Objective:To generate Epstein-Barr virus(EBV) Latent Membrane Protein 2A(LMP2A) recombinant adenovirus,and provide for further investigation on the therapy vaccine against EBV associated malignancies.Methods:Full length cDNA of encoding LMP2A of EBV had been amplified by reverse transcription-PCR and cloned into pGEM-T vector.The encoding cDNA of LMP2A was inserted into E1,E3-substituted adenovirus vector pAX1CW,then the LMP2A recombinant adenovirus vector was contransfected into 293 cells togetherwith EcoT221 digested Ad5-TPC.The LMP2A recombinant adenovirus was generated by homologous recombination,and primarily identificated by ClaI enzyme digestion.The expression of LMP2A on CV1 cells infected with recombinant adenovirus analyzed by fluorescence-activated cell sorting(FACS) and confocal microscope.Results:The replication-deficient LMP2A recombinant adenovirus was generated efficiently with the titers of 2.3?10 8 pfu/ml.The LMP2A could be seen on CV1 cells membrane with confocal microscope 48 h post infected with recombinant adenovirus and the percentage of CV1 cells expressing LMP2A was 94.4% by means of FACS analysis.Conclusion:These suggested that LMP2A could be expressed efficiently by recombinant adenovirus mediated transfer,and it was the foundation of further researching in its function and developing the suitable genetic engineering vaccine against EBV associated malignancies.

BACKGROUND: The postnatal sexual health of women is crucial in reproductive health, due to lack of sexual knowledge and sexual heath care service as well as conventional idea, the medical staff and postnatal women themselves pay little attention and discussion on the sexual health after childbirth.OBJECTIVE; To investigate the prevalence and types of women's sexual problems after childbirth, identify the factors associated with dyspareunia and investigate the status quo of postnatal sexual health care service. DESIGN: Cross-sectional study and follow-up survey within six months after delivery.SETTING: Department of Reproductive Medicine, Chongqing University of Medical Sciences. PARTICIPANTS: A total of 798 primiparous women delivering a live birth in Department of Obstetrics at the First Affiliated Hospital of Chongqing University of Medical Sciences between November 2000 and July 2001, and 79 husbands who accompanied their wives during the follow-ups were adopted in this study.METHODS: The primiparous women after 6 months of childbirth and husbands conducted routine postnatal follow-ups and were informed of the follow-up content. The interview was used in this cross-sectional study and the questionnaires were finished under the help of health professional. And some advice, consultations and treatments can be given in time. MAIN OUTCOME MEASURES; ①main change of postnatal sexual life;②postnatal sexual problems and related factors. RESULTS: Totally 460 out of 798 primiparous women in accordance with the inclusive and exclusive criterions received follow-up surveys after telephone or letter notification. ①Main change of postnatal sexual life: Of the 460 respondents, 94.74% had resumed sexual activity within six months after delivery. Compared with one year pre-pregnancy, the number of sexual activity decreased obviously within 3-6 months after delivery, and 66.0％ of postnatal women decreased the sexual desire.②Postnatal sexual problems and related factors: Sexual morbidity increased significantly after the childbirth: In the first three months after delivery, 70.6％ of women experienced sexual problem(s), declining to 55.6％ during the 4th-6th months, and reduced to 34.2％ at the 6th month, but not reaching pre-pregnancy levels of 7.17％. The sexual problems of women after delivery included dyspareunia, vaginal dryness, orgasm disorder and vaginal relaxation, etc.4. And dyspareunia was the most common type. Between the 4th and 6th month after delivery, dyspareunia was significantly associated with the effect of traditional sexual ideas, experience of dyspareunia before pregnancy and current breastfeeding (P ＜ 0.05), while the association of dyspareunia with delivery mode was not significant.③Sexual care service and sexual counseling after delivery: Only 20.8％ of women received information about sexual health from health care personnel within 42 days after delivery, and consultation rate of sexual intercourse problems was 8.0％.④Husbands' comprehension of postnatal problems in women: Only 59.2％ of partners whose wives had postnatal sexual problem(s) knew that their wives had ever experienced sexual problem(s) and 8.1％ of partners had sought medical help. CONCLUSION; Sexual health problems are very common in women after childbirth, taking up for 70.6％. And the quality of postnatal sexual health is unrelated with delivery mode, but postnatal health care service are often overlooked, additionally lack of professional counseling and treatment. The current practices of encouraging the husband's supports after the childbirth may well influence the prevention and treatment of postnatal sexual problems in women.

ObjectiveTo investigate the clinicopathologic features,therapy,and prognosis of primary small cell carcinoma of the pancreas.MethodsDatabases including Chinese Journal Full-text Database,VIP Database for Chinese Technical Periodicals,Medline/Pubmed,and OVID were searched electronically up to April 2012.A systematic review was performed together with one case in our hospital.ResultsTwenty-eight articles fulfilling the criteria consisting of 46 patients with pathologically confirmed diagnosis of primary small cell carcinoma of the pancreas were studied,together with 1 patient in our Drum Tower Hospital,finally 47patients were included.The results of this systematic review showed:( 1 ) Primary small cell carcinoma of the pancreas was more common in men with a median age of 62.The most common clinical presentations were abdominal pain,jaundice and weight loss.Para-neoplastic syndrome was rarely observed.(2)Most cases were found to have abnormally elevated serum levels of neuron-specific enolase.CT displayed heterogeneous,and marked enhancing masses in most cases.The conclusive diagnosis depended on histological confirmation.(3)63.8％ of the cases were found to be associated with metastasis at the time of diagnosis.The overall median survival time was 28 weeks.(4) There was no consensus on the treatment of primary small cell carcinoma of the pancreas. Chemotherapy was currently considered as the treatment of choice among the systematic management for these patients.ConclusionsPrimary small cell carcinoma of the pancreas was a rare and aggressive neuroendocrine tumor with a poor prognosis.

Objective To investigate the regulatory effects of PTEN-PI3K/AKT pathway on the apoptosis of gastric cancer MKN28 cells and the possible mechanisms.Methods The specific sequence of PTEN was transfected to MKN28 cells by eukaryotic expression vector (transfection group),and then vacant vector (negative control group) and PBS (blank control group) were transfected to the MKN28 cells,respectively.The effects of PTEN-PI3K/AKT pathway on the apoptosis of MKN28 cells and the expressions of PI3K,AKT,Caspase-3 and Caspase-9 were investigated.The growth curve and apoptosis of the MKN28 cells were detected by MTT assay and TUNEL staining,respectively,and the protein expression was detected by the Western blot.MKN28 cells which did not transfected by the PTEN were processed by inhibitor of PI3K (LY294002) (treated group),and MKN28 cells in the control group were processed by PBS.The expressions of apoptosis protein and apoptosis-related protein were detected after inhibition of PI3 K.All data were analyzed using the t test.Results The model of over-expression of PTEN was obtained and transfected into MKN28 cells.The survival of MKN28 cells in the transfection group was significantly inhibited in a time-dependant manner ( r =0.938,P ＜ 0.05 ).The mean apoptotic rate of the transfection rate was 27.86％ ± 4.78％,which was significantly higher than 0.01％ ± 0.01％ of the negative control group ( t =9.527,P ＜ 0.05 ).The protein expression of PI3K in the transfection group was 0.25 ± 0.03,which was significantly lower than 0.93 ± 0.16 of the blank control group and 0.96 ± 0.15 of the negative control group (t =7.235,8.883,P ＜ 0.05 ).The protein expression of P-AKT in the transfection group was 0.21 ± 0.03,which was significantly lower than 0.93 ± 0.13 of the blank control group and 0.91 ± 0.12 of the negative control group (t =9.347,9.802,P ＜ 0.05 ).The expressions of Caspase-3 and Caspase-9 of the transfection group were 0.86 ± 0.11 and 0.87 ± 0.12,which were significantly higher than 0.16 ± 0.03 and 0.18 ± 0.04 of the negative control group,and 0.15 ± 0.02 and 0.16 ± 0.03 of the blank control group ( t =10.634,10.999,9.448,9.942,P ＜0.05).The apoptotic rate of the MKN28 cells of the treated group was 28.60％ ± 4.50％,which was significantly higher than 0.12％ ± 0.06％ of the control group (t =10.961,P ＜ 0.05 ).The protein expression of PI3K and P-AKT of the treated group were 0.18 ± 0.02 and 0.11 ± 0.01,which were significantly lower than 0.93 ± 0.14 and 0.90 ± 0.12 of the control group (t =9.186,11.363,P ＜ 0.05 ).The protein expression of PTEN of the treated group was 1.15 ± 0.15,which was significantly higher than 0.21 ± 0.08 of the control group (t =9.577,P ＜ 0.05 ).The relative expressions of Caspase-3 and Caspase-9 of the treated group were 0.86 ± 0.12 and 0.88 ± 0.11,which were significantly higher than 0.25 ± 0.02 and 0.21 ± 0.03 of the control group (t =8.685,10.178,P ＜ 0.05).Conclusions Over-expression of PTEN may enhance the apoptosis of gastric cancer cells through inhibition of PI3K/AKT pathway.Inhibition of PI3K can enhance the expression of PTEN.PI3K and PTEN regulate the apoptosis of gastric cancer cells.

Peacekeeping is a military operation under certain conditions.In the special operations,the military personnel will have a special emotional response,mainly tension.It includes physical and psychological stress,based on the cognition and evaluation of the environment and forms of military activities.The psychological problems of peacekeepers caused by peacekeeping operation have been identified as one of the major reasons for non-combat attrition.Thus,researches on mental health of peacekeepers should not be ignored.So,we reviewed the interaction between peacekeeping operations and peacekeepers'mental health,and a few psychological intervention techniques to provide theoretical and practical basis for serving peacekeepers with mental health.

Objective To determine sexual function in 612 male addicts with methadone maintenance therapy (MMT),analyze its cause and explore the treatment strategies.Methods Self-made questionnaire and International Index of Erectile Function-5 ( IIEF-5 ) were used in 612male addicts receiving MMT.ResultsThe number of addicts who felt worse sexual function was significantly more than those who felt better sexual function (P ＜0.01 ).About 88.6％ addicts were dissatisfied with their current sexual function,90.7％ of whom were willing to continuously receive MMT.The number of addicts suffering from hyposexuality and erectile dysfunction (ED) was significantly increased at post-MMT than at pre-MMT ( P ＜ 0.01 ),while the number of addicts with normal sexuality and inhibitive ejaculation decreased significantly (P ＜0.01 ).Both the dose of methadone and the age of subjects were negatively correlated with IIEF-5 scores.Correlation between the duration of MMT and IIEF-5 scores was not found.ConclusionMMT might deteriorate sexual function in male addicts.To improve the compliance of MMT and sexual function in male addicts,the dose of methadone should be adjusted to minimally effective one.

Objective To explore clinical features and antimicrobial resistance of Klebsiella pneumoniae (K.pneumoniae) lower respiratory tract infection(LRTI) in children.Methods Clinical data of 107 children with K.pneumoniae LRTI confirmed by sputum culture from January to December 2015 were analyzed retrospectively.Results 62.62% of children with LRTI were aged less than 6 months and 64.49% episodes occurred in autumn and winter.All cases had cough and 39 had fever, the main complications were type I respiratory failure, type Ⅱ respiratory failure, cardiac insufficiency, and electrolyte disturbance, 39 cases(36.45%) had complications involving two systems, 5 cases(4.67%)had complications involving three systems,47 cases (43.93%) met the diagnostic criteria of severe pneumonia.43 cases (40.19%) had primary underlying diseases, the major were congenital heart disease, preterm and low birth weight, and malnutrition.Children with imipenem-resistant bacteria infection were more prone to develop extrapulmonary complications than those with non-resistant pathogenic infection.The resistance rate of K.pneumoniae to amikacin was the lowest(9.35%).90 cases were recovered and markedly effective, 11 cases were effective, 4 cases were not healed and voluntarily discharged from hospital, and 2 cases died.Conclusion Children aged less than 6 months and with underlying diseases are prone to develop LRTI, and complications are more.

Increasing the production and secretion of endogenous opioid peptide by immune cell can significantly induce myocardial protective effects against ischemia-reperfusion injury. Gene therapy is promising to increase endogenous enkephalin (ENK). However, classical viral and plasmid vectors for gene delivery are hampered by immunogenicity, gene recombination, oncogene activation, the production of antibacterial antibody and changes in physiological gene expression. Minimalistic immunologically defined gene expression (MIDGE) can overcome all the deficients of viral and plasmid vectors. The exon of rat's preproenkephalin (PPENK) gene was amplified by PCR and the fragments were cloned into pEGFP-N1 plasmids. The recombined plasmids were digested with enzymes to obtain a linear vector contained promoter, preproenkephalin gene, RNA stable sequences and oligodesoxy nucleotides (ODNs) added to both ends of the gene vector to protect gene vector from exonuclease degradation. A nuclear localization sequence (NLS) was attached to an ODN to ensure the effective transport to the nucleus and transgene expression. Flow cytometry, laser confocal microscopy and Western blotting demonstrated that PPENK-MIDGE-NLS can transfect leukocyte of rat in vivo, increase the expression of proenkephalin (PENK) in tissue, and the transfection efficiency depends on gene vector's dosage. These results indicate that PPENK-MIDGE-NLS could be an innovative method to protect and treatment of myocardial ischemia-reperfusion injury.
Animals ; Cloning, Molecular ; Enkephalins ; genetics ; Gene Expression ; Genetic Therapy ; Genetic Vectors ; Leukocytes ; Plasmids ; Promoter Regions, Genetic ; Protein Precursors ; genetics ; Rats ; Transfection ; Transgenes

Objective To investigate the role of long noncoding RNA-AJ227913 in the pathogenesis of primary gout arthritis (GA). Methods The subjects were divided into three groups:30 acute gout patients (AGA), 30 non-acute gout patients (NAGA), 30 healthy controlsand 30 hyperuricemia patients (HUA). Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to examine the expression of AJ227913 in peripheral blood mononuclear cells(PBMCs) from four groups. 100 μg/ml monosodium urate (MSU) was used to stimulate the peripheral blood of NAGA and healthy controls patients. Then the expression ofAJ227913 was detected by RT-qPCR. Kruskal-Wallis test, Mann-Whitney test, Spearman correlations were used for statistical analysis. Results The expression level of AJ227913 in the AGA group (0.0557 ±0.0156) was higher than that in the NAGA group (0.0223±0.018) and healthy controls group (0.0038±0.0013). There was significant difference between the NAGA group and healthy controls group (P>0.05). Compared with the control group, the expression of AJ227913 in NAGA group which were stimulated by MSU was significantly increased. The Spearman correlation analysis found that the AJ227913 expression levels in GA groups were correlated with UREA (r=0.608, P<0.01), CREA (r=0.337, P<0.05), CYSC (r=0.422, P<0.01). Conclusion Altered expression of AJ227913 may be involved in the inflammatory process of GA and the balance of uricacid.

It has gradually become a consensus in the industry that the traditional Chinese medicine gypsum should be decocted first, but the understanding of decocting method is not completely unified in the works of doctors since ancient times, and there are occasional disputes about whether it is necessary to decocting first. In this study, the phase determination, physical and chemical characterization, qualitative and quantitative analysis of inorganic and organic components of the decoctions of herbal pairs and the whole prescription Maxingshigan decoction with gypsum as the center, and the pre-decoctions and co-decoctions of them were carried out to explore the scientific connotation of the pre-decoctions of gypsum. Results show that decoction phases were different between the co-decoctions and pre-decoctions of licorice-gypsum (Gancao-Shigao, GC-SG), ephedra-gypsum (Mahuang-Shigao, MH-SG) and almond-gypsum (Xingren-Shigao, XR-SG). The results of the micromorphology, particle size and zeta potential of herbal pairs and prescription (Quanfang, QF) showed that the supramolecular particles in pre-decoctions were smaller, more uniform and more stable than the co-decoctions. The results of organic components analysis showed that different cooking methods did not change the organic composition and content. ICP-OES results showed that the content of inorganic components in pre-decoctions was higher than in co-decoctions for the same boiling time of gypsum. The IR results showed that the pre-decoctions had stronger chemical functional group effect than the co-decoctions. To sum up, compared with the co-decoction, the pre-decoction of gypsum has different phase state and chemical composition interaction, and the difference of inorganic composition is an important material basis affecting the change of phase state compared with the co-decoction. It indicates that the material basis of traditional Chinese medicine decoction is indeed different whether gypsum is decocted first or not, which can provide a basis for the clinical application of decocted gypsum.