ObjectiveTo understand the whole genome characteristics and the information for genetic evolution in the two coxsackievirus A2 (CVA2) strains isolated from patients with herpangina in Shanghai, and to provide a scientific basis for the prevention and treatment of herpetic angina. MethodsTwo CAV2 strains isolated from patients with herpetic angina in Shanghai were performed whole genome sequencing and analysis for phylogenetics, nucleotide homology, and evolution. ResultsA phylogenetic analysis of the VP1 region revealed that the two Shanghai strains both belonged to CVA2 genotype D, with the highest homology to OL357660, a strain from Yunnan. The average nucleotide identity (ANI) of the whole genome between the two Shanghai strains was 98.88%, and the ANI of the whole genome comparisons to other CVA2 genotype D strains and CVA2 genotypes A-C strains ranged from 84.64% to 97.42% and from 79.21% to 84.20%, respectively. The two Shanghai strains had low homology in the 3D region compared to the existing CVA2 strains. The phylogenetic analysis and sliding window nucleotide similarity analysis indicated that the two Shanghai strains and the Yunnan OL357660 strain might constitute a new genetic lineage. ConclusionThe two CVA2 strains isolated for the first time in Shanghai are assigned to genotype D (GenBank: PQ130039 and PQ130040), which is identical to the existing subtype prevalent in China. As represented by the Shanghai strains, a new CVA2 genetic lineage is been identified. This study has enriched the data on genetic evolution and genetic variation of CVA2 in Shanghai, indicating the requirement to strengthen surveillance for the epidemiological pattern of CVA2.

Objective This study aims to analyze the factors influencing per capita current health expenditure in Guizhou Province from 2016 to 2022 using the gray correlation analysis method.Methods Based on the"SHA2011"accounting results of current health expenditure in Guizhou Province,as well as data from the"Guizhou Statistical Yearbook"and"Guizhou Health Statistical Yearbook",the gray correlation analysis method was used to analyze the factors influencing per capita current health expenditure in Guizhou Province from 2016 to 2022.Results The factors with the highest correlation to per capita current health expenditure in Guizhou Province were health expenditure(0.829),followed by the number of health technical personnel per thousand people(0.715),the number of practicing(assistant)physicians per thousand people(0.705),and per capita GDP(0.704).The factor with the lowest correlation was the proportion of the tertiary industry to GDP(0.543).Conclusion Health expenditure investment has the strongest correlation with per capita current health expenditure in Guizhou Province.Health re-source investment and health service capacity are the main influencing factors of per capita current health expenditure in Guizhou Province.At the same time,the impact of economic and social factors on current health expenditure should be fully recognized.

Objective This study aims to analyze the factors influencing per capita current health expenditure in Guizhou Province from 2016 to 2022 using the gray correlation analysis method.Methods Based on the"SHA2011"accounting results of current health expenditure in Guizhou Province,as well as data from the"Guizhou Statistical Yearbook"and"Guizhou Health Statistical Yearbook",the gray correlation analysis method was used to analyze the factors influencing per capita current health expenditure in Guizhou Province from 2016 to 2022.Results The factors with the highest correlation to per capita current health expenditure in Guizhou Province were health expenditure(0.829),followed by the number of health technical personnel per thousand people(0.715),the number of practicing(assistant)physicians per thousand people(0.705),and per capita GDP(0.704).The factor with the lowest correlation was the proportion of the tertiary industry to GDP(0.543).Conclusion Health expenditure investment has the strongest correlation with per capita current health expenditure in Guizhou Province.Health re-source investment and health service capacity are the main influencing factors of per capita current health expenditure in Guizhou Province.At the same time,the impact of economic and social factors on current health expenditure should be fully recognized.

Objective To investigate the difference in visual quality between oval and round corneal flaps in femtosecond excimer laser in situ keratomileusis(FS-LASIK).Methods In a prospective randomized controlled clinical study,patients who voluntarily underwent FS-LASIK surgery in Wuhan University Affiliated Aier Ophthalmology Hospital from June 2022 to February 2023 were randomly divided into oval valve group and round valve group,with 36 patients(72 eyes)in each group.During FS-LASIK,oval and round corneal flaps were made,and excimer laser surgery was performed in parallel.Observe intraoperative and postoperative complications.The unaided visual acuity(UCVA)and spherical equivalent(SE)at 1 day,1 week,1 month and 3 months after surgery were compared between the two groups at 3 months after surgery,and the modulation transfer function(MTF),Strel ratio(SR)and total higher-order aberration at 3 months after surgery were compared between the two groups.Results All patients had successfully completed the surgery without serious complications.There was no significant difference between the oval valve group and the round valve group in UCVA at 1 day,1 week after surgery,1 month after surgery,3 months after surgery.There was no significant difference between the two groups on the postoperative SE at 1 day,1 week after surgery,1 month after surgery,and 3 months after surgery.There was no significant difference in MTF,SR and total higher-order aberration between the two groups at 3 months after surgery.Conclusion Ideal visual acuity can be achieved with either oval or round corneal flap during FS-LASIK,and there is no significant difference in visual quality between the two in the early postoperativeperiod.

ObjectiveTo determine the genomic characteristics of a subgenus B human adenovirus strain isolated in Shanghai in 2021. MethodsAn adenovirus type 55 strain was isolated and identified from a patient with acute hemorrhagic conjunctivitis （AHC）. Complete genome of the strain was obtained using the next-generation sequencing （NGS）. Phylogenetic trees were reconstructed based on the sequences of Hexon， Fiber， Penton and complete genome to genomically characterize this strain. ResultsPhylogenetic analysis based on the complete genome classified this strain （MH2021001） into subgenus B， subspecies B2 of HAdV-55. Hexon gene of MH2021001 had close phylogenetic relationship with HAdV-11， while Fiber and Penton genes had close relationship with HAdV-14. The MH2021001 showed high nucleotide identity with currently prevalent HAdV⁃55 strains （>99.90%）. The complete genome had 99.96% nucleotide identity to the 73-GD_CHN_2016 strain isolated in Guangdong. In addition， the amino acid sequence of MH2021001 had several substitutions in regions coding for E1B， L4， E3 and L5. ConclusionThis strain has been classified to HAdV-B55. No recombination event is identified in the complete genome. Due to multiple amino acid substitutions， the biological characteristics of the strain need to be further identified.

Objective To analyze the biomechanical feasibility of two-point fixation by distal radius plate for the treatment of SandersⅢ calcaneal fractures. Methods The three-dimensional (3D) finite element musculoskeletal foot model was established based on CT and MRI images, which comprised bones, muscles, plantar fascia, ligaments and soft tissues. After validation, the SandersⅢ calcaneal fracture models fixed by distal radial plate (two-point fixation) and calcaneal plate (three-point fixation) were established, so as to compare the biomechanical characteristics of two calcaneal models. Results The maximum stress of the two-point fixation and three-point fixation model was 324.70 and 407.90 MPa, respectively. The maximum displacements of the two models were 2.498 and 2.541 mm, respectively. There was no significant difference in the posterior articular surface displacement between the two models. In both models, the Bohler’s angle and Gissane’s angle were within the normal range. Conclusions The two-point fixation by distal radial plate can satisfy the biomechanical stability of calcaneal fracture treatment. Compared with traditional steel plate, the two-point fixation shows the advantage of smaller surgical trauma, more uniform overall stress distribution, early weight-bearing rehabilitation after surgery, which is a novel treatment recommended for treating calcaneal fractures.

Objective:To investigate the factors affecting the outcome of surgical treatment for femoral intertrochanteric fractures.Methods:A retrospective case series study was conducted for 424 patients with intertrochanteric fractures admitted to four hospitals in Shanghai from January 2014 to December 2016. There were 200 males and 224 females, aged 45-98 years [(77.8±10.5)years]. Normal union was observed in 326 patients (normal union group), and abnormal union was founded in 98 patients including nonunion in 2 patients (abnormal union group). Data were recorded including the age, gender, injury side, AO classification, Evans-Jesen classification, thickness of femoral lateral wall, medial support, tip-apex distance, and screw position within the femoral head. Fracture healing was assessed. Univariate analysis was conducted to examine differences of the above factors between two groups, and Logistic regression was used to screen the main factors affecting fracture union. At the latest follow-up, the hip joint function was estimated according to the Oxford score system.Results:All patients were followed up for 1-3 years [(2.2±0.3)years]. Univariate analysis showed significant differences between two groups regarding the AO classification, Evans-Jesen classification, medial support, tip-apex distance, and screw position within the femoral head ( P<0.05). Multivariate regression analysis revealed that the simple fracture ( OR=1.030), medial support ( OR=0.395), tip-apex distance ≤25 mm ( OR=0.266) and inferior screw position ( OR=0.986) were significantly related to fracture union. The hip function in normal union group was (42.6±4.5)points, better than (35.4±3.2)points in abnormal union group ( P<0.05). Conclusion:The stability of intertrochanteric fracture is the most important factor for fracture union. The medial support, tip-apex distance ≤25 mm and inferior screw position are helpful to promote fracture healing and recovery of hip joint function.

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 construct mitochondrial antiviral signaling ( MAVS) gene or Nuclear factor kappa B1 ( NFκB1) gene knockout Madin-Darby canine kidney (MDCK) cells, and identify the cell growth and proliferation characteristics of influenza virus (Flu) in these cells. Methods:MAVS or NFκB1 knockout MDCK cells were established using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated 9 (Cas9) technique. Flu was inoculated into these cells, then the hemagglutination (HA) titers and TCID 50 were determined and compared with the original MDCK cells. Results:MDCK cells knocked out of MAVS (MDCK- MAVS-) or NFκB1 (MDCK- NFκB1 -) were obtained stably. The two cells were both adherent epithelioid cells as well as wild type MDCK cells. The HA titers showed no obvious difference among these cells after inoculating Flu. Nevertheless, the TCID 50 titers were respectively increased 9.5 times in MDCK- MAVS-cell culture and 10 times in MDCK- NFκB1 - cell culture, compared to the wild type MDCK cells. Conclusions:The infectivities of Flu were both increased in MDCK- MAVS-cells and MDCK- NFκB1 - cells, illustrating their potential in Flu culture.

Objective: To analyze the changes in the expression of hypoxia inducible factor-1α (HIF-1α) and inflammatory cytokines and to investigate the role of HIF-1α in regulating the production of inflammatory cytokines during influenza A (H1N1) virus infection. Methods: BALB/c mice were injected with H1N1 virus to establish the mouse model of H1N1 virus infection. Fifteen BALB/c mice were randomly divided into three groups: control group, H1N1 virus group and H1N1 virus+ HIF-1α inhibitor group. Inflammatory cytokines (IL-6, TNF-α, IL-1β and IL-10) in samples of serum and lung tissues were detected by Luminex and ELISA. Levels of HIF-1α in serum and lung tissue samples were detected by Western blot and ELISA, respectively. Results: Compared with the control group, the levels of inflammatory cytokines in serum (IL-6, TNF-α, IL-1β and IL-10) and lung tissues (IL-6 and TNF-α) and the expression of HIF-1α in serum and lung tissues in the H1N1 virus group were significantly increased. The levels of HIF-1α, IL-6, TNF-α IL-1β and IL-10 in lung tissues in H1N1 virus+ HIF-1α inhibitor group were significantly lower than those of the H1N1 virus group. Conclusion During H1N1 virus infection, the levels of inflammatory cytokines and HIF-1α were significantly increased. The production of inflammatory cytokines was significantly reduced after inhibiting HIF-1α expression, suggesting that HIF-1α might promote the production of inflammatory cytokines.