Objective To obtain tree shrew Vasorin(VASN)recombinant protein through prokaryotic expression and purification,prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein,and preliminarily evaluate its application value.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the full-length sequence of tree shrew VASN gene in vitro.The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid.The recombinant plasmid was subjected to double digestion with BamH Ⅰ and Sal Ⅰfor identification,and its correctness was further verified by sequencing.The recombinant plasmid with correct sequencing was transformed into BL21(DE3)competent cells,and isopropyl β-D-thiogalactoside(IPTG)was used to induce expression of VASN recombinant protein.Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and the VASN recombinant protein was purified by KCI.Purified recombinant protein was used to immunize BALB/c mice for four times,and serum antibody titer was detected by enzyme-linked immunosorbent assay(ELISA).Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells.Hypoxanthine-aminopterin-thymidine(HAT)culture medium was first used to screen hybridoma cells.ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies,and monoclonal hybridoma cell lines were obtained by limiting dilution method.VASN monoclonal antibodies were prepared in large quantities by ascites induction method,purified using rProtein G,and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting.Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed.The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence.Recombinant protein VASN mainly existed in the form of inclusion bodies,and the purity after purification reached 90％,meeting the requirements of subsequent immunization experiments.After four immunizations with recombinant protein VASN,mouse serum antibody titer reached 1:729 000.Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification,and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x107 L/mol.Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein,but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein,human VASN-leucine rich repeat recombinant protein,or bovine serum albumin.Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart,liver,spleen,lung,kidney and muscle,with clear bands and clean background.Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen,lung,and tree shrew immortalized fibroblasts with high VASN mRNA expression levels,and the detection results were positive.Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared.This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts,spleen tissue,and lung tissue,providing an important tool for further research on the function of VASN in tree shrew models.

Objective To obtain tree shrew Vasorin(VASN)recombinant protein through prokaryotic expression and purification,prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein,and preliminarily evaluate its application value.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the full-length sequence of tree shrew VASN gene in vitro.The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid.The recombinant plasmid was subjected to double digestion with BamH Ⅰ and Sal Ⅰfor identification,and its correctness was further verified by sequencing.The recombinant plasmid with correct sequencing was transformed into BL21(DE3)competent cells,and isopropyl β-D-thiogalactoside(IPTG)was used to induce expression of VASN recombinant protein.Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and the VASN recombinant protein was purified by KCI.Purified recombinant protein was used to immunize BALB/c mice for four times,and serum antibody titer was detected by enzyme-linked immunosorbent assay(ELISA).Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells.Hypoxanthine-aminopterin-thymidine(HAT)culture medium was first used to screen hybridoma cells.ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies,and monoclonal hybridoma cell lines were obtained by limiting dilution method.VASN monoclonal antibodies were prepared in large quantities by ascites induction method,purified using rProtein G,and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting.Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed.The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence.Recombinant protein VASN mainly existed in the form of inclusion bodies,and the purity after purification reached 90％,meeting the requirements of subsequent immunization experiments.After four immunizations with recombinant protein VASN,mouse serum antibody titer reached 1:729 000.Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification,and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x107 L/mol.Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein,but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein,human VASN-leucine rich repeat recombinant protein,or bovine serum albumin.Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart,liver,spleen,lung,kidney and muscle,with clear bands and clean background.Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen,lung,and tree shrew immortalized fibroblasts with high VASN mRNA expression levels,and the detection results were positive.Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared.This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts,spleen tissue,and lung tissue,providing an important tool for further research on the function of VASN in tree shrew models.

Objective To analyze the clinical characteristics of co-infection of Talaromyces marneffei(TM)and non-tuberculous Mycobacterium(NTM)in human immunodeficiency virus(HIV)-negative patients.Methods Clinical data of 8 HIV-negative patients with co-infection of TM and NTM in a hospital from 2019 to 2022 were co-llected.Clinical manifestations,auxiliary examination,treatment and prognosis were retrospectively analyzed.Results Among the 8 patients,5 were females and 3 were males,with an average age of(52.25±12.31)years old.All patients presented TM and NTM disseminated infection.The major involved organs were lung(100％),lymph nodes(87.5％),and skin(75.0％).Clinical symptoms mainly included cough and expectoration(87.5％),fatigue(62.5％),joint and lumbosacral pains(62.5％),fever(50.0％),as well as skin and soft tissue abscess(50.0％),etc.Anti-interferon-γ(INF-γ)autoantibodies were detected in 4 patients and the results were positive.All 8 patients(100％)had pulmonary lesions,with chest CT mainly showing spots,patches,and striped shadows in both lungs.Among them,7 cases(87.5％)had increased and enlarged mediastinal lymph nodes,4 cases(50.0％)had pleural thickening and pleural effusion,2 cases each(25.0％for each)were accompanied by pulmonary mass shadows,bronchial stenosis,as well as increased and enlarged hilar lymph nodes.One case each(12.5％for each)had pulmonary cavity formation,bronchiectasis,and pericardial effusion.Conclusion The co-infection of TM and NTM in non-HIV patients presents disseminated infection,with multiple clinical symptoms.Chest imaging shows a wide variety of pulmonary lesions.It is prone to miss diagnosis in clinic,and the effect is not ideal after treatment for single pathogen infection.

Objective:To analyze the characteristics of anonymous online dating and related factors of not being tested for HIV among men who have sex with men and to provide the basis for developing targeted HIV prevention strategies and intervention measures.Methods:Using a cross-sectional study design, respondents were recruited with the assistance of MSM social organizations from April to July 2023 in eight cities in Shandong Province. Snowball sampling, activity venues, and networks were employed, targeting a sample size of 400 people per city. A face-to-face questionnaire survey collected data on sociodemographic characteristics, drug use, sexual intercourse, and online informed dating. Blood samples were also collected for HIV and syphilis antibody testing.Results:In the past six months, 2 787 MSM sought homosexual partners through the Internet, and 78.26% (2 181/2 787) chose to make anonymous online dating. Before having homosexual sex with online sexual partners, 10.41% (290/2 787) of them did not ask each other about HIV infection status and 33.37% (930/2 787) of them not being tested for HIV. There were statistically significant differences in the distribution of age, marital status, household registration, education level, sexual orientation, ways to find male sexual partners, not asking online sexual partners about their HIV status before sexual intercourse, no mutual HIV testing with online sexual partners before sexual intercourse, and syphilis antibody between anonymous and non-anonymous online dating persons (all P<0.05). The results of multivariate logistic regression analysis showed that <25 years old (a OR=1.43, 95% CI: 1.15-1.77) and ≥35 years old age group (a OR=1.61, 95% CI: 1.29-2.01), people with no AIDS knowledge (a OR=2.37, 95% CI: 1.25-4.49), drug users (a OR=1.38, 95% CI: 1.17-1.63), online dating anonymous (a OR=2.19, 95% CI: 1.76-2.73), HIV antibody positive (a OR=3.61, 95% CI: 2.22-5.88) and no intervention services in the last 1 year (a OR=1.58, 95% CI: 1.05-2.38) were the related factors of MSM not being tested for HIV before having sexual intercourse through online dating. Conclusions:The phenomenon of anonymous online dating was common among MSM in Shandong Province, and the proportion of MSM not being tested for HIV before having sexual intercourse through anonymous online dating was relatively high. It is necessary to strengthen health publicity and promote informed dating in MSM.

Objective:To understand the related factors of new-type drugs use and high-risk behaviors in online-dating men who have sex with men (MSM) in Shandong Province, and provide evidence for the development of targeted HIV prevention and intervention strategies.Methods:A cross-sectional study was conducted from April to July 2023 in eight sentinel surveillance sites across cities in Shandong, recruiting MSM participants. Each city enrolled a sample of 400 individuals. Face-to-face questionnaire surveys were conducted to collect data on socio-demographic characteristics, drug use and sexual behaviors, risk perception, and online-dating practices of the MSM, and χ2 test was used to compare the differences between online-dating MSM who used new-type drugs and those who didn't use. Logistic regression model was used to analyze the related factors of group sex behavior and the lack of HIV detection before sexual behavior in the online-dating MSM. Statistical analysis was performed by using software SPSS 29.0. Results:A total of 3 235 MSM were surveyed, in whom 2 787 (86.15%) used online-dating platforms to find partners in the past six months. The majority were those who were aged 25-34 years (39.18%, 1 092/2 787), unmarried/divorced/widowed (71.51%, 1 993/2 787), had an education level of college or above (69.36%, 1 933/2 787), and primarily identified as homosexual gays (77.97%, 2 173/2 787), 37.78% (1 053/2 787) believed that they were at low risk for HIV infection and 10.88% (303/2 786) believed that using new-type drugs was less harmful and 57.59% (1 605/2 787) reported new-type drugs use. Multivariate logistic regression analysis indicated that MSM who used new-type drugs had higher risk for group sex in the past six months (a OR=1.31, 95% CI: 1.06-1.63) and sexual activity without pre-HIV testing (a OR=1.57, 95% CI:1.14-2.15) compared with those who didn't use new-type drugs.MSM who believed that the risk of using new-type drugs was general, higher and very high had higher risks for group sex in the past six months (a OR=1.54, 95% CI: 1.09-2.18) and sexual activity withou pre-HIV testing (a OR=2.02, 95% CI:1.34-3.05) compared with those who believed using new-type drugs was less harmful. Conclusions:The use of new-type drugs was relatively common in the online-dating MSM in Shandong, with widespread high-risk behaviors and poor awareness of risks to health. A gap between knowledge awareness and behavior exists. It is necessary to strengthen the health education and promotion of HIV informed dating practices in MSM.

Objective:To investigate the safety and efficacy of submucous radiofrequency ablation of the inferior turbinate, and to provide a clinical basis for the surgical treatment of allergic rhinitis in children. Methods:Patients with obstructive sleep apnea syndrome and allergic rhinitis who were admitted to the Department of Otolaryngology Head and Neck Surgery of Shanghai Children's Hospital from January 2021 to December 2023 and underwent bilateral submucous radiofrequency ablation of the inferior turbinate and radiofrequency ablation of the adenoid tonsil were included in the study. Observational and statistical indexes were used to evaluate the curative effect. Results:A total of 51 cases were included in this study, and 43 cases were followed up for half a year. Submucous radiofrequency ablation of the inferior turbinate plus radiofrequency ablation of the adenoid tonsil achieved a good effect （total effective rate 93%）, and there was a statistically significant difference in the preoperative and postoperative symptoms of the children（P<0.05）. There were no complications such as bleeding, Eustachian tube injury, nasal adhesion, or nasal dryness. Conclusion:Under the premise of strict control of surgical indications, children with allergic rhinitis can be treated surgically. Inferior turbinate submucous radiofrequency ablation is more minimally invasive, effective, and safe, and can be used in clinical practice.
Humans ; Turbinates/surgery* ; Rhinitis, Allergic/surgery* ; Radiofrequency Ablation/methods* ; Child ; Male ; Female ; Sleep Apnea, Obstructive/surgery* ; Treatment Outcome ; Catheter Ablation ; Adolescent

Objective:To analyze the characteristics of anonymous online dating and related factors of not being tested for HIV among men who have sex with men and to provide the basis for developing targeted HIV prevention strategies and intervention measures.Methods:Using a cross-sectional study design, respondents were recruited with the assistance of MSM social organizations from April to July 2023 in eight cities in Shandong Province. Snowball sampling, activity venues, and networks were employed, targeting a sample size of 400 people per city. A face-to-face questionnaire survey collected data on sociodemographic characteristics, drug use, sexual intercourse, and online informed dating. Blood samples were also collected for HIV and syphilis antibody testing.Results:In the past six months, 2 787 MSM sought homosexual partners through the Internet, and 78.26% (2 181/2 787) chose to make anonymous online dating. Before having homosexual sex with online sexual partners, 10.41% (290/2 787) of them did not ask each other about HIV infection status and 33.37% (930/2 787) of them not being tested for HIV. There were statistically significant differences in the distribution of age, marital status, household registration, education level, sexual orientation, ways to find male sexual partners, not asking online sexual partners about their HIV status before sexual intercourse, no mutual HIV testing with online sexual partners before sexual intercourse, and syphilis antibody between anonymous and non-anonymous online dating persons (all P<0.05). The results of multivariate logistic regression analysis showed that <25 years old (a OR=1.43, 95% CI: 1.15-1.77) and ≥35 years old age group (a OR=1.61, 95% CI: 1.29-2.01), people with no AIDS knowledge (a OR=2.37, 95% CI: 1.25-4.49), drug users (a OR=1.38, 95% CI: 1.17-1.63), online dating anonymous (a OR=2.19, 95% CI: 1.76-2.73), HIV antibody positive (a OR=3.61, 95% CI: 2.22-5.88) and no intervention services in the last 1 year (a OR=1.58, 95% CI: 1.05-2.38) were the related factors of MSM not being tested for HIV before having sexual intercourse through online dating. Conclusions:The phenomenon of anonymous online dating was common among MSM in Shandong Province, and the proportion of MSM not being tested for HIV before having sexual intercourse through anonymous online dating was relatively high. It is necessary to strengthen health publicity and promote informed dating in MSM.

Objective:To understand the related factors of new-type drugs use and high-risk behaviors in online-dating men who have sex with men (MSM) in Shandong Province, and provide evidence for the development of targeted HIV prevention and intervention strategies.Methods:A cross-sectional study was conducted from April to July 2023 in eight sentinel surveillance sites across cities in Shandong, recruiting MSM participants. Each city enrolled a sample of 400 individuals. Face-to-face questionnaire surveys were conducted to collect data on socio-demographic characteristics, drug use and sexual behaviors, risk perception, and online-dating practices of the MSM, and χ2 test was used to compare the differences between online-dating MSM who used new-type drugs and those who didn't use. Logistic regression model was used to analyze the related factors of group sex behavior and the lack of HIV detection before sexual behavior in the online-dating MSM. Statistical analysis was performed by using software SPSS 29.0. Results:A total of 3 235 MSM were surveyed, in whom 2 787 (86.15%) used online-dating platforms to find partners in the past six months. The majority were those who were aged 25-34 years (39.18%, 1 092/2 787), unmarried/divorced/widowed (71.51%, 1 993/2 787), had an education level of college or above (69.36%, 1 933/2 787), and primarily identified as homosexual gays (77.97%, 2 173/2 787), 37.78% (1 053/2 787) believed that they were at low risk for HIV infection and 10.88% (303/2 786) believed that using new-type drugs was less harmful and 57.59% (1 605/2 787) reported new-type drugs use. Multivariate logistic regression analysis indicated that MSM who used new-type drugs had higher risk for group sex in the past six months (a OR=1.31, 95% CI: 1.06-1.63) and sexual activity without pre-HIV testing (a OR=1.57, 95% CI:1.14-2.15) compared with those who didn't use new-type drugs.MSM who believed that the risk of using new-type drugs was general, higher and very high had higher risks for group sex in the past six months (a OR=1.54, 95% CI: 1.09-2.18) and sexual activity withou pre-HIV testing (a OR=2.02, 95% CI:1.34-3.05) compared with those who believed using new-type drugs was less harmful. Conclusions:The use of new-type drugs was relatively common in the online-dating MSM in Shandong, with widespread high-risk behaviors and poor awareness of risks to health. A gap between knowledge awareness and behavior exists. It is necessary to strengthen the health education and promotion of HIV informed dating practices in MSM.

Objective To translate the Urinary Incontinence Awareness and Attitude Scale(URINAS)and test its reliability and validity.Methods The Chinese version of the URINAS was developed by Brislin's translation model for translation,back translation,cultural adaptation,and pilot investigation.A convenience sampling method was used to select 384 urinary incontinence patients who visited a tertiary hospital in Nanchang,Jiangxi Province from June 2024 to October 2024 for investigation,in order to evaluate the reliability and validity of the scale.Results The Chinese version of the URINAS consisted of 5 dimensions with 26 entries.The Cronbach's alpha coefficient of this scale was 0.843;the folded half reliability was 0.917;the retest reliability was 0.852.The content validity of the scale at the level of the entries ranged from 0.846 to 1.000,and that at the level of the scale was 0.979.A total of 5 metrics were extracted by exploratory factor analysis,and the cumulative variance contribution rate of 74.286％.The results of the validation factor analysis showed a chi-square/degree of freedom of 2.268,a root mean square error of approximation of 0.064,a standardized fit index of 0.916,a Tucker-Lewis index of 0.923,a comparative fit index of 0.906,and a goodness-of-fit index of 0.922.Conclusion The URINAS has good reliability and validity,and can better reflect the level of patients' cognition and attitude towards urinary incontinence,thus providing theoretical basis for the development of corresponding intervention programs for such patients.

Objective To analyze the clinical characteristics of co-infection of Talaromyces marneffei(TM)and non-tuberculous Mycobacterium(NTM)in human immunodeficiency virus(HIV)-negative patients.Methods Clinical data of 8 HIV-negative patients with co-infection of TM and NTM in a hospital from 2019 to 2022 were co-llected.Clinical manifestations,auxiliary examination,treatment and prognosis were retrospectively analyzed.Results Among the 8 patients,5 were females and 3 were males,with an average age of(52.25±12.31)years old.All patients presented TM and NTM disseminated infection.The major involved organs were lung(100％),lymph nodes(87.5％),and skin(75.0％).Clinical symptoms mainly included cough and expectoration(87.5％),fatigue(62.5％),joint and lumbosacral pains(62.5％),fever(50.0％),as well as skin and soft tissue abscess(50.0％),etc.Anti-interferon-γ(INF-γ)autoantibodies were detected in 4 patients and the results were positive.All 8 patients(100％)had pulmonary lesions,with chest CT mainly showing spots,patches,and striped shadows in both lungs.Among them,7 cases(87.5％)had increased and enlarged mediastinal lymph nodes,4 cases(50.0％)had pleural thickening and pleural effusion,2 cases each(25.0％for each)were accompanied by pulmonary mass shadows,bronchial stenosis,as well as increased and enlarged hilar lymph nodes.One case each(12.5％for each)had pulmonary cavity formation,bronchiectasis,and pericardial effusion.Conclusion The co-infection of TM and NTM in non-HIV patients presents disseminated infection,with multiple clinical symptoms.Chest imaging shows a wide variety of pulmonary lesions.It is prone to miss diagnosis in clinic,and the effect is not ideal after treatment for single pathogen infection.