Objective To study the effects of valproic acid (VPA) to the expression of SMN2 mRNA in neuron-like cells (NLCs) derived from patients with spinal muscular atrophy (SMA). Methods Polymerase chain reaction-restriction fragment length polymerphism (PCR-RFLP) was performed to select the patients with SMA. Mesenchymal stem cells (MSCs) derived from patients were induced into NLCs which were set as the model of neurons. The transcripts of SMN2 by reverse transcription-polymerase chain reaction (RT-PCR) combined with sequenceing were detected. Semi-quantitative RT-PCR analysis was performed to detect the changes of SMN2 mRNA expression between before and after the NLCs were treated by VPA. Results Two bands (266 bp and 212 bp) were found in the gel picture of RT-PCR, which were respectively the products of full length transcript (fl-SMN mRNA) and skipping exon 7 (SMN?7 mRNA). NLCs had significantly increased fl-SMN mRNA and SMN?7 mRNA levels as compared with the untreated cells after treatment with VPA, and shown a dose effect(0.210?0.035,0.282?0.041,0.351?0.020,0.450?0.052,0.553?0.035,P

Objective To study the antitumor effects in vitro and impact on colon cancer in rats of endostatin transfected bifidobacterium oral powder preparation(ETB-2).Methods Growth inhibitory effect of the cecropins on normal human gastric epithelial cell line(GES-1) and human colon adenocarcinoma cell line(LS-174T) was observed using a microculturetetrazolium(MTT) colorimetric methods.Male Wistar rats were divided into 4 groups.All treatments were completed in a course of 18 weeks and the experiment was finished at week 33.Results The cecropins showed selective cytotoxic activity against the human colon adenocarcinoma cell line.There was a significant lower in incidence of colon tumors in rats(70%vs100%,P0.05).Conclusion ETB-2 has significant preventive effect on colon cancer induced by DMH in rats.

Objective:To explore the personnel arrangement in the outpatient pharmacy by calculating working hour to provide ref-erence for the rational staffing in hospital. Methods:The daily work content and working hour of 18 pharmacists in the outpatient phar-macy of a large general hospital from January to March in 2013 were following-up observed and recorded using the working hour meas-urement. The data were input the EXcellsoftware to establish the database, and the workload in various positions was collected and sorted. The obtained relative parameters were used to calculate the needed worker number on the basis of manpower planning model. Results:The research confirmed the mean operation time for 9 work programs in the outpatient pharmacy, and the time for drug dispen-sing and distributing was detailed. The needed number of pharmacists was 13. 29 according to the calculation, plus the officer-in-charge and sanitation workers, the total number was 15. 29(approx. 16). Conclusion:The working hour measurement can scientifically de-termine the time for each job, and the workload should be used as the foundation for configuring personnel qualification and the number in outpatient pharmacy.

Objective:To explore the inhibition of microRNA (miRNA, miR)-5590-3p on the expression of transforming growth factor beta typeⅡreceptor (TGFBR2) gene and its effect on the invasion and proliferation of gastric cancer cell line HS-746T.Methods:The gastric cancer cell line was cultured in vitro. Real-time quantitative polymerase chain reaction (qRT-PCR) was used to analyze the expression of miR-5590-3p in gastric cancer tissues and cell lines. The miR-NC and miR-5590-3p mimic were transfected into gastric cancer cell line HS-746T by lipofection, and named as miR-NC group and miR-5590-3p group, respectively. qRT-PCR was used to measure transfection effects. Transwell assay and cell counting kit-8 (CCK-8) assay were used to detect cell invasion and proliferation after transfection. The bioinformatics software predicts and the dual luciferase reporter gene system validates the target gene of miR-5590-3p. qRT-PCR and western blot were used to measure the expression of TGFBR2 and its downstream proteins in the transfected cells. Results:The expression of miR-5590-3p in gastric cancer tissues was significantly lower than that in adjacent tissues ( P<0.01). The expression of miR-5590-3p in gastric cancer cell lines was significantly lower than that in normal gastric mucosal epithelial cells ( P<0.05), and lowest in HS-746T cells ( P<0.01). After transfection, the expression of miR-5590-3p in miR-5590-3p group (11.76±0.21) was significantly higher than that in miR-NC group (1.06±0.21), with statistically significant difference ( P<0.01). The number of invasive cells in miR-NC group and miR-5590-3p group were (101.20±15.47) and (26.53±6.53), respectively, and the invasion ability of miR-5590-3p group was significantly decreased ( P<0.01). Compared with the miR-NC group, the cell proliferation ability of the miR-5590-3p group was significantly decreased ( P<0.05). Bioinformatics software showed that the target gene for miR-5590-3p is TGFBR2. The dual luciferase reporter gene system confirmed that miR-5590-3p can target the TGFBR2 gene ( P<0.01). Western blot results showed that compared with miR-NC group, the expression of TGFBR2 in HS-746T cells in miR-5590-3p group was significantly decreased ( P<0.01), the expression of ZEB-1 and ZEB-2, and the expression of CDK1 and cyclin B proteins were decreased as well. Conclusions:miR-5590-3p can inhibit the invasion and proliferation of gastric cancer cell HS-746T by targeting and regulating the expression of TGFBR2 gene.

Objective To evaluate long-term outcome of simultaneous kissing sirolimus-eluting stent (SKS) technique versus single sirolimus-eluting stent (SSS) technique for percutaneous treatment of true coronary bifurcation lesions in large-size vessels.Methods This randomized study assigned 190 patients with a coronary bifurcation lesion to simultaneous kissing stenting (SKS) in both main and side branches and 190 patients to main vessel stenting only (SSS).The endpoints included restenosis,death,non-fatal myocardial infarction,target-lesion revascularization (TLR),stent thrombosis,success rate of percutaneous coronary intervention (PCI) and the operation duration.Results During 1-year follow-up,the SKS group and the SSS group had similar incidences of overall re stenosis [30 ( 15.8 ％ ) vs.24 ( 15.2 ％ ),x2=0.000,P＜0.05],mainbranch restenosis [20 ( 10.5％ ) vs.16 ( 10.1％ ),x2=0.003,P ＞ 0.05];side-branch restenosis [13 ( 6.8％ )vs.23 ( 14.6％ );x2=4.73,P＜0.05];death [2 ( 1.1％ ) vs.1 ( 0.6％ ),x2=0.026,P ＞ 0.05],non-fatal myocardial infarction [4 (2.1％ ) vs.2 ( 1.3％ ),x2=0.034,P ＞ 0.05],TLR [23 ( 12.1％ ) vs.20 ( 12.7％ ),x2=0.000,P ＞ 0.05] and stent thrombosis [4 (2.1％ ) vs.2 ( 1.3 ％ ),x2=0.034,P ＞ 0.05] and a shorter operation duration[(20 ± 8) min vs.(45 ± 9) min,t=1.98,P＜0.05] than the SSS group.Conclusion For true coronary bifurcation lesions in large-size vessels,SKS and SSS have similar long-term outcomes.The SKS group has a higher success rate of PCI and shorter operation duration.

Objective:To explore the effect of microRNA (miRNA)-3126-5p on the proliferation and migration of colorectal cancer cells by inhibiting the expression of LIM and SH3 protein 1 ( LASP1). Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of miR-3126-5p in colorectal cancer cell lines (HT-29, HCT116, LoVo, SW480) and normal intestinal mucosal epithelial cells (HIEC). The cell line with the lowest expression level was selected as the experimental object. The experiment was divided into 2 groups: the negative control group (transfected with miR-NC) and the miR-3126-5p group (transfected with miR-3126-5p). Cells of each group were collected 48h after transfection. qRT-PCR method was used to detect the expression level of miR-3126-5p in each group. The MTS method and the scratch healing experiment were used to detect the proliferation level and migration ability of the cells in each group. The bioinformatics software microRNA.org and the dual-luciferase reporter gene experiment were used to predict and verify the target genes of miR-3126-5p, respectively. qRT-PCR and Western blot were used to detect the expression levels of target genes in each group of cells. Measurement data were expressed as mean±standard deviation ( Mean± SD), t test was used for comparison between two groups, and one-way analysis of variance was used for comparison between multiple groups. Results:Compared with normal intestinal mucosal epithelial cells (HIEC), the expression level of colorectal cancer cell line miR-3126-5p was significantly reduced ( P<0.05), and the cell line with the lowest expression level was HCT116 cells ( P<0.01). The expression of miR-3126-5p in HCT116 cells in the negative control group and miR-3126-5p group were (1.05±0.16) and (7.91±1.26) respectively, and the difference was statistically significant ( t=5.40, P<0.01). Compared with the negative control group, the proliferation ability of HCT116 cells in the miR-3126-5p group was significantly reduced ( t=4.52, P<0.05), and the migration ability was significantly reduced ( P<0.01). microRNA.org shows that miR-3126-5p has complementary binding sites with LIM and SH3 protein 1 ( LASP1) gene mRNA. miR-3126-5p can target LASP1 mRNA ( P<0.01). Compared with the negative control group, the expression of LASP1 gene in HCT116 cells of the miR-3126-5p group was significantly reduced ( t=4.56, P<0.01). Conclusion:The expression of miR-3126-5p in colorectal cancer cell lines is low, and miR-3126-5p can reduce the proliferation and migration ability of colorectal cancer HCT116 cells by inhibiting the expression of the target gene LASP1.

Objective:To investigate the effect of long non-coding RNA (lncRNA) HAGLR on the proliferation and migration of gastric cancer cells by inhibiting the expression of microRNA (miRNA, miR)-93-5p.Methods:Real-time quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of HAGLR in gastric cancer cell lines (HS-746T, BGC823, SGC7901, MGC803) and normal gastric mucosal epithelial cells (GES-1). Selected the cell line with the lowest HAGLR expression and transfected with the negative control plasmid (negative control group) or HAGLR-high-expression plasmid (HAGLR group) respectively. The MTS method and the scratch healing test were used to detect the proliferation and migration ability of the cells after transfection. The bioinformatics software miRcode database was used to predict the target gene of HAGLR, and the dual luciferase reporter gene experiment was used to verify the binding of HAGLR to the target gene. qRT-PCR was used to detect the expression of the target gene. Western blot was used to detect the expression of Hippo signaling pathway. The software SPSS 21.0 was used to conduct statistical analysis. The t test was used for comparison between two groups, and the one-way analysis of variance was used for comparison between multiple groups. Results:Compared with GES-1 cells, the expression level of HAGLR in gastric cancer cell lines was lower (all P<0.05), and the cell line with the lowest HAGLR expression was SGC7901 cells ( P<0.01). The HAGLR expression in SGC7901 cells in the HAGLR group and the negative control group were 1.03±0.13 and 9.75±1.10, respectively. The expression level of HAGLR in the negative control group was significantly lower than that in the HAGLR group ( t=7.87, P<0.01). Compared with the negative control group, the absorbance of SGC7901 cells in the HAGLR group was significantly reduced ( P<0.05), and the scratch healing rate was significantly reduced ( P<0.01). The miRcode database showd that HAGLR and miR-93-5p have complementary binding sites. The dual luciferase reporter gene experiment showed that HAGLR can complement miR-93-5p ( P<0.01). Compared with the negative control group, the expression of miR-93-5p in SGC7901 cells in the HAGLR group was significantly reduced ( P<0.01), and the expression of Hippo signaling pathway protein was significantly reduced (all P<0.01). Conclusions:HAGLR is low expressed in gastric cancer cell lines. HAGLR inhibits the proliferation and migration of gastric cancer SGC7901 cells by negatively regulating miR-93-5p.

AIM: To explore the pathological features of airway inflammation in patients with eosinophilic bronchitis(EB) and compared to those with cough variant asthma(CVA).METHODS: Flexible fibre optic bronchoscopy was performed in 11 patients with EB,10 with CVA,14 with bronchial asthma and 10 normal controls.The mean thickness of the basement membrane was measured by light microscopy.Using immunohistochemical and special staining,the localization and density of inflammatory cells(eosinophils,mast cells,T lymphocytes) were detected in bronchial submucosa in EB and CVA patients.RESULTS: The mean thickness of the basement membrane was significantly increased in the subjects with EB [2.92 ?m(2.10-6.50 ?m)],CVA [5.64 ?m(3.23-8.48 ?m)] and bronchial asthma [9.08 ?m(6.61-11.99 ?m)] rather than that in the normal controls [2.08 ?m(1.62-3.40 ?m)].There were also significant differences among the three groups.The number of mast cells and eosinophils in the bronchial submucosal from subjects with EB [75 cells/mm~2(35-112 cells/mm~2),7 cells/mm~2(0-31(cells/mm~2))] was substantially decreased than those in subjects with CVA [148 cells/mm~2(34-200 cells/mm~2),114 cells/mm~2((1-768 cells/mm~2));P

Objective The aim of this study was to determine whether multi?parameters MRI of tongue carcinoma have the potential to predict cervical lymph node metastases. Methods A total of 46 patients with tongue carcinoma, who underwent MRI scan preoperatively, were investigated retrospectively and were divided into cervical lymph node (LN) metastases group (unilateral LN+, n=16;bilateral LN+, n=14) and no cervical lymph node metastases group (LN-, n=16) according to their pathological grading. Of the 40 patients with tongue carcinoma underwent plain and contrast MRI scan, 6 patients have plain MRI scan, and 32 have DWI examination.The ADC value, tumor length, tumor thickness, sublingual distance between tumor and sublingual space, and para?lingual distance between tumor and tongue midlinedetermined from MRI, were preoperatively estimated and compared with the pathological findings of cervical lymph nodes. A unpaired t test was used to analyze normal distributed continuous data, and a Mann?Whitney U test was used to analyze abnormally distributeddata. The ROC was used to evaluate the efficacy of MRI in predicting the metastasis of cervical lymph nodes. Results The indexes of ADC value, tumor length, tumor thickness, and para?lingual distance between tumor and tongue midline, which all showed significant difference between LN+group and LN-group (all P<0.05), and the index of sublingual distance between tumor and sublingual space showed no significantly association with LN+ (P>0.05). The index of ADC value showed significant difference between unilateral LN+group and bilateral LN+group (P<0.05), and the other indexes, which all showed no significantly association with bilateral LN+ (all P>0.05). The ROC curve analysis of the ADC value, tumor length, tumor thickness, and para?lingual distance between tumor and tongue midline of the neck lymph node metastasis were carried out, with the cutoff set as 1.13×10?3 mm2/s, 31.08 mm, 17.33 mm and-2.26 mm. The corresponding area under curve(AUC), sensitivity, and specificity were 0.878, 90.9%and 90.0%; 0.822, 83.3% and 81.3%; 0.834, 86.7% and 81.3%; 0.794, 86.7% and 75.0%, respectively. The ROC curve analysis of the ADC of the bilateral neck lymph node metastasis was also carried out, with the cutoff of ADC value set as 1.07×10?3 mm2/s, the corresponding AUC, sensitivity, and specificity were 0.806, 80.0%and 75.0%. Conclusion The ADC value, tumor length ,tumor thickness and para?lingual distance between tumor and tongue midline,determined from MR imaging, all can be used as independent factors in predicting cervical lymph node metastasis, where ADC value may be helpful to predict bilateral neck lymph node metastasis.

Objective To predict the core targets and action pathways of Hedysari Radix based on UPLC-MS/MS and network pharmacology methods,and to verify the results of network pharmacology by molecular docking and molecular dynamics techniques.This article aims to investigate immune regulation mechanism of effective components absorbed into blood from Hedysari Radix.Methods Qualitative quantification of effective components absorbed into blood from Hedysari Radix were operated by using UPLC-MS/MS technique.The corresponding targets of effective components absorbed into blood from Hedysari Radix were screened by TCMSP and HERB databases.Targets of immune-related disease were obtained through DisGeNET,OMIM,TTD,and MalaCards databases.The network of"components absorbed into blood from Hedysari Radix-immune-related diseases"was then constructed.GO and KEGG enrichment analysis and mapped the PPI network were performed.Molecular docking and molecular dynamics techniques were applied for validation.Results A total of 8 prototype components absorbed into blood,synergistically acting on 101 targets,were identified by UPLC-MS/MS.They mediated 538 biological processes including immune response,positive regulation of gene expression,receptor binding,and cytokine activity.Meanuhile,116 signaling pathways,such as HIF-1,Toll-like receptor,JAK-STAT,T cell receptor,PI3K-Akt,and FoxO etc.were involved.The core targets were MAPK14,PTGS2,MMP9,PPARG,CCND1,etc..The results of molecular docking showed that formononetin and calycosin had strong docking binding activity with MAPK14.And molecular dynamics simulations further demonstrated that the binding between MAPK14 and formononetin or calycosin had good structural stability and binding affinity.Conclusion The results of serum pharmacochemistry,network pharmacology and molecular dynamics were verified to reveal the material basis and mechanism of Hedysari Radix in regulating immunity.The aim of this study is to provide scientific basis for its immunomodulatory mechanism.