Objective:To investigate the feasibility of combination of en-bloc resection of bladder tumor (ERBT) with the NBI(narrow band imaging) flexible cystoscopy, immunotherapy and chemotherapy in bladder-preserving treatments(called as TMT) for patients with stage T 2 bladder carcinoma. Methods:We retrospectively reviewed and analyzed a series of 16 patients with pT 2N 0M 0 pathologically confirmed. All patients are male with a median age of 63yr(56, 73yr). The American Association of Anesthesiologists scored ≤Ⅱ in 12 cases and Ⅲ in 4 cases. There were 9 cases with smoking history, 5 cases with hypertension, 3 cases with diabetes, and 2 cases with heart disease. The results of preoperative tissue biopsy were all urothelial carcinoma. All patients were taken lithotomy position and performed ERBT with NBI imaging technique and thulium-laser energy platform under general anesthesia. The tumor was brown and the surrounding normal mucosa was cyan in color. The procedure was ensured with a minimal tumor margin of 1cm and minimal rection depth to the deep musculi, and with the acquirement for the en-bloc specimen of which the basal site was marked afterwards, the patients all took a full length of chemoimmunotherapy (four cycles of Tislelizumab combined with Gemcitabine and cisplatin regimens) followed by a secondary ERBT. The perioperative data from sequential operations including complications were comprehensively analyzed for evaluating the therapeutic outcome and safety. All patients received a follow-up to detect efficacy and safety of the treatment with the primary end point of pCR, downgrading rate and objective response rate. Results:All operations were successfully completed. There was no obturator reflex, severe bleeding or grade Ⅱ bladder perforation, only 4 patients got a grade Ⅰ bladder perforation. The postoperative 30-day complication occured in 2 cases(1 pt with hematuresis, 1 pt with bladder irritation). The pathologic complete response rate and tumor downstaging rate were 100%. One patient recurred during a median follow-up of 13.0 months (6, 36 months).Conclusions:The En Bloc Resection of Bladder Tumor with the narrow band imaging (NBI) flexible cystoscopy has several remarkable advantages, including a better intra-operative visual, a lower complication rate and tumor residual. The combination of ERBT with immunotherapy and chemotherapy lead to affirmative curative effect and the feasibility for clinical application is relatively high.

Objective:To investigate the prevalence of lymphocytic choriomeningitis virus (LCMV) and hantavirus (HV) specific antibodies among cross-border migrant workers for assessment of the risk of rodents-borne virus infection.Methods:From 2019 to 2020, a survey was conducted on cross border migrant workers engaged in outdoor activities, and serum samples were collected, LCMV specific IgG antibody was detected by an indirect ELISA and Western blot based on recombinant nucleoprotein, and indirect immunofluorescence assay (IFA) based on recombinant expressed glycoprotein. HV IgG antibody in serum was detected by a commercial indirect IgG ELISA kit and IFA based on hantavirus infected Vero cells.Results:A total of 139 cross-border workers, aged 25~57, were surveyed; 64% (89/139) had working experience in multiple countries, involving 26 countries, including 14 countries in Asia and 12 countries in Africa; 11.51% (16/139) of serum samples were tested positive for LCMV antibodies, and the positive samples were verified by Western blot and IFA. The antibody detection rate was slightly higher than the published infection rate from other similar studies. And, HV antibodies were detected from one serum sample (0.72%, 1/139) by ELISA and IFA. However, it was still uncertain when and where the viral infections were acquired.Conclusions:Through this serological cross-sectional preliminary analysis, the infection status and existing risks of LCMV and HV viruses among cross border migrant workers were revealed, which suggested the necessity of strengthening the prevention and control of rodents borne diseases in outdoor engineering sites.

Objective:To establish a simple, rapid and low-cost 2019 novel coronavirus (2019-nCoV) neutralizing antibody detection method.Methods:The 2019-nCoV RBD specific immunoglobulin G (RBD-IgG) detection method was established based on the principle of quantum dot immunochromatography(QDs), and the detection was evaluated by using of sera from coronavirus disease 2019 (COVID-19) convalescent patients ( N = 97), vaccinated donors ( N = 82) and healthy donors ( N = 299). The suitability of fingertip blood was evaluated by matching blood samples with peripheral blood ( N=54). Results:The 2019-nCoV RBD-IgG detection method based on QDs was successfully established. The detection result of QDSs had strong correlation ( Spearman r > 0.73, P < 0.000 1) and good consistency ( Kappa=0.93, P < 0.01) with the result of micro-neutralization test(MNT). The sensitivity and specificity were 92% and 99%, respectively. There was high correlation ( Spearman r =0.932 6, P < 0.000 1) and no significant difference ( P=0.102 6) between result of fingertip blood and peripheral blood. Fingertip blood can be used as a surrogate sample for testing. Conclusions:The 2019-nCoV neutralizing antibody detection method established in this study can provide an immediate, efficient and low-cost method selection for the assessment of herd immunity status, and provide technical support for the herd immunity monitoring of 2019-nCoV vaccinated population and the prevention and control of the epidemic.

Objective:To establish a quick on-site emergency detection method for severe fever with thrombocytopenia syndrome virus (SFTSV), dengue virus (DENV), and hantaan virus (HTNV).Methods:This research was based on the traditional TaqMan fluorescent probe technology, using the domestic rapid one-step quantitative RT-PCR kit, combined with the Magnetic induction cycler (Mic) qPCR instrument. The detection limit, specificity and repeatability of this method were evaluated by simulated samples, other virus infected samples and normal human blood samples.Results:Compared with the traditional RT-PCR assay, the required time of this method was greatly shortened, and the detection can be completed within 35 minutes. The limit of quantitation for SFTSV, DENV and HTNV are less than 100copies/PCR. No nonspecific amplification was found in the simulated negative samples and other virus infected samples. All the simulated positive sample for verification could be detected, and coefficient of variation Ct value of each group was less than 4%. Conclusions:The rapid fluorescence quantitative RT-PCR assays have certain application prospects for on-site emergency detection, and provide important technical supports and new directions for the prevention and control of common hemorrhagic fever viruses.

Objective:To evaluate deep learning in improving the diagnostic rate of adenomatous and non-adenomatous polyps.Methods:Non-magnifying narrow band imaging (NBI) polyp images obtained from Endoscopy Center of Renmin Hospital, Wuhan University were divided into three datasets. Dataset 1 (2 699 adenomatous and 1 846 non-adenomatous non-magnifying NBI polyp images from January 2018 to October 2020) was used for model training and validation of the diagnosis system. Dataset 2 (288 adenomatous and 210 non-adenomatous non-magnifying NBI polyp images from January 2018 to October 2020) was used to compare the accuracy of polyp classification between the system and endoscopists. At the same time, the accuracy of 4 trainees in polyp classification with and without the assistance of this system was compared. Dataset 3 (203 adenomatous and 141 non-adenomatous non-magnifying NBI polyp images from November 2020 to January 2021) was used to prospectively test the system.Results:The accuracy of the system in polyp classification was 90.16% (449/498) in dataset 2, superior to that of endoscopists. With the assistance of the system, the accuracy of colorectal polyp diagnosis was significantly improved. In the prospective study, the accuracy of the system was 89.53% (308/344).Conclusion:The colorectal polyp classification system based on deep learning can significantly improve the accuracy of trainees in polyp classification.

Objective: To establish a method for the simultaneous identification of Zika, Chikungunya and Mayaro viruses. Methods: The complete genome sequences of Zika, Chikungunya and Mayaro virus were retrieved from Global Shared Database for comparative analysis, estimate its conservative region and determine the target gene location, specific primers and probes were designed, then a triplex real-time RT-PCR assay was developed. The specificity, sensitivity and repeatability of the assay were assessed by viral nucleic acid of Zika virus, Chikungunya virus a, in vitro transcriptional RNA of Mayaro virus, normal human serum and related virus simulation sample. Results: The result showed that the established method could detect Zika virus, Chikungunya virus, as well as simulated Mayaro virus samples, the limit of detection (LOD) of Zika and Chikungunya virus was 16.22 Copy/PCR and 12.02 Copy/PCR, respectively, the LOD for simulated Mayaro virus RNA was 2.82 Copy/PCR, no significant difference was detected between the triplex and monoplex assays. No cross reaction was found in the detection of dengue virus, Hantavirus, severe fever with thrombocytopenia syndrome (SFTS) virus, yellow fever virus and influenza virus, and 100 healthy adults blood samples, the specificity of the method was 100%. The repeatability result showed that the standard deviation of all three detections were blow 0.5 and the coefficient of variation was less than 2% by selecting viral nucleic acids or transcribed RNA with high, medium and low concentration gradients. Conclusions A triplex real-time RT-PCR assay for detection of Zika, Chikungunya and Mayaro virus has been established with an acceptable specificity, sensitivity and repeatability.

Objective: To establish a fluorescent bead-based multiplex assay for the simultaneous detection of seven viral diseases endemic in Africa. Methods: The genomic sequences of the viral pathogens causing Rift valley fever, Yellow fever, Marburg virus disease, Ebola virus disease, Lassa fever, Crimean-Congo hemorrhagic fever and Chikungunya fever were compared, PCR detection target fragments were selected, and amplification primers and hybrid probes were designed. The reference samples of related pathogens were prepared by chemical synthesis of DNA and in vitro transcription RNA. The sensitivity and stability of the detection method were evaluated. The specificity was evaluated by testing 30 samples of suspected dengue fever, and hantavirus diseases, and 32 healthy human blood samples. Results: The fluorescent bead-based multiplex assay could specifically detect the corresponding pathogen, the detection limit was at a range of 10²-10⁵ copies/ μl, the specificity was 100%, and the intra-assay coefficient of variation was below 12%, and the inter-assay coefficient of variation was below 15%. Conclusions A fluorescent bead-based multiplex PCR assay for the simultaneous detection of seven viral diseases endemic in Africa was established, which may provide a new choice for the screening of suspected infectious diseases.

Objective: To establish a multiplexed immunoassay for detection of IgG antibodies against viral hemorrhagic fever epidemic in Africa. Methods: The recombinant antigens of hemorrhagic fever viruses (HFVs) epidemic in Africa were expressed and purified, and then coupled with the fluorescent microspheres. The coupling effects were evaluated by monoplexed detection of rabbit immune sera. Blood specimens were collected from people from Africa with fever, and multiplexed detection of IgG antibodies to HFVs was performed. Comparison of multiplexed assay and ELISA was performed by paired χ² test using SPSS software. Results: Both the purity and concentration of HFVs recombinant antigen met the standards for coupling and detection, and the antigens were successfully coupled with fluorescent microspheres. Seventy-eight sera samples of people from Africa with fever were multiplex detected. Among these, one was tested positive for LASV-specific IgG, one was tested positive for LUJV-specific IgG, 4 were tested positive for DENV-specific IgG and 6 tested positive for YFV-specific IgG. There was no statistically significant difference compared with ELISA, and the two method were highly correlated. Conclusions A multiplexed luminex-based immunoassay for detection of IgG antibodies to viral hemorrhagic fever epidemic in Africa was established, which laid the foundation for the differential diagnosis.

Objective: To establish a method for detection of chikungunya virus(CHIKV) antigen. Methods: CHIKV virus like particle(VLP), that contains all structural proteins, was prepared by baculovirus expression system. Mice and rabbits were immunized with the VLP to develop antibodies against CHIKV. A double antibody sandwich ELISA was established for detection of CHIKV antigens. The concentrations of the antibodies used and the reaction conditions were optimized. The detection limit and repeatability of the ELISA was evaluated. Results: The sensitivity and specificity was estimated by 10 mimicking CHIKV sera, 90 health person sera, 40 other virus infected sera. It was show that the specificity of DAS-ELISA was 100%, the detection limit was 10 TCID₅₀, the coefficients of variation (CV) within plate was <5%, the CV of different plates was <10%. Conclusions The double antibody sandwich ELISA established in this study can be used to detect the CHIKV antigen in acute phase serum of patient and provide a method for detection of CHIKV.