Objective To evaluate the influence of C-reactive protein (CRP),insulin resistance (IR) and epicardial fat volume (EFV) on the extent of coronary atherosclerosis in patients with different body mass index(BMl).Methods One hundred and three patients with coronary artery disease were involved in current study who underwent 64-slice dual source CT and percutaneous coronary angiography.Measurements of height,weight,waist circumference (WC) were recorded,and BMI was calculated.All patients were divided into obesity group (n =45) and non-obesity group (n =58) based on BMI.EFV were calculated through 64-slice dual source CT.Blood samples were collected for biochemical examination.Gensini score were adopted to quantify the severity of coronary artery stenosis.The relationship between Gensini score and EFV,CRP and homeostasis model assessment-insulin resistance(HOMA-IR) index were statistical analyzed by SPSS16.0 software.Results The level of CRP,WC,EFV and BMI in obesity group were (11.0 ± 5.8) mg/L,(96.1 ± 7.0) cm,(122.7 ± 43.3) cm3,(27.9 ± 2.9) kg/m2 respectively,significantly higher than those in non-obesity group ((6.5 ± 3.4) mg/L,(86.4 ± 7.6) cm,(92.9 ± 39.5) cm3,(22.4 ± 1.9) kg/m2) and the differences were significant (t =2.24,6.74,3.64,11.74,and P ＜ 0.05).CRP were positively correlated with EFV (r =0.404,0.364,P ＜0.05) in both obesity and non-obesity group,While HOMA-IR were only associated with BMI in obese group(r =0.322,P ＜0.05).Gensini score in non-obesity groups were positively related with EFV and CRP (r =0.358,0.315,P ＜ 0.05),while in obesity groups were positively related with EFV,CRP and HOMA-IR(r =0.348,0.297,0.384; P ＜ 0.05).The associations between Gensini score and CRP were not significant in obesity group after adjusting BMI and WC.Multiple linear regression analysis showed that EFV and diabetes mellitus were independent risk factors of patient Gensini score.Conclusion Coronary atherosclerosis is positively related with EFV and CRP in all patients.While,coronary atherosclerosis is influenced by BMI,WC and HOMA-IR in obese group.EFV is an independent risk factor of coronary atherosclerosis.

Aim To develop an in vitro high throughput drug screening system based on reporter gene assay for identification of novel compounds with PXR, FXR and LXRα agonist activity. Methods The expressions of exogenous PXR, FXR and LXRαgene in HEK293, HepG2 and LS174T cells were examined by Real-Time quantity PCR. pSG5-hPXR and pGL3-XREM-CYP3A4, pEGFP-N3-hFXR and EcRE-TK-Luc, pCMX-FLAG-hLXRα and pGL3-XREM-CYP3A4 were cotransfected into cells and the optimal ratio of three plasmids was determined. The dose-response relationship between the positive drug and the fold induction was determined. The specificity of the model was ex-amined, and the repeatability was also determined by Z′ value. Results ① The PXR, FXR and LXRα mRNA expression in HEK293 cell is low among three different cells. ②reporter gene vector and expression plasmid ratio of 1∶ 1, 2∶ 1 and 2∶ 1 were proved to be suitable for highest relative luciferase activity for PXR, FXR or LXRα agonist screening model. ③ The relative luciferase activity was induced by Rif, CDCA or T0901317 in a dose-dependent manner. ④Only Rif, CDCA or T0901317 could significantly increase the relative luciferase activity in PXR,FXR or LXRα agonist screening model, no effect of other nuclear re-ceptors agonist was observed, and the values of Z′-factor for PXR, FXR and LXRαagonist screening model were 0. 58, 0. 66 and 0. 63, respectively. Conclusion An in vitro PXR, FXR and LXRα agonist high-throughput screening models are devel-oped with acceptable specificity and repeatability, and the mod-els can be used to screen PXR, FXR and LXRα agonist.

Recently the methylotrophic Pichia pastoris,with many advantages such as high expression,stable genetics and protein secretion,has been used extensively as a host expressing heterologous proteins.The optimal seed medium among seven media MM,MD,MGY,BMGY,YMPD,YMPGy and MGyB is BMGY with addition of 4mL/L PTM1 The high density synthesized medium in shake flask culture is as follows: Glycerol 4%(NH 4) 2SO 4 10g/L,CaSO 4 0.93g/L,K 2SO 4 18 2g/L,MgSO 4?7H 2O 14 9g/L,add 0.1mol/L PBS(pH=6 0) to 1 0 liter.The harvested yeast cell optical density (OD 600 ) reached 65 or more after 26 hours cultivation.By analysis of SDS\|PAGE,the results of Pichia pastoris culture by methanol inducement for 72 hours showed that the expression of recombinant human serum albumin had been achieved by methanol inducement after 12 hours,and the mount of targeted protein reached the summit after 24 hours inducement by methanol.The high density synthesized medium in shake flask culture in this experiment,which is similar to the mediums of batch fermentation and batch\|fed fermentation,is benefit to direct the P.pastoris fermentation in the fermentor.

Abstract: Objective　To study severe fever with thrombocytopenia syndrome virus (SFTSV) in tick samples from different species and genera in Suizhou City, Hubei Province, China, and to explore the phylogenetic relationship between ticks and patients sources of viruses at the molecular evolutionary level. Methods In 2016 and 2017, over a continuous two-year period, 1 158 ticks were collected from Suizhou, Hubei, and their species and genera were identified. Meanwhile, 86 serum samples were collected to detect SFTSV RNA by real-time quantitative reverse transcription-PCR. All viral RNA-positive supernatants of tick homogenates were inoculated into Vero cells for viral isolation, and full genome sequencing of isolated strains was conducted. Phylogenetic tree research on SFTSV strains from ticks and cases was performed using the bootstrapped maximum-likelihood (1 000 iterations) method with Molecular Evolutionary Genetics Analysis (MEGA) software, ver. 11.0 to provide confidence estimates. Results Haemaphysalis longicornis, Ixodes sinensis, and Rhipicephalus microplus were the dominant species (95.34%) in Suizhou City, Hubei Province, China. Tick samples were pooled according to their species and developmental stage, yielding 832 pools, of which 4 were positive for SFTSV by qRT-PCR. The overall minimum infection rate (MIR) in the region was 0.35%. One SFTSV strain named HB 2016-P35, was successfully isolated from Haemaphysalis longicornis and demonstrated high homology to 16 previously reported patient-derived viruses in Hubei Province, especially to the human strain HB 2017-49 from the same region, with a genome similarity of 99.9%. In addition, the molecular phylogenetic analysis revealed five distinct SFTSV genotypes in Hubei, covering almost all currently known SFTSV genotypes. Conclusions Some areas of Suizhou City, Hubei Province, demonstrate a relatively low level of SFTSV carrying and transmission by ticks. The new SFTSV strain isolated from ticks exhibits similar genotype characteristics and high sequence homology with viruses carried by cases in surrounding cases. The study suggests that tick-to-human transmission is most likely the pathway for human infection with SFTSV, highlighting the need for continual and long-term monitoring of tick carriage of SFTSV in endemic areas.

AIMTo study the mechanism of protective effect of GABA against hypoxic injury in rat hippocampal slices.

METHODSThe hippocampal slices from adult rats and extracellular recording technique were used to observe the effect of GABA on the evoked population spikes in rat hippocampal slices after hypoxia in vitro.

RESULTSGABA can significantly delay the disappearance of PV, but have no effect on PS. When the receptor antagonist of GABA (bicuculline) and the inhibitor of Cl- channel (NPPB) were given, the protect effect could be suppressed.

CONCLUSIONGABA increases hypoxic tolerance of hippocampal slices. The mechanism of the effect of GABA may be involved in the elevation of chloride influx through GABA receptor.

Animals ; Chloride Channels ; physiology ; Evoked Potentials ; Hippocampus ; physiopathology ; Hypoxia ; drug therapy ; physiopathology ; In Vitro Techniques ; Male ; Neuroprotective Agents ; therapeutic use ; Rats ; Rats, Wistar ; gamma-Aminobutyric Acid ; therapeutic use

OBJECTIVETo investigate the inhibiting action of transforming growth factor-beta type R II receptor(TGF-beta R II) antibody to fibroblasts.

METHODSFour groups of cultured skin fibroblasts were treated with different dosages of TGF-beta R II antibody. Examinations were performed with HYP, MTT, microscope, and flow cytometry techniques.

RESULTSTGF-beta R II antibody resulted in significant inhibition of fibroblasts(P < 0.01).

CONCLUSIONTGF-beta R II antibody has inhibiting effects on the proliferation of skin fibroblasts and with a dose-dependent manner.

Adolescent ; Antibodies, Monoclonal ; pharmacology ; Cell Division ; drug effects ; Cells, Cultured ; Child ; Fibroblasts ; cytology ; drug effects ; ultrastructure ; Flow Cytometry ; Humans ; Microscopy, Electron ; Protein-Serine-Threonine Kinases ; Receptors, Transforming Growth Factor beta ; analysis ; immunology

Objective According to the clinical and gene mutation characteristics of ACADVL-related very long chain acyl-CoA dehydrogenase deficiency(VLCADD),the types that contribute to the gene mutation of ACADVL were summarized.Methods By analyzing clinical,laboratory and genetic data of 1 case with ACADVL-related very long chain acyl-CoA dehydrogenase deficiency diagnosed from Department of Pediatrics,the Affiliated Hospital of Inner Mongolia Medical University in August 2016,based on the agreement signed by both the litde patient's parents and the hospital,plus the high-throughput sequencing analysis and PCR sequencing test for the 2 generation genes,some presented mutation sites were analyzed and concluded,in addition to taking "ACADVL" as key words to search the databases from CNKI,Wanfang(updated in 2016) as well as PubMed and related documents from On-line Mendal Inheritant databases of Man (OMIM) and HGMD.Results Through physical examination,VLCADD was diagnosed.After being given Levocamitine and the diet likemedium-chain fatty acid food for a week,the metabolism returned to normal.Tracking him for 3 months,his hepatitis obviously rebounded,within the reach of 3 cm under the right rib and 1 cm under the xiphoid.The exome sequencing study (trios) was identified the novel heterozygous mutation according to the statistics below A CAD VL (N M_000018.3) Exon7:c.608 C ＞ T;p.(Pro203 Leu) (heterozygous) and A CAD VL (NM _000018.3) Exon18:c.1748C ＞ T;p.(Ser583Leu) (heterozygous) in ACADVL.Relevant literature reported suggest these two mutations from both the parents are pathogenic genes,which can account for the reason why the boy got ill.However,these two mutations had not been reported in ACADVL-related VLCADD so far.Up to now,73 types of mutations from documents index were related to the VLCADD,but the clinical case included 75 kinds of gene mutations.Conclusions The apparent symptoms of the boy with the gene mutation were reflected in abnormal heart rates,hepatomegaly and hypoglycemia.VLCADD was diagnosed through genetic testing,and systematic treatment can partly control the development of the disease.In conclusion,the findings (exon 7 and 18) show that according to the genetic tests,disease-causing genes from both parents are new mutations of ACADVL and they are pathogenic.

BACKGROUND: Multiple techniques are commonly used for posterior cruciate ligament (PCL) reconstruction. However, the optimum method regarding the fixation of PCL reconstruction after PCL tears remains debatable. The purpose of this study was to compare the biomechanical properties among three different tibial fixation procedures for transtibial single-bundle PCL reconstruction. METHODS: Thirty-six porcine tibias and porcine extensor tendons were randomized into three fixation study groups: the interference screw fixation (IS) group, the transtibial tubercle fixation (TTF) group, and TTF + IS group (n = 12 in each group). The structural properties of the three fixation groups were tested under cyclic loading and load-to-failure. The slippage after the cyclic loading test and the stiffness and ultimate failure load after load-to-failure testing were recorded. RESULTS: After 1000 cycles of cyclic testing, no significant difference was observed in graft slippage among the three groups. For load-to-failure testing, the TTF + IS group showed a higher ultimate failure load than the TTF group and the IS group (876.34 ± 58.78 N vs. 660.92 ± 77.74 N [P < 0.001] vs. 556.49 ± 65.33 N [P < 0.001]). The stiffness in the TTF group was significantly lower than that in the IS group and the TTF + IS group (92.77 ± 20.16 N/mm in the TTF group vs. 120.27 ± 15.66 N/m in the IS group [P = 0.001] and 131.79 ± 17.95 N/mm in the TTF + IS group [P < 0.001]). No significant difference in the mean stiffness was found between the IS group and the TTF + IS group (P = 0.127). CONCLUSIONS In this biomechanical study, supplementary fixation with transtibial tubercle sutures increased the ultimate failure load during load-to-failure testing for PCL reconstruction.
Animals ; Biomechanical Phenomena ; Posterior Cruciate Ligament Reconstruction ; Sutures ; Swine ; Tendons/surgery* ; Tibia/surgery*

Objective To evaluate the application of multi-parameter three-dimension arterial spin labeling (3D-ASL) in observing the brain perfusion of patients with transient ischemic attack (TIA). Methods A total of 42 TIA patients, admitted to our hospital from July 2014 to March 2017, were included in this study. All subjects underwent conventional MRI, diffusion-weighted imaging (DWI), magnetic resonance angiography (MRA) and 3D-ASL scanning. Abnormal signals, and cerebral arterial stenosis or occlusion were observed under MRI, DWI and MRA; cerebral blood flow (CBF) map was drew after analyzing the 3D-ASL imaging, and abnormal reperfusion of ASL-CBF was qualitatively and quantitatively analyzed. The detection rate of abnormal reperfusion in TIA patients by 3D-ASL (PLD=1.5 s, PLD=2.5 s) and MRA were compared. Results Forty-two TIA patients showed no positive findings on conventional MRI and DWI maps, of which 18 patients showed different degrees of cerebral artery stenosis on MRA maps. Twenty-seven patients (PLD=1.5 s, 64.29％) and 21 (PLD=2.5 s, 50％) on ASL-CBF maps showed different sizes and degrees of abnormal hypoperfusion, and significant difference was found in detection rate of hypoperfusion by 3D-ASL (PLD=1.5 s and PLD=2.5 s, χ2=23.333, P=0.000). The detection rates of hypoperfusion by 3D-ASL (PLD=1.5 s and PLD=2.5 s) were 中华神经医学杂志2017年12月第16卷 第12期 Chin J Neuromed, December 2017, Vol.16, No.12 significantly higher than that by MRA (χ2=17.500, P=0.000; χ2=31.500, P=0.000). Conclusions The 3D-ASL can quantitatively analyze the degrees of perfusion of patients with TIA. 3D-ASL can comprehensively reflect the perfusion status in patients with TIA, and short PLD 3D-ASL is more sensitive than long PLD ASL in finding TIA, while long PLD 3D-ASL can reflect the perfusion status more truly.

Objective To investigate the expression level of IL-32 in serum and its correlation with serum biochemical indices of liver function test and HBV DNA load in patients with HBV-related liver failure.Methods Fifty-five patients with HBV-related liver failure (severe hepatitis group) and twenty normal cases (control group) were enrolled in the study.Total RNA in PBMCs was extracted by using TRIzol.IL-32 mRNA level was assayed by using Real-time PCR.IL-32 protein level in serum was detected by ELSIA method.The correlation between IL-32 and ALT,AST,TBIL,HBV DNA load was analyzed using pearson's correlation analysis,respectively.Results Serum IL-32 expression level in severe hepatitis group was higher than that of control group.Moreover,the difference between them was statistically significant (P ＜0.05).Serum IL-32 level was positively correlated with serum ALT,AST,TBIL,respectively (P ＜ 0.05),but was not correlated with HBV DNA load (P ＞ 0.05).Conclusion Serum IL-32 expression level was increased in patients with HBV-related liver failure and was associated with the severity of inflammation.We,therefore,believe that IL-32 might be involved in the pathogenesis of HBV-related liver failure.