroperitoneal puncture and surgical intervention required during hospitalization was greater, the lengths of ICU stay and total hospital stay were longer, and the mortality rate was higher. APACHEⅡ score, abdominal pressure and lactic acid were independent risk factors for HTG-SAP concurrent AKI, and the prediction model established based on these three factors had higher value in predicting HGT-SAP concurrent AKI

A splicing mutation in VPS4B can cause dentin dysplasia type I (DD-I), a hereditary autosomal-dominant disorder characterized by rootless teeth, the etiology of which is genetically heterogeneous. In our study, dental follicle cells (DFCs) were isolated and cultured from a patient with DD-I and compared with those from an age-matched, healthy control. In a previous study, this DD-I patient was confirmed to have a loss-of-function splicing mutation in VPS4B (IVS7 + 46C > G). The results from this study showed that the isolated DFCs were vimentin-positive and CK14-negative, indicating that the isolated cells were derived from the mesenchyme. DFCs harboring the VPS4B mutation had a significantly higher proliferation rate from day 3 to day 8 than control DFCs, indicating that VPS4B is involved in cell proliferation. The cells were then replenished with osteogenic medium to investigate how the VPS4B mutation affected osteogenic differentiation. Induction of osteogenesis, detected by alizarin red and alkaline phosphatase staining in vitro, was decreased in the DFCs from the DD-I patient compared to the control DFCs. Furthermore, we also found that the VPS4B mutation in the DD-I patient downregulated the expression of osteoblast-related genes, such as ALP, BSP, OCN, RUNX2, and their encoded proteins. These outcomes confirmed that the DD-I-associated VPS4B mutation could decrease the capacity of DFCs to differentiate during the mineralization process and may also impair physiological root formation and bone remodeling. This might provide valuable insights and implications for exploring the pathological mechanisms underlying DD-I root development.
ATPases Associated with Diverse Cellular Activities ; genetics ; Case-Control Studies ; Cell Differentiation ; genetics ; Cells, Cultured ; Dental Sac ; cytology ; Dentin Dysplasia ; genetics ; pathology ; physiopathology ; Endosomal Sorting Complexes Required for Transport ; genetics ; Humans ; Mutation ; genetics ; Osteogenesis ; genetics ; RNA Splicing ; genetics