Objective To compare and analyze the differences and characteristics of three strain mouse models in-fected by influenza virus aerosol inhalation, and provide the reference for choosing the appropriate infection model in the re-search of pathogenesis of influenza and the development of vaccines and drugs.Method C57BL/6, BALB/c and ICR mice were infected with A/Puerto Rico/8/34 (H1N1) virus strain by aerosol inhalation.The symptoms and body weight of mice were observed every day.At 3, 7, 14 days after infection, the mice were sacrificed.The lungs of mice were weighed, then virus assay and pathological observation were carried out.Results The three strains of mice were infected.The sur-vival rate in the C57BL/6 mice was lower than those in the BALB/c and ICR mice.The lung index and viral load of C57BL/6 mice were significantly higher than those of ICR mice ( P<0.05) at 3 days after infection.The pathological changes of C57BL/6 mice were also more obvious than other two strains.Compared with other two mouse strains, the weight recovery of BALB/c mice was the slowest.The survival rate in BALB/c mice was higher than that of C57BL/6 mice and lower than that of ICR mice.The lung index and viral load were not significantly different among the three strains of in-fected mice.The pathological changes among the three strains of infected mice were similar, but the degrees of pathological changes in the BALB/c mice were milder than in the C57BL/6 mice and worse than in the ICR mice.Compared with other two mouse strains, the process of disease is similar, but the body weight, mortality, lung index, viral load, and the micro-scopic pathological changes were lighter in the ICR mice than in the other two strain mice.Conclusions The three strain mouse models can be established by influenza virus aerosol inhalation, but showing different characteristics.Appropriate strain mice can be chosen to build model according to different research purpose in the experiment.

Objective To develop mycobacterial inducible expression vectors which permit to overexpress Mycobacterium tuberculosis (Mtb) immunodominant antigen, and to analyze its immunogenicity after purification by affinity chromatography. Methods The regulatory region of M. smegmatis (Ms) acet-amidase(pACE) was obtained by PCR amplification, and was used as promoter to construct the mycobacteri-al inducible expression vectors, pMF series. The coding gene of Mtb chimeric antigen Ag856A2 which is a recombinant Ag85A with 2 copies of ESAT-6 inserted in its Acc Ⅰ site and showed excellent immunogenicity in the animal experiments we described previously, was cloned into the pMF vector series, and was induced to express by the addition of acetamide. The recombinant protein expressed in the Ms was purified by the Ni~(2+)-NTA affinity chromatography, the resulted homologous recombinant antigen was added into the spleen cells separated from BCG vaccinated mice, and the immunogenicity was analyzed by the IFN-γ ELISPOT as-say. Results The mycobacterial inducible expression vectors, pMF series was constructed successfully, target antigen could be. induced to express in the Ms by the addition of 0.02% acetamide, and could be puri-fied by the Ni~(2+) -NTA affinity chromatography due to the addition of 6×His tag in the vector pMF406. Fur-thermore, the mycobactefial homologous antigen could induce more IFN-γ secretion than the heterogonous one. Conclusion The mycobacterial inducible expression system based on the regulatory region of Ms acet-amidase as promoter could permit the Mtb target antigen of interest overexpression and purification, and the immunogenicity of the homologous antigen from Ms is better than that of be expressed from E. coli, which may be more potential for immunological detection of tuberculosis.

Objective To evaluate the value of enzyme-linked immunospot assay (TB ELISPOT) combined with serum latex agglutination test (LA) for diagnosis of pulmonary tuberculosis plus pulmonary cryptococcosis.Methods Serum and biopsy specimens of 76 patients, who were suspected of pulmonary tuberculosis and/or pulmonary cryptococcosis based on clinical and imaging features, were collected from March 2006 to September 2008 in Shanghai Public Health Clinical Center. TB ELISPOT assay, LA and histopathological examination were performed in all the patients. Results Histopathological and pathogenic examination confirmed pulmonary cryptococcosis in 15 cases and pulmonary tuberculosis in 22 cases, pulmonary tuberculosis plus pulmonary cryptococcosis in 8 cases. The sensitivity and specificity of TB ELISPOT were 91% and 94.4%. The sensitivity and specificity of LA were both 100%. TB ELISPOT assay and LA test were both positive in the 8 cases of pulmonary tuberculosis plus pulmonary cryptococcosis.Conclusions The value of enzyme-linked immunospot assay combined with serum latex agglutination test is high for diagnosis of pulmonary tuberculosis plus pulmonary cryptococcosis.

Objective This study explored the effect of the multidisciplinary-team collaborative nursing model in physical examinations of people with critical conditions (based on test results).Methods We selected 962 patients with critical conditions based on test-result values found from February to April 2018 as the general process group treated through the routine nursing model,and we also selected 1009 patients with critical conditions based on test-result values found from May to July 2018 as the multidisciplinary collaborating group using a team nursing model.The multidisciplinary collaborative nursing team members included health management center nurses,outpatient nurses,resident nurses,and ward nurses.We compared visiting rates,hospitalization rates,average visiting times,and overall satisfaction after the patients received notification of their abnormal results.There were 488 male patients (50.7％) and 474 female patients (49.3％) in the general process group,with an average age of 51.9 ± 14.9 years;there were 537 male patients (53.2％) and 472 female patients (46.8％) in the multidisciplinary collaboration group,with an average age of 51.0 ± 13.0.Results For the multidisciplinary collaboration group and the general process group,respectively,the visiting rate was 53.0％ and 44.7％ (x2=13.65);the hospitalization rate was 26.7％ and 20.9％ (x2=4.38);overall satisfaction was 97.9％ and 95.9％ (x2=6.49);and the average visiting time was 4 days and 6 days (Z=5.04).The differences were statistically significant (P＜0.05).By category,the visiting rate for radiology and ultrasound patients among the multidisciplinary collaboration group was significantly higher than in the general process group (64.4％ vs.50.8％ for radiology,45.9％ vs.37.3％ for ultrasound,x2=7.65,7.11,P＜0.05).The hospitalization rate for ultrasound patients in the multidisciplinary collaboration group was significantly higher than in the general process group (12.5％ vs.6.4％,x2=10.17,P＜0.05).The average visiting time of ultrasound,abnormal blood pressure,and laboratory testing patients was significantly lower in the general process group (4 days vs.6 days,4 d vs.7 days,4 days vs.5 days,Z=3.37,1.97,2.62,P＜0.05).The overall satisfaction of radiology patients was significantly higher than in the general process group (98.6％ vs.94.3％,x2=5.39,P＜0.05).Conclusion The multidisciplinary team collaborative nursing model improves the rate of visiting and hospitalization of patients with critical conditions after physical examination,shortening their stays,helping patients get timely diagnoses and treatment,and improving patient satisfaction,making the model worth popularizing and applying more broadly.

Objective: To analyze the drug resistance of clinical isolates of Candida tropicalis in patients with infectious diseases, and preliminarily study their molecular characteristics. Methods: 95 strains of Candida tropicalis were isolated from the fungal culture specimens of 87 patients with infectious diseases in Shanghai Public Health Clinical Center from 2012 to 2015. Meanwhile, basic clinical data of patients were collected. The drug resistance of the strains to fungal drugs was analyzed by ATB FUNGUS 3 drug sensitivity test strips. All strains were classified by Multilocus sequence typing(MLST). Then, homology analysis was conducted by MEGA 5.2 software, and the evolutionary tree was mapped by using UPGMA method. Results: Patients distribution of strains was rendered as following: 31 strains from TB patients, 21 strains from HIV/AIDS patients, 19 strains from patients with liver disease, and 24 strains from rare cause infection or fever patients. The drug resistance rate to five antifungal drugs commonly used in clinical (amphotericin B, 5-fluorine cytosine, fluconazole, itraconazole, voriconazole) were 2.11% (2 strains), 0, 26.32% (25 strains), 26.32% (25 strains), and 26.32% (25 strains) respectively. Among the 25 azole-resistant strains: 14 strains were from rare cause infection or fever patients, 8 strains were from HIV/AIDS patients, and 3 strains were from tuberculosis patients. In MLST, 72 sequence types (ST types) were produced, 70 of which were new types. Evolutionary tree analysis showed that 95 strains of clinical strains distribute as three large clusters. 24 azole resistant strains (96.0%) were located in CLUSER Ⅰ. Conclusion The isolated Candida tropicalis were mainly resistant to azole drugs. MLST typing indicates that they was closely related to their genetic background.

Objective:To understand the genotyping and molecular epidemiological characteristics of Norovirus (NoV) GⅡ in adult acute gastroenteritis outpatients in Shanghai from 2015 to 2016.Methods:A total of 912 stool specimens from adult patients with acute diarrhea from March 2015 to December 2016 (431 in 2015 and 481 in 2016) were collected. The Norovirus GⅡ type was detected by one-step quantitative reverse transcription PCR assay and the RdRp region of open reading frame (ORF) 1 and 5′end of ORF2 were amplified. The region was sequenced and classified.Results:From March 2015 to December 2016, NoV GⅡ were detected in 17.76% (162/912) of the samples, 15.08% (65 /431) in 2015 and 20.17% (97/481) in 2016. Based on sequence analysis of the RdRp and capsid sequences, 145 identified NoV strains were divided into 10 genotypes: GⅡ.Pe_GⅡ.4 Sydney 2012 (60), GⅡ.P17_GⅡ.17 (45), GⅡ.P16_GⅡ.13 (10), GⅡ.P12_GⅡ.3 (10), GⅡ.P7_GⅡ.6 (9), GⅡ.P21_GⅡ.21 (5), GⅡ.P16_GⅡ.4 Sydney 2012 (2), GⅡ.Pe_GⅡ.17 (2), GⅡ.P2_GⅡ.2 (1), and GⅡ.P7_GⅡ.14 (1).Conclusions:The main epidemic NoV GⅡ genotype in Shanghai was GⅡ.P17_GⅡ.17 in the spring of 2015. GⅡ.Pe_GⅡ.4 Sydney 2012 and GⅡ.P17_GⅡ.17 were identified as predominant genotypes during the winter of 2015 and spring of 2016. The most common genotype was GⅡ.Pe_GⅡ.4 Sydney 2012 in autumn of 2016. Continuous NoV outbreak surveillance is important for identifying changing trends in genotype distribution and emerging new strains.

Objective:To explore the therapeutic mechanism of electroacupuncture(EA)in treating Parkinson disease(PD)using Tandem mass tag(TMT)quantitative proteomics technology. Methods:Forty-eight PD patients were randomly divided into a control group and an observation group,with 24 patients in each group.The control group received routine drug treatment,while the observation group received EA in addition to the routine drug treatment.EA was administered for 30 min per session,3 times a week,for a total of 12 weeks.Nine patients from each group were randomly selected to provide peripheral blood serum samples before and after treatment for TMT quantitative proteomics analysis.Differentially expressed proteins between the two groups were compared,and bioinformatics analysis was performed.The screened differentially expressed proteins were validated using enzyme-linked immunosorbent assay(ELISA). Results:In the observation group,scores on the unified Parkinson disease rating scale(UPDRS),UPDRS Ⅱ,and UPDRS Ⅲ were significantly reduced after treatment(P<0.05).In the control group,these scores tended to increase,but the changes were not statistically significant(P>0.05).After treatment,the UPDRS and UPDRS Ⅲ scores in the observation group were significantly lower than those in the control group(P<0.05).The observation group showed 62 differentially expressed proteins,while the control group had 36.Compared to the control group,the observation group had 142 differentially expressed proteins.These proteins were primarily involved in the cyclic adenosine monophosphate(cAMP)signaling pathway,T helper(Th)1 and Th2 cell differentiation,ATP-binding cassette transporter,vascular endothelial growth factor signaling pathway,and high-affinity immunoglobulin E receptor(FcεRI)signaling pathway.ELISA verification indicated that after EA treatment,the levels of α-Synuclein(αSyn)and heat shock protein beta 1(HSPB1)in the observation group were significantly lower than those in the control group(P<0.05),while the regulator of G-protein signaling 10(RGS10)level was significantly higher(P<0.05). Conclusion:EA,combined with routine drug therapy,can significantly improve clinical symptoms of PD,potentially through the regulation of the cAMP signaling pathway and the contents of differentially expressed proteins of αSyn,HSPB1,and RGS10.

OBJECTIVETo determine the function of twin-arginine translocation system (Tat) and gene cluster in Vibrio strains and to analyze the homology of tat gene cluster among different Vibrio spp. strains based on N16961 and tatABC mutant strains N169-dtat.

METHODSDifferent serotypes of biotype strains of Vibrio spp. were selected to detect the transcription of 4 genes of Tat transport system and upstream ubi aarF gene and downstream cyt551 gene by the total RNA reverse transcription and homologicity of the gene cluster by sequencing analysis.

RESULTSOur results showed that the 4 genes of tat cluster (tatA, tatB, tatC, and tatE) were intragenic and co-transcribed. We found that ubi aarF gene could be co-transcribed with tatA, tatB, but not with tatC. The electron transport chain and energy metabolism-related genes, cytochrome C551 peroxidase gene, and 4 genes located at upstream of tatABC operon were not transcribed with tatABC. Although the co-transcription between ubi aarF and tatAB was blocked in N169-dtat strain, they were still transcribed separately. Homologous analysis of genes of tat cluster in different types of Vibrio cholerae showed that tat gene cluster was a very conservative.

CONCLUSIONThe ubi and aarF gene might be co-transcribed with genes of tat cluster in Vibrio cholerae, which and the close relationship showed that they might play a key function in Vibrio cholerae.

Objective To investigate the species distribution characteristics of nontuberculous mycobacteria(NTM)in acquired immunodeficiency syndrome(AIDS)patients in Shanghai,and to provide evidences for clinical treatment.Methods A total of 775 Mycobacteria strains were isolated from patients(including 129 isolates from AIDS patients and 646 isolates from HIV-negative patients)admitted to Shanghai Public Health Clinical Center during 2015.All the species were identified by the sequence analysis of 16S rRNA gene and hsp65 gene.Differences in the species distribution were compared between patients with and without HIV infection.CD4+T lymphocyte count was detected by flow cytometry and its relation with mycobacteria infection was also analyzed.Chi-square test was used for comparison between groups.Results The ratio of NTM isolation from HIV-negative patients was 15.79%(102/646),while that was 46.51% in AIDS patients(60/129),and the difference was statistically significant(x2=61.38,P <0.01).Among the 60 NTM strains isolated from AIDS patients,mycobacterium avium-intracelluare complex(MAC)and Mycobacterium gordonae were the predominant species(43.33% and 20.00%,respectively).Moreover,five Mycobacterium colombiense strains,which were relatively rare,were also obtained.A total of 102 NTM were identified from HIV-negative patients,of which Mycobacterium gordonae(32 isolates,31.37%)and MAC(30 isolates,29.41%)were the most frequently isolated.In addition,the positive rates of NTM and MAC were significantly higher in patients with CD4+T lymphocyte counts ≤50 cells/μL(58.33% and 76.92%,respectively)than those with CD4+T lymphocyte counts>50 cells/μL(20.00% and 15.38%,respectively)(x2=4.048 and 6.524,respectively,both P<0.05).Four out of five Mycobacterium colombiense infected patients died of disseminated infections,except the remaining one whose CD4+T lymphocyte counts>50 cells/μL.Conclusions The prevalence of NTM isolation is significant higher in AIDS patients than HIV-negative patients in Shanghai,and the most prevalent NTM species is MAC.The NTM infection in AIDS patients is related with low CD4+T lymphocyte counts.

Objective:To investigate the impact of residual cholesterol (RC) on the progression of arteriosclerosis in individuals receiving physical examination.Methods:A cross-sectional study. Participants aged 18 years and above who underwent arteriosclerosis testing at the Health Management Center of Zigong Fourth People′s Hospital from January to December 2023 were selected as the subjects of the study. A total of 9 016 subjects were included in this study, of whom 6 213 were considered to have arteriosclerosis. The physical examination conclusions, basic information (age, gender, body mass index, waist circumference, blood pressure, history of hypertension, history of smoking and drinking), and biochemical indicators (lipids, fasting blood glucose, liver and kidney function) were extracted in those individuals. Based on the examination results, the subjects were categorized into arteriosclerosis and normal groups. The RC levels of the participants were calculated using a formula and then the subjects were categorized into binary and quartile RC groups. Additionally, four regression models were used to analyze the impact of RC levels on the progression of arteriosclerosis while adjusting for various confounding factors.Results:The RC level was (0.63±0.44) mmol/L in the normal group and (0.76±0.61) mmol/L in the arteriosclerosis group. Based on level of RC, the normal group was divided into two subgroups: 652 individuals with elevated RC level and 2 241 with normal RC level. In the arteriosclerosis group, there were 2 069 individuals with elevated RC level and 4 144 with normal RC level. Grouped according to quartiles of RC level, the number of individuals with RC in the Q1-Q4 interval in the normal group was 838 (28.97%), 752 (25.99%), 760 (26.27%), and 543 (18.77%), respectively, showing a gradual decreasing trend. The number of individuals with RC in the Q1-Q4 interval in the arteriosclerosis group was 1 414 (22.76%), 1 438 (23.15%), 1 589 (25.58%), and 1 771 (28.51%), respectively, showing a gradual increasing trend. The difference between the groups was statistically significant ( P<0.05). After adjusting for various factors by four regression models, it was found that elevated RC levels increased the risk of arteriosclerosis progression, with a odds ratio ( OR) of 1.381, 1.242, 1.233, and 1.214, respectively. Additionally, individuals in the Q4 RC level quartile had 1.502, 1.318, 1.311, and 1.284-times higher risk of arteriosclerosis progression when compared to those in the Q1 quartile. Conclusion:The impact of RC on the progression of arteriosclerosis tend to stabilize and remain consistent, indicating that elevated RC is an independent risk factor for the progression of arteriosclerosis.