Aim To investigate the effect of transduct-ed-hemeoxygenase-1 ( HO-1 ) protein on brain ischemi-a-reperfusion ( I/R ) rat hippocampal neurons injury. Methods 11 R ( arginine residues )-fused HO-1 pro-tein was established and 50 male Mongolian gerbils were randomly divided into 5 groups ( n=10 ):I/R ( control group ) , I/R + 5 mg · kg-1 saline group ( group S ) , I/R + 5 mg · kg-1 11 R group ( group R), I/R + 5mg·kg-1 11R-HO-1 group (group H1) and I/R + 25 mg · kg-1 11 R-HO-1 group ( group H2). For I/R experiments, ischemia was induced for 5 min by occluding the common carotid arteries bilater-ally with aneurysm clips under Ketamine anesthesia. The experiment was conducted after the neurons were intraperitoneally injected with 5mg·kg-1 saline,11R , 11R-HO-1,or 25mg · kg-1 11R-HO-1 for 3 h. The rats were killed after 24h of reperfusion. Hippocampus was removed immediately for determination of cAMP level, neuronal apoptotic rate, and expression of HO-1 and Caspase-3 protein, mitochondria was observed un-der electron microscope. Results Among group C, group S and group R,there were no differences in the expressions of HO-1, Caspase-3 protein, cAMP level , neuronal apoptotic rate and mitochondria damage ( P>0. 05). Compared with group C, group S and group R, the expression of HO-1 protein was up-regulated, the expression of Caspase-3 protein was down-regula-ted, cAMP level increased, the apoptotic rate was sig-nificantly decreased and mitochondria damage de-creased in group H1 ( P < 0. 01 ) . Compared with group H1 , the expression of HO-1 protein was up-regu-lated, the expression of Caspase-3 protein was down-regulated, cAMP level increased, the apoptotic rate was significantly decreased and mitochondria damage decreased in group H2 ( P <0. 01 ) . Conclusion Transducted-HO-1 protein can attenuate brain ischemi-a-reperfusion rat hippocampal neurons injury.

Objective To investigate the effects of estrogen ( E2) on the resistance to anoikis and a possible role of extracellular signal-regulated kinse ( ERK)-focal adhesion kinse ( FAK) signaling in the effect of estrogen to under-stand its underlying mechanism .Methods Poly-Hema-coated culture of human breast cancer cell line MCF-7 was used to induce anoikis .Cells were treated with E2 and/or pretreated with MEK or FAK inhibitors .Western blot was used to assess the phosphorylation of ERK and FAK , trypan blue staining and cell counting were employed to evaluate cell viability , and Hoechst staining was used to check apoptosis .Results Suspension culture greatly re-duced cell survival (P<0.01), and exposure of MCF-7 cells to E2 (10 nmol/L) led to a significantly increased resistance to anoikis and survival ( P<0.05 ) as compared to DMSO .Meanwhile , E2 induced increased phospho-rylation of both ERK and FAK .Pharmacological inhibition of MEK with U 0126 ( 10 μmol/L ) reduced E2-in-creased cell survival by 57.48%(P<0.01) and E2-decreased anoikis;Treatment with FAK inhibitor (10μmol/L) attenuated E2-enhanced cell survival by 53.59% ( P<0.01 ) and E2-reduced apoptosis .Conclusions E2 con-tributes to the enhanced cell viability and increased resistance to anoikis in MCF-7 breast cancer cells , and ERK-FAK signaling may be involved in the E 2-stimulated survival during suspension culture of MCF-7 cells.

Hydrogen spectrum nuclear magnetic resonance (1 H-NMR ) is a commonly used method for metabolomics and has been applied in the study of liver cirrhosis and liver cancer,however,study on serum metabolites in patients with primary biliary cholangitis (PBC)is rare.Aims:To investigate the capability of 1 H-NMR for screening serum metabolites of PBC.Methods:Twenty PBC patients,20 HBV-related cirrhosis patients and 20 healthy controls were detected by 1 H-NMR.The 1 H-NMR spectra data were processed by principal component analysis (PCA)and orthogonal partial least squares-discriminant analysis (OPLS-DA)so as to create the diagnostic model.Based on the correlation coefficient P (corr),VIP value and non-paired t test of OPLS-DA model,the differential metabolites between normal group and the two cirrhosis groups were screened.Results:OPLS-DA model could effectively distinguish healthy controls and PBC patients,model interpretation ability and prediction ability were 81.9% and 44.8%,respectively (P=0.0293), glutamine,folic acid,urocanic acid,4-ethylbenzoic acid were differential metabolites.OPLS-DA model could also effectively distinguish healthy controls and HBV-related cirrhosis patients, urocanic acid, 1-methylhistamine, 1-methyladenine,glucose,L-acetylcarnitine were differential metabolites.OPLS-DA model could not effectively distinguish PBC patients and HBV-related cirrhosis patients.Conclusions:Serum glutamine and folic acid may be the potential biomarkers of PBC,which may be closely related to the immune damage mechanism and prognosis of PBC;1 H-NMR combined with OPLS-DA diagnostic model are expected to become a new method for studying liver cirrhosis.

Objective To evaluate the effect of sustained silencing Forkhead box Ml (F0XM1) gene by short-hairpin RNA (shRNA) expression vector on cell growth of hepatocelluar carcinoma (HCC) in vitro.Methods Four shRNA expression vectors targeting different sequences of human F0XM1 mRNA were constructed.The expression vector with the best interfering effect and the negative control plasmid were used to transfect HCC cell line QGY-7703, stably transfected cell clones were selected by neomycin (G418).Cell growth was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and colony formation was assessed by clonogenic assay.Cell apoptosis was detected by double staining with APC conjugated Annexin V and PI.Results F0XM1 protein was detected with different levels in all these studied human cell lines.The expression vector shRNA-1026 exhibited excellent interference effect after transient transfection, which showed 38.5% and 53.2% reduction of FOXM1 mRNA and protein level respectively.The growth of QGY-7703 cells was inhibited after stable inhibition of FOXM1 expression by shRNA-1026, which was indicated by decreased absorbance value of the test group after culture for 48, 72 and 96 h compared to control group (t = 10.830,3.578 and 5.734 respectively, P ＜ 0.05).Stable inhibition of F0XM1 also led to reduced colony formation ( t = 5.336, P ＜ 0.05 ) and increased apoptosis of QGY-7703 cells in comparison to control cells (t = 6.827, P ＜ 0.05 ).Conclusions Stable silencing F0XM1 gene by shRNA suppresses the growth of HCC cells in vitro.

Objective: To investigate the protective effects of matrine combined with glycyrrhizic acid on chronic liver injury induced by carbon tetrachloride, and explore the protective mechanism from the points of energy metabolism and CYP enzyme.Methods: The chronic hepatic injury model of rats was induced by CCl4.The changes of activity of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum were measured to observe the protective effect of the two drugs and their combination.The contents of glutamate dehydrogenase (GLDH) in serum and adenine nucleoside three phosphate (ATP), adenosine diphosphate (ADP) and adenine monophosphate (AMP) in liver tissue were determined to evaluate the regulation effect on hepatic energy metabolism and mitochondrial function.The levels of CYP1A2, CYP2E1 mRNA and protein in liver tissue were detected by real-time PCR and Western Blot to evaluate the two drugs and their combination on the regulation function of liver CYP enzyme.Results: Matrine (72.8 mg×kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease the serum activities of ALT and ALT in chronic hepatic injury model, and the combination (matrine 36.4 mg·kg-1+glycyrrhizic acid 21.7 mg·kg-1) had the most significant protective effect (P＜0.05).Matrine (72.8 mg·kg-1)and glycyrrhizic acid(43.4 mg·kg-1)could decrease GLDH in serum,and restore the content of ATP in liver (P＜0.05).Matrine (72.8 mg·kg-1) had no effect on the expression of CYP1A2 and CYP2E1mRNA, and glycyrrhizin (43.4 mg·kg-1) could inhibit the expression of CYP1A2, CYP2E1mRNA and protein (P＜0.05).Conclusion: Matrine combined with glycyrrhizin has obvious regulation effect on mitochondrial function and liver protective effect in chronic hepatic injury model.

OBJECTIVE: To explore biomechanical characteristics of minimally invasive different screw fixations in treating Sanders typeⅡcalcaneal fractures. METHODS: Dicom data of calcaneus by CT scan were input into Mimics 21.0 software and Ansys15.0 software to construct three-dimensional finite element digital model of calcaneus;this model was input into UG NX 10.0 software, and calcaneus was cut according to Sanders classification to establish Sanders typeⅡ calcaneus model with posterior articular surface collapse;then simulated minimally invasive screw internal fixation after calcaneal fracture:a screw from posterior articular surface was used to outside-in fix sustentaculum tali, other 4 screws were used to fix calcaneus by different methods through calcaneal tuberosity, and 4 different calcaneal models were obtained. Under the same conditions, 4 types of internal fixation models were loaded respectively, and nonlinear finite element analysis was performed to calculate the stress distribution of different internal fixation models. RESULTS: Under the same condition of loading, the model 3 had smaller displacement value, maximum calcaneus displacement value and maximum equivalent stress value of the screw than other three internal fixation models, and the stress was more dispersed. CONCLUSION In minimally invasive screw internal fixation of calcaneus fracture, after 1 sustentaculum tali screw fixation, 2 screws crossed fix posterior articular surface from calcaneal tuberosity, 2 screws fix parallelly calcaneocuboid joint from calcaneal tuberosity are more suitable for biomechanical requirements, and could provide basic theory for clinical treatment.
Bone Screws ; Calcaneus/surgery* ; Finite Element Analysis ; Fracture Fixation, Internal ; Fractures, Bone/surgery* ; Humans ; Treatment Outcome

Objective: To investigate the use of conventional MR imaging to guide treatment in patients with cholecystolithiasis and diffuse inflammatory thickening of gallbladder wall. Methods: The clinical data of patients who were treated in the Ningbo Huamei Hospital, University of the Chinese Academy of Sciences between January 2017 and January 2018 were analyzed. These patients were divided into two groups: patients with acute cholecystitis (n=139) and patients with viral hepatitis combined with cholecystolithiasis (n=67). Differences in the imaging signs in standardized upper abdominal contrast enhanced MRI examinations were retrospectively analyzed. Results: The imaging signs, including stone location, continuity of gallbladder mucosa, exudation in peri-gallbladder space, edema of intrahepatic portal area showed significant differences between the two groups (all P<0.05). On stratification analysis, the type of thickened gallbladder wall, background of liver parenchyma and extent of edema in intrahepatic catchment area also showed significant differences (all P<0.05). The imaging signs, including non-gallbladder neck ductal stones, concentric thickening of gallbladder wall, continuous mucous membrane in gallbladder and no peri-gallbladder space exudation but diffuse edema of intrahepatic catchment area supported the diagnosis of viral hepatitis combined with gallstones. The imaging signs, including discontinuity of gallbladder mucosa, exudation of peri-gallbladder space, diffuse edema of gallbladder wall without a cirrhotic background and edema in intrahepatic portal area supported the diagnosis of acute calculous cholecystitis of gallbladder. Conclusions Routine upper abdominal contrast enhanced MRI plays an important role in demonstrating the underlying cause of gallbladder wall diffuse edema thickening in patients with gallstones. It provides an important reference for the choice of clinical treatment pathway.

This experimental study was aimed to construct the recombinant bisbicistronic eukaryotic expression vector containing endocrine and exocrine protein (EECP) gene associated with breast cancer and enhanced green fluorescent protein (EGFP) gene. And then we transfected it into breast cancer cells MCF-7 to detect the expression of EECP protein and study preliminary biological function of EECP gene. The EECP sequence was cloned to pBluescript II SK (+) plasmid. After restriction endonuclease reaction of pBluescript II SK(+) plasmid, the EECP fragment was cloned to pIRES2-EGFP vector forming a recombinant eukaryotic expression vector named pEECP-IRES2-EGFP. The potential vector was identified by restriction endonuclease digestion and sequencing. Correct plasmid was extracted and transfected into breast cancer cells MCF-7. The expression of EECP protein was detected by western blot analysis. Its biological function was studied by MTT and Flow-cytometry. It turns out that the recombinant eukaryotic expression vector containing EECP gene and EGFP gene was constructed successfully, and it could transfect MCF-7 cells efficiently. It can get higher expression of EECP protein and higher cell proliferation, thus providing an important and convenient tool for studying the function of EECP gene in vitro and in vivo.
Base Sequence ; Breast Neoplasms ; genetics ; pathology ; Female ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; MCF-7 Cells ; Molecular Sequence Data ; Proteins ; analysis ; genetics ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Ribosomes ; chemistry ; metabolism

Objective:To investigate the current state of standardized training in trauma surgeons in medical institutions in Shanghai.Methods:Questionnaire surveys, focus discussions and expert conferences were conducted to investigate the professional background, status of standardized training on trauma care knowledge, training needs and suggestions in 236 trauma surgeons in 25 Shanghai medical institutions.Results:The investigation revealed that only 59.3% (140/236) of the trauma surgeons had received a standardized training on trauma care knowledge. Of them, those who participated in the training by an international, domestic and local medical institution accounted for 26.4% (37/140), 47.1% (66/140) and 84.3%(118/140), respectively. Only 83 surgeons (70.3%) received regular training and most of them 66.3% (55/83) did in a low frequency of 1-2 times a year. Only 12.7% (15/118) of the surgeons received a training course of ≥48 classes (45 min per class) in the past 3 years. The training courses by medical institutions were mostly lectures on theoretical knowledge and training of a single skill while the simulated scenario drills accounted for less than 48.3%(57/118). The surgeons who had received a standardized training on trauma care were more likely to use their training knowledge to grade and score the patients with trauma, significantly more than those who had not( P<0.05). Conclusions:The coverage, time or frequency of standardized training on trauma care was not enough in Shanghai. The standardized training of trauma care surgeons should be strengthened in all medical institutions and be supervised by the management administration. A set of standardized materials and assessment standards for trauma care training should be formulated as soon as possible which fits the domestic conditions in China. Training bases should be established in Shanghai for standardized care of severe trauma and trauma specialists. A training system for trauma specialists should be established to promote traumatology in shanghai.

Objective: To analyze guanine nucleotide-binding protein subunit beta-2-like 1 (GNB2L1) expression based on bioinformatics, so as to evaluate its role and its relationship with survival rate during the occurrence and development of hepatocellular carcinoma. Methods: GEPIA, UALCAN and HPA databases were used to analyze the expression level of GNB2L1 and its relationship with HCC survival rate. Mutations in the GNB2L1 gene and their impact on survival were analyzed using the cBioPortal database. LinkedOmics database was used to analyze GNB2L1-related genes in HCC. Gene Ontology (GO) functional annotation and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis were performed simultaneously. STEING database was used to construct the GNB2L1 protein interaction network. TIMER database was used to analyze the relationship between GNB2L1 gene expression and immune infiltration in hepatocellular carcinoma. Differential expression of GNB2L1 in plasma platelets of HCC patients and healthy controls was analyzed using mRNA-based sequencing technology. Data between groups were compared using an independent-samples t-test. Results: GNB2L1 expression level was significantly increased in HCC tissues (P<0.05), and its expression was significantly correlated with body weight, classification and stage (P<0.05). The overall survival rate was higher in GNB2L1 low expression group (P<0.001). GNB2L1 and its related genes were related to biological process regulation, metabolic process, protein binding, oxidative phosphorylation, JAK-STAT signaling pathway, Ras signaling pathway and so on. GNB2L1 had interaction with RPS12, RPS11 and RPL19, and participated in multiple biological processes such as liver regeneration and positive regulation of endogenous apoptotic signaling pathway. GNB2L1 expression was significantly positively correlated with the infiltration degree of various immune cells in HCC (P<0.05). Cox regression analysis showed that GNB2L1 was an independent risk factor for lower survival rate in patients with HCC [Hazard ratio (95% confidence interval)=1.456 (1.034~2.051), P=0.031]. GNB2L1expression levels were significantly higher in platelets of HCC patients than that of healthy controls (10.40±1.36 vs. 9.58±0.51, t=2.194, P=0.037). Conclusion: GNB2L1 has high expression and close relationship to survival rate in HCC. Therefore, GNB2L1 may be a potential biomarker of HCC.
Humans ; Carcinoma, Hepatocellular/pathology* ; Computational Biology ; Liver Neoplasms/pathology* ; Protein Subunits/metabolism* ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; RNA, Messenger ; Guanine Nucleotides ; Gene Expression ; Biomarkers, Tumor/genetics*