This study established an indirect ELISA method for antibody detection based on human rotavirus group A(RVA)VP6 protein,to provide technical support for RVA antibody detection.An ELISA method for detecting RVA antibodies was established through a checkerboard assay for optimization of the reaction conditions with VP6 protein of RVA as the diagnostic antigen.Serum samples from healthy children and patients infected with RVA were collected and detected with the ELISA method.The optimal work-ing conditions for ELISA involved coating the ELISA plate with VP6 protein at a concentration of 2 μg/mL and diluting the serum at a ratio of 1∶250.The critical value of negative and positive samples was 0.137.The ELISA method had good specificity,sensitivity,and repeatability,and showed a 95%consistency rate with the western blotting antibody detection method.We tested 370 serum samples collected from children,which showed an antibody positivity rate of 81.1%.We additionally tested the acute and recovery phase sera from 15 patients infected with RVA,and observed a significantly higher RVA antibody titer in the recovery phase serum than the acute infection phase serum.The ELISA antibody detection method showed good specificity,sensitivity,and reproducibility,and therefore can be used for RVA antibody detection and auxiliary diagnosis of RVA infection.

This study expressed the VP6 protein of human group A rotavirus in a prokaryotic expression system and prepared mouse polyclonal antibodies to this protein.The human rotavirus VP6 gene was amplified through RT-PCR from fecal samples infected with group A rotavirus and cloned into the pET-32a vector to construct the recombinant pET-32a-VP6 prokaryotic expression plas-mid.The recombinant plasmid was transformed into Escherichia coli BL21(DE3).VP6 protein was expressed through induction with isopropyl-β-D-thiopyranoside(IPTG),purified with nickel nitrilotriacetic acid(Ni-NTA)affinity chromatography,and identified through SDS-PAGE and western blotting.Polyclonal antibodies to the VP6 protein were obtained by immunization of BALB/c mice with VP6 protein.Antibody titers were determined through indirect ELISA.The recombinant expression plasmid pET-32a-VP6 was con-structed,and VP6 protein was expressed primarily in inclusion bodies.Polyclonal antibodies to VP6 protein with a titer of 1∶32 000 were obtained by immunization of mice with VP6 protein purified by affinity chromatography.The antibodies specifically reacted with VP6 protein.The VP6 protein of human group A rotavirus was expressed and purified,and mouse polyclonal antibodies to this protein were prepared,thus providing a foundation for the preparation of a diagnostic antigen for human group A rotavirus and the establishment of serological detection methods.

This study established an indirect ELISA method for antibody detection based on human rotavirus group A(RVA)VP6 protein,to provide technical support for RVA antibody detection.An ELISA method for detecting RVA antibodies was established through a checkerboard assay for optimization of the reaction conditions with VP6 protein of RVA as the diagnostic antigen.Serum samples from healthy children and patients infected with RVA were collected and detected with the ELISA method.The optimal work-ing conditions for ELISA involved coating the ELISA plate with VP6 protein at a concentration of 2 μg/mL and diluting the serum at a ratio of 1∶250.The critical value of negative and positive samples was 0.137.The ELISA method had good specificity,sensitivity,and repeatability,and showed a 95%consistency rate with the western blotting antibody detection method.We tested 370 serum samples collected from children,which showed an antibody positivity rate of 81.1%.We additionally tested the acute and recovery phase sera from 15 patients infected with RVA,and observed a significantly higher RVA antibody titer in the recovery phase serum than the acute infection phase serum.The ELISA antibody detection method showed good specificity,sensitivity,and reproducibility,and therefore can be used for RVA antibody detection and auxiliary diagnosis of RVA infection.

This study expressed the VP6 protein of human group A rotavirus in a prokaryotic expression system and prepared mouse polyclonal antibodies to this protein.The human rotavirus VP6 gene was amplified through RT-PCR from fecal samples infected with group A rotavirus and cloned into the pET-32a vector to construct the recombinant pET-32a-VP6 prokaryotic expression plas-mid.The recombinant plasmid was transformed into Escherichia coli BL21(DE3).VP6 protein was expressed through induction with isopropyl-β-D-thiopyranoside(IPTG),purified with nickel nitrilotriacetic acid(Ni-NTA)affinity chromatography,and identified through SDS-PAGE and western blotting.Polyclonal antibodies to the VP6 protein were obtained by immunization of BALB/c mice with VP6 protein.Antibody titers were determined through indirect ELISA.The recombinant expression plasmid pET-32a-VP6 was con-structed,and VP6 protein was expressed primarily in inclusion bodies.Polyclonal antibodies to VP6 protein with a titer of 1∶32 000 were obtained by immunization of mice with VP6 protein purified by affinity chromatography.The antibodies specifically reacted with VP6 protein.The VP6 protein of human group A rotavirus was expressed and purified,and mouse polyclonal antibodies to this protein were prepared,thus providing a foundation for the preparation of a diagnostic antigen for human group A rotavirus and the establishment of serological detection methods.

OBJECTIVE To calculate the cost of centralized dispensing of four categories of drugs （ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions） in pharmacy intravenous admixture service （PIVAS）， and provide reference for setting charging standards for relevant departments. METHODS The operating costs of PIVAS in 12 medical institutions from Shaanxi province were collected through questionnaire survey， including labor costs， medical and health material costs， fixed asset depreciation and repair costs， water and electricity costs， and management costs. The operation time allocation coefficient method and workload allocation coefficient method were comprehensively used to allocate the above costs， and the unit preparation costs of four categories of drugs were calculated. RESULTS The average annual total costs of dispensing ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions in Shaanxi province were （2 195 900.25±1 680 893.73） yuan， （746 341.59±725 839.39） yuan， （331 420.15±183 258.83） yuan， and （330 322.68±277 281.70） yuan， respectively， with labor costs accounting for the highest proportion， averaging 85.49%. The costs of dispensing a set of ordinary drugs， antibacterial drugs， and hazardous drugs were 5.89， 7.60， and 14.37 yuan， respectively； the cost of dispensing one bag of parenteral nutrition solution was 32.15 yuan （excluding the cost of disposable intravenous nutrition bags）. CONCLUSIONS The cost calculation method and data of different types of intravenous drugs obtained in this study can provide reference for relevant departments to formulate and adjust PIVAS fee standards.

OBJECTIVE To calculate the cost of centralized dispensing of four categories of drugs （ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions） in pharmacy intravenous admixture service （PIVAS）， and provide reference for setting charging standards for relevant departments. METHODS The operating costs of PIVAS in 12 medical institutions from Shaanxi province were collected through questionnaire survey， including labor costs， medical and health material costs， fixed asset depreciation and repair costs， water and electricity costs， and management costs. The operation time allocation coefficient method and workload allocation coefficient method were comprehensively used to allocate the above costs， and the unit preparation costs of four categories of drugs were calculated. RESULTS The average annual total costs of dispensing ordinary drugs， antibacterial drugs， hazardous drugs， and parenteral nutrition solutions in Shaanxi province were （2 195 900.25±1 680 893.73） yuan， （746 341.59±725 839.39） yuan， （331 420.15±183 258.83） yuan， and （330 322.68±277 281.70） yuan， respectively， with labor costs accounting for the highest proportion， averaging 85.49%. The costs of dispensing a set of ordinary drugs， antibacterial drugs， and hazardous drugs were 5.89， 7.60， and 14.37 yuan， respectively； the cost of dispensing one bag of parenteral nutrition solution was 32.15 yuan （excluding the cost of disposable intravenous nutrition bags）. CONCLUSIONS The cost calculation method and data of different types of intravenous drugs obtained in this study can provide reference for relevant departments to formulate and adjust PIVAS fee standards.

Objective:To evaluate the risk of HBV recurrence after liver transplantation in patients with end-stage hepatitis B related liver disease, and to explore the indications for antiviral therapy withdrawal.Methods:The data of HBV DNA, cccDNA in liver puncture tissues and peripheral blood in 31 patients after liver transplantation was retrospectively analyzed.Results:Among the 31 patients, 15 (48%) had detectable and quantified HBV DNA in liver biopsy tissue, while their HBV related serological indicators were negative, suggesting an occult HBV infection in some patients. The study found 15 out of 19 cases who were taking Entecavir were cccDNA negative (78.9%), compared to 5 out of 12 cases (41.6%) in Lamivudine regiment ( P=0.03). Conclusions:Hidden HBV infection can be detected by amplifying cccDNA and HBV DNA in liver puncture tissue by using ddPCR. Entecavir is superior to lamivudine in the clearance of cccDNA.

We conducted a prospective study to assess the non-inferiority of adjuvant chemotherapy alone versus adjuvant concurrent chemoradiotherapy (CCRT) as an alternative strategy for patients with early-stage (FIGO 2009 stage IB-IIA) cervical cancer having risk factors after surgery. The condition was assessed in terms of prognosis, adverse effects, and quality of life. This randomized trial involved nine centers across China. Eligible patients were randomized to receive adjuvant chemotherapy or CCRT after surgery. The primary end-point was progression-free survival (PFS). From December 2012 to December 2014, 337 patients were subjected to randomization. Final analysis included 329 patients, including 165 in the adjuvant chemotherapy group and 164 in the adjuvant CCRT group. The median follow-up was 72.1 months. The three-year PFS rates were both 91.9%, and the five-year OS was 90.6% versus 90.0% in adjuvant chemotherapy and CCRT groups, respectively. No significant differences were observed in the PFS or OS between groups. The adjusted HR for PFS was 0.854 (95% confidence interval 0.415-1.757; P = 0.667) favoring adjuvant chemotherapy, excluding the predefined non-inferiority boundary of 1.9. The chemotherapy group showed a tendency toward good quality of life. In comparison with post-operative adjuvant CCRT, adjuvant chemotherapy treatment showed non-inferior efficacy in patients with early-stage cervical cancer having pathological risk factors. Adjuvant chemotherapy alone is a favorable alternative post-operative treatment.
Female ; Humans ; Uterine Cervical Neoplasms/drug therapy* ; Prospective Studies ; Quality of Life ; Neoplasm Staging ; Chemoradiotherapy ; Chemotherapy, Adjuvant/adverse effects* ; Adjuvants, Immunologic ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use* ; Retrospective Studies

Background: The use of molecular categorisation is shifting paradigm towards the use of molecular information to refine risk stratification in endometrial cancer (EC). To date, evidence to support molecular-guided therapies is limited to retrospective studies and secondary molecular analyses of patients receiving standard treatment. The PROBEAT study is the first randomized phase III trial to evaluate tailored adjuvant treatment based on WHO-endorsed molecular classification in Chinese EC patients. It is expected to provide a clinical decision-making tool for adjuvant treatment of patients with high-intermediate risk (HIR) or intermediate risk (IR) EC to better optimise and personalise patient care and increase relapse-free survival. Methods The PROBEAT trial is a prospective, multicentre study led by Women’s Hospital of Zhejiang University Gynaecologic Oncology Group. Recruitment started on January 24, 2022, and 590 patients with HIR or IR endometrioid EC are expected to be recruited from 13 clinical centres in China. All tumor tissues will be classified into four molecular subtypes (POLEmut, MMRd, p53abn, or NSMP) based on WHO-endorsed molecular classification. Patients will be randomly assigned at a 2:1 ratio to either experimental arm and will receive molecular profile-based adjuvant treatment (observation in the POLEmut subgroup, vaginal brachytherapy in the MMRd or NSMP subgroup, or chemoradiotherapy in the p53abn subgroup) or to standard arm and will receive preferred adjuvant radiotherapy as recommended by the recent National Comprehensive Cancer Network guidelines version 1 (2022). The primary outcome is 3-year rates of recurrence. Secondary outcomes are relapse-free survival, overall survival, adverse events and health-related cancer-specific quality of life.

Objective:Diet balance index (DBI_16) was used to evaluate the dietary status of smoking adults in Tianjin, and the relationship between DBI_16 and serum uric acid was analyzed.Methods:A total of 1 478 inhabitants aged 18 and above were enrolled. The dietary status was obtained with a 3-day dietary recall and condiment weighing method. And their fasting venous blood was collected to detect uric acid. Food intake and DBI_16 scores of smokers and non-smokers were described, and the relationship among smoking, diet quality distance (DQD-DBI), and serum uric acid was analyzed.Results:The intake of vegetables, fruits, and milk of smokers in Tianjin was lower than while the salt, oil, cereals, and aquatic products were higher than that of non-smokers. The DBI_16 scores of vegetables and fruits, food types, milk, and beans of smokers were lower than those of non-smokers in Tianjin, and the scores of pure energy foods and condiments were higher than those of the non-smokers. The DQD-DBI, high bound score and low bound score of smokers in Tianjin were 42.0, 14.0, and 29.0 respectively, which were all higher than those of non-smokers. The main problems appeared as moderately inadequate intake (accounting for 67.0%), low, excessive intake (accounting for 70.9%), and moderate imbalance of intake (accounting for 67.2%). The serum uric acid of smokers was higher than in the non-smokers, and there the same result appeared under the conditions of "not suitable" in DQD-DBI.Conclusions:In Tianjin, the dietary imbalance was more evident in smokers than the non-smokers, and the serum uric acid was significantly higher than the non-smokers. Strategies as strengthening the nutrition education and intervention targeted for smokers were in urgent need.