Objective:To discuss the necessity of the opening of a pharmacist section for outpatients for risk assessment and com-munication of drugs in pregnancy by surveying the use situation and risk reasons of the drugs was in pregnancy. Methods:Referring to the risk classification of medicines in pregnancy formulated by FDA and integrating various factors in pregnant women, such as drug dosage, administration time, genetic factors, prenatal care and potential diseases et al, pharmacists established the section for outpa-tients for risk assessment and communication of drugs in pregnancy. Results:The visiting number of pregnant women was gradually in-creased after the establishment by reviewing the clinical data from the hospital information system. The new pharmacy service mode pro-vided by pharmacists for obstetrical patients was positively recognized by physicians and patients. Conclusion:The section for outpa-tients for risk assessment and communication of drugs in pregnancy should be established in order to promote the pharmaceutical knowl-edge in pregnancy and improve the medication safety.

ObjectiveTo explore the relationship of marriage quality with individual characteristics and copy style among the epilepsy patients.MethodsThe Chinese Marriage Quality Questionnaire,and Eysenck Personality Questionnaire Short Form Scale-Chinese edition (EPQ-RSC),and the Questionnaire of Simple Way to Coping were assessed used to assess 97 cases of the aduh married epilepsy patients and 93 cases of the control group 1 (epilepsy patients spouses) and 91 cases of the control group 2 (healthy crowds ).ResultsThe total scores of the marriage quality of epilepsy patients ( 295.78 ± 40.37 ) was lower than ones in the control group 2 ( ( 354.85 ± 28.11 ),P =0.000) and higher than that of the control group 1 ( (278.55 ± 42.23 ),P =0.014),the difference was all statistically significant(P＜ 0.05).The total scores of marriage quality of the epilepsy patients was negative correlated with the sex,and the age,and the marriage age and the severity,and the duration,and psychoticism( the Pearson correlation was respectively:-0.204,-0.436,-0.243,-0.691,-0.568,-0.635 ),and was positive correlated with which the formal schooling,and the attack types (symptoms epilepsy),and the introversion or extroversion,and the hidness,and the positive( the Pearson correlation was respectively 0.317,0.750,0.267,0.386,0.188) the difference was all statistically significant (P ＜ 0.05).Those seven variables that the attack type,and the hidness,and the severity,and the sex,and the education,and the psychoticism,and the positive could be predict to marriage quality of epilepsy patients,Adjusted R square was 0.903,After adjustment was 0.021 (P ＜ 0.05 ).ConclusionThe lever of marriage quality of the epilepsy patients is lower,and correlate with severity of epilepsy disease,and personality,copy style,and can be do comprehensive intervention from those.

0.05).The accuracy of Borrmann type classification in 14 cases of advanced gastric carcinoma undergone gastrectomy was 92.8%.Conclusion The gastric carcinoma detection rate with NHSCT is similar to that with fibro-gastroscopic or double-contrast barium examination.The direct and indirect signs of gastric carcinoma can be found and the Borrmann type classification can be made by NHSCT.However,the non-contrast enhancement scanning is limited for the early gastric carcinoma detection,and can be improved by contrast enhancement scanning.

Objective To investigate the role of Wnt/β-catenin pathway in regulating allergic airway inflammation in asthmatic mice.Methods We induced dendritic cells (DCs) from bone marrow of BALB/c mice,and then treated the cells with LiCl and PKF118-310,separately.We observed the morphological features of DCs under light microscope.Mixed lymphocyte reaction (MLR) was used to observe the functional changes of DCs.Western blot was used to detect the expressions of GSK-3β and β-catenin at the protein level.We established a mouse asthma model by using ovalbumin (OVA),and then treated these mice with LiCl and PKF118-310.The total number of cells and eosinophil percentage in BALF were determined.The lungs of mice were observed by HE staining to evaluate the degree of allergic inflammation.The cytokines in BALF and spleen cells supernatant were assayed by enzyme-linked immunoassay (ELISA),and the total IgE in the serum was also measured by ELISA.The protein expression levels of GSK-3β and β-catenin in lung tissue were assayed by Western blot.Results ① The DCs treated with LiCl promoted the proliferation of allogeneic T lymphocytes in MLR more weakly than those treated with PKF118-310 (P＜0.01).② The GSK-3β protein expression level of DCs treated with LiCl was significantly lower than DCs treated with PKF118-310.In contrast,the β-catenin protein expression of DCs treated with LiCl was higher than that of DCs treated with PKF118-310 (P ＜ 0.01).③ The total number of cells and eosinophil percentage in BALF were significantly increased in the experimental group compared with those in the control group (P＜0.01).There was also a significant difference between LiCl group and PKF118-310 group (P＜0.01).④ In the three experimental groups,the severity of inflammation in the lungs of LiCl group was weaker than that in PKF118-310 group (P＜0.05).⑤ Compared with that in the normal control group,IL-4 in BALF and spleen cell culture supernatant of the experimental group was significantly higher while IFN-γ was the opposite (P＜0.01).LiCl group had the lowest level of IL-4 and the highest level of IFN-γ;PKF groups was the opposite (P＜0.05).⑥ The total IgE in serum was significantly increased in the experimental group compared with the control group (P＜0.01).There was also a significant difference between LiCl group and PKF118-310 group (P＜0.05).⑦ GSK-3β protein expression was significantly lower in LiCl group than in PKF118-310 group (P＜0.05),while β-catenin protein expression was significantly higher in LiCl group than in PKF118-310 group (P＜0.05).Conclusion LiCl and PKF118-310 can affect the severity of asthma by regulating Wnt/β-catenin signal pathway and the expressions of GSK-3β andβ-catenin protein,which provides a new direction for asthma treatment.

BACKGROUNDTo study the effects of extraneous p53 antisense RNA on malignant growth and sensitivity to cisplatin of human lung cancer cell line.

METHODS801D cell line with p53 deletion and mutation at 248 codon was selected as a parent cell line. An 1.8 kb human p53 full length cDNA was inserted into a mammalian expression vector PEGFP to construct a p53 antisense RNA recombined plasmid PEGFP-p53(AS) and GFP gene at plasmid was a report gene to monitor extraneous gene expression. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohitochemical stain of p53 monoclonal antibody. The inhibition growth efficacy of extraneous p53 in vitro was determined by clonogenic survival assay. Sensitivity of cells to cisplatin was examined with MTT assay. FCM analysis was performed to measure the effect of p53 antisense RNA on cell cycle.

RESULTSTwo cell lines, PEGFP-p53(AS)-801D and PEGFP-801D, were established after transfection of 801-D cells by lipofection and selection. Presence of extraneous p53 gene in PEGFP-p53(AS)-801D was proved by PCR and expression of extraneous p53 was estimated when green fluorescence in those cells was found out under the fluorescent microscopy. Mutated p53 protein in parent cell line 801D was positive and in PEGFP-p53(AS)-801D was negative with immunochemical stain. The inhibition rate of colony formation was 61% for PEGFP-p53(AS)-801D (P < 0.001). The sensitivity of PEGFP-p53(AS)-801D cells to cisplatin was increased. FCM analysis showed that the cell line was arrested at G1 phase.

CONCLUSIONSp53 mutation at 248 code plays an important role on malignant growth and resistance to cisplatin of human lung cancer cell line 801D. Malignant growth of cells with p53 deletion and mutation at 248 codon can be inhibited by extraneous p53 antisense RNA, and simultaneously the sensitivity to cisplatin is also increased.

BACKGROUND AND OBJECTIVEThe p53 as a transcription factor in cell stress was activated to regulate cell cycle and programmed cell death to inhibit tumor growth. Usually, p53 is kept in non-activated state through various mechanisms, including the action of p53 C-terminal negative regulatory sequences. The purpose of the study is to prepare the two types p53 recombinant adenoviruses that carry full-length p53 as well as deletion of negative regulatory sequences at p53 C-terminus and to detect exogenous GFP expression in human lung cancer cell infected-virus by FCM scatter plot.

METHODSUsing pAdEasy-Track vector system the p53 recombinant plasmids was constructed and the homologous recombinants in E. coli was produced. The three kinds of recombinant adenovirus in L293 cells was generated, sequencing proved. Exogenous GFP expression in human lung cancer 801D cells infected-virus was detected by FCM scatter plot.

RESULTSp53 recombinant adenoviruses named Ad-p53(wtp), Ad-p53(del) and Ad-(empty carrier) were produced. Results of sequences indicate that the Ad-p53(del) was deletion of 111 bases before stop codon TGA and of 3 untranslated region at p53, the Ad-p53(wtp) no loss of any p53 base, the Ad-(empty carrier) no p53 sequence. FCM scatter plot indicate the percentage of 801D cells expressed GFP with three kinds of viral infection was almost same and was increased with the virus density. 801D contains ratio of cells with different fluorescence intensity.

CONCLUSIONThe preparation of recombinant adenovirus, Ad-p53(del), pA-p53(wtp) and Ad-(empty carrier). The cells expressed-GFP can be quantitatively detected by FCM scatter plot. It was provide that the reliability of the virus system and accurate method for selecting viruses density to infecting cells.

Adenoviridae ; genetics ; Animals ; Flow Cytometry ; methods ; Genes, p53 ; Green Fluorescent Proteins ; genetics ; Humans ; Mice ; Recombination, Genetic

BACKGROUNDTo study the inhibition effects of both extraneous right sense and antisense p53 on malignant phenotype of human lung cancer cell line.

METHODSThe named 801D cell line with p53 deletion and mutation at 248 code was selected as a model in vitro. The recombined plasmid pEGFP-p53(RS) and pEGFP-p53(AS) were constructed. The extraneous gene was detected by PCR. The p53 mutation protein was examined by immunohistochemical stain of p53 antibody. The inhibition effect of extraneous p53 on tumor growth in vitro were determined by clonogenic survival assay. FCM analysis was carried out in cells. The inhibition effect on malignant growth of extraneous p53 in vivo was observed by heteroplastic transplant on nude mouse.

RESULTSThe transfected cell lines, pEGFP-p53(AS)-801D, pEGFP-p53(RS)-801D and pEGFP-801D were established. Presence of extraneous p53 and neo genes in pEGFP-p53(AS)-801D and pEGFP-p53(RS)-801D was proved by PCR and green fluorescence was found out in those cells under the microscope. Mutant protein in pEGFP-p53(AS)-801D was negative by immunohistochemical stain. The malignant growth of these transfected cell lines was inhibited comparing with parents in vivo and in vitro. Inhibition rate of colony formation was 62.0% for pEGFP-p53(AS)-801D and 80.8% for pEGFP-p53(RS)-801D. The tumorigenicity in nude mice was suppressed. Inhibition effects of extraneous right sense p53 on malignant growth of 801D was more distinct. FCM analysis showed that pEGFP-p53(AS)-801D cells were arrested at G1 phase.

CONCLUSIONSThe transfected cell lines with extraneous right sense and antisense p53 appear that malignant growth can be inhibited in vivo and in vitro.

The oral microbiota is associated with oral diseases and digestive systemic diseases. Nevertheless, the causal relationship between them has not been completely elucidated, and colonisation of the gut by oral bacteria is not clear due to the limitations of existing research models. The aim of this study was to develop a human oral microbiota-associated (HOMA) mouse model and to investigate the ecological invasion into the gut. By transplanting human saliva into germ-free (GF) mice, a HOMA mouse model was first constructed. 16S rRNA gene sequencing was used to reveal the biogeography of oral bacteria along the cephalocaudal axis of the digestive tract. In the HOMA mice, 84.78% of the detected genus-level taxa were specific to the donor. Principal component analysis (PCA) revealed that the donor oral microbiota clustered with those of the HOMA mice and were distinct from those of specific pathogen-free (SPF) mice. In HOMA mice, OTU counts decreased from the stomach and small intestine to the distal gut. The distal gut was dominated by Streptococcus, Veillonella, Haemophilus, Fusobacterium, Trichococcus and Actinomyces. HOMA mice and human microbiota-associated (HMA) mice along with the GF mice were then cohoused. Microbial communities of cohoused mice clustered together and were significantly separated from those of HOMA mice and HMA mice. The Source Tracker analysis and network analysis revealed more significant ecological invasion from oral bacteria in the small intestines, compared to the distal gut, of cohoused mice. In conclusion, a HOMA mouse model was successfully established. By overcoming the physical and microbial barrier, oral bacteria colonised the gut and profiled the gut microbiota, especially in the small intestine.
Animals ; Bacteria ; Gastrointestinal Microbiome ; Germ-Free Life ; Humans ; Mice ; Microbiota ; RNA, Ribosomal, 16S

Urea transporters (UT) play a vital role in the mechanism of urine concentration and are recognized as novel targets for the development of salt-sparing diuretics. Thus, UT inhibitors are promising for development as novel diuretics. In the present study, a novel UT inhibitor with a diarylamide scaffold was discovered by high-throughput screening. Optimization of the inhibitor led to the identification of a promising preclinical candidate,