Objective To amplify the 16S RNA fragments of 7 clinically isolated strains of Brucella spp. by PCR-RFLP technique, so as to provide experimental basis for the studies on diagnostics, genetics and epidemiology of Brucella spp. Methods According to the gene sequence of ATCC 25840 standard strain in GenBank, special primers for the 16S RNA conservative area in the Brucella spp. were designed. DNA extraction and PCR amplification of the 16S RNA fragments were performed with the 7 isolated strains. PCR products were then sequenced and RFLP analysis was conducted with appropriate restricted enzymes to study the homology and the mutation sites in those strains. Meanwhile, the clinical data of infected patients were retrospectively analyzed to evaluate the relationship between the clinical features and genotypes of Brucella infection. Results The amplified target fragments were about 1500bp in length and consistent with what was expected. The sequencing and homology analysis showed a 98.88% homology and 11 mutation sites among the 7 isolated strains. Four genotypes were identified by RFLP. Retrospective analysis of the clinical data indicated that no obvious relationship existed between the genotypes and the clinical features. Conclusions Amplifying 16S RNA fragments by PCR technique is a feasible method to make an early diagnosis of Brucella infection. The 7 clinically isolated strains are different in genotypes and 16S RNA fragment is a highly conservative fragment in bacterial genome with some mutations. The research provides evidence for the genetics and epidemiology of brucellosis.

Invasion of glioma is a complex process with multiple steps and multiple factors,including the inhibition of tumor cell adhesion,the degradation of extracellular matrix,the promotion of tumor cells migration and angiogenesis.The abnormal activation of Wnt/β-catenin signaling pathway is closely related to the invasion of glioma.The study of Wnt/β-catenin pathway affecting invasion mechanism of glioma will provide new ideas and targets for the treatment of glioma.

Objective To explore the effect of psychological rehabilitation on patients with limb motor dysfunction. Methods 136 cases of hospitalized patients who needed rehabilitation therapy and training were divided into observation group and control group. The control group received conventional rehabilitation, and the observation group received psychological rehabilitation (psychotherapy and medicine) in addition. Symptom Checklist-90 (SCL-90), Fugl-Meyer Assessment (FMA) and modified Barthel Index (MBI) were assessed before and 8 weeks after treatment. Results Most of the 136 patients were involved in psychological problems, including anxiety, depression, somatization,fear, hostility. There was no difference in the scores of SCL-90, FMA and MBI between 2 groups before treatment (P>0.05). After 8 weeks treatment, the scores of SCL-90 were lower, except psychotism (P>0.05), in the observation group than in the control group (P<0.05),the scores of FMA and MBI were higher in the observation group than in the control group (P<0.01). Conclusion The psychological rehabilitation plays an important role in rehabilitation of limb motor dysfunction.

Objective To evaluate the clinical significance of the multiple cytokines in the serum of SARS patients and explore the rel ationship between the immune-reactivity and pathological damages. Methods 12 different serum cytokines have been detected in 4 groups ( inchoation, metaphase, convalescence of SARS patients and Healthy control) by using biochips technique(RANDOX) and to study the changes of each cytokine level in SARS patients. Results Compared with healthy group, IL-6 , IL-8 , IL-10 ,IFN? increased obviously and IL-1? ,IL-2,IL-4,VEGF,EGF,MCP-1, TNF?decreased obviously. Whereas IL-1?has no statistic changes among different stages of SARS. Conclusion There were obvious changes of multiple cytokines in different phase of SARS pathological process, especially in the early phase. It is further support the hypothesis that over-reaction of the immune system initiated the pathological injuries of the patients.

Objective To monitor the constituents and resistant tendency of bacterial pathogens isolated from diarrheal patients in our hospital form 1994 to 2005 to offer the basis for guiding epidemiologic study,vaccination research and clinical treatment. Methods Enteric pathogenic bacteria were cultured and identified to species,group and serotype with biochemical and serologic methods and the susceptibility of bacteria to antimicrobial agents were tested. Results Enteric pathogenic bacteria were isolated predominantly in male patients and mainly in children and youngsters. It reached a peak from July to September every year. Shigella spp.(75.11%) was the most frequendy isolated pathogens and followed by Vibrio spp.(12.7%),Salmonella spp.(6.28%),Aeromonas spp.(4.43%) and Escherichia coli(1.25%).During the period from 1994 to 2005,diarrheal pathogens had a trend of decrease especially Shigella spp.and Salmonella spp.. Of the 6329 isolates of Shigella spp., 75.62% was S. flexneri and S.soanei,S.dysenteriae and S. boydii constituted 23.98%,0.22% and 0.01% respectively.The sensitivity of different species,group or serotype to different antimicrobial agents was not the same.S.flexneri and Aeromonas spp. were highly resistant to most of antibiotics. However, S.sonnei and Vibrio spp.had good susceptibility to antibiotics tested except trimethoprim/sulfamethoxazole and ampicillin. Conclusion There are many species and serotypes of enteric pathogenic bacteria causing infective diarrhea and the distribution changes gradually in Beijing. The resistance rate of enteric pathogenic bacteria to antibiotics is not the same in different species and serotypes.so strict surveillance iS always needed.

Objective To evaluate inhibitory effects of the Chlamydiaphage phiCPG1 capsid protein Vp1 on Chlamydia psittaci strain guinea pig inclusion conjunctivitis (GPIC) and Chlamydia trachomatis (Ct) serovar E,and to provide new ideas for the treatment of Ct infection.Methods The Chlamydiaphage phiCPG1 capsid protein Vp1 was expressed in Escherichia coli BL21 transfected with the recombinant plasmid Vp1-pET30a (+),identified by Western blot analysis and purified by using dialysis bags.Bicinchonininc acid (BCA) assay was performed to determine the concentration of Vp1 protein.GPIC and Ct serovar E strains were both classified into 4 groups to be firstly incubated with Vp1 protein (Vp1 group),Tris-glycine solution (Tris group),S protein (S group) or Dulbecco's Modified Eagle Medium (DMEM,DMEM group) at room temperature for 3 hours,then were used to infect Hela cells followed by 72-hour (GPIC) or 48-hour (Ct serovar E) culture with the presence of Vp 1 protein (Vp 1 group),Tris-glycine solution (Tris group),S protein (S group) or DMEM (DMEM group).Subsequently,immunofluorescence staining was conducted to observe and count chlamydial inclusions.Results The number of GPIC inclusions was significantly different between the 4 groups after 72-hour culture (F=476.632,P＜ 0.05),and lower in the Vp1 group (5.0 ± 1.5) than in the Tris group (24 ± 1.2,P＜ 0.05),S group (25 ± 1.7,P＜ 0.05) and DMEM group (25 ± 1.5,P＜ 0.05),but insignificantly different between the latter 3 groups (P ＞ 0.05).Compared with the DMEM group,the Vp1 group showed a significant decrease of 80.2％ ± 3.99％ and 77.2％ ± 1.79％ in the number of GPIC and Ct serovar E inclusions respectively,with no significant difference in the inhibitory effect of Vp1 on GPIC versus Ct serovar E (t =2.057,P ＞ 0.05).Conclusion The phiCPG1 capsid protein Vp1 can obviously inhibit GPIC and Ct serovar E infections to a similar degree.

Objective To discuss the diagnostic value of the examination of serum NAG, AFU, PAB, LAP, ASTm, GLDH,ADA and AFP in patients suffering from liver diseases.Methods Serum of 274 hepatitis cases and 30 healthy cases are examined with auto biochemical analyzer and analyzed statistically.Results The mean values of LAP, ASTm, GLDH, ADA and AFU in acute hepatitis patients are higher than health′s significantly, AUC of AFU,LAP and ASTm are 0.842,0.816 and 0.782 separately, positive rate of AFU,LAP and ASTm are 84.2%,95% and 80% separately; The mean values of ADA、AFU and NAG in liver cirrhosis patients are higher than health′s significantly while the mean value of PAB is lower significantly, AUC of ADA is 0.689, positive rate of ADA is 89.5%; The mean values of ADA and NAG in severe hepatitis patients are higher than health′s significantly while the mean values of PAB and AFU are lower significantly, AUC of PAB and AFU all is 0.861, positive rate of PAB and AFU is 100% and 52.1%; The mean values of LAP,AFP,NAG,ADA and AFU in liver cancer patients are higher than health′s significantly while the mean value of PAB is lower significantly, AUC of LAP and AFU is 0.697 and 0.653 separately, positive rate of LAP and AFU are 74% and 79.5% separately.Conclusions AFU、LAP and ASTm are valuable markers for diagnosing of acute hepatitis, ADA is a valuable marker for diagnosing of liver cirrhosis, PAB and AFU are valuable markers for diagnosing of severe hepatitis, LAP and AFP are valuable markers for diagnosing of liver cancer.

Objective:To explore the potential facilitators and barriers to healthy behavior among stroke patients.Methods:Semi-structured interviews were conducted among 16 stroke patients from September 2022 to March 2023 using an objective sampling method.The interview guide was developed using the theoretical domain framework(TDF). Interviews were transcribed and refined the theme using directed content and induction analysis.Using the TDF as the initial coding framework, the themes were then merged into the most relevant domains.Finally, the correspondence between theoretical domains and the healthy behavior of stroke patients was determined based on the frequency and relationship between themes.Results:This study identified nine theoretical domains that affected the healthy behavior of stroke patients: knowledge, skills, motivation and goals, social influences, social/professional role and identity, environment context and resources, belief about capability, consequence belief and behavioral regulation.Conclusion:The healthy behavior of stroke patients is complex and influenced by several factors.The nine theoretical domains identified in this study will provide recommendations for future healthy behavior interventions for stroke patients.

Objective:To construct an evaluation indicator system for the health management effects in high-risk stroke population.Methods:From March to May 2020, based on health ecology theory, social cognitive theory and knowledge, attitude/belief, practice theory, the first draft of the evaluation index system for health management effects of high-risk stroke groups was drawn up by literature review and group discussion. Through two rounds of Delphi expert consultation, the evaluation index system of health management effects of high-risk stroke groups was established, and the weight of each index was determined by analytic hierarchy process (AHP). A total of 22 experts were invited to participate in expert consultation. Twenty-two questionnaires were sent out in the first round, and 20 questionnaires were recovered, of which 20 were valid (90.9%). In the second round of correspondence, 20 questionnaires were sent out and 19 questionnaires were recovered, of which 19 were valid (95.0%).Results:The authority coefficients of the two rounds of expert consultation (Cr) was 0.933 and 0.937, respectively. The Kendall coordination coefficients W of importance in the second round was significantly higher than that in the first round (0.299 vs 0.172) ( P<0.001). The mean of importance score (Mj) of each index was 4.10-5.00, coefficient of variation (CV) was 0-0.235, and full mark ratio (Kj) was 0.26-1.00. Finally, an evaluation index system of the health management effects for high-risk stroke population was constructed, which included 3 first-level indicators (individual characteristics, behavior style, environmental support), 12 second-level indicators and 58 third-level indicators. Conclusions:The evaluation index system of the health management effects for high-risk stroke population is established in this study, which provides scientific quantitative indicators and evaluation tools. The enthusiasm, authority and coordination of consultation experts are strong, which indicates that the indicator system is feasible.

Objective To construct active phages against Chlamydia trachomatis,and to evaluate its effect on Chlamydia trachomatis.Methods The M13 phage was recombined with the IN5 sequences encoding the capsid protein VP1 of chlamydiophage phiCPG1,and then the recombinant M13-IN5 phage was obtained.PCR amplification,enzyme digestion and sequencing were performed to verify whether the target fragment was inserted into the phage successfully.The viability of the phage was evaluated by plaque formation assay.Cell counting kit-8 (CCK8) assay was conducted to evaluate the effect of M13 phage and recombinant M13-IN5 phage at the titer of 1011 plaque-forming units (PFU)/ml on the proliferation of Hela cells,and Hela cells uninfected with chlamydia served as the blank control group.Western blot analysis was performed to determine the expression of the IN5 loop protein in the recombinant M13-IN5 phage,M13 phage and Escherichia coli ER2738 at exponential growth phase.Cultured standard Chlamydia trachomatis serovar E strain was treated with M13 phage and recombinant M13-IN5 phage at the titer of 1011 PFU/ml separately,and chlamydia control group without the treatment with phages was set up.After 36-hour infection,confocal microscopy was performed to detect the location of the M13 phage and the recombinant M13-IN5 phage.Moreover,iodine staining was conducted to count inclusion bodies at 36,48,60 and 72 hours separately after infection.Statistical analysis was carried out by a two-sample t-test for comparisons between two groups,one-way analysis of variance (ANOVA) for intergroup comparison,and Bonferroni test for multiple comparisons.Results The bioactive recombinant M13 phage containing the IN5 loop gene was constructed successfully,and Western blot analysis confirmed that the recombinant phage expressed IN5 loop/p Ⅲ fusion protein with a high titer of 3.05 × 1011 PFU/ml.As CCK8 assay showed,there was no significant difference in proliferation of Hela cells among the blank control group,M 13 phage group and recombinant M13-IN5 phage group (A450 values:3.63 ± 0.01,3.55 ± 0.02,3.70 ± 0.01,respectively,F =12.0,P ＞ 0.05).Confocal microscopy showed overlap between the phage fluorescence and chlamydial inclusion body fluorescence.The M13-IN5 phage group and M13 phage group both showed significantly decreased number of inclusion bodies compared with the control group (both P ＜ 0.05) at 36 and 72 hours after chlamydial infection,and the number of inclusion bodies was significantly lower in the M 13-IN5 phage group than in the M13 phage group (P ＞ 0.05).After 48,and 60 hours of chlamydial infection,the number of inclusion bodies did not differ among the M13 phage group,M13-IN5 phage group and control group (both P ＞ 0.05).Conclusions The recombinant M13-IN5 phage was bioactive and could successfully express the IN5 loop protein.In the in vitro experiments,the recombinant phage could enter into chlamydia inclusion bodies,and markedly inhibited the infection of Chlamydia trachomatis.