Objective:To investigate the effect of nuclear receptor subfamily 2 group F member 2(NR2F2)on the biological behaviors of human ovarian cancer SKOV3 cells,and to clarify its molecular mechauism and provide the new idea for treatment of ovarian cancer.Methods:Gene Expression Profiling Interactive Analysis(GEPIA)Database analyse the expression level of NR2F2 gene in ovarian tissue,and analyse its correlation with clinical prognosis of ovarian cancer patients.The human ovarian cancer SKOV3 cells were divided into control group and NR2F2 over-expression(NR2F2 OE)group,which were transfected with mCherry control virus and NR2F2 OE over-expression virus,respectively,when the cell deusity reached 70％,and the stable transfection SKOV3 cell lines were screened with puromycin(puro)48h lafter.Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting methods were used to detect the transfection efficiencies of the cells;RT-qPCR method was used to detect the expression levels of NR2F2 and sex-determining region Y-box 2(SOX2)mRNA in the cells in two groups;Western blotting method was used to detect the expression levels of NR2F2,ATP-binding cassette superfamily G member 2(ABCG2),and programmed cell death 1-ligand 1(PD-L1)protcins in the cells in two groups.CCK-8 assay was used to detect the proliferation activities of the cells in two groups;Wound assay was used to detect the migration rates of the cells in two groups;Transwell chamber assay was used to detect the number of transmembrane cells;Spheroidization assay was used to detect the numbers of spheroids in the cells;peripheral blood mononuclear cells(PBMCs)-mediated tumor cell killing assay was used to detect the relative densities of surviving tumor cells;CCK-8 assay was used to detect the half maximal inhibitory concentration(IC50)of paclitaxel(PTX)and carboplatin(CBP).Results:Compared with normal ovarian tissue,the expression level of NR2F2 gene in ovarian tumor tissue was decreased(P＜0.05),and decreased with the improvement of clinical pathological grading of ovarian tumor.The patients with higher expression level of NR2F2 gene had better clincal prognosis.The SKOV3 cells with NR2F2 over-expresson were successfully constructed,and the expression levels of NR2F2 mRNA and protein in the cells in NR2F2 OE group were increased compared with control group(P＜0.001).The CCK-8 assay results showed that compared with control group,the proliferation activities of the cells in NR2F2 OE group were decreased at different time points(1,2,3,and 4 d)(P＜0.05 or P＜0.01).The cell wound assay results showed that compared with control group,the migration rate of the cells in NR2F2 OE group was decreased(P＜0.001).The Transwell assay results showed that compared with control group,the number of transmembrane cells in NR2F2 OE group was decreased(P＜0.01).Compared with control group,the number of the spheroids in NR2F2 OE group was decreased(P＜0.05),and the expression levels of SOX2 mRNA(P＜0.01)and protein(P＜0.001)were increased.Compared with control group,the relative density of surviving tumor cells in NR2F2 OE group was decreased,but the difference was not significant(P＜0.05),and the expression level of PD-L1 protein was decreased(P＜0.05).Compared with control group,the proliferation activities of cells in NR2F2 OE group were decreased(P＜0.05),and the drug sensitivities of the cells to PTX and CBP were enhanced(P＜0.05);the IC50 of PTX was significantly reduced,while the IC50of CBP could not be calculated due to excessively high drug concentration;the expression level of ABCG2 protein was decreased(P＜0.05).Conclusion:The over-expression of NR2F2 may inhibit the proliferation,migration,and invasion of the human ovarian cancer SKOV3 cells,decrease the expression levels of SOX2,PD-L1 and ABCG2 proteins,suppress the stemness and immune evasion ability of the SKOV3 cells,and enhance the sensitivities of the SKOV3 cells to PTX and CBP.

Objective:To discuss the effect of long non-coding RNA(lncRNA)LINC00973 on the migration,invasion,and distant metastasis of epithelial ovarian cancer,and to clarify its molecular mechanism.Methods:The human ovarian cancer SKOV3 and OVCAR3 cells were divided into EF1a-FH empty vector control group(control group),LINC00973 overexpression group(LINC00973 OE group),U6-shRNA empty vector control group(SHV group),and LINC00973 knockdown group(LINC00973 KD group),and were transfected with lentivirus containing nonsense sequence(pLent-EF1a-FH-CMV-copGFP-P2A-Puro),LINC00973 overexpression,nonsense sequence(pLent-U6-shRNA-CMV-copGFP-P2A-Puro)and LINC00973 shRNA,respectively,followed by puromycin screening to obtain stably transfected SKOV3 and OVCAR3 cells.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the mRNA expression levels of target genes in the cells in various groups;wound healing assay was used to detect the migration rate of the cells in various groups;Transwell chamber assay was used to detect the number of transmembrane cells in various groups;The mice were divided into control group(WT group),LINC00973 OE group,and LINC00973 KD group,with 4 mice in each group.SKOV3 wild-type cells,LINC00973 OE cells,and LINC00973 KD cells were intraperitoneally injected into the mice in various groups,respectively,to establish the epithelial ovarian cancer intraperitoneal implantation and metastasis model;HE staining was used to observe the morphology of the colon and liver tissues of the mice in various groups;RNA-secquencing(RNA-seq)was used to analyze the differentially expressed genes between SHV and LINC00973 KD groups in the SKOV3 cell line;RT-qPCR method was used to detect the mRNA expression levels of LINC00973 in the normal ovarian epithelial cells IOSE-80 and epithelial ovarian cancer cells SKOV3,A2780 and OVCAR3,the mRNA expression levels of LINC00973,Vimentin,Snail family transcriptional repressor 1(Snail),Twist family basic helix-loop-helix transcription factor 1(Twist),zinc finger E-box binding homeobox 1(ZEB1),zinc finger E-box binding homeobox 2(ZEB2),CXCL8 and matrix metalloproteinase(MMP)16 in the cells in various groups,and the mRNA expression levels of LINC00973,Vimentin and Twist in liver and colon tissues of the mice in various groups.Results:Compared with normal ovarian epithelial cells IOSE-80,the expression level of LINC00973 mRNA in the epithelial ovarian cancer cells SKOV3,OVCAR3 and A2780 was significantly increased(P<0.01),with the highest expression level of LINC00973 in SKOV3 and OVCAR3 cells,which were therefore selected for subsequent experiments.In SKOV3 and OVCAR3 cells,compared with control group,the expression level of LINC00973 mRNA in the cells in LINC00973 OE group was increased(P<0.01);compared with SHV group,the expression level of LINC00973 mRNA in the cells in LINC00973 KD group was decreased(P<0.05 or P<0.01),indicating successful construction of LINC00973 overexpression and knockdown cell lines.In SKOV3 cells,compared with control group,the mRNA expression levels of Vimentin and Twist in LINC00973 OE group were increased(P<0.05 or P<0.01),while no significant difference was observed in Snail mRNA expression level(P>0.05);compared with SHV group,the mRNA expression levels of Vimentin,Snail and Twist in LINC00973 KD group were decreased(P<0.01).In OVCAR3 cells,compared with control group,the mRNA expression levels of Vimentin,Snail and Twist in LINC00973 OE group were increased(P<0.01);compared with SHV group,the expression levels of Vimentin,Snail,and mRNA Twist in LINC00973 KD group were decreased(P<0.01).The wound healing assay results showed that compared with control group,the wound healing rates of the SKOV3 and OVCAR3 cells in LINC00973 OE group were significantly increased(P<0.01);compared with SHV group,the wound healing rates of the cells in LINC00973 KD group were significantly decreased(P<0.01).The Transwell chamber assay results showed that compared with control group,the numbers of transmembrane cells of the SKOV3 and OVCAR3 cells in LINC00973 OE group were significantly increased(P<0.01);compared with SHV group,the numbers of transmembrane cells in LINC00973 KD group were significantly decreased(P<0.01).Compared with WT group,the number of peritoneal nodules in LINC00973 OE group was increased,with rough liver surface and multiple nodules formed on mesentery and colon surface,and the expression levels of LINC00973,Vimentin,and Twist mRNA in colon tissue were increased(P<0.01);compared with WT group,no nodules were formed in the peritoneal cavity of LINC00973 KD group,with smooth liver surface,no nodules in liver tissue,and decreased expression levels of LINC00973,Vimentin,and Twist mRNA,and no nodules were observed on mesentery and colon surface.The HE staining results showed that compared with WT group,the multiple lesions were observed in liver and colon tissues in LINC00973 OE group,manifested as uneven cell size,irregular shape,unclear cell boundaries,increased nuclear division,and uneven red staining in cytoplasm,while in LINC00973 KD group,the cells in liver and colon tissues were arranged neatly with regular shape,and uniform distribution of nuclei and cytoplasm.The RNA-seq results showed that compared with SHV group,no key signaling pathways related to tumor metastasis were enriched in LINC00973 KD group,and the transcription levels of metastasis-related genes CXCL8,MMP16,ZEB1,and ZEB2 were decreased.The RT-qPCR results showed that compared with control group,the expression levels of ZEB1,ZEB2,CXCL8,and MMP16 mRNA in the cells in LINC00973 OE group were significantly increased(P<0.01);compared with SHV group,the expression levels of ZEB1,ZEB2,CXCL8,and MMP16 mRNA in the cells in LINC00973 KD group were significantly decreased(P<0.01).Conclusion:LINC00973 can up-regulate the expression of metastasis-related factors Vimentin,Snail,Twist,ZEB1,ZEB2,CXCL8,and MMP16,and promote the migration,invasion,and distant metastasis of epithelial ovarian cancer.

Objective To investigate whether the population pharmacokinetics (PPK)models can optimize the initial dosage of individualized Valproic acid (VPA)in children with epilepsy. Methods The epileptic children without taking VPA previously were recruited from October 2015 to May 2017 at the Department of Pediatrics,Peking University First Hospital,and they were divided into the PPK model group and the traditional empirical method group by randomized method. The initial VPA dosages for the PPK model group were calculated by PPK model,whereas those of the traditional empirical method group were dosed at 20-25 mg/(kg·d)regularly. The steady-state serum trough concentrations of VPA were extracted,and then the number and percentage of the patients whose serum trough concen-trations of VPA were 50-100 mg/L in the 2 groups were analyzed and compared with prospectively randomized me-thod. Results Totally 65 epileptic children were recruited and they were randomly divided into the traditional empirical method group (32 cases)and the PPK model group (33 cases). Twenty-seven children in the traditional empirical method group were observed,and 12 children had local epilepsy attack and 15 had generalized seizures;whereas among 29 cases in the PPK model group,there were 12 local attack of epilepsy and 17 had generalized seizures. VPA add-on therapy was administrated in 9 cases and 15 cases in the traditional empirical method group and the PPK model group, respectively. There were 5 cases,21 cases and 1 case with VPA serum concentrations of ＜50 mg/L,50-100 mg/L and＞100 mg/L in the traditional empirical group;while there were 9 cases,20 cases and 0 case in the PPK model group. The VPA serum concentrations of 21 cases (77. 8%,21/27 cases)in the traditional empirical method group and 20 ca-ses (69. 0%,20/29 cases)in the PPK model group were 50-100 mg/L,respectively,and the difference was not sta-tistically significant(P＞0. 05). Conclusion Although the study doesn't suggest that the established PPK model of VPA in Chinese epileptic children is superior to the traditional empirical method,the PPK model might be potentially valuable for optimized individualized dosage adjustment for those with serum trough concentrations not in the reasonable range by the traditional empirical method and with clinical seizure or brain firing activities.

OBJECTIVE:To promote the realization of information management for hospital pharmacy. METHODS:The practice of pharmacy information management pathway in our hospital was introduced in respects of drug supply,rational drug use monitoring,pharmacy information dissemination,etc. The effects of information management were evaluated in respects of work efficiency,the rate of drug dispensing error and rational drug use index,etc. RESULTS:The information management pathway of our hospital included drug purchase,warehousing and keeping system,drug distribution management system,reasonable drug use monitoring information module,etc. It realized automatic generation and issue of purchase plan,acceptance bar code of drug warehousing,real-time drug stocking and location changing,drug guarantee delivery in advance,window drug delivery and check barcode scanning,drug distribution quality information management,whole process temperature and humidity monitoring, unreasonable prescription after the event,in the event,in advance,early warning of drug use,real-time monitoring of drug use and administration of drugs for chronic diseases,etc. After the implementation of information management,the time of drug warehousing invoice input and drug stocking were shortened.Among drug dispensing error,the percentage of type error decreased from 0.005 2% during Jun.-Dec. 2015 to 0.001 6% during Jan.-Jun. 2016;that of number error decreased from 0.006 9% to 0.001 6%;that of other error dropped to zero. During 2014-2016,the ratio of drug cost to treatment cost decreased gradually;those of inpatient were 26.62%,24.91%,24.36%,respectively;those of outpatient were 44.06%,42.10%,41.32%, respectively. Utilization rates of antibiotics in the inpatients decreased gradually,and were 44.82%,44.14%,43.91%, respectively. CONCLUSIONS:Information construction plays an important role on hospital pharmacy,can effectively improve work efficiency,reduces medication error and promotes rational drug use.

Objectives: To investigate the effect of β-catenin interacting protein 1 (ICAT) silence in Wnt signaling pathway and sterol drug NSC67657 induced cell differentiation of HL60 cell. Methods: HL-60 cells were treated with NSC67657, the cell surface antigen CD14 expression was detected by flow cytometry. Lentivirus LV-ICAT-RNAi vector was constructed and infected HL-60 cells. Then the ICAT gene and protein expression were analyzed using real-time qPCR and Western blot technique. Furthermore, Co-immunoprecipitation assay was used to confirm the interaction of β-catenin/ICAT proteins, and Western blot was employed to compare the expressions of Wnt signaling pathway downstream targets Cyclin D1, TCF-1 and c-Jun between Lentivirus LV-ICAT-RNAi vector infected HL-60 (HL-60i) cells and un-infected HL-60 (HL-60v) cells. The cellular differentiation of HL-60i and HL-60v cells treated with NSC67657 for 24 h was evaluated by Wright’s staining, transmission electron microscopy and flow cytometry analysis. Results: HL-60 cells could be induced to differentiate into monocytes by 10 μmol/L NSC67657. The CD14 positive cells could reach to (92.30±5.14) % after NSC67657 treatment for 5d. The co- immunoprecipitation assay demonstrated that ICAT protein did interact with β-catenin protein, and the absorbance of protein electrophoresis bands increased in differentiated cells. The expressions of Wnt signaling pathway downstream target proteins in HL-60i cells were higher than that in HL-60v cells when they were treated by 10 μmol/L NSC67657, but lower than NSC67657 untreated cells. CD14 positive HL-60i cells were significantly lower than that of HL-60v cells[ (8.33±3.14) % vs (19.08±4.73) %]when treated with NSC67657, but still higher than that of uninfected and untreated HL60 cells[ (0.60±0.03) %] (F=119.24, P=0.010) . The results of cellular morphology and ultrastructure observation were also in accord with that of cell surface antigen analysis. Conclusions ICAT does participate in HL-60 cells monocytic differentiation induced by NSC67657, and Wnt/β-catenin signaling pathway might play a bridge role.

Objective To explore the clinical significance of detecting serum levels of IL-6 and TNF-α in patients with intracranial hemorrhage.Methods The serum levels of IL-6 and TNF-α in different period were detected in patients with intracranial hemorrhage, by enzyme-linked immunosorbent assays (ELISA), and compared with those of healthy subjects (the control group).ResultsThe serum levels of IL-6, TNF-α in the severe and slight patients of study group on 1st, 3rd, 5th and 7th were signiifcantly higher than those in the control group (all withP<0.05). The serum levels of IL-6, TNF-α in the severe patients of study group were signiifcantly higher than those in slight patients of study group (all withP<0.05) on 5th and 7th. The serum levels of IL-6 and TNF-α in dead cases on 5th, 7th days admission were significantly higher than those in survival cases (P<0.05). The serum levels of IL-6 was positively correlated with TNF-α (r=0.721,P<0.05).Conclusion The detection of dynamically serum levels of IL-6 and TNF-α is of great clinical value for assessing the disease development therapeutic efifcacy and prognosis of brain injury patient with intracranial hemorrhage.

[ABSTRACT]OBJECTIVETo investigate the expression levels of IL-25 and IL-33 mRNA in the nasal mucosa of allergic rhinitis(AR) mice.METHODSBalb/c mice were used for establishing the animal model of allergic rhinitis with oval bumin sensitization as AR group, at the same time, the physiological saline as the control group. IL-25 and IL-33 mRNA in nasal mucosa of the two groups were detected by real-time quantitative PCR.RESULTSThe expression of IL-25 and IL-33 mRNA could be detected in both the control and AR group. The expression level of IL-25 and IL-33 mRNA in AR group were significantly higher than that in control group, the difference was statistically significant (P<0.05).CONCLUSIONIL-25 and IL-33 were involved in the development of allergic rhinitis. This result will be helpful for the further understanding of the pathogenesis of allergic rhinitis, and provide a theoretical basis for the treatment of allergic rhinitis.

Objective In order to investigate the impact of interactive online teaching in Laboratory Diagnosis, a randomized experimental study was conducted among the medical students in Grade 2009.Methods A total of 65 subjects were randomized into the experimental group (N=31) and the control group (N=34).In addition to the standard in-class presentation/discussion and school-wide refined online courses offered in the control group, interactive online teaching was added to experimental group.After completion of the course, the impact of interactive online teaching in the experimental group was compared to that in the control group through a final exam and a question-naire concentrating on motivation of study, problem solving and team working skills.Statistical signifi-cance of the comparison was evaluated using t-test.Results The results from the final exam showed that the average score in the experimental group was higher than the control group (87.20 ± 5.25 vs.82.69 ± 5.76), It is statistically significant (P=0.002).The average scores of short answer and case study questions in the experimental group was higher than the control group (26.16 ± 1.57 vs.25.31 ± 1.73, 18.94 ± 1.05 vs.15.53 ± 1.15), It is statistically significant (P=0.043,P=0.000).The satisfaction evaluation of interest in learning, timely answers to questions, learning, analytical skills, teamwork culture reflected by the questionnaire indicated statistically higher (P=0.041,0.000,0.000,0.000,0.000) average scores in the experimental group as compared to its counterparts in the control group (4.30 ± 0.65 vs.3.97 ± 0.62, 4.63 ± 0.49 vs.3.25 ± 0.88, 4.43 ± 0.57 vs.3.49 ± 0.65, 4.46 ± 0.57 vs.3.37 ± 0.73, 4.33 ± 0.66 vs.3.68 ± 0.58).Conclusion Interactive online teaching helps to improve the study performance of medical students in Laboratory Diagnosis.Incorporating interactive online teaching into the current course regarding Laboratory Diagnosis merits recommendation.

Objective To evaluate the limit of blank (LoB),limit of detection (LoD),limit of quantitation(LoQ)and functional sensitivity (FS)of prealbumin (PA)detected by the Roche Modular P automatic biochemical analyzer.Methods According to the EP17A file of the American Clinical and Laboratory Standards Institute (CLSI),saline as the blank sample and a series of low con-centration samples were detected by the Roche Modular P automatic biochemical analyzer for determining LoB,LoD and LoQ.And FS was determined based on the domestic universal method .Results LoB of PA was 16.35 mg/L,LoD was 18.23 mg/L,LoQ was temporarily unable to evaluate and FS was 25.00 mg/L.The report scope and the report mode in clinic were affirmed by combi-ning with the low value of the reportable scope.Conclusion LoD of PA detected by the Roche Modular P automatic biochemical an-alyzer is established,which provides more valuable information for clinical diagnosis and treatment.Conducting the comparison of different evaluation methods determines the advantages and limitation of the practical application of different methods.

Objective To investigate the effect of leptin on the secretion of chemokine in THP1 cells and explore its related mechanism, and to provide basis for research on the role of leptin in immune response.Methods The expressions of Ob-Rb and Ob-Rt in THP1 cells were detected by RT-PCR and flow cytometry (FCM).The THP1 cells at logarithm growth phase were selected and randomly divided into blank control group and different concentrations(10,50,100,200μg· L-1 )of leptin groups.Transwell chamber assay was performed to detect the number of invated THP1 cells.The THP1 cells were divided into blank control group and 100μg·L-1 leptin group.Western blotting method was carried out to detect the expressions of p-AKT,p-ERK 1/2,and p-STAT3 in THP1 cells.The THP1 cells were divided into blank control group and 100μg·L-1 leptin group,100μg·L-1 leptin+ DMSO group,100μg·L-1 leptin+50μmol·L-1 AG490 group,100μg·L-1 leptin+10μmol·L-1 LY294002 group and 100μg·L-1 leptin+ 10 mol·L-1 PD980590 group.RT-PCR and Western blotting methods were performed to detect the expression of IL-8.Results Ob-Rb and Ob-Rt were highly expressed in THP1 cells. Compared with blank control group,the number of invated THP1 cells was significantly increased in 50,100,and 200μg·L-1 leptin groups (P<0.05).Compared with blank control group,the expressions of p-STAT3,p-ERK 1/2 and p-AKT in THP1 cells were up-regulated in 100 ug·L-1 leptin group(P<0.05).Compared with blank control group,the mRNA and protein expressions of IL8 in THP1 cells in 50,100,and 200μg·L-1 leptin groups were remarkably increased(P<0.05);compared with 100μg·L-1 leptin group,the expressions of IL-8 in THP1 cells in 100μg·L-1 leptin+10 mol·L-1 PD980590 group and 100μg·L-1 leptin+10μmol·L-1 LY2 94002 group were decreased(P<0.05),while the expression of IL-8 in 100μg·L-1 leptin+ 50μmol·L-1 AG490 group had no change(P>0.05).Conclusion leptin can up-regulate the expression of chemokine in THP1 cells,which might be associated with PI3K-AKT and MAPK/ERK 1/2 signaling pathways.