Objective To get phagc's capsid Vp3 gene and protein of guinea pig inclusion conjunctivitis (GPIC) chlamydia. Methods The genome DNA was extracted from the φCPG1 phage.The full sequence of Vp3 gene was amplified by polymerase chain reaction (PCR) from the above genome DNA. The Vp3 gene was digested by restriction endonuclease and then inserted into prokaryotic plasmid vector pET30a (+). The recombinant plasmid was transformed into E. coil BL21, and was identified by restriction endonuclease, PCR and sequencing. The E. coil BL21 with expected recombinant plasmid was induced and the expressed recombinant Vp3 protein was identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot, then purified by agarose gel. Results The recombinant gene was sequenced and proved to be 447 bp which was consistent with the φCPG1 Vp3 gene sequence in GenBank. A 25 000 capsid protein was expressed and confirmed by SDS-PAGE and Western blot. The purified protein was obtained. Conclusion The capsid Vp3 protein of φCPG1 is successfully expressed and purified, which is helpful for the further study on its mechanism and clinical applications.

Objective To express recombinant Vp1 protein of chlamydiaphage φCPG1, prepare monoclonal antibody against Vp1 protein and utilize it to screen clinical isolates of Chlamydia trachomatis. Methods The Vp1 protein was obtained by prokaryotic expression, and monoclonal antibody against this protein was prepared by hybridoma technique. ELISA and Western blot were used to identify monoclonal antibodies. Then,the monoclonal antibody was prepared in quantity by injecting hybridoma cells into the abdominal cavity of BALB/C mice, and purified by using protein G affinity chromatography. Clinical isolates of Chlamydia trachomatis were screened for the chlamydiaphage by immumofluorescence assay using the prepared monoclonal antibody.Results Purified Vp1 protein was obtained. The monoclonal antibody against Vp1 protein was gained after 3times of sub-clone and consistently identified as IgG1. Three hybridoma cell strains that stably secreted monoclonal antibody were generated. Chromosome analysis of hybridoma cells showed that the mean number of chromosome was 96, most of them were telocentric and a few were submetacentric. The titer of purified monoclonal antibody was more than 1: 12 800. Twenty clinical isolates were screened by using the monoclonal antibody and no positive results were obtained. Conclusions The monoclonal antibody against Vp1 protein of chlamydiaphage φCPG1 is successfully prepared, while no chlamydiaphage is detected by immumofluorescence assay using the prepared antibody in 20 clinical isolates of Ct.

Since initiation of training on laboratory of molecular biology in my school, we accumulated experience in teaching process especially among international students and application of pedagogy as well as technology. Strengths, weakness and opportunities are analyzed to improve teaching quality.

Objective To compare the infectivity of several transformed Chlamydia trachomatis (Ct) mouse pneumonitis (Mopn) strains to host cells, and to identify cell invasion-related virulence genes in Chlamydial plasmids. Methods Several Ct strains, including wild-type Ct Mopn strain(WT strain), plasmid-free Ct strain(CMUT3 strain), Ct Mopn strain transformed with the shuttle vector carrying pGFP and the complete C. muridarum (CM) plasmid (pGFP::CM strain)and Ct Mopn strains transformed with shuttle vectors carrying pGFP and mutant CM plasmids with in-frame deletions of Pgp3, 4, 5 or 7 (pGFP::CM△Pgp3, 4, 5, 7 strains), were cultured, amplified and collected. After the concentrations of Ct were determined, each of these strains was divided into four groups to be inoculated at a same amount(1.5 × 104 inclusion forming units(IFU)) followed by four different treatments respectively: centrifugalization +DEAE group treated with centrifugalization followed by ion-exchange chromatography on diethylaminoethyl(DEAE)-cellulose columns, centrifugalization group treated with centrifugalization only, DEAE group treated with chromatography on DEAE-cellulose columns only, control group receiving no treatment. After additional culture for 20 - 24 hours, indirect immunofluorescence assay was performed to count the number of chlamydial inclusions. At 20, 40 and 60 hours after infection, the growth rate and area of chlamydial plaques were assessed after three continuous passages. Lugol′s iodine staining was conducted to observe glycogen synthesis in bacterial inclusions. Results The inclusion number in the centrifugalization + DEAE group, centrifugalization group, DEAE group and control group was 10.20 ± 1.30, 6.80 ± 0.44, 3.00 ± 1.22 and 0.80 ± 0.45 respectively for the pGFP::CM△Pgp4 strain, 6.40 ± 0.89, 3.80 ± 0.83, 1.60 ± 0.89 and 0.60 ± 0.54 respectively for the CMUT3 strain. Under same experiment conditions, the pGFP::CM△Pgp4 strain and CMUT3 strain showed similar infectivity, and formed less inclusions compared with the other Ct strains (all P < 0.01). The number of inclusions formed by the same Ct strains were significantly different among the 4 groups(F = 845.310, P <0.01), and were highest in the centrifugalization + DEAE group for all the strains. The pGFP::CM△Pgp4 strain showed significantly lower growth rate and area of plaques with an abnormality in glycogen accumulation compared with the other strains at 20, 40 and 60 hours after infection. Conclusion The plasmid-encoding gene Pgp4 may be a cell invasion-associated virulence gene in chlamydial plasmids.

To observe the influence of the placental apoptosis on the expression of Bax,Bcl-2, Fas, FasLand TNF-α during the second trimester of pregnancy, mice of experimental group were intraperitoneal injected with 100 purified Toxoplasma gondii tachyzoites added in 0.2mL of PBS, while those of the control group were injected with 0.2 mL of sterile PBS (0.01 mol/L, pH 7.4) in the 8-th day of pregnancy. During the 12, 14, 16 and 18-th days of pregnancy, 5 mice both in experimental and control group were randomly killed and the expression levels of the apoptosis-related proteins Bax, Bcl-2, Fas, FasL and TNF-α in the placental tissues were determined by means of immunohistochemical methods. It was showed that the apoptosis-related protein expressed both in villus and decidua of the placenta, most of which were expressed in syneytiotrophoblast (ST). The positive cells with expression of Bax, Fas, FasL and TNF-α increased along with the increase of the pregnant days in both the experimental group and the control group, and the positive cells with expression of Bcl-2 decreased along with the increase of the pregnant days. It was also demonstrated that the positive cells with expression of Bax, Fas, FasL and TNF-α of the experimental group showed a higher percentage of expression than that of the control group on the same pregnant days, but the positive cells with Bcl-2 expression of the experimental group were fewer than that of the control group. It is concluded that the expression of apoptosis-related protein Bax, Bcl-2, Fas, FasL and TNF-α in the placenta were altered when the pregnant mice were infected with Toxoplasma gondii during the second trimester, which may induce the apoptosis through the endogenic and ectogenic pathway.

Objective To analyze the effects of five proteins secreted by Chlamydia trachomatis on the phagocytosis of macrophages and dendritic cells derived from bone marrow cells of C3H/HeJ mice. Methods Glutathione S-transferase ( GST)-CT311, GST-GIgA, GST-cHtrA, GST-OmcBc and GST-Pgp3 proteins were prepared through an Escherichia coli prokaryotic expression system and purified by GST Mag-Beads. Chlamydia membrane protein GST-IncA was also prepared as a control. Proteins of interest were ob-tained by cleaving off GST-tag with PreScission protease. Macrophages (MΦ) and dendritic cells (DC) were prepared from bone marrow cells of C3H/HeJ mice and pretreated with either 100 μg/ml or 500 μg/ml of the above proteins. LPS was used as a control to testify the specificity of the proteins' functions. Four hours after pretreatment,fluorescent beads were added to culture media to evaluate the changes in phagocytosis with direct immunofluorescence assay. Results LPS and low concentration (100 μg/ml) of these proteins had no significant influence on the phagocytosis of DC and MΦ,while high concentration (500 μg/ml) of Pgp3, cHtrA and CT311 could significantly promote the phagocytosis of DC and MΦ. Conclusion Pgp3, cHtrA and CT311 can promote the in vitro phagocytosis of DC and MΦ,which may facilitate the in vivo dissemina-tion of Chlamyida trachomatis.

Objective:To analyze the incidence of human papillomavirus (HPV) infection in 1 902 patients and to evaluate the efficacy of drug treatment in 266 patients, aiming to provide reference for the treatment of HPV infection.Methods:The subtypes of HPV isolated from 1 902 patients aged 15-86 years visiting the venereology outpatient clinic of Tianjin Medical University General Hospital from October 2019 to May 2021 were identified by polymerase chain reaction-reverse dot blot hybridization. Drug treatment efficacy in 266 patients of them was retrospectively analyzed.Results:The overall incidence of HPV infection in the 1 902 patients was as high as 53.84% (1 024/1 902). It was 52.60% (689/1 310) in males and 56.59% (335/592) in females. There was no significant difference in the incidence between males and females ( P>0.05). The most common HPV genotype in males and females was HPV6 [15.27% (200/1 310) and 21.96% (130/592)], followed by HPV16 [10.61% (139/1 310) and 9.46% (56/592)], HPV11 [9.31% (122/1 310) and 8.61% (51/592)], HPV52 [6.79% (89/1 310) and 8.95% (53/592)] and HPV43 [5.64% (87/1 310) and 8.45% (50/592)]. The majority of HPV-positive patients were aged between 20 and 39 years. There were 476 cases (25.03%, 476/1 902) of single-type infection and 548 cases (28.81%, 548/1 902) of multiple infection. The incidence of multiple infection was higher than that of single-type infection ( P<0.05). The incidence of multiple infection in females was higher than that in males ( P<0.05). Among the 266 patients, 106 were treated with Paiteling, a traditional Chinese medicine (TCM) preparation, and 68 of them tested negative (64.15%) after treatment. Fifty-eight patients were treated with recombinant human interferon α2b and 22 of them (37.93%) tested negative after treatment. Twenty out of the 56 subjects treated with imiquimod tested negative after treatment. Eight out of the 46 patients without treatment also turned negative. Conclusions:The incidence of HPV infection in the 1 902 patients visiting the venereology outpatient clinic was very high, and most of them were young adults. Multiple infection was more common than single-type infection. Topical application of drugs such as Paiteling, recombinant human interferon α2b and imiquimod was effective in treating HPV infection.

Objective To explore the effect of self-care agency on frailty syndrome in hospitalized elderly patients.Methods A total of 102 hospitalized elderly patients were recruited by convenience sampling method and investigated by cross-sectional survey. The prevalence of frailty was evaluated by frailty phenotype, and socio-demographic characteristics scale and exercise of self-care agency scale were used to evaluated these patients.Results The prevalence of frailty syndrome were 31.4%(32/102). The total score of self-care agency scale was (104.38±20.20). Univariate analysis showed that the prevalence of frailty syndrome in elderly was statistically different in the age, household income, career, the number of chronic disease, the use of medication and self-assessment of health (P<0.05). Compared with no-frailty elderly, the self-care agency, especially self-worth of frailty syndrome patients reduced significantly(P<0.05). Multiple Logistic regression indicated that self-care skills (OR=0.775, 95%CI: 0.641-0.937) and self-worth (OR=0.585,95%CI: 0.424-0.807) were significant protective factors of frailty syndrome.Conclusions The poorer self-care skills and self-worth of the elderly, the higher prevalence of frailty syndrome. Raising the level of self-care agency actively can help to prevent or improve frailty syndrome.

Objective To investigate the relationship between health literacy and cognitive function in elderly inpatients.Methods A cross-sectional survey was conducted among 216 elderly people who were admitted to a class Ⅲ grade A hospital by convenience sampling. Social demographic characteristics scale, questionnaire on Chinese citizens' health literacy and minimum mental state examination were used in this survey.Results The health literacy score of elderly inpatients was (64.33±12.79) and the cognitive function score was(26.52±2.27). The scores of health literacy and cognitive function had moderate correlation in hospitalized elderly patients (r=0.52,P<0.05). The single factor analysis showed that the health literacy of the elderly inpatients was statistical different in the education level,the average monthly family income,occupation, chronic disease and cognitive function (P<0.05). The results of multiple linear regression analysis showed that educational level,cognitive function,occupational and chronic diseases were independent factors of health literacy among the elderly inpatients.Conclusions The cognitive function of elderly inpatients varies greatly, and the higher level of cognitive function is a protective factor for the health literacy of elderly inpatients. It is an important approach to improve health literacy among elderly patients in hospital by lowering the burden of cognitive function to communicate health information.

In order to meet the urgent needs of social high-level nursing talents in the times and reality, scholars at home and abroad pay more and more attention to the development of professional spirit of nursing students. By using systematic analysis, this paper summarized five kinds of widely used professional spirit teaching methods, such as role model teaching, reflective learning teaching, PBL teaching, behavior-oriented teaching and hidden curriculum teaching so as to break the traditional teaching mode and explore scientific and effective teaching methods.