OBJECTIVE:To provide reference for the standardization of online pharmacies pharmacist service. METHODS:Lit-erature research method,online entity investigation method and the method of mysterious customer were adopted to investigate 10 well-known online pharmacies,then described the analysis of current online pharmacies pharmacist service in China. RESULTS:There were 5 online pharmacies showing pharmacist service,but only 1 pharmacy could provide pharmacist registration certificate;on average each online pharmacy was allocated with 1.9 licensed pharmacists;only 2 online pharmacies provided 24 hours of phar-macist service;8 online pharmacies provided more than 2 consulting method;messagebox would pop actively when clicking the“Buy”button in 6 online pharmacies;the correct rate of pharmacist’s guidance was less than 60%,only 20% pharmacists could provide right explanation. CONCLUSIONS:The reasons of the lack of online pharmacies pharmacist service include online pharma-cies imperfect laws and regulations and supervision,lack of licensed pharmacists quantity and quality is not high enough,online pharmacies,pharmacists consulting system is not mature. Online pharmacies can improve relevant laws and regulations,strengthen the government supervision,reasonable and effective use of licensed pharmacists resources,perfect the pharmacist online consult-ing service water system and improve the licensed pharmacists to improve pharmacists service function status.

Objective To investigate the surgical collaboration in laparoscopic live donor nephrectomy.Method A retrospective analysis was made on the clinical data of 18 cases of laparoscopic live donor nephrectomy.Result All laparoscopic live donor nephrectomy were completed successfully,without severe complications caused by surgical collaboration.Conclusion The key points of successful operation are delicate mental care,sufficient preoperative preparation,good cooperation during the operation and related health education.

CG. And that was why SSP failed to determine the allele.Conclusion The difficulty in HLA genotyping by SSP resulted from the primers, which involved unknown sequence of Exon 1 at locus B in the studied sample.

Objective:To investigate influence of living locations on the personality characteristics of the only child and none-only children in an air force academy.Methods:1921 cadets were tested by the Chinese version of the MBTI-G.Results:Scores of the only child cadets from cities on the Sensing-Intuition and Thinking-Feeling dimensions were higher than those of none-only children group from cities and those of the only child group from countries.The scores of them on the SN scale were 93.0?18.5,90.4?17.6 and 90.1?17.7(t=2.25,P=0.025;t=1.99,P= 0.048).The scores of them on the TF scale were 94.6?22.6,89.1?21.1 and 88.4?21.0(t=4.02,P

Objective To construct a recombinant lentivirus vector containing the human NIS gene and HIF-1α with the myosin light chain-2v(MLC-2v) as a promoter and to investigate the specific expression and feasibility of NIS as a reporter gene in cardiomyocytes.Methods The target gene HIF-1α and NIS were subcloned into the lentivirus (Lv)-elongation factor (EF)1-HIF-1α-internal ribosome entry site (IRES)-NIS and Lv-MLC-HIF-1α-IRES-NIS lentivirus vectors.The recombinated vectors were transfected into Hela cells by lipofectamine 2000.The expression of HIF-1α and NIS in the transfected Hela cells was detected by indirect immunofluorescence and Western blot.The H9C2 cells were exposed to different multiplicities of infection (MOI; 5,10,20,40) with packaged virus particles.The infection efficiency was detected by Western blot.MOI 20 was used for H9C2,NIH-3T3 and L6 cell lines and the specificity of the MLC-2v promoter was detected by the count of NIS protein in the 3 different cell lines with Western blot.The function and features of NIS protein were evaluated by dynamic iodine uptake and NaClO4 iodine uptake inhibition tests in vitro.Two-sample t test was used to analyze the data.Results The two recombinant lentivirus vectors were constructed successfully.The HIF-1α protein was expressed in the cytoplasm and the NIS protein was expressed on the cell membrane in Hela cells.The grey levels of NIS and HIF-1α proteins in the positive control were 69.8 and 71.9,respectively,which were 109.4 and 92.7 after being prompted by EF1,and 141.9 and 132.4 by MLC-2v.The expression of these proteins was much higher by EF1 promoter than that by MLC-2v promoter.The optimal MOI for the Lv-MLC-HIF-1α-IRES-NIS virus to infect H9C2 cells was 20.With the MOI of 20,the grey levels of NIS protein promoted by EF1 were 23.4,29.8 and 28.6 for H9C2,NIH-3T3 and L6 cells infected with Lv-EF1-HIF-1α-IRES-NIS virus,respectively.The expression of NIS protein promoted by MLC-2v was much higher in H9C2 cells than the other two cell lines.The grey level of NIS protein was 157.9 in H9C2 cells,178.8 in L6 cells and 217.3 in NIH-3T3 cells.The NIS protein expressed in infected H9C2 cells showed high radioiodine uptake.The peak of iodine uptake was 4 287.2 counts · min-1 at 40 min which was 16.85 times of the control group (254.4 counts · min-1) (t=5.34,P＜ 0.01).The inhibition rate of iodine uptake was up to 85.5％ (3 666.4/4 287.2,t=21.3,P＜0.01) by NaClO4.Conclusions MLC-2v promoter allows specific expression of the external gene HIF-1α and NIS in myocardium.The cardiomyocytes transfected with NIS gene acquires the function of iodine uptake.Therefore,NIS may have a potential to be the reporter gene to monitor the external gene therapy in ischemic cardiomyopathy.

Objective To investigate the photo-protective mechanisms of hydroxychloroquine and epigallocatechin gallate (EGCG) on HaCaT cells damaged from UVB irradiation. Methods Subconfluent HaCaT cells were irradiated with different doses of UVB irradiation and treated with the above listed agents. The mRNA expression levels of p53, p21, c-fos and GADPH genes were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Results UVB irradiation induced mRNA expression of p53, p21 and c-fos in cultured HaCaT cells, which were alleviated by hydroxychloroquine and EGCG treatment in UVB irradiation group. Conclusions The photo-protective effects of hydroxychloroquine and EGCG on HaCaT cells by UVB irradiation might be related to inhibition of the expression of p53,p21 and c-fos genes.

Arrhythmias, Cardiac ; physiopathology ; Child ; Electrocardiography ; Humans ; Male ; Tachycardia, Ventricular ; physiopathology

Objective To evaluate the effect of Rab23 on the proliferation of a squamous cell carcinoma cell line Sa3,and to investigate its mechanisms.Methods Cultured Sa3 cells were classified into four groups:normal control group transfected with green fluorescent protein (GFP),Rab23-overexpressing group transfected with a GFP-labelled Rab23-overexpressing plasmid,Rab23-silencing group transfected with a plasmid carrying a Rab23-targeting shRNA,empty vector group transfected with an empty vector.After additional culture for different durations,plate colony formation assay and flow cytometry were performed to evaluate the proliferative activity of Sa3 cells,and Western blot was conducted to detect the expression of Erl/phosphorylated-Erk in Sa3 cells.Statistical analysis was carried out by t test,one-way analysis of variance and Bonferroni's multiple comparison test.Results Stable Sa3 cell lines with overexpression or silencing of Rab23 were established by plasmid construction and lentivirus-mediated transfection.The plate colony formation assay showed that the colony formation rate was significantly lower in the Rab23-overexpressing group than in the normal control group (2.3％ ± 0.2％ vs.3.6％ ± 0.3％,P ＜ 0.05),but higher in the Rab23-silencing group than in the empty vector group (4.1％ ± 0.2％ vs.1.8％ ± 0.03％,P ＜ 0.01).Rab23 overexpression induced G1 phase arrest in Sa3 cells.The proliferation index was significantly decreased in the Rab23-overexpressing group compared with the normal control group (0.581 ± 0.035 vs.0.698 ± 0.018,P ＜ 0.05),but increased in the Rab23-silencing group compared with the empty vector group (0.567 ± 0.015 vs.0.444 ± 0.014,P ＜ 0.01).As Western blot showed,there were no significant changes in the expression of Erk in the Rab23-silencing or-overexpressing group compared with the normal control group,whereas the expression of p-Erk was attenuated in the Rab23-overexpressing group compared with the normal control group,but enhanced in the Rab23-silencing group compared with the empty vector group.Conclusions Rab23 could inhibit the proliferation of Sa3 cells,which may be associated with the Erk pathway.

Objective To construct a recombinant lentiviral expression vector containing NIS and EGFP gene,and to explore the feasibility of NIS gene for monitoring the bone marrow derived mesenchymal stem cells (BMSCs) migration to the intracranial glioma.Methods The NIS and EGFP gene fragments were subcloned into lentiviral vector pLVX-puro,then packaged and amplified in HEK293T cells to obtain recombinant lentivirus pLVX-CMV-NIS-EGFP.pLVX-CMV-0-EGFP was constructed as control.BMSCs were isolated,cultured,and transfected by lentivirus.The antibiotic-resistant transfected BMSCs (BMSCs-NIS-EGFP and BMSCs-EGFP) were selected.The expression of NIS gene was examined by Western blot.Functional NIS activity was confirmed by the uptake of 125I and the inhibition effect of NaClO4.The nude mice intracranial glioma models were established.MicroSPECT was performed at 24 h post BMSCs-NIS-EGFP injection via the tail vein.Results pLVX-CMV-NIS-EGFP and pLVX-CMV-0-EGFP were successfully constructed and packaged.BMSCs were successfully isolated and cultured.Stable cell lines BMSCs-NIS-EGFP and BMSCs-EGFP were constructed after lentivirus transfection and puromycin selection.The expression of NIS gene was detected by Western blot in BMSCs-NIS-EGFP,but not in BMSCs-EGFP.BMSCs-NIS-EGFP showed significantly more uptake of 125I (nearly 10 times than the uptake in BMSCs-EGFP) and the uptake could be significantly inhibited by NaClO4.The nude mice intracranial glioma models were successfully established and the BMSCs-NIS-EGFP in glioma foci could be visualized by microSPECT imaging at 24 h post injection.Conclusions A recombinant lentivirus containing NIS gene could be successfully constructed for monitoring BMSCs migration towards intracranial glioma.It might provide evidence on the research of BMSCs and NIS gene mediated therapy for glioma.

Objective To explore the thrombomodulin(TM) expreesion levels changes in plasma and placenta in patients with early onset severe preeclampsia. Methods Sixty cases of severe preeclampsia women who delivered in the affiliated Xuzhou Maternity and Child Health Care Hospital of Xuzhou Medical College were enrolled in the study from June 2012 to February 2014, including 30 patients with early onset severe preeclampsia (early onset group), and 30 patients with late onset severe preeclampsia (late onset group). Healthy pregnant women were divided into two control groups according to gestational weeks at delivery: early control group ( n=23, at 28-33+6 weeks), and late control group (n=30, delivered after 34 weeks). ELISA was used to detect the levels of TM in plasma. Immunohistochemistry SP was applied to detect the TM protein expression on placenta. TM mRNA was determined by reverse transcription polymerase chain reaction (RT)-PCR technique. Results (1) TM level in plasma in early onset group and late onset group were (90.8±6.9) and (87.5±7.0)μg/L, and TM level in plasma in early control group and late control group were (37.7 ± 2.3) and (37.7 ± 2.5)μg/L. Plasma TM level in early onset group was higher than that in late onset group, early control group and late control group. The TM level had no statistically significant compare of early-onset group to late onset group.(P>0.05). The plasma TM level in early onset group was significantly higher than that in early control group (P<0.05), and the plasma TM level in late onset group was significantly higher than that in late control group (P<0.05). (2)TM expressed mainly in the membrane and cytoplasm of placental syncytiotrophoblasts and endothelial cells. The expression of TM protein in early onset group was 47%(14/30), significantly lower than that in late onset group, early control group and late control group (P<0.05), in which the positive rate were 90%(27/30), 91%(21/23) and 93%(28/30) respectively (P<0.05).There was no difference between late onset group and late control group (P>0.05). There was no difference between early control group and late control group (P>0.05). (3) TM mRNA expression in early onset group, late onset group, early control group and late control group were 0.14±0.06, 0.89 ± 0.23, 0.88 ± 0.22 and 0.93 ± 0.19, respectively. The expression of TM mRNA in early onset group was significantly lower than that in late onset group, early control group and late control group (P<0.05), and the difference between early control group and late control group was not statistically significant (P>0.05). There was no difference between late onset group and late control group (P>0.05). There was no difference between early control group and late control group (P>0.05). Conclusions Decreased expression of TM in placenta may be associated with the pathogenesis of early onset severe preeclampsia, there may be different pathogenesis in early onset and late onset severe preeclampsia.