ObjectiveTo investigate the relationship between carotid intima-media thickness (IMT)and plaque extent of carotid artery with coronary heart disease (CHD).Methods 131 inpatients were divided into 3 groups,in which 54 cases of coronary heart disease group,46 cases of risk group with coronary heart disease,31 cases of control group.The carotid wall IMT,plaque length and thickness was measured by Color Doppler ultrasound.The obtained data using SPSS 17.0 software for statistical processing.ResultsCarotid intimal thickening and the incidence had no significant difference between the group of coronary heart disease and risk group with coronary heart disease risk( all P ＞ 0.05),carotid artery IMT and incidence rate between the above two groups and the control group had statistical differene( t =3.26,3.48,all P ＜ 0.05 ),and the three groups of carotid artery plaque score and plaque classification were statistically significant( F =4.28,P ＜ 0.05 ).ConclusionCarotid IMT and plaque formation was the independent risk factor of CHD,and it could predict the occurrence and development of CHD,especially in carotid artery plaque specificity is higher,in the primary hospital could be used as the auxiliary examination method of CHD.

Objective To investigate the expression changes of aspartic proteinase (cathepsin D) and cysteine proteinase (cathepsin K) in photoaged fibroblasts. Methods The senescence of human fibroblasts was induced via culture in the presence of 8-methoxypsralen (MOP) of 50 mg/L in darkness for 24 hours followed by irradiation with UVA of 80 kJ/m~2. Then, aged fibroblasts were confirmed by senescence-associated β-galactosidase (SA-β-gal) staining. Real-time RT-PCR and Western blot were carried out to detect the mRNA and protein expressions of cathepsin D and cathepsin K in photoaged and normal control fibroblasts, respectively. Results Western blot showed a significant difference between photoaged and control fibroblasts in the grey scale of cathepsin D and cathepsin K (3.25 ± 0.33 vs 14.18 ± 2.25, f = 30.61, P < 0.01; 2.39 ± 0.66 vs 29.38 ± 4.62, t = 12.63, P< 0.01). The △Ct values for cathepsin D and cathepsin K mRNA were 2.79 ± 0.17 and -0.92 ± 0.06, respectively, in photoaged fibroblasts, significantly lower than those in the control fibroblasts (4.54 ± 0.34, 2.57 ± 0.13, t = 20.78, 28.50, respectively, both P < 0.01). According to the value of 2~(-△△Ct), the expression of cathepsin D and cathepsin K mRNA decreased 0.24 ± 0.021 and 0.09 ± 0.005 folds, respectively, in photoaged fibroblasts compared with the control fibroblasts. Conclusion The expression of cathepsin D and cathepsin K is decreased in photoaged fibroblasts.

Background: The onset and progression of type 1 diabetes mellitus (T1DM) is closely related to autoimmunity. Effective monitoring of the immune system and developing targeted therapies are frontier fields in T1DM treatment. Currently, the most available tissue that reflects the immune system is peripheral blood mononuclear cells (PBMCs). Thus, the aim of this study was to identify key PBMC biomarkers of T1DM. Methods: Common differentially expressed genes (DEGs) were screened from the Gene Expression Omnibus (GEO) datasets GSE9006, GSE72377, and GSE55098, and PBMC mRNA expression in T1DM patients was compared with that in healthy participants by GEO2R. Gene Ontology, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and protein-protein interaction (PPI) network analyses of DEGs were performed using the Cytoscape, DAVID, and STRING databases. The vital hub genes were validated by reverse transcription-polymerase chain reaction using clinical samples. The disease-gene-drug interaction network was built using the Comparative Toxicogenomics Database (CTD) and Drug Gene Interaction Database (DGIdb). Results: We found that various biological functions or pathways related to the immune system and glucose metabolism changed in PBMCs from T1DM patients. In the PPI network, the DEGs of module 1 were significantly enriched in processes including inflammatory and immune responses and in pathways of proteoglycans in cancer. Moreover, we focused on four vital hub genes, namely, chitinase-3-like protein 1 (CHI3L1), C-X-C motif chemokine ligand 1 (CXCL1), matrix metallopeptidase 9 (MMP9), and granzyme B (GZMB), and confirmed them in clinical PBMC samples. Furthermore, the disease-gene-drug interaction network revealed the potential of key genes as reference markers in T1DM. Conclusion These results provide new insight into T1DM pathogenesis and novel biomarkers that could be widely representative reference indicators or potential therapeutic targets for clinical applications.

Diagnosis, Differential ; Female ; Humans ; Laparotomy ; methods ; Pregnancy ; Pregnancy, Ectopic ; diagnosis ; surgery ; Retroperitoneal Space ; diagnostic imaging ; Tomography, X-Ray Computed ; Ultrasonography

Objective: To investigate the clinical manifestations, imaging features, clinicopathologic features, and differential diagnosis of solitary fibrous tumors/anginoblastomas (SFT/HPCs) originating in the central nervous system. Methods: Sixty cases of SFT/HPCs originating in the central nervous system were collected at Nanjing Jinling Hospital, from January 1, 2008 to December 31, 2016. The clinical data, imaging data, histomorphologic changes and immunohistochemical finding were analyzed in the sixty cases. Results: The 60 cases included 26 males and 34 females, aged 14 to 85 (median 49) years. The main clinical manifestations were headache, dizziness with nausea and vomiting. Radiologically, the tumors were large, enhancing, solid and cystic masses attached to the dura. Histopathologically, the neoplasms were composed of spindle cells with oval nuclei, inconspicuous nucleoli and moderate amount of eosinophilic cytoplasm arranged in fascicles with areas of hyalinized stroma, myxoid changes and a staghorn vascular pattern. Immunohistochemically, tumor cells of all cases were positive for vimentin (100.0%, 60/60), STAT6 (98.3%, 59/60), CD34 (61.7%, 37/60), and the tumor cells were typically positive for CD99, bcl-2, EMA and SSTR2 as well.Negative for S-100 protein, SOX10, E-cadherin, GFAP. Ki-67 index ranged from 1% to 50%. Forty cases were followed up for 6 to 82 months with average of 40 months, 30 patients were alive and 10 patients died. Conclusions Central nervous system SFT/HPCs can be aggressive and relapses may occur several years after diagnosis. STAT6 is highly sensitive and specific for the diagnosis. Complete tumor resection is optional treatment followed by radiotherapy and chemotherapy. There is a correlation between the prognosis and the location of the disease, the histological grade, Ki-67 index, and fusion gene variants.

Objective To investigate the changes of coagulatory function in septic rats induced by cecal ligation and puncture(CLP). Methods Cecal ligation and puncture(CLP)were performed to induce sepsis in SD rats. Coagulation indexes were detected at 8,16 and 48 h after operation, and histopathological changes of the lung, kidney, liver and spleen were examined using HE staining. Results The 12-day survival rate of the CLP-induced septic rats was 30%,with an acute onset and high mortality. In the acute phase of disease development of the CLP rats, the activated partial thromboplastin time(APTT)was prolonged(P<0.05)at 8 h,the prothrombin time(PT)was prolonged at 16 h (P<0.05), the factor XII activity in the endogenous coagulation pathway and the factor VII activity in the extrinsic coagulation pathway showed a transient inhibition, the thrombin time(TT)was prolonged at 48 h(P<0.01), and the content of fibrinogen(FIB)was increased gradually from 16 h(P<0.001). Among the other important coagulation and anticoagulation indexes,the number of platelets(PLT)was decreased gradually from 8 h(P<0.01),while the number of vWF:Ag increased gradually from 8 h(P<0.001). The D-dimer amount gradually increased from 16 h(P<0.05),and the amount of PS:Ag significantly decreased until 48 h(P<0.001). However, there was no significant change in the antithrombin-III(AT-Ⅲ)content. The histopathological examination showed that there are different degrees of damages in the lung,kidney,liver and spleen tissues,but no obvious venous thrombosis and bleeding were found. Conclusions In the acute phase,there is coagulatory dysfunction in the septic rats,however,no histopathological changes such as venous thrombosis and bleeding were observed in the lung,kidney,liver and spleen tissues due to coagulatory dysfunction.

Objective:To evaluate the ability of Sysmex urine automatic analyzer UF-5000 to detect renal tubular epithelial cells, and to explore the value of detection of renal tubular epithelial cells in renal tubular injury of diabetes mellitus.Methods:Case control study. 452 urine samples were collected from the third Xiangya Third Hospital of Central South University from October 2018 to April 2019 (252 in the control group, 113 in diabetes without renal injury group and 87 in diabetes with renal injury group). All samples were detected by both UF-5000 and microscopic examination, established reference range for normal population, then contrasted the coincidence rate and uniformity of the two methods, to evaluate the ability of urine automatic analyzer UF-5000 to detect renal tubular epithelial cells, and the diagnostic value of tubular epithelial cells for renal tubular injury in diabetic patients. All statistical analyses were performed using SPSS17.0, Kappa consistency analysis, ROC curve analysis, Kruskal-Wallis test and Chi-square test were used.Results:The reference range of renal tubular epithelial cells by Sysmex urine automatic analyzer UF-5000 is 0-1.7/μl. The results of the two methods were analyzed by Kappa consistency analysis. The Kappa value was 0.699, P>0.05, which meant highly consistent. ROC curve analysis showed when cut-off value was 1.7/μl. The sensitivity, specificity and area under ROC curve were 0.791, 0.817 and 0.861 respectively. The median of renal tubular epithelial cells was 0.4/μl, 2.0/μl and 2.3/μl in the healthy control group, the diabetes without renal injury group and the diabetes with renal injury group, respectively; the positive rate of renal tubular epithelial cells in the three groups were 2.78%, 56.64% and 75.86% respectively. Compared with the control group, the median and positive rate of renal tubular epithelial cells in the diabetes without renal injury group and the diabetes with renal injury group were significant different; there was also significant difference in the positive rate of renal tubular epithelial cells between the two groups. Conclusion:Compared with the control group, the positive rate of urine renal tubular epithelial cells indiabetes without renal injury group is significantly higher, which is helpful to detect renal tubular injury, to carry out early intervention and to prolong the time of progression to chronic kidney disease.

Objective To investigate the role of HK2 and VDAC1 in diacetylmorphine-induced cardiomyocyte apoptosis.Methods A dose-escalation method was used to establish a rat model of diacetylmorphine addiction.Forty SD rats were randomly divided into three groups,the normal group(n=10)was injected with an equal amount of saline subcutaneously,the model group(n=15)was injected with 5 mg/kg of diacetylmorphine for the first time,and then the dose was increased by 2.5 mg/(kg·d)day by day for 20 days,and the group of model +10 D(n=15)continued to increase the dose based on the model group up to the 10th day.Lactate dehydrogenase(LDH)and glutamic oxaloacetic transaminase(GOT)were detected by ELISA;HE staining was used to observe the pathological changes of myocardial tissues in each group;TUNEL staining was used to detect apoptosis in myocardial tissues in each group;and immunohistochemistry,RT-q-analysis,and immunochemistry were used to detect apoptosis in myocardial tissues in each group.Immunohistochemistry,RT-qPCR and Western bl-ot were used to detect the mRNA and protein expression of HK2,VDAC1 and apoptosis-related factors.Results HE staining revealed that myocardial tissues exhibited different degrees of damage with the prolongation of diacetylmorphine intervention.Compared with the normal group,serum LDH,GOT content and myocardial apoptosis rate increased in the model group,mRNA and protein levels of HK2 and anti-apoptotic factor Bcl-2 decreased,mRNA and protein levels of VDAC1 and pro-apoptotic factors Bax and Caspase-3 increased,and the protein level of Clevead Caspase-3 increased;in the model +10 D group the above indexes,there was a statistically significant difference(P<0.05).Conclusion Diacetylmorphine can cause cardiomyocyte apoptosis,and VDAC1 may be involved in the process of cardiomyocyte apoptosis caused by diacetylmorphine.

Intestinal failure (IF) is defined as the critical reduction of functional intestines below the minimum needed to absorb nutrients and fluids, so that intravenous supplementation with parenteral nutrition (PN) is required to maintain health and/or growth. Although the benefits are evident, patients receiving PN can suffer from serious cholestasis due to lack of enteral feeding and small intestinal bacterial overgrowth (SIBO). One such complication that may arise is intestinal failure-associated liver disease (IFALD). Evidences from recent studies suggest that alterations in the intestinal microbiota, as well as intraluminal bile acid driven signaling, may play a critical role in both hepatic and intestinal injury. Since Marshall first proposed the concept of the gut-liver axis in 1998, the role of gut-liver axis disorders in the development of IFALD has received considerable attention. The conversation between gut and liver is the key to maintain liver metabolism and intestinal homeostasis, which influences each other and is reciprocal causation. However, as a "forgotten organ" , intestinal microbiota on the pathogenesis of IFALD has not been well reflected. As such, we propose, for the first time, the concept of gut-microbiota-liver axis to emphasize the importance of intestinal microbiota in the interaction of gut-liver axis. Analysis and research on gut-microbiota-liver axis will be of great significance for understanding the pathogenesis of IFALD and improving the prevention and treatment measures.
Bacterial Infections/physiopathology* ; Bile Acids and Salts/physiology* ; Cholestasis/physiopathology* ; Enteral Nutrition ; Gastrointestinal Microbiome/physiology* ; Humans ; Intestinal Diseases/physiopathology* ; Intestines/physiopathology* ; Liver/physiopathology* ; Liver Diseases/physiopathology* ; Parenteral Nutrition/adverse effects* ; Short Bowel Syndrome/physiopathology* ; Signal Transduction

OBJECTIVETo investigate the effects of microRNA(miRNA)-29b on the proliferation and migration of breast cancer cells and its molecular mechanism.

METHODSThe recombinant lentiviral expression vector (lenti-miRNA-29b) was constructed and transfected into 293T cells to obtain lentivirus particles that were used to infect breast cancer MCF-7 cells. Transfection efficiency of lenti-miRNA-29b in MCF-7 cells was identified by the expression of green fluorescent protein (GFP). The expression of miRNA-29b was detected by real-time PCR. The cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The bioinformatics softwares were used to predict and screen the downstream target genes regulated by miRNA-29b, which were verified by double luciferase reporter gene assay, RT-PCR and Western blot. The effects of screened target gene RTKN on the growth and migration of MCF-7 cells were verified by RTKN siRNA.

RESULTSRecombinant lentiviral expression vector of miRNA-29b were successfully constructed. About 90% and 60% of the breast cancer cells showed green fluorescence in lenti-miRNA-29b and lenti-miRNA-NC groups, respectively. The expression of miRNA-29b in lenti-miRNA-29b group increased significantly compared with the lenti-miRNA-NC group and blank control group (all<0.05); the proliferation and migration ability of MCF-7 cells significantly reduced compared with the control group (all<0.05). The screening with bioinformatics softwares found that the 3'UTR coding region RTKN had the binding site to miRNA-29b; the dual luciferase reporter gene assay showed that the luciferase activity decreased significantly after the MCF-7 cells were co-transfected with wild type RTKN-WT-3'UTR and miRNA-29b mimics report gene vector (<0.05). The RTKN proteins in MCF-7 cells were significantly decreased after transfection with siRNA-RTKN, and the proliferation and migration ability of MCF-7 cells were significantly reduced (all<0.05).

CONCLUSIONSMiRNA-29b can inhibit the proliferation, invasion and metastasis of breast cancer cells by inhibiting the expression of RTKN.