Objective To study the current clinical application and advancement of microbioecological preparation.Methods Literatures about microbioecological preparation published in China and abroad were collected and reviewed.Results The microbioecological preparation has been widely used at present.It is used to rebuild a balanced microbial population in human body,particularly in intestinal,to promote the stability of internal environment,control dysbacteriosis and to treat a variety of gastrointestinal diseases associated with ectopic microbial population.Conclusion Although microbioecological preparation has been widely used in clinical settings,its effect yet should be further supported and evaluated both by large sample research in randomized double-blind control trails and evidence-based medicine.

Aim Subcortical ischemic vascular demen-tia ( SIVD ) induced by chronic hypoperfusion due to small-artery disease is a common cause of vascular de-mentia ( VaD) , which is recognized as the second most prevalent type of dementia. The aim of this study was to determine whether carnosine played a protective role in cognitive impairment induced by permanent occlu-sion of the right unilateral common carotid arteries ( rUCCAO ) in SIVD. Methods Adult male mice ( C57BL/6 strain ) were subjected to rUCCAO, and treated with carnosine or saline. Locomotor test, open field test, hot plate test, freezing test and Morris water maze were performed after rUCCAO. Results There were no differences among rUCCAO group, carnosine group and sham group for total distance traveled in lo-comotor test. In the open field test, carnosine (200, 500 mg · kg-1 ) significantly revised the decrease of latency spent in the center induced by SIVD . There were no differences between rUCCAO and sham groups for the pain threshold. In freezing test, rUCCAO in-duced a significant reduction in content memory, which was completely reversed by treatment of carnosine. In Morris water maze training trials, rUCCAO-treated mice showed prolonged escape latency in acquisition phase, carnosine ( 200, 500 mg · kg-1 ) markedly shortened the escape latency. Conclusion These data suggest that carnosine has a neuroprotective effect on cognitive impairment induced by rUCCAO in mice.

Objective To analyze the predictive value of platelet related indicators for patent ductus arteriosus (PDA) in extremely low birth weight infants (ELBW). Methods The data of 79 ELBW infants born from June 2013 to June 2016 were retrospective analyzed. There were 48 cases without PDA (nPDA group) and 31 cases with PDA (PDA group). Among 31 cases with PDA, there were 17 cases of non-haemodynamically significant PDA (nhsPDA group) and 14 cases of haemodynamically significant PDA (hsPDA group). The clinical feature and platelet related indicators among nPDA group, PDA group, nhsPDA group and hsPDA group were compared. Multivariate logistic regression was used to analyze the effects of various factors on the occurrence of PDA. ROC curve analysis was performed to evaluate the early predictive value of platelet related indicators for PDA. Results Compared with the nPDA group, the PDA group had a smaller gestational age, a higher proportion of male infants, and a smaller platelet distribution width (PDW), and there were statistically significant differences in all of those (P all<0.05). Multivariate logistic regression analysis indicated that the risk of PDA was increased as the PDW was decreased (OR=1.26, 95%CI: 1.05~1.52). The ROC curve analysis showed that the best diagnostic value of PDW was 13.4 GSD, and the sensitivity of early prediction of PDA was about 67.74%, and the specificity was 68.75%. Compared with nhsPDA group, hsPDA group had a smaller gestation age, lower cesarean section rate, and there were statistically significant differences (P all<0.05). There was no significant difference in platelet related indicators between hsPDA group and nhsPDA group (P>0.05). Conclusion PDW has certain early predictive value for PDA in ELBW. ELBW infants with PDW<13.4 GSD need to be watched closely for the occurrence of PDA.

Objective Design and synthesize short hairpin RNA (shRNA) expression vector of RNA for specific silencing of heparanase (HPA) gene,screened plasmid which silence effects is the best.Observe the function of ceil invasion after inhibiting the expression of HPA in cervical carcinoma cell lines (HeLa).Methods The genomic sequence of HPA gene was retrieved from GenBank database.Designed four pairs of specific oligonucleotide sequences and a negative control according to the shRNA design principles.They were inserted into the vector pYr-1.1,vectors,and transfected into HeLa cells via lipofectamine.Reverse transcription (RT)-PCR and immunofluorescence were employed to detect the expression of HPA gene in the transfected cells at the mRNA and protein levels,respectively.The plasmid were screened and transfected into HeLa cells,then transwell small room stromal invasion experiment were employed to observe the cervical carcinoma cell invasion.Results RT-PCR results of transfected HeLa cells shown that the mRNA amplification multiples were 0.54 ±0.05 in the HPA-592 group,0.89 ±0.18 in HPA-995 group,0.82 ±0.22 in the HPA-1351 group,0.91 ±0.47 in HPA-1658 group.While,they were 1.31 ±0.72 and 1.09 ±0.16 in negative control and blank control group,respectively.Green fluorescence was visible in the cytoplasm,which indicated that the HPA protein was expressed in the cytoplasm,of them the weakest green fluorescence in the HPA-592 group.The relative numbers of invasive cells among the HeLa cells were as follows:182 ±6 in the blank control group,258 ± 17 in the negative control group,and 44 ± 4 in the HPA-592-specific interference group(P ＜ 0.01).Conclusion Successfully screened shRNA vector targeting human HPA,efficiently inhibit expression of HPA gene when transfected into HeLa cells,and significantly reduced the invasion capacity of cervical carcinoma cells.

Objective To detect the expression of heparanase (HPA) in cervical cancer cells and investigate the impact of quercetin on the expression of HPA,and the molecular mechanism that quercetin inhibits the growth of cervical cancer cells.Methods The experimental groups included cervical cancer cell lines (HeLa and Caski) exposed to different concentrations of quercetin (20,40 and 80 μmol/L) in the culture medium.The control groups included a negative control group,which was cultured with RPMI 1640 only,and a positive control group,in which cervical cancer cells were transfected with HPA small interference RNA (siRNA) to silence HPA expression.The cellular expression levels of HPA were detected with fluorescence quantitative real-time PCR and western blot analysis at 24,48 and 72 hours after treatment.Results (1) HPA was significantly expressed in both cervical cancer cell lines (HeLa and Caski),and it exists both nucleus and cytoplasm.(2)The real-time PCR shows as follows:as the quercetin concentration increased (20,40 and 80 μmol/L),the mRNA expression level of HPA decreased (P ＜0.01),in which the inhibition of HPA expression was concentration dependent.In addition,the inhibition of HPA expression was also time dependent.As time growth,the expression level of HPA mRNA (24,48 and 72 hours) in HeLa and Caski cells decreased (P ＜ 0.01).Compared with negative control group,the expression level of HPA mRNA decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ; Compared with positive control group,the expression level of HPA mRNA expressed no obvious difference in quercetin (80 μmol/L) group (P ＞ 0.05) in HeLa cells,while it was opposite in Caski cells(P ＜0.01).(3)The result of western blot shown that,as the quercetin concentration increased(20,40 and 80 μmol/L)and time growth (24,48 and 72 hours),the expression level of HPA protein decreased (P ＜ 0.01),and the inhibition of HPA protein expression was concentration and time dependent.Compared with negative control group,the expression level of HPA protein decreased in different concentrations of quercetin (40 and 80 μmol/L) in both HeLa and Caski cells (all P ＜ 0.05) ;Conpared with positive control group,the expression level of HPA protein expressed no obvious difference in quercetin (80 μmol/L) group (all P ＞ 0.05) in both HeLa cells and Caski cells (all P＞0.05).Conclusion Quercetin could inhibits the expression of HPA in cervical carcinoma cell lines,which inhibition is concentration and time dependent.

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Aim To investigate the roles of intracellu-lar reactive oxygen species ( ROS ) and Nrf2 pathway in shikonin-induced A549 cell apoptosis. Methods The cytotoxicity was analyzed by MTT assay. The ap-optosis of A549 cells was analyzed by both cellular morphological and biochemical methods. The relative changes of the redox marks ( ROS/GSH) were studied by fluorescence assay in the shikonin-treated A549 cells in accompany with the changes of the intracellular redox homeostasis by GSH/GSSG ratio. ROS inhibitor was also employed in the treatment to find the role of ROS in shikonin-induced A549 cell apoptosis. Real-time PCR analysis and ELISA assay were performed as well to determine the role of Nrf2 pathway in the shiko-nin-induced A549 cell apoptosis. Results The IC50 of shikonin on A549 cells was 3. 2 mg·L-1 . The cellu-lar redox homeostasis shifted toward oxidation signifi-cantly in shikonin treatment in a time-dependent man-ner. The expression of the Nrf2 pathway related genes was up-regulated by shikonin ( 3 . 2 mg · L-1 , 8 h ) . The expression of the anti-apoptotic genes was down-regulated , and proapoptotic genes were up-regulated by shikonin (3. 2 mg·L-1, 24h). Futhermore, the inhi-bition of intracellular ROS alleviated the cytotoxicity of shikonin in A549 cells. Conclusion The critical role of shikonin-induced redox imblance in A549 cell, coped with the secondary produced ROS and Nrf2 path-way antioxidants, result in A549 cell apoptosis.

A sensitive, rapid and accurate liquid chromatography-tandem mass spectrometric (LC-MS/MS) method with pre-column derivatization was developed for the simultaneous determination of erdosteine and its thiol-containing active metabolite in human plasma. Paracetamol and captopril were chosen as the internal standard of erdosteine and its active metabolite, respectively. Aliquots of 100 microL plasma sample were derivatized by 2-bromine-3'-methoxy acetophenone, then separated on an Agilent XDB-C18 (50 mm x 4.6 mm ID, 1.8 microm) column using 0.1% formic acid methanol--0.1% formic acid 5 mmol x L(-1) ammonium acetate as mobile phase, in a gradient mode. Detection of erdosteine and its active metabolite were achieved by ESI MS/MS in the positive ion mode. The linear calibration curves for erdosteine and its active metabolite were obtained in the concentration ranges of 5-3 000 ng x mL(-1) and 5-10 000 ng x mL(-1), respectively. The lower limit of quantification of erdosteine and its active metabolite were both 5.00 ng x mL(-1). The pharmacokinetic results of erdosteine and its thiol-containing active metabolite showed that the area under curve (AUC) of the thiol-containing active metabolite was 6.2 times of that of erdosteine after a single oral dose of 600 mg erdosteine tables in 32 healthy volunteers, The mean residence time (MRT) of the thiol-containing active metabolite was (7.51 +/- 0.788) h, which provided a pharmacokinetic basis for the rational dosage regimen.

Objectiv e Renal ischemia-reperfusion ( I/R) may cause myocardial injury and dexmedetomidine ( DEX) is a new alpha-2 adrenergic agonist with the effects of antisympathia , seda-tion, and analgesia.This study was to investigate the effect of DEX on the myocardial tissue of rats at different time points after renal I/R. Methods Forty male Wistar rats were randomized into 5 groups of equal number,sham operation, 60 min renal ischemia and 3 h reperfusion (I/R1), 120 min ischemia and 3 h reperfusion (I/R2 ), 60 min ischemia and DEX+3 h reperfusion (D1), 120 min ischemia and DEX+3 h reperfusion ( D2) .Renal I/R was induced by removal of the right kidney and ligation of the left re-nal artery and vein followed by 3 hours of reperfusion.Meanwhile, intraperitoneal injection of DEX at 50μg/kg was given to the ani-mals in groups D1 and D2 at 60 at 120 min respectively after ischemia.After 3 hours of reperfusion, blood samples were collected for measurement of the concentrations of serum creatinine (Cr) and blood urea nitrogen (BUN), and renal and myocardial tissues harvested for observation of pathological changes under the light microscope and determination of the expressions of TNF-αand IL-10 by ELISA.Results Significant increases were observed in the concentrations of serum Cr and BUN , the expressions of TNF-αand IL-10 in the renal tissues and those in the myocardial tissues in groups I /R1([84.67 ±9.62] μmol/L, [8.55 ±1.08] mmol /L), I/R2 ([167.11 ±18.81] μmol/L, [13.42 ±1.25] mmol/L), D1 ([69.67 ±9.52] μmol/L, [7.56 ±0.70] mmol/L), and D2 ([114.29 ±12.50] μmol/L, [10.27 ±0.78] mmol/L), as compared with the sham operation group ([53.20 ±9.21] μmol/L, [3.75 ±0.78] mmol/L), (all P <0.05).Significant decreases was observed in the sham operation group as compared with other groups in the expressions of TNF-αand IL-10 (P<0.05).Significant decreases was observed in the D1 and D2 groups compared with other groups in the expressions of TNF-α, but increasing in IL-10.②Injury was reduced in the D1 and D2 groups compared with other groups.③The horizontal stripes of myocardial tissue disappeared in I/R1 and I/R2 decreases of inflammatory cells was observed in D1 and D2 groups compared with others. Conclusion Dexmedetomidine can attenuate myocar-dial injury induced by renal ischemia-reperfusion in rats and its inhibitory effect on inflammatory factors may be involved in the mechanism.

OBJECTIVE:To study the correlation between antibacterials amount and drug resistance of Echerichia coli,and to provide reference for clinical use of antibacterials. METHODS:Retrospective review was used to calculate DDDs of antibacterials and resistance rate of Escherichia coli to 11 kinds of antibacterials each quarter. The correlation analysis was carried out using the SPSS 13.0 statistical software. RESULTS:The resistance rates of E. coli to piperacillin/tazobactam,cefoperazone/sulbactam and le-vofloxacin were with upward trends,and the others showed downward trends. The resistance rates of E. coli to meropenem and imi-penem/cilastatin appeared in 2014,increasing from 0 to 8.8% and 9.4%,respectively. DDDs of them were significantly correlated to drug resistance of E. coli,showing positive correlation(r=0.915,0.793,P<0.01). DDDs of piperacillin/tazobactam was signif-icantly correlated to resistance rate of E. coli(r=0.807,P<0.01),while that of ceftazidime was negatively correlated to resistance rate of E. coli(r=-0.672,P<0.05). There was no statistical significance in resistance rate of E. coli to other 7 kinds of antibacte-rials. CONCLUSIONS:There are some correlations between the DDDs and resistance rates. We should strengthen the monitoring of bacterial resistance and the management of rational application of antibacterials.