Object To study the effects of SINI TANG (SNT) on the rat aortic rings pre-contracted by high K + and phenylephrine (Phe). Methods The effects of SNT on the aortic rings in the presence of 60 mmol/L KCl and Phe (1?10 -9 -1?10 -4 mmol/L) were observed and t heir me chanisms were studied after treatment with Propranolol (Pro, 3?10 -6 mmol/ L) and Bay K8644 (BK, 1?10 -5 mmol/L) as tool drugs. Results SNT inhibited the contraction induced by cumulative Phe and decreased the maximum tension (T max ); Pro couldn't influe nce the effects of SNT. SNT attenuated the amplitude of contractile effect of hi gh K +; BK couldn't reverse the effects of SNT. Conclusion SNT can shift the dose-response curve to the right and decrease the T max . It shows that SNT is a kind of noncompe titive antagonism. SNT decreases the effect of high K + against contraction of the artery. BK, a L-type Ca 2+ channels activator, couldn't recover the inhibition induced by SNT. The results suggest that SNT inhibit ? 1 recep tor, while calcium channel may not be involved in attenuating the effect of SNT on high K +-induced contraction.

Objective　To investigate the mechanisms of Pinacidil prec onditioning protection of hemorrhagic shock rats. 　Methods　The rats were divided into 3 groups: a normal group ( n=10), a control group (n=40) and a preconditioning group (n=40). Pinacidil preconditio ning was processed 24 h before making the hemorrhagic shock model. The cPLA 2 expression in myocardium was observed at different time points with western blot; Pinacidil preconditioning was processed 24, 48 and 72 h before making th e hemorrhagic shock model to observe hsp70 expression in myocardium and liver tissue with western blot. 　Results　Pinacidil preconditioning could increase hsp70 expres sion and decrease cPLA2 expression. 　Conclusions　Pinacidil preconditioning protects "shock cell" by inducing hsp70 overexpression and inhibiting cPLA2 expression, which is re sponsible for protecting myocardial and liver tissues of the hemorrhagic shock rats.

Objective:To explore the relationship between DRD2 - 141C Ins/Del polymorphism and heroin craving. Methods:380 heroin addicts who were under abstinence were given the cue- elicited heroin craving experiment. And then we detected the polymorphism of DRD2- 141CIns/Del for them by using PCR- RFLP. We compared the relationship between genotypes of DRD2- 141CIns/Del and heroin craving before and after cue exposure. Results:No significant differ- ence has been found between heroin craving before and after cue- exposure in three genotypes of DRD2- 141CIns/Del. Conclusion:D2 receptor gene - 141CIns/Del polymorphisms may have no association with the susceptibility of heroin crav- ing.

Objective To investigate the antimicrobial resistant and transmission mechanisms of carbapenem-resistant K.pneumonia (CR-KP) infection of newborns.Methods A retrospective study was conducted on totally 37 non-repetitive CR-KP which were isolated from patients hospitalized between April 2011 and October 2013.Resistance genes were identified by PCR and sequencing.Plasmid was analyzed by pulsed-field gel electrophoresis (PFGE).Conjugation experiments were performed to determine the transferability of beta-lactamase.Multilocus sequence typing (MLST) was used to determine the genotypes and homology of these isolates.Out-membrane proteins were examined by PCR and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE).Results Thirty-seven CR-KP isolates were tested.The resistant rates of imipenem, meropenem, ertapenem were 89.2％ (33/37), 83.8％ (31/37) ,97.3％ (36/ 37), respectively.All the 37 CR-KP exhibited 100％ (37/37) sensitivity to tigecycline, colistin, levofloxacin and amikacin, while resistance to most of the other antibiotics.By PCR, 67.6％ (25/37) isolates were blaNDM-1 positive, 35.1％ (13/37) isolates were blaIMP-4 positive and 2.7％ (1/37) isolate were blaIMP-8 positive, including two isolates carrying both blaNDM-1 and blaIMP-4.PFGE results showed that the isolates carried 2-4 plasmids and both blaNDM-1 and blaIMP-4 were transferable by plasmids.MLST assigned them to sequence type (ST) 20, ST17, ST54, ST705, ST290,which showed that there were infectious outbreaks caused by NDM-1-producing and IMP-4-producing respectively among newborns.SDS-PAGE result indicated that there was no absence of outer membrane proteins OmpK35 and OmpK36.Conclusions The main resistant mechanisms of CR-KP causing infection in newborns were those the isolates carried carbapenemase of blaNDM-1 or blaIMP-4 and the K.pneumonia with two kinds of carbapemenase were detected.

Objective To establish a quantitative detection method for Mycobacterium tuberculosis by immunomagnetic capture combined with PCR-ELISA detection system with double internal standards(IMC-PCR-ELISA) .Methods The immunomagnetic (Dynabeads? ) which could specifically capture Mycobacterium tuberculosis were prepared .According to Mtp40 and IS6110 gene sequence of Mycobacterium Tuberculosis ,2 pairs of primers(upstream primer was modified with Biotin at 5′end) ,2 same-length mutant fragments with PCR amplified fragments ,and 3 capture probes(modified with digoxigenin at 3′end) were designed .Myco-bacterium tuberculosis were captured by immunomagnetic ,then detected by PCR-ELISA with double internal standards .Results The IMC-PCR-ELISA method could yield quantitative results in about 4 h with a detection limit at 5 × 103 copies/mL .There was a fine linear relationship between the copies of Mtp40(IS6110)in fact and in the calculation through formula when the concentrations of low internal standards were 30-70 copies/mL and the concentrations of high internal standards were 8 000-12 000 copies/mL (r2 =0 .998) .No nonspecific amplification was observed .Conclusion A rapid and quantitative method for the detection of Myco-bacterium tuberculosis was established successfully .The IMC-PCR-ELISA method was rapid ,sensitive ,secific and quantitative .

Objective To compare the difference of three methods testing the antibiotic susceptibility of mucoid Pseudomonas aeruginosa in order to provide accurate and reliable antibiotic susceptibility result for clinic.Methods A total of 630 mucoid Pseudomonas aeruginosa were collected from Linyi People′s Hospital during January 2015 to December 2016.They mainly come from respiratory medicine and the most common specimen source was sputum.All specimens were examined in 2 h.The strains isolated from the same patient were discarded.Antibiotic susceptibility was tested by the automatic microorganism analyzer VITEK2 compact, E-test, which was reference method, and K-B disk.The results of three methods were analyzed and compared by χ2 test.Results The result of E-test showed that antibiotic sensitivity of 630 mucoid Pseudomonas aeruginosa was above 52.7% except for Cefepime (39.2%).The result of K-B disk was compared with E-test, the antibiotic sensitivity of mucoid Pseudomonas aeruginosa to imipenem (72.4% vs 52.7%) and amikacin (48.6% vs 71.1%)had significant difference (χ2=8.283 7 and 10.533 8, P<0.05).The result of VITEK2 compact showed that the antibiotic susceptibility of mucoid Pseudomonas aeruginosa to imipenem(70.8% vs 52.7%), cefepime(60.8% vs 39.2%), gentamicin (87.6% vs 74.1%)and levofloxacin(81.3% vs 65.4%) was significant higher than the result of E-test (χ2=6.935 2,9.331 2,5.885 6 and 6.466 5, P<0.05).For tobramycin, piperacillin/tazobactam and ciprofloxacin, the result of three methods is more consistent.Compared to VITEK2 compact, the consistency between K-B disk and E-test was higher.The rate of very major error and major error were between 0.0%-4.8% (Amikacin 12.2%) and minor error was 4.6%-20.3%.Conclusions The drug sensitivity of mucoid Pseudomonas aeruginosa is different between various methods.The result of K-B disk and E-test using blood MH is more reliable than VITEK2 compact.

Objective To study the expression and the function of DKK2 and to explore its potential mechanisms in breast cancer.Methods The expression of DKK2 was detected by RT-PCR in normal breast tissues and breast cancer cells.we have transfected DKK2 into breast cancer cell lines MDA-MB-231 and MCF-7.The cells before transfection were used as control group and marked with Vector.The cells after transfection were used as experimental group and marked with DKK2.Furthermore by qRT-PCR and Western-blot,the expression of DKK2,Notch signaling pathway and related factors were analyzed.We also detected the function of DKK2 by cloning assay,Transwell assay and proliferation assay.Results No expression of DKK2 was found in breast cancer cell lines MDA-MB-231 or MCF-7,with relatively high expression in normal breast tissue.The number of apoptotic cells was 2.57±1.18 before transfectionin in cell line MDA-MB-231,and 49.53±8.27 after transfection.The difference was statistically significant between the two groups (P＜0.005).The relative colony formation rate of MDA-MB-231 cells and MCF7 cells after transfection accounted for 20.44％ and 15.21％,respectively.The difference was statistically significant by t test.The number of apoptosis cells in MB231/DKK2 group was 49.53± 8.27 and that in MB231 / Vector group was 2.57±1.18.The difference was statistically significant (P＜0.005).The number of migrated cells in MB231/DKK2 group was 112.0±8.1 and that in MB231/Vector group was 178.0±12.0.The difference was statistically significant (P＜0.005).The mRNA expression of Notch 1 in group MB231/Vector was recorded as 1.The mRNA expression of JAG1 in MB231/DKK2 group was 0.2891.The difference was also statistically significant (P＜0.005).Conclusions Restored expression of DKK2 in silenced breast cells suppresses breast cancer cell proliferation and migration through repressing Notch signaling.DKK2-Notch signaling pathway may be its potential molecular mechanism to function in breast cancer.DKK2 may be one of the target genes for early diagnosis and treatment of breast cancer.

OBJECTIVETo compare the gene expression status of NB4 cells before and after arsenic sulfide treatment by cDNA microarray.

METHODSTwo cDNA probes were made from mRNA of untreated or arsenic sulfide treated NB4 cells. The cells were labelled with Cy3 or Cy5 fluorescence dyes individually, hybridized with cDNA microarray, and scanned for fluorescent intensity. The altered gene expression was screened through the analysis of difference in gene expression profile.

RESULTSThirty four genes related to apoptosis, cell cycle and others expressed different after the treatment of arsenic sulfide, 28/34 were up-regulated, 6/34 down-regulated.

CONCLUSIONABC50, PNAS-2 and cyclin G(2) might take part in the process of NB4 cell apoptosis induced by arsenic sulfide.

Arsenicals ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Oligonucleotide Array Sequence Analysis ; Sulfides ; pharmacology ; Tumor Cells, Cultured ; drug effects ; metabolism

Objective: To evaluate the ability of matrix-assisted laser desorption/ionization-time of flight mass spectrometry(MALDI-TOF MS) in the homology analysis of Carbapenems-resistant klebsiella pneumonia. Methods: Twenty-one non-duplicated strains of Carbapenems-resistant klebsiella pneumoniae were isolated from Shandong Provincial Hospital affiliated to Shandong University during April 2011 and October 2013 in this study. Twenty isolates were from neonatal unit and one from cardiac surgery. The homology analysis of Carbapenems-resistant klebsiella pneumoniae was performed with pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST) and MALDI-TOF MS respectively. Results: The result of PFGE was consistent with MLST. The twenty-one CRKPN strains were divided into three groups by MALDI-TOF MS according to their relationship, 18 of them belonged to II group, and the homology was higher than 75%. From the analysis of protein mass spectra of 18 strains, the protein peaks were roughly the same. Thus, it was concluded that their relationship was close, and the results were basically consistent with the results of PFGE and MLST. The H13 strain with low homology (<60%) was different from the above strains, especially in the molecular weight 4365, 5381 and 6289.The PFGE analysis showed that the homology between H13 and other strains was 61%, and the MLST classification result was ST54. Conclusions MALDI-TOF MS can be used to identify CRKPN accurately and analyze its homology analysis more conveniently than other methods in clinical laboratory. MALDI-TOF MS has the potential to be used as an easy and rapid epidemiology typing tool for nosocomial infection investigation caused by drug-resistant bacteria.(Chin J Lab Med, 2018, 41: 589-595)

Objective To investigate the antibiotic resistance of clinical isolates in Shandong Provincial Hospital during 2016.Method The antimicrobial susceptibility of isolates was tested by using VITEK 2-Compact system or disk diffusion method.All the data were analyzed by WHONET 5.6 software according to CLSI 2016 breakpoints.Results A total of 4 810 non-duplicate clinical strains were collected during 2016,of which gram-negative bacilli and gram-positive cocci accounted for 70.2％ and 29.8％,respectively.The most common specimen source was respiratory tract (38.1％),followed by skin and soft tissue (21.1％) and urine (17.5％).Escherichia coli was the most frequently isolated bacteria,followed by Staphylococcus aureus,Pseudomonas aeruginosa,Klebsiella and Enterococcus,accounting for 24.3％,10.2％,9.9％,9.8％ and 9.6％,respectively.The prevalence of ESBLs-positive E.coli and K.pneumoniae was 55.0％ and 37.9％,respectively.Carbapenem-resistant E.coli and K.pneumoniae accounted for 1.6％ and 4.2％,respectively.The percentage of P aeruginosa resistant to imipenem and meropenem was 24.3％ and 21.9％,respectively.More than 40％ of the A.baumannii strains were resistant to all the antibiotics tested except minocycline (33.2％).The prevalence of beta-lactamase positive strains was 70.1％ in H.influenzae and 97.5％ in M.catarrhalis.The prevalence of methicillin-resistant S.aureus (MRSA) was 31.4％.No staphylococcal strains were found resistant to vancomycin,teicoplanin or linezolid.E.faecium strains showed higher resistance rate to most antibiotics tested than E.faecalis.One strain of E.faecium was resistant to both vancomycin and teicoplanin.A total of 191 strains of S.pneumoniae were isolated,of which 143 (74.9％) isolates were from pediatric wards.None of the non-meningitis strains was resistant to penicillin.Other hemolytic Streptococcus strains were sensitive to penicillin,cephalosporins and vancomycin.Conclusions Bacterial resistance is still on rise.We should pay more attention to strengthening antimicrobial resistance surveillance and improving rational use of antimicrobial agents.