Objective To analyze the genetic characteristics and variations of influenza A (H3N2) viruses in Ma'anshan from 2022 to 2024, and to provide a scientific basis for local influenza prevention and control. Methods From April 2022 to March 2024, influenza-like illness (ILI) specimens were collected from three national influenza surveillance sentinel hospitals in Ma’anshan. Samples positive for influenza by real-time PCR were subjected to virus culture and identification. A total of 40 representative A/H3N2 strains with hemagglutination titers ≥8 were selected for whole-genome sequencing. Genetic evolution, homology, amino acid variations, and glycosylation sites were analyzed. Results All H3N2 representative strains from the 2022–2023 influenza season belonged to clade 3C.2a1b.2a.1a.1, while those from the 2023–2024 season fell into clade 3C.2a1b.2a.2a.3a.1. The nucleotide and amino acid sequence similarities of HA and NA between the 40 representative strains and the vaccine strain A/Darwin/6/2021 were all above 97.35%. Compared with the vaccine strain, amino acid mutations were identified in antigenic sites A, B, C, and E, as well as in receptor-binding sites of the HA protein. An I222V substitution was detected in the NA protein. The HA protein contained four additional glycosylation sites compared to the vaccine strain, while the glycosylation pattern of the NA protein remained consistent. Conclusion No antigenic drift was observed in the influenza A/H3N2 viruses in Ma'anshan City from 2022 to 2024, but genetic changes such as branching variations, key amino acid substitutions, and an increase in HA glycosylation sites were observed. These findings underscore the importance of sustained molecular surveillance of local influenza viruses.

Objective:To analyze the genetic characteristics of influenza B virus Victoria lineage in Ma′anshan from 2019 to 2022.Methods:Throat swabs were collect from cases with influenza-like illness in three sentinel hospitals and from influenza outbreak events in Ma′anshan city from 2019 to 2022. Real-time fluorescence-based PCR was performed to detect the nucleic acids of influenza viruses in the swab samples. Statistical analysis was conducted using the influenza year as the detection cycle. MDCK cells and chicken embryos were used for virus isolation, and 31 strains of B/Victoria lineage were selected for whole-genome sequencing and genetic analysis.Results:A total of 11 258 throat swabs were collected from ILI cases, and 8.90% (1 002/11 258) of them tested positive for B/Victoria. Except for the period from the winter of 2020 to the spring of 2021, during which no epidemic outbreak of influenza was observed, there were peaks in the prevalence of B/Victoria influenza in winter and spring of the other years, with the positive rates ranging from 12.65% (54/427) to 46.68% (176/377) during peak seasons. Compared with the recommended vaccine strains, the homology of the HA gene nucleotides of the viral strains isolated from 2019 to 2022 ranged from 98.40% to 99.26%, and the homology of amino acids ranged from 96.21% to 98.80%. All isolated strains carried mutations in the 190-helix of the hemagglutinin protein; the strains isolated from 2019 to 2020 had two amino acid insertions at position 160-loop; and the strains isolated from 2021 and 2022 had mutations at positions 120-loop and 150-loop. No drug-resistant mutations were detected in the neuraminidase gene.Conclusions:The B/Victoria strains isolated in Ma′anshan city from 2019 to 2022 have key site variations as compared with the recommended vaccine strains, suggesting a decrease in vaccine immune efficacy. Therefore, it is necessary to strengthen the surveillance of viral mutations to provide reference for updating vaccine strains and formulating epidemic prevention and control measures.

Objective To analyze the correlation between ferroptosis and post operative cognitive dysfunction(POCD)in elderly patients with fractures.Methods A total of 120 elderly patients with fracture were divided into the control group and the POCD group according to whether POCD occurred,with 60 cases in each group.Basic data of the two groups were collected.The levels of nerve injury indicators[brain myelin basic protein(MBP),glial fibrillary acidic protein(GFAP)],and fractures markers[glutathione peroxidase 4(GPX4),cyclooxygenase 2(COX2),long-chain acyl-CoA synthase 4(ACSL4)]were detected by enzyme-linked immunosorbent assay at 6 h after surgery.Logistic regression was used to analyze risk factors of POCD in elderly patients with fracture.The correlations between MBP,GFAP and GPX4,COX2 and ACSL4 were analyzed by Pearson correlation analysis.Receiver operating characteristic(ROC)curves were used to analyze predictive values of MBP,GFAP,GPX4,COX2 and ACSL4 levels to POCD.Results The age,proportion of general anesthesia,anesthesia time,intraoperative blood loss and MBP,GFAP,COX2,ACSL4 levels were higher in the POCD group than those in the control group(P＜0.05),while GPX4 was lower than that in the control group(P＜0.05).The levels of MBP and GFAP were negatively correlated with levels of GPX4 in elderly patients with fractures,and positively correlated with levels of COX2 and ACSL4(P＜0.05).Advanced age,general anesthesia,long duration of anesthesia,increased levels of MBP,GFAP,COX2 and ACSL4,and decreased level of GPX4 were independent risk factors for POCD in elderly fracture patients(P＜0.05).The critical values of GPX4,COX2 and ACSL4 for predicting POCD in elderly patients with fractures were GPX4≤23.05 μg/L,COX2≥20.35 μg/L and ACSL4≥237.85 μg/L,and the AUC were 0.869,0.736 and 0.841.The sensitivity was 76.67%,68.33%and 88.33%,and the specificity was 86.67%,78.33%and 75.00%,respectively.The diagnostic efficacy of GFAP,COX2 and ACSL4 was higher than that of MBP and GFAP.Conclusion The incidence of POCD in elderly patients with fracture is associated with ferroptosis,and levels of GPX4,COX2 and ACSL4 have certain predictive value for the incidence of POCD after surgery in elderly patients with fracture.

Objective To analyze the correlation between ferroptosis and post operative cognitive dysfunction(POCD)in elderly patients with fractures.Methods A total of 120 elderly patients with fracture were divided into the control group and the POCD group according to whether POCD occurred,with 60 cases in each group.Basic data of the two groups were collected.The levels of nerve injury indicators[brain myelin basic protein(MBP),glial fibrillary acidic protein(GFAP)],and fractures markers[glutathione peroxidase 4(GPX4),cyclooxygenase 2(COX2),long-chain acyl-CoA synthase 4(ACSL4)]were detected by enzyme-linked immunosorbent assay at 6 h after surgery.Logistic regression was used to analyze risk factors of POCD in elderly patients with fracture.The correlations between MBP,GFAP and GPX4,COX2 and ACSL4 were analyzed by Pearson correlation analysis.Receiver operating characteristic(ROC)curves were used to analyze predictive values of MBP,GFAP,GPX4,COX2 and ACSL4 levels to POCD.Results The age,proportion of general anesthesia,anesthesia time,intraoperative blood loss and MBP,GFAP,COX2,ACSL4 levels were higher in the POCD group than those in the control group(P＜0.05),while GPX4 was lower than that in the control group(P＜0.05).The levels of MBP and GFAP were negatively correlated with levels of GPX4 in elderly patients with fractures,and positively correlated with levels of COX2 and ACSL4(P＜0.05).Advanced age,general anesthesia,long duration of anesthesia,increased levels of MBP,GFAP,COX2 and ACSL4,and decreased level of GPX4 were independent risk factors for POCD in elderly fracture patients(P＜0.05).The critical values of GPX4,COX2 and ACSL4 for predicting POCD in elderly patients with fractures were GPX4≤23.05 μg/L,COX2≥20.35 μg/L and ACSL4≥237.85 μg/L,and the AUC were 0.869,0.736 and 0.841.The sensitivity was 76.67%,68.33%and 88.33%,and the specificity was 86.67%,78.33%and 75.00%,respectively.The diagnostic efficacy of GFAP,COX2 and ACSL4 was higher than that of MBP and GFAP.Conclusion The incidence of POCD in elderly patients with fracture is associated with ferroptosis,and levels of GPX4,COX2 and ACSL4 have certain predictive value for the incidence of POCD after surgery in elderly patients with fracture.

Objective:To analyze the genetic characteristics of influenza B virus Victoria lineage in Ma′anshan from 2019 to 2022.Methods:Throat swabs were collect from cases with influenza-like illness in three sentinel hospitals and from influenza outbreak events in Ma′anshan city from 2019 to 2022. Real-time fluorescence-based PCR was performed to detect the nucleic acids of influenza viruses in the swab samples. Statistical analysis was conducted using the influenza year as the detection cycle. MDCK cells and chicken embryos were used for virus isolation, and 31 strains of B/Victoria lineage were selected for whole-genome sequencing and genetic analysis.Results:A total of 11 258 throat swabs were collected from ILI cases, and 8.90% (1 002/11 258) of them tested positive for B/Victoria. Except for the period from the winter of 2020 to the spring of 2021, during which no epidemic outbreak of influenza was observed, there were peaks in the prevalence of B/Victoria influenza in winter and spring of the other years, with the positive rates ranging from 12.65% (54/427) to 46.68% (176/377) during peak seasons. Compared with the recommended vaccine strains, the homology of the HA gene nucleotides of the viral strains isolated from 2019 to 2022 ranged from 98.40% to 99.26%, and the homology of amino acids ranged from 96.21% to 98.80%. All isolated strains carried mutations in the 190-helix of the hemagglutinin protein; the strains isolated from 2019 to 2020 had two amino acid insertions at position 160-loop; and the strains isolated from 2021 and 2022 had mutations at positions 120-loop and 150-loop. No drug-resistant mutations were detected in the neuraminidase gene.Conclusions:The B/Victoria strains isolated in Ma′anshan city from 2019 to 2022 have key site variations as compared with the recommended vaccine strains, suggesting a decrease in vaccine immune efficacy. Therefore, it is necessary to strengthen the surveillance of viral mutations to provide reference for updating vaccine strains and formulating epidemic prevention and control measures.

Objective:To discuss the clinical efficacy of recombinant human growth hormone(rhGH)in the treatment of the pediatric patients with growth hormone deficiency(GHD)and idiopathic short stature(ISS),and to clarify its clinical application value in the pediatric patients with short stature of different etiologies.Methods:The clinical data of 132 children with short stature who treated with rhGH from January 2018 to January 2023 were collected.They were divided into GHD group(n=70)and ISS group(n=62)based on different etiologies.The bone age,target height(TH),body mass index(BMI),height standard deviation score(HtSDS),changes in height standard deviation scores(ΔHtSDS)before treatment and 6 months after treatment,and growth velocity(GV)of the pediatric patients were calculated.Propensity score matching(PSM)and inverse probability of treatment weighting(IPTW)were used to balance the confounding factors between the pediatric patients in two groups and the efficacy and safety of the pediatric patients in two groups were evaluated.Results:There were significant differences in whether children were full-term,bone age,bone age maturity,and TH of the pediatric patients between two groups(P<0.05).Compared with before treatment,the height and HtSDS of the pediatric patients in both GHD and ISS groups were significantly increased after treated for 6 months(P<0.05).Before matched by PSM,there were significant differences in full-term,bone age,bone age maturity,and TH of the pediatric patients between two groups(P<0.05).After matched by PSM,there were no significant differences in gender,region,term birth status,mode of delivery,feeding method,age,bone age,height,BMI,TH,and pretreatment HtSDS of the pediatric patients between two groups(P>0.05);the standardized mean difference(SMD)differences of covariates except for region were<0.2.After weighted by IPTW,there were no significant differences in gender,region,term birth status,mode of delivery,feeding method,age,bone age,height,BMI,TH,and pretreatment HtSDS of the pediatric patients between two groups(P>0.05);all SMD of covariates except for term birth status were<0.2.Before balancing covariates,after meatched by PSM matching,and after weighted by IPTW weighting compared with GHD group,the GV and ΔHtSDS of the pediatric patients in ISS group were slightly increased,but the difference was not significant(P>0.05).In terms of adverse reactions,2 cases(2.68%)of fasting hyperglycemia and 7 cases(10.00%)of hypothyroidism occurred in GHD group;3 cases(4.84%)of fasting hyperglycemia and 2 cases(3.23%)of hypothyroidism occurred in ISS group.Conclusion:rhGH can promote the height increase in the patients with GHD and ISS,and there is no significant difference in the height-increasing efficacy between GHD and ISS children.The incidence of adverse reactions is relatively low during treatment,indicating good overall safety.

Objective:To evaluate the effect of intravenous infusion of lidocaine on the efficacy of conventional treatment for rheumatoid arthritis.Methods:Forty-four patients with rheumatoid arthritis of either sex, aged 32-85 yr, weighing 40-76 kg, who were admitted to the Department of Pain and Nephrology in our hospital from September 2019 to September 2020, were divided into 2 groups ( n=22 each) according to the random number table method: control group (C group) and lidocaine group (L group). Both groups received conventional treatment.When visual analogue scale (VAS) score ≥5, glucocorticoid (GC) and non-steroidal anti-inflammatory drugs (NSAIDs) were taken orally to maintain the VAS score ≤4.In group L, 0.2% lidocaine hydrochloride injection 3 mg/kg (diluted with 0.9% sodium chloride injection 500 ml) was intravenously infused at a rate of 25 ml/h for 2 h, once a day, for 5 consecutive days, based on the conventional treatment.The VAS score, 28-joint Disease Activity Score (DAS28 score), simplified disease activity index score (SDAI score), consumption of GC and NSAIDs and adverse reactions were recorded before treatment (T 1) and at 1, 4 and 8 weeks after treatment (T 2-4). The temperature of the pain area of the affected joint was evaluated through infrared thermal imaging at T 1 and T 2. Results:Compared with the baseline at T 1, VAS score, DAS28 score and SDAI score were significantly decreased at each time point, and the temperature of the pain area of the affected joint at T 2 was decreased in the two groups ( P<0.05). There were no significant differences in VAS score, DAS28 score and SDAI score at each time point between two groups ( P>0.05). Compared with group C, the consumption of GC and NSAIDs was significantly decreased, and the temperature of the pain area of the dorsum of both hands and the dorsum of right foot at T 2 and incidence of adverse reactions were decreased in group L ( P<0.05). Conclusions:Intravenous infusion of lidocaine can optimize the efficacy of conventional treatment for rheumatoid arthritis.

Objective:To explore the correlation between the indexes of tongue coating exfoliated cells and the Traditional Chinese Medicine (TCM) syndromes of patients with chronic gastritis.Methods:One hundred and forty-eight patients with chronic gastritis in our hospital from March 2017 to May 2018 were selected and divided into 4 groups, including 36 patients with spleen and stomach weakness syndrome, 39 patients with liver-stomach discordance syndrome, 32 patients with stomach yin deficiency syndrome, and spleen and 41patients with stomach damp-heat syndrome according to the TCM classification. In addition, another 50 healthy people without cold and heat deficiency syndrome were selected as healthy control group. The maturity index (MI) and mature value (MV) of tongue coating exfoliated cells, chemical indicators and cell cycle of tongue coating exfoliated cells were detected. The spearman rank correlation method was used to analyze the indicators of tongue coating exfoliated cells and TCM syndromes.Results:Compared with the healthy control group, the subjects with spleen and stomach weakness syndrome, liver-stomach discord syndrome, stomach-yin deficiency syndrome, spleen-stomach damp-heat syndrome showed significantly higher percentage of intermediate cells [(14.85 ± 4.03) % vs. (26.47 ± 3.94) %, (22.32 ± 5.41) %, (31.47 ± 3.28) %, (35.62 ± 3.96) %, P<0.05], significantly lower percentage of surface cells [(85.15 ± 5.33) % vs. (73.53 ± 6.47) %, (77.68 ± 5.38) %, (68.53 ± 4.20) %, (64.38 ± 4.39) %, P<0.05], and significantly lower percentage of MV value [(92.61 ± 3.74) % vs. (83.52 ± 3.10) %, (87.64 ± 2.95) %, (79.38 ± 3.21) %, (75.63 ± 2.83) %, P<0.05]. Compared with the healthy control group, the ACP, LDH, SDH, and -SH in tongue coating exfoliated cells of subjects with spleen and stomach weakness syndrome, liver and stomach discordance syndrome, and stomach yin deficiency syndrome were all significantly decreased ( P<0.05), while ACP, LDH, SDH, and -SH in subjects with spleen and stomach damp-heat syndrome were all significantly increased ( P<0.05). the percentage of cells in G1 phase of subjects with gastric yin deficiency and spleen and stomach damp-heat syndrome were significantly decreased ( P<0.05), while the percentage of S phase cells were significantly increased ( P<0.05). Spearman rank correlation analysis showed that each syndrome type was correlated with ACP, LDH, SDH, -SH ( r values were 0.608, 0.712, 0.704, 0.631, respectively, all Ps<0.05). Conclusion:Patients with different TCM syndromes of chronic gastritis may have different morphological changes of tongue coating exfoliated cells, large differences in cell cycle, and different levels of cell biochemical indicators.

Objective To investigate the effects of lincRNA-cox2 on the polarization of murine RAW264. 7 macrophages by analyzing the expression of lincRNA-cox2 in RAW264. 7 macrophages of M1 and M2 phenotypes. Methods Murine RAW264. 7 cells were induced by IFN-γand LPS to polarize to M1 phenotype, and were induced by IL-4 to polarize to M2 phenotype. The expression of lincRNA-cox2 in M1 and M2 macrophages were analyzed by real-time quantitative PCR ( RT-PCR) . We designed and synthesized siRNA oligo for lincRNA-cox2 and unrelated sequences. Then the siRNA oligo and NC oligo were transfected into RAW264. 7 cells by LipofectmineTM 2000. The transfected RAW264. 7 cells were induced by IFN-γand LPS or by IL-4 to polarize to M1 or M2 macrophages. Enzyme linked immunosorbent assay ( ELISA) was performed to measure the secretion of IL-10 and IL-12 induced in different conditions. The expression of in-ducible nitric oxide synthase ( iNOS ) , TNF-α, arginase 1 ( Arg-1 ) and found in inflammatory zone 1 (Fizz1) at mRNA level were detected by RT-PCR. The M1 macrophages were transfected with siRNAs to knock down the expression of lincRNA-cox2 for analyzing the biological effects of lincRNA-cox2 on the polar-ization of macrophages. Results The relative expression of lincRNA-cox2 in M1 macrophages was signifi-cantly higher than that in RAW264. 7 cells and M2 macrophages. Compared with the control group, the RAW264. 7 cells transfected with lincRNA-cox2-siRNA showed decreased secretion of IL-12 and inhibited expression of iNOS and TNF-αat mRNA level after IFN-γand LPS induction, but increased secretion of IL-10 and enhanced expression of Arg1 and Fizz1 at mRNA level after IL-4 induction. Transfecting the M1 mac-rophages with lincRNA-cox2-siRNA inhibited the secretion of IL-12, but promoted the secretion of IL-10. Conclusion This study indicated that lincRNA-cox2 was involved in the regulation of macrophage pheno-types by promoting the polarization to M1 macrophages and inhibiting the polarization to M2 macrophages.

Objective To construct a lentiviral vector-based short hairpin RNA (shRNA) targeting the gene encoding tryptophan-aspartate containing coat protein ( TACO) and to evaluate its inhibitory effect on the expression of TACO , and to further elucidate its effects on the phagocytosing and intracellular killing of My-cobacterium tuberculosis (M.tb) by macrophages and the possible mechanisms.Methods Three shRNA frag-ments targeting TACO gene and a scrambling control shRNA fragments were designed and cloned into the lentivi -ral vector pSicoR .The recombinant lentiviral vectors were identified by sequencing analysis and then packed in 293T cells.Real-time RT-PCR and Western blot assay were performed to evaluate the gene-silencing efficiency of the recombinant lentiviral vectors among RAW 264.7 cells transfected with the concentrated lentivirus .The most effective lentivirus was screened out to transfect the RAW 264.7 cells for 48 hours, followed by infection those cells with M.tb strains.The entry and intracellular survival of M .tb strains in RAW264.7 cells were de-termined by bacterial culture at indicated time points .The colocalization of M .tb and lysosomes was detected by immunofluorescence staining .The cyto-ID autophagy kit was used to detect the cellular autophagy and the auto-phagy-associated protein LC 3 was determined by Western blot assay .Results The recombinant lentiviral vec-tors were successfully constructed and confirmed by sequencing analysis .Decreased expression of TACO in RAW264 .7 cells was detected after transfecting the cells with the lentiviral vector-based shRNA vectors targeting TACO gene for 48 hours.The most effective lentivirus , LV-pSRT1, decreased the expression of TACO by 85.24%and 69.00%at the mRNA and protein levels, respectively.The bacterial loads in LV-pSRT1 trans-fected RAW264.7 cells were significantly decreased at the time point of 0 h after M.tb infection as compared with those in the control lentivirus treated cells (5.50×104 vs 8.1 ×104, P<0.05).Compared with the RAW264 .7 cells transfected with control lentivirus , the survival rate of intracellular M .tb strains in LV-pSRT1 transfected cells was significantly decreased at the time point of 48 h (134.54% vs 213.58%, P<0.05) and 72 h (148.18%vs 262.96%, P<0.05) considering the bacterial load at the time point of 0 h as the standard. The immunofluorescence staining demonstrated that the colocalization of M .tb strains with lysosomes was signifi-cantly enhanced in LV-pSRT1 transfected cells as compared with that in control lentivirus treated cells (75.67%vs 10.66%, P<0.05).Moreover, significantly enhanced autophagy and relative expression of LC 3Ⅱ protein were observed in RAW264.7 cells with TACO gene knockdown (16.20%vs 8.50%, P<0.05;0.51 vs 0.34, P<0.05).Conclusion The lentiviral vector-based shRNA targeting TACO gene could effectively knockdown the expression of TACO protein , decrease the entry and increase the intracellular killing of M .tb strains in mac-rophages.The enhanced intracellular killing of M .tb strains by macrophages was associated with the increased fusion of M.tb-containing phagosome and lysosome .