Clinical Application of Serum miRNA-126,miRNA-155 Detection in Evaluation of Plaque Property in the Carotid Atherosclerotic Desease / 现代检验医学杂志

Objective To investigate the clinical application of serum miRNA-126,miRNA-155 detection in evaluation of plaque property in the carotid atherosclerotic(CAS)desease.Methods A total of 75 patients with the CAS from May 2015 to May 2017 in the Xianyang Central Hospital and Shiquan Country Chinese Traditional Medicine was chosen,consisted of 35 cases of vulvernable plaque group and 40 cases of stable plaque group.Meanwhile,39 cases of healthy physical examines at the same time were regarded as the control group.The expression levels of serum miRNA-126,miRNA-155 in the groups were detected using the real-time reverse transcription-polymerase chain reaction technique.The largest carotid artery plaque thickness(MAPT)and intima-media thickness(IMT)in the groups were measured using the cervical enhancement CT.Re-sults The results of MAPT and IMT were(3.27±1.01 mm,1.93±0.51 mm)in the vulvernable plaque group and(2.50 ±0.79 mm,1.60±0.26 mm)in the stable plaque group.The carotid artery largest plaque thickness and intima-media thick-ness was higher in the vulvernable plaque group than in the stable plaque group(t=9.76,7.86,P<0.01),and there were significant differenes between the two groups.The expression levels of serum miRNA-126and miRNA-155 were(0.22 ± 0.06,0.87±0.18)in the vulvernable plaque group,(0.50±0.12,0.47±0.10)in the stable plaque group and(0.90±0.19, 0.19±0.05)in the control group.MiRNA-155 expression levels significantly increased in stable plaque group and vulvern-able plaque group compared with in the control group,which increased in the vulvernable plaque group compared with in the stable plaque group,and miRNA-126 expression levels markedly decreased,the differences were statistically significant(F=119.3,102.9,P<0.01).In the vulvernable plaque group,miRNA-126 expression negatively correlated with miRNA-155(r=0.912,P<0.01).miRNA-126 expression levels were inversely associated with the carotid artery largest plaque thickness and the intima-media thickness(r=-0.913,-0.893,P<0.01).While miRNA-155 expression levels were positively corre-lated with them(r=0.899,0.907,P<0.01).Conclusion Serum miRNA-155,miRNA-126 detection can be applied to pre-diction of CAS plaques rupture,and may become a useful warning marker of ischemic stroke events.

Pathogens Distribution and Drug Susceptibility Analysis of Adults Patients by Automatic Blood Culture / 现代检验医学杂志

Objective To study the distribution of pathogens,the positive time and drug resistance of pathogenic bacteria by blood culture of adult patients,in order to provide the basis for the early clinical discovery and treatment of bacteremia. Methods 3 537 specimens of adult blood culture were collected from July 2016 to December 2016,then identified the posi-tive bacteria strains,and analysed the antimicrobial susceptibility.Results In 3 537 specimens of adult blood culture,485 positive samples were detected,and the positive rate was 13.7%(485/3 537).Including 203 cases(41.9%)of both aerobic and anaerobic positive bottles,220 cases(45.3%)of aerobic positive bottles,and 62 cases(12.8%)of anaerobic positive bottles.About pathogens,229 specimens were gram-negative bacteria strains,accounting for 47.2%.The great majority of bacteria was E.Coli,Klebsiella pneumoniae,Pseudomonas aeruginosa,and they all showed sensitivity to imipenem.202 specimens were gram-positive bacteria strains,accounting for 41.7%,mainly on Staphylococcus aureus and Staphylococcus epidermidis,they all showed sensitive to vancomycin.54 strains were Fungi,accounting for 11.1%.For analysis of 203 ca-ses of aerobic and anaerobic both positive bottles,the results showed that:there were 121 cases of gram-negative bacteria strains,95(78.5%)specimens anaerobic jar to positive time earlier than aerobic bottle to positive time,26(21.5%)speci-mens aerobic bottle jar to positive time earlier than anaerobic bottles to positive time.78 cases of gram positive bacteria,an-aerobic jar to earlier than aerobic bottle to positive time had 45 strains,accounting for 57.7%.Aerobic bottle to positive time earlier than anaerobic bottles to positive time had 33 strains,accounting for 42.3%.Fungi,a total of 4 strains,50% each. Positive pathogens were mainly distributed in I,emergency surgery,respiratory medicine department.Conclusion Pathogen-ic bacteria isolated from the adult blood culture was given priority to gram-negative bacteria,pathogenic bacteria species and drug susceptibility difference was obviously.Clinicians should be combined with blood culture and drug susceptibility results of use of antimicrobial drugs to patients.

Diagnosis Value of CEA,FDP and DD in the Pleural Effusion Specimens for Benign and Malignant Pleural Effusion / 现代检验医学杂志

Objective To investigate the significance of expressions of carcinoembryonic antigen(CEA),fibrinogen degrada-tion products(FDP)and D-dimer(DD)in the pleural effusion specimens for differential diagnosis between the benign and malignant pleural effusion.Methods 40 patients with benign pleural effusion patients and 30 patients with malignant pleural effusion were divided into benign or malignant group.The levels of CEA in pleural effusion was detected by electrochemilu-minescence,and FDP and DD were measured by turbidimetry.The value of diagnosis of three separate indicators and combine detection was compared by ROC analysis.Results The levels of CEA,FDP and DD in the pleural effusion specimens of ma-lignanat group were significantly higher than the benign group(t=2.523~3.889,all P<0.01).The sensitivity of combined detection of CEA,FDP and DD was 76.7%,and the rate of correct diagnosis was 82.9%.The sensitivity and diagnostic dffeiciency of combined detection were higher than separate indicators(χ2=1.036~3.324,all P<0.05).Conclusion Com-bined detection of CEA,FDP and DD is helpful to differential diagnosis of benign and malignant pleural effusion.

Role of up-regulation of THEM4 on collagen secretion in human renal tubular epithelial cells treated with high glucose / 中国药理学通报

Aim To investigate the effect of thioester-ase superfamily member 4(THEM4)expression on col-lagen secretion in human renal proximal tubular epithe-lial cells (HKC)treated with high glucose. Methods In order to examine the direct effect of THEM4 ex-pression vector on PI3K/ Akt pathway and collagen se-cretion,pYr-ads-4-THEM4 expression vector was con-structed and transfected into the HKC with lipo-fectamine 2000 in vitro. HKC cells were randomly di-vided into four groups:normal glucose group (Con-trol),high glucose group (HG),high glucose plus pYr-ads-4-THEM4 vector group (HG + THEM4 vec-tor) and high glucose plus pYr-adshuttle-4 vector group (HG + V vector). After 48 h with HG stimula-tion,the cells were collected for extraction of protein and phospho-Akt (Ser 473),THEM4,TGF-β1 andα-SMA protein expression were examined by Western blot and immunofluorescence staining respectively. Col Ⅰ and Col Ⅲ were detected using the competitive sandwich ELISA kit according to the manufacturer's instructions. Results High glucose inhibited THEM4 expression,and induced increased phospho-Akt (Ser 473),TGF-β1,α-SMA and secreted ColⅠand secre-ted Col Ⅲ in HKC cells. Up-regulation of THEM4 re-versed high glucose-induced decreased THEM4,in-creased phospho-Akt (Ser 473),TGF-β1,α-SMA, secreted Col Ⅰ and secreted Col Ⅲ in HKC cells. Conclusion The up-regulation of THEM4 may de-crease Col Ⅰ and Col Ⅲ secretion by inhibiting the phosphorylation of Akt and down-regulating the expres-sion of TGF-β1 and α-SMA in high glucose-induced HKC cells.

A simple method for recombinant protein purification using self-assembling peptide-tagged tobacco etch virus protease.

Recombinant protein purification remains to be a major challenge in biotechnology and medicine. In this paper we report a simple method for recombinant protein purification using self-assembling peptide-tagged tobacco etch virus protease (TEVp). After construction of an N-terminal ELK16 peptide fusion expression vector, we expressed ELK16-TEVp fusion protein in E. coli. SDS-PAGE analysis showed that ELK16-TEVp was expressed as active protein aggregates which could be purified to 91% purity with 92% recovery by centrifugation in the presence 0.5% Triton X-100. By using His-tagged bovine interferon-γ (His-BoIFN-γ) as the substrate, we demonstrated that EKL16-TEVp had a protease activity of 1.3 × 10(4) units/mg protein with almost 100% cleavage efficiency under the optimized conditions. More importantly, EKL16-TEVp could be removed from the cleavage reaction by single-step centrifugation. After removing the His-tag by nickel-conjugated agarose bead absorption, the recombinant BoIFN-γ (rBoIFN-γ) was purified to 98.3% purity with 63% recovery. The rBoIFN-γ had an antiviral activity of 1.6 × 10(3) units/mg protein against vesicular stomatitis virus. These data suggest that ELK16-TEVp may become a universal tool for recombinant protein purification.

Single-step purification of recombinant proteins using elastin-like peptide-mediated inverse transition cycling and self-processing module from Neisseria meningitides FrpC.

Purification of recombinant proteins is a major task and challenge in biotechnology and medicine. In this paper we report a novel single-step recombinant protein purification system which was based on elastin-like peptide (ELP)-mediated reversible phase transition and FrpC self-processing module (SPM)-mediated cleavage. After construction of a SPM-ELP fusion expression vector, we cloned the coding sequence for green fluorescent protein (GFP), the Fc portion of porcine IgG (pFc) or human ß defensin 3 (HBD3) into the vector, transformed the construct into Escherichia coli, and induced the fusion protein expression with IPTG. The target-SPM-ELP fusion proteins GFP-SPM-ELP, Fc-SPM-ELP and HBD3-SPM-ELP were expressed in a soluble form and efficiently purified from the clarified cell extracts by two rounds of inverse transition cycling (ITC). Under the optimized conditions, the SPM-mediated cleavage efficiencies for the three fusion proteins ranged from 92% to 93%. After an additional round of ITC, the target proteins GFP, pFc and HBD3 were recovered with purities ranging from 90% to 100% and yields ranging from 1.1 to 36mg/L in shake flasks. The endotoxin levels in all of the three target proteins were <0.03EU/mg. The three target proteins were functionally active with the expected molecular weights. These experimental results confirmed the high specificity and efficiency of SPM-mediated cleavage, and suggested the applicability of SPM-ELP fusion system for purification of recombinant proteins.

Effects of Tongguan Capsule on post-myocardial infarction ventricular remodeling and cardiac function in rats.

OBJECTIVE: To observe the effects of Tongguan Capsule (TGC) on post-myocardial infarction ventricular remodeling and heart function in rats. METHODS: A rat model of acute myocardial infarction (AMI) was established by coronary ligation. Experimental rats were randomized to 4 groups including three model groups (Group A: captopril 5 mg/kg * day, n=7; Group B: TGC 10 g/kg * day, n=7; and Group C: placebo, n=8), and a sham-control group (Group D: blank control, n=6). Animals were treated for 4 weeks. The cardiac function of rats was assessed at the end of the experiment based on left ventricular ejection fraction (LVEF) and left ventricular short axis fractional shortening (LVFS) detected by colored echocardiography; meanwhile, the condition of ventricular remodeling was observed through the levels of left ventricular mass (LVM), plasma aldosterone (ALD), myocardial angiotensin II (Ang II) and myocardial collagen measurements. RESULTS: At the end of the experiment, LVEF and LVFS in Group A and B were improved significantly, while those in Group C were unchanged, the LVEF in Group A, B, C, and D was 0.57+/-0.46, 0.61+/-0.08, 0.36+/-0.55 and 0.76+/-0.02, respectively; and their LVFS was 0.31+/-0.52, 0.34+/-0.04, 0.23+/-0.57 and 0.45+/-0.03, respectively. The difference was statistically significant when comparing the two indexes in Group A and B with those in Group C and D (P<0.05). LVM, levels of plasma ALD and myocardial Ang II were lower in Group A and B than in Group C, but a comparison between Group A and B showed an insignificant difference in lowering LVM and ALD, while the lowering of Ang II was more significant in Group B than in Group A (754.7 +/- 18.7 pg/mL vs 952.6+/-17.6 pg/mL, P<0.05). Morphological examination showed that in Group A and B the swollen myocardial cells had shrunk, with regularly arranged myocardial fibers and decreased collagen proliferation, but the improvements in Group B were more significant. CONCLUSION: TGC could markedly improve the post-infarction ventricular remodeling and cardiac function in rats, showing that the efficacy was better than or equal to that of captopril.