Objective:To analyze the genetic characteristics of coxsackievirus A16 (CVA16) related to hand, foot and mouth disease (HFMD) in Jiaxing from 2021 to 2023, and understand the biological and molecular evolutionary characteristics of CVA16 in this city.Methods:Real-time fluorescent quantitative PCR was used to detect enterovirus and its types in collected HFMD case samples. At the same time, virus isolation and cultivation were performed on positive samples using RD cells. Then high-throughput sequencing of the whole genome was performed on 23 strains of CVA16 identified from the isolated samples. Finally we obtained its whole genome sequence. DNAStar, MEGA 6.0, Simplot 3.5.1 and other bioinformatics software were used to compare and analyze the sequences, construct the phylogenetic tree of VP1 region, and realize the genotype composition. And these software were also used to analyze the homology of the whole genome nucleotide sequence and the encoded amino acids, while know well the amino acid mutation sites and gene recombination in the main regions.Results:The results showed that among the 1 836 HFMD specimens tested from 2021 to 2023, 1 432 (78.00%, 1 432/1 836) were positive for enterovirus general genes, of which 263 were positive for CVA16, accounting for 18.37% (263/1 432) of the confirmed positive cases. Twenty-three CVA16 strains were sequenced and all of them were B1 subtype, of which 6 strains belong to B1b and 17 strains belong to B1a, with B1a being dominant. B1a strains in Jiaxing showed genetic relatedness to strains isolated in Beijing, Yunnan, Guangzhou, Jiangsu and other places in 2018 to 2023, as well as strains from Vietnam, Thailand, and Australia from 2015 to 2017. B1b strains exhibited consistent amino acid mutations of L23M in the VP1 region and V217I in the VP2 region, while B1a strains exhibited amino acid mutations of S14N/D, T164K, and V251I in the VP1 region and R41H in the VP3 region. Compared with the prototype strain, the 23 strains of CVA16 in Jiaxing accumulated the largest number of amino acid mutations in the coding region, with 28 and 50 mutations in the VP1 and 3D regions, respectively. The recombination patterns of B1a and B1b strains were slightly different, with B1a type showing recombination with CVA8 in the 5′-UTR region instead of CVA4, B1a was similar to enterovirus A71 in most of P2 and P3 regions, and recombined with CVA5 in the 3D region of P3.Conclusions:The prevalent strain of CVA16 in Jiaxing may share a common trend of co-circulation and evolution with those in other provinces. The recombination mainly occurs in the 5′-UTR region and non-structural protein coding regions of P2 and P3. Continuous molecular surveillance of CVA16 is in need, and whole-genome sequencing can help understand the genetic variation, evolution, and recombination of strains.This information will provide a more robust basis for the monitoring and early warning, vaccine development, and prevention and control efforts against HFMD.

Objective:To analyze the genetic characteristics of coxsackievirus A16 (CVA16) related to hand, foot and mouth disease (HFMD) in Jiaxing from 2021 to 2023, and understand the biological and molecular evolutionary characteristics of CVA16 in this city.Methods:Real-time fluorescent quantitative PCR was used to detect enterovirus and its types in collected HFMD case samples. At the same time, virus isolation and cultivation were performed on positive samples using RD cells. Then high-throughput sequencing of the whole genome was performed on 23 strains of CVA16 identified from the isolated samples. Finally we obtained its whole genome sequence. DNAStar, MEGA 6.0, Simplot 3.5.1 and other bioinformatics software were used to compare and analyze the sequences, construct the phylogenetic tree of VP1 region, and realize the genotype composition. And these software were also used to analyze the homology of the whole genome nucleotide sequence and the encoded amino acids, while know well the amino acid mutation sites and gene recombination in the main regions.Results:The results showed that among the 1 836 HFMD specimens tested from 2021 to 2023, 1 432 (78.00%, 1 432/1 836) were positive for enterovirus general genes, of which 263 were positive for CVA16, accounting for 18.37% (263/1 432) of the confirmed positive cases. Twenty-three CVA16 strains were sequenced and all of them were B1 subtype, of which 6 strains belong to B1b and 17 strains belong to B1a, with B1a being dominant. B1a strains in Jiaxing showed genetic relatedness to strains isolated in Beijing, Yunnan, Guangzhou, Jiangsu and other places in 2018 to 2023, as well as strains from Vietnam, Thailand, and Australia from 2015 to 2017. B1b strains exhibited consistent amino acid mutations of L23M in the VP1 region and V217I in the VP2 region, while B1a strains exhibited amino acid mutations of S14N/D, T164K, and V251I in the VP1 region and R41H in the VP3 region. Compared with the prototype strain, the 23 strains of CVA16 in Jiaxing accumulated the largest number of amino acid mutations in the coding region, with 28 and 50 mutations in the VP1 and 3D regions, respectively. The recombination patterns of B1a and B1b strains were slightly different, with B1a type showing recombination with CVA8 in the 5′-UTR region instead of CVA4, B1a was similar to enterovirus A71 in most of P2 and P3 regions, and recombined with CVA5 in the 3D region of P3.Conclusions:The prevalent strain of CVA16 in Jiaxing may share a common trend of co-circulation and evolution with those in other provinces. The recombination mainly occurs in the 5′-UTR region and non-structural protein coding regions of P2 and P3. Continuous molecular surveillance of CVA16 is in need, and whole-genome sequencing can help understand the genetic variation, evolution, and recombination of strains.This information will provide a more robust basis for the monitoring and early warning, vaccine development, and prevention and control efforts against HFMD.

Objective:To explore the application effect of portable device of airway humidification on patients with head and neck tumor who underwent tracheostomy.Methods:A total of 100 patients with head and neck tumors who underwent tracheostomy at Department of Head and Neck Surgery of Cancer Hospital of Chinese Academy of Medical Sciences and Peking Union Medical College from October 2021 to May 2023 were selected as the study subjects.They were randomly divided into the control group and the observation group by using a random number table method,with 50 cases in each group.The control group received conventional nebulization inhalation for airway humidification,and the observation group adopted portable device of airway humidification with 0.45%sodium chloride solution to conduct sustained airway humidification.The effect of airway humidification,occurrence of phlegm scabs,injury and bleeding of airway,tracheal tube obstruction,and comfort level of patients after surgery were observed and compared between the two groups.Results:The satisfaction rates of the patients of observation group at the 1st,2nd,3rd,4th,and 5th day after surgery were 84.0%,88.0%,92.0%,96.0%and 96.0%for humidification of wet sputum,which were better than those of control group,and the differences were statistically significant(x2=46.240,64.103,70.560,67.919,74.227,P<0.05),respectively.The formations of phlegm scabs were respectively 1,5,1 and 1 cases at the 2nd,3rd,4th,5th d after surgery in observation group,which were less than those in control group,and the differences were statistically significant(x2=7.111,23.926,21.760,17.344,P<0.05),respectively.The numbers of airway injury bleeding and overall tracheal tube obstruction at the 2nd d after surgery in the observation group were respectively 2 cases and 1 case,which were significantly less than those in the control group,and the differences were statistically significant(x2=4.000,5.982,P<0.05),respectively.The comfort levels of airway humidification and suction phlegm,and overall comfort evaluation were respectively(17.96±0.75)scores,(16.06±1.05)scores and(34.04±1.63)scores in the observation group,which all were lower than those in the control group,and the differences were statistically significant(t=9.843,17.298,15.792,P<0.05),respectively.Conclusion:The portable device of airway humidification has a significant effect on airway humidification of patients with head and neck tumors who undergo tracheostomy,which can reduce the occurrence of phlegm scabs,and reduce the number of overall tracheal tube obstruction,and improve comfort level of patient with head and neck tumors who undergo tracheostomy.

Objective:To explore the value of detecting plasma donor-derived free DNA (dd-cfDNA) fraction in distinguishing antibody mediated-rejection (ABMR) and T cell-mediated rejection (TCMR) of renal allografts.Methods:Patients with acute rejection confirmed by allograft biopsy in the First Affiliated Hospital of Medical College of Zhejiang University from December 1, 2017 to July 18, 2019 were retrospectively included. Based on pathological classification of Banff renal allograft rejection in 2017, the patients were divided into ABMR group and TCMR group, and the latter was subdivided into TCMR Ⅰ subgroup and TCMR Ⅱ subgroup. The second generation sequencing and target region capture were used to detect candidates' peripheral blood dd-cfDNA. The demographic and clinicopathological data of the two groups were compared. The receiver operating characteristic curve (ROC) was used to evaluate the differential value of plasma dd-cfDNA and serum creatinine levels in two kinds of acute renal allograft rejection.Results:A total of 60 patients with acute rejection of renal transplantation were enrolled in this study, including 42 patients in TCMR group and 18 patients in ABMR group. The plasma dd-cfDNA percentage (%) in the ABMR group was significantly higher than that in the TCMR group [2.33(1.19, 4.30)% vs 0.98(0.50, 1.82)%, P=0.001]. The absolute value of dd-cfDNA in ABMR group was obviously higher than that in TCMR group [0.94(0.60, 2.27) ng/ml vs 0.43(0.20, 0.96) ng/ml, P=0.003]. ROC analysis to discriminate TCMR from ABMR showed that, the area under the curve ( AUC) of dd-cfDNA% was 0.76(95% CI 0.64-0.88), when the threshold was 1.11%, the sensitivity and specificity were 88.89% and 59.52%, respectively; the AUC of absolute value of dd-cfDNA was 0.74(95% CI 0.61-0.86), when the threshold was 0.53 ng/ml, the sensitivity was 88.89% and the specificity was 54.76%. TCMR subgroups were further analyzed, there was no significant difference between TCMR subgroups on the absolute value and percentage of dd-cfDNA (both P>0.05); dd-cfDNA% in ABMR group was apparently higher than that in TCMRⅠ subgroups ( P=0.008) and TCMRⅡsubgroup ( P=0.030). The absolute value of dd-cfDNA in ABMR group was significantly higher than that in TCMRⅠsubgroups ( P=0.003). Conclusion:Plasma dd-cfDNA level may help to distinguish between ABMR and TCMR rejection.

Objective:To explore the effect of decalcified bone matrix (DBM) rich in biological activity on surgical-grade medical calcium sulfate, and to observe the change of different content of DBM on the physical and chemical properties of calcium sulfate, which provide theoretical basis for the preparation of calcium sulfate bone cement with osteogenic and injectable properties.Methods:DBM with weight content of 0, 5%, 10%, 20%, 30%, 40% was fully mixed with CSH. Dissolve 0.3 g of methyl cellulose in 10 ml of deionized water to prepare a 3% methyl cellulose solution. Methylcellulose solution was added according to the liquid-solid ratio of 0.4. The mixture was evenly stirred to form slurry, then the degradation rate, compressive strength, setting time and and pH value of calcium sulfate in vitrowas measured.Results:The initial setting time and final setting time of calcium sulfate were 4.96±0.20 and 5.83±0.12 min respectively. With the increase of DBM content, the initial setting time and final setting time increased significantly ( F=49.275, P<0.05; F=124.859, P<0.05). The compressive strength of pure calcium sulfate is 23.33±6.35 MPa; when the content is 40%, the compressive strength is only 3.33 MPa. With the increase of DBM content, the compressive strength first increased and then decreased; the content of 5%, 10%, 20% DBM had little effect on the compressive strength ( P>0.05), while the compressive strength of 30% and 40% groups decreased significantly ( t=3.259, P<0.05). DBM with different contents can significantly change the degradation rate of calcium sulfate complex. When the content of DBM is 30% and 40%, the complete degradation time in vivo is only 10 d, while the degradation rate of calcium sulfate is 63% in 30 d. At any time point in vitro degradation, DBM had no significant effect on the pH value of calcium sulfate complex culture medium, and the change law was consistent with that of pure calcium sulfate. Conclusion:With the increase of DBM content, the degradation rate is gradually accelerated, the compressive strength is reduced, and the setting time is prolonged, which is not conducive to the preparation of injectable calcium sulfate cement.

Objective To study the effect of multidisciplinary rehabilitation nursing on the number of acute exacerbation, quality of life and pulmonary function in patients with chronic obstructive pulmonary disease (COPD). Methods A total of 1088 cases of COPD patients admitted to department of respiratory medicine from February 2015 to July 2016 were divided into the study group and the control group by random digits table method, 544 cases in each group. The control group was given COPD related health education and routine care. The study group was given comprehensive rehabilitation of multidisciplinary lung care. After 1 years, the mental status, dyspnea grade improvement, acute exacerbation and 6 min walk test distance (6MWD) were compared between the two groups. Results One year after intervention, 6MWD and the times of acute exacerbation in the study group were (450.32 ± 67.21)m and (0.37 ± 0.28) times, and (394.12 ± 63.17)m and (2.08 ± 1.65) times in the control group, the study group was better than the control group (t=2.064, 2.029, P<0.05). One year after intervention, the scores of mental state, function, symptom were 1.76 ± 1.02, 1.69 ± 1.01, 1.61 ± 0.79 in the study group respectively, and 2.10 ± 1.08, 2.08 ± 2.03, 1.79 ± 0.80 in the control group,the difference was statistically significant (t=2.051, 2.017, 2.103, P<0.05). Conclusions After comprehensive nursing intervention of multidisciplinary rehabilitation, COPD patients can not only have obvious improvement in lung function, but also reduce the frequency of acute exacerbation of COPD.