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 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.

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.

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.

Objective To observe the clinical efficacy of midazolam intravenous therapy for children with convulsive status epilepticus (CSE).Methods 133 admitted CSE children were randomly divided into treatment group (n =6 8) and control group (n =6 5).Based on symptomatic treatment,the control group was given diazepam plus phenobarbital intravenous injection,the treatment group was given midazolam injection plus intravenous infusion scheme.The clinical efficacy and adverse drug reactions were observed and compared between the two groups.Results After treatment for 3h,the total effective rate of the treatment group was 91.2％,which was significantly higher than 76.9％ of control group(x2 =5.078,P =0.024).Among children with markedly effective and effective effect,the mean onset time (49.3 ± 10.4)min and seizure control time (112.1 + 24.7)min of the treatment group were significantly lower than those of control group (73.8 + 15.4) min,(157.2 ± 38.4) min,the differences were statistically significant(u =9.619,7.191,P =0.000).15 ineffective cases of control group were transferred into midazolam intravenous therapy,the total effective rate after 3h was 73.3％ (11/15).1 case died in both two groups.In control group,the proportion of complications such as muscle tension descending,heart rate and blood pressure variation,respiratory depression,et al.was 49.2％,which was significantly higher than 30.9％ of the treatment group (x2 =4.668,P =0.31).Conclusion Compared with diazepam plus phenobarbital scheme,midazolam intravenous administration in treatment of children with CSE takes effect faster,and with higher safety.With the increasing of midazolam dosage,alert should be taken to drug influence on respiration and heart rate.

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.

AimPurpose-The aim of this study is utilizing the highthrough genechip data to Compare the difference of the pharmacological pathways among the Qingkailing effective components Baicalin(BA),Jasminoidin(JA),cholic acid(CA) and Concha margaritiferausta(CM)in the treatment process of cerebral ischemia.Methods The focal cerebral ischemia-reperfusion model mice were randomly divided into groups of Baicalin(BA),Jasminoidin(JA),cholic acid(CA),Concha margaritiferausta(CM)and model group(M),15 mice for each group,24 hours later total RNA were abstracted from the hippocampus,we selected 374 gene expression profile related to cerebral ischemia,made cDNA chip marked by Cy3/Cy5,detect the variation of different components,Then apply Arraytrack software to select differentiate expressed genes between BA and M,JA and M,CA and M,CM and M by T-tests,select genes with P<0.05,Fold change>1.5,according GeneGO software to find the top two pathways of each components.Results the number of differentiate expressed genes between BA,JA,CA,CM and M is separately 46,50,54 and 30,according to the top two pathways of GeneGo display JA,CA,CM all participate Apoptosis and survival_TNFR1 signaling pathway,besides BA participate in regulating G-protein signaling and Development_A2A receptor signaling while CA in Neurophysiological process_NMDA-dependent postsynaptic long-term potentiation in CA1 hippocampal.Conclusion Qingkailing effective components take diversity Pharmacological characteristics,BA mainly for anti-apoptosis,JA mainly for inhibit apoptosis and promote ischemic brain protection,etc,CA focused on inhibiting calcium influx,and anti-neuron variability.But CM has no good results on this.

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