Objective To investigate the expression of GJB4 gene in pancreatic cancer tissue and its correlation with clinicopathology.Methods The expression levels of GJB4 mRNA in pancreatic cancer and adjacent cancer tissues were analyzed using bioinformatics to analyze the Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)RNA sequencing datasets.A total of 120 pancreatic cancer samples and 40 adjacent cancer samples from the Pathology Department of The Second Affiliated Hospital of Kunming Medical University from January 2019 to December 2023 were collected and sorted.Immunohistochemistry staining method was used to detect the expression difference of GJB4 protein between the two groups.RT-qPCR method was used to detect the expression levels of GJB4 in four pancreatic cancer cell lines.Univariate and multivariate Cox regression and Kaplan-Meier curves were used to analyze the clinical pathological data of GJB4 and pancreatic cancer patients.DAVID functional annotation bioinformatics and GSEA enrichment analysis were used to explore the relevant pathways of GJB4 in pancreatic cancer.Results The expression level of GJB4 mRNA in pancreatic cancer was higher than that in adjacent tissues,and the high expression of GJB4 was significantly associated with poor prognosis of patients(P<0.05).Immunohistochemical analysis showed that GJB4 protein was brown-yellow granular in pancreatic cancer tissues,mainly expressed in cytoplasm and cell membrane,and GJB4 protein expression was up-regulated in pancreatic cancer(P<0.05).The RT-qPCR test results showed that out of 4 pancreatic cancer cell lines,3 of them had upregulated expression(P<0.05).COX regression analysis showed that GJB4 gene was an independent risk factor in the prognosis of pancreatic cancer patients.The results of GO enrichment analysis showed that GJB4 was related to the transmembrane transport,ion channel,signal release and membrane potential regulation of pancreatic cancer.GSEA analysis showed that GJB4 was enriched in the Wnt/β-catenin signaling pathway.Conclusion In pancreatic cancer,the high expression level of GJB4 is closely related to the clinicopathological features of the patients,which may predict the poor prognosis of the patients.GJB4 may be involved in regulating the Wnt/β-catenin signaling pathway of pancreatic cancer,and is expected to be one of the potential biomarkers of pancreatic cancer prognosis.

Objective:To analyze the changes in the phylogenetic and antigenic characteristics of the hemagglutinin (HA) gene of influenza virus A(H3N2) [A(H3N2)] during the 2022-2024 influenza seasons in Beijing.Methods:The data of influenza-like cases and A(H3N2) strains from 17 network laboratories and their corresponding sentinel hospitals were collected during the 2022-2024 influenza seasons. The HA genes were amplified and sequenced after extracting nucleic acids of the chosen virus strains. BioEdit, the nucleotide and amino acid sequence identity were conducted, and the maximum likelihood method in MEGA 5.0 software was used to construct the phylogenetic tree of HA genes. Web Logo displayed the amino acid mutation, and the N-glycosylation sites of HA online were analyzed using the NetNGlyc1.0 Server online. The Datamonkey platform was utilized to analyze the positive selection pressure sites of the HA protein.Results:The 2022-2024 influenza season includes 2022-2023 and 2023-2024. During the influenza seasons of 2022-2024, the positive rates of A(H3N2) nucleic acid were 10.35% (2 127/20 543) and 10.47% (4 386/41 876), respectively. In the 2022-2023 influenza season, there were two peaks in the A(H3N2). The comparison of HA genes between all A(H3N2) strains studied with the 2022-2024 vaccine strain (A/Darwin/9/2021) revealed that all of the strains studied have the two amino acid mutations involving 186 and 225 receptor binding sites. There were 31 amino acid substitutions in the 2022-2023 influenza season, of which 18 variant sites involved antigenic determinants. There were 35 amino acid mutations during the 2023-2024 influenza season, of which 14 were related to antigenic determinants. There were changes in the genetic evolutionary subclades of A(H3N2) strains in two influenza seasons: from 2022 to 2023, three evolutionary subclades were co-prevalent together, with the 3C.2a1b.2a.2a.3a.1 accounting for 76.67% (23/30), the 3C.2a1b.2a.1a accounting for 20.00% (6/30), the 3C.2a1b.2a.2a.1 accounting for 3.33% (1/30); from 2023 to 2024, two subclades were prevalent, with 3C.2a1b.2a.2a.3a.1 accounting for 95.12% (39/41) and 3C.2a1b.2a.2a.1 accounting for 4.88% (2/41). The glycosylation site changes of the HA protein of A(H3N2) have been enhanced from 2023 to 2024. The 145 amino acid position of the HA protein of the A(H3N2) was the positive selection site for stress selection site analysis.Conclusions:The evolutionary subclades of the HA gene of A(H3N2) in Beijing showed changes from 2022 to 2024, and the glycosylation site polymorphism of the HA protein of A(H3N2) significantly increased from 2023 to 2024. Continuous monitoring of HA mutations in the A(H3N2) is crucial, providing a basis for developing influenza prevention and control strategies, as well as new strategic support for screening influenza vaccine components, vaccine design, and discovery of drug targets.

Objective:To evaluate the preservation efficacy of 7 non-inactivating virus preservation solutions.Methods:Equal amounts of H1N1 virus were added to 7 commercially available non-inactivating virus preservation solutions, and the samples were stored at -20 ℃, 4 ℃, 25 ℃ and 37 ℃ for 1 hour, 6 hours, 1 day, 3 days, and 5 days. The viral nucleic acid in each simulated sample under different storage conditions was measured using real-time quantitative PCR. The hemagglutination (HA) titer was determined through viral isolation culture and hemagglutination assay, comparing the differences in viral growth activity across different storage solutions and conditions.Results:Except for solution E, the other solutions effectively protected viral nucleic acid at the 4 storage temperatures. In terms of viral activity, solutions A, B, C, and D effectively maintained viral viability. A and B showing the best performance, E and F showed poorer performance, and G performed the worst.Conclusions:Most non-inactivating virus preservation solutions effectively protect viral nucleic acid, but there are significant differences in their ability to maintain viral viability. To ensure optimal virus preservation, it is recommended that medical institutions evaluate the effectiveness of preservation solutions before use.

A 38-year-old male patient with ankylosing spondylitis received subcutaneous injection of adalimumab 40 mg once every 2 weeks. After 21 months of medication, the patient developed fever, fatigue, swelling, and pain in the right neck lymph node and throat. Laboratory tests showed that the tuberculin test was strong positive, mycobacterium tuberculosis γ-interferon release test was 1 911.98 ng/L, and erythrocyte sedimentation rate was 27 mm/1 h. The biopsy of right neck lymph node showed granulomatous inflammation of the lymph node. The patient was diagnosed with cervical lymph node tuberculosis, which was considered to be related to adalimumab. The drug was stopped and anti-tuberculosis treatments were given. The next day, the patient′s temperature returned to normal. After 5 days, the swelling and pain of cervical lymph nodes and throat, and the fatigue were relieved gradually. After 45 days, the above symptoms in the patient disappeared.

Objective:To investigate the related factors of cognitive impairment in middle-aged and old-aged patients with type 2 diabetes mellitus(T2DM).Methods:A total of 970 patients with T2DM(585 middle-aged group and 385 old-aged group)were selected from residents of a large community in Beijing from September to December 2018.The Mini-Mental State Examination(MMSE)was used to assess the cognitive func-tion.Multivariate logistic regression was used to analyze the related factors.Results:The detection rates of cognitive impairment were 12.0％and 13.5％in middle-aged and old-aged patients with T2DM,respectively.Among mid-dle-aged patients with T2DM,work(OR=0.22,95％CI:0.03-0.77)and education at the junior college or un-dergraduate level and above(OR=0.18,95％CI:0.04-0.55)were protective factors for cognitive impair-ment.Myocardial infarction(OR=4.13,95％CI:1.26-13.63)was a risk factor for cognitive impairment.Among old-aged patients with T2DM,drinking tea 1-2 times a week(OR=0.11,95％CI:0.01-0.58)and education at the junior college or undergraduate level and above(OR=0.19,95％CI:0.05-0.54)were protective factors for cognitive impairment.Stroke(OR=3.64,95％CI:1.55-8.39)and good sleep self-assessment(OR=2.75,95％CI:1.13-7.35)were risk factors for cognitive impairment.Conclusion:Cognitive impairment in middle-aged pa-tients with T2DM is related to work,education level and myocardial infarction,and cognitive impairment in old-aged patients with T2DM is related to lifestyle,education level and stroke.

Objective:To analyze the changes in the phylogenetic and antigenic characteristics of the hemagglutinin (HA) gene of influenza virus A(H3N2) [A(H3N2)] during the 2022-2024 influenza seasons in Beijing.Methods:The data of influenza-like cases and A(H3N2) strains from 17 network laboratories and their corresponding sentinel hospitals were collected during the 2022-2024 influenza seasons. The HA genes were amplified and sequenced after extracting nucleic acids of the chosen virus strains. BioEdit, the nucleotide and amino acid sequence identity were conducted, and the maximum likelihood method in MEGA 5.0 software was used to construct the phylogenetic tree of HA genes. Web Logo displayed the amino acid mutation, and the N-glycosylation sites of HA online were analyzed using the NetNGlyc1.0 Server online. The Datamonkey platform was utilized to analyze the positive selection pressure sites of the HA protein.Results:The 2022-2024 influenza season includes 2022-2023 and 2023-2024. During the influenza seasons of 2022-2024, the positive rates of A(H3N2) nucleic acid were 10.35% (2 127/20 543) and 10.47% (4 386/41 876), respectively. In the 2022-2023 influenza season, there were two peaks in the A(H3N2). The comparison of HA genes between all A(H3N2) strains studied with the 2022-2024 vaccine strain (A/Darwin/9/2021) revealed that all of the strains studied have the two amino acid mutations involving 186 and 225 receptor binding sites. There were 31 amino acid substitutions in the 2022-2023 influenza season, of which 18 variant sites involved antigenic determinants. There were 35 amino acid mutations during the 2023-2024 influenza season, of which 14 were related to antigenic determinants. There were changes in the genetic evolutionary subclades of A(H3N2) strains in two influenza seasons: from 2022 to 2023, three evolutionary subclades were co-prevalent together, with the 3C.2a1b.2a.2a.3a.1 accounting for 76.67% (23/30), the 3C.2a1b.2a.1a accounting for 20.00% (6/30), the 3C.2a1b.2a.2a.1 accounting for 3.33% (1/30); from 2023 to 2024, two subclades were prevalent, with 3C.2a1b.2a.2a.3a.1 accounting for 95.12% (39/41) and 3C.2a1b.2a.2a.1 accounting for 4.88% (2/41). The glycosylation site changes of the HA protein of A(H3N2) have been enhanced from 2023 to 2024. The 145 amino acid position of the HA protein of the A(H3N2) was the positive selection site for stress selection site analysis.Conclusions:The evolutionary subclades of the HA gene of A(H3N2) in Beijing showed changes from 2022 to 2024, and the glycosylation site polymorphism of the HA protein of A(H3N2) significantly increased from 2023 to 2024. Continuous monitoring of HA mutations in the A(H3N2) is crucial, providing a basis for developing influenza prevention and control strategies, as well as new strategic support for screening influenza vaccine components, vaccine design, and discovery of drug targets.

Objective:To document any effect of repeated high-frequency repetitive transcranial magnetic stimulation (rTMS) using double-cone coils on the lower limb motor function of stroke survivors.Methods:A total of 40 stroke survivors were randomly divided into an rTMS group and a sham stimulation group, each of 20. The rTMS group received rTMS at 10Hz with a double-cone coil, while a coil that produced sound but no magnetic stimulation was used with the sham group. The treatments were administered daily, five times a week, for three weeks. Before as well as after 1, 2 and 3 weeks of treatment, lower limb motor function, balance, and the root mean square (RMS) and median frequency (MF) of the rectus femoris and tibialis anterior muscles were evaluated using the Fugl-Meyer lower extremity assessment (FMA-LE), the Berg Balance Scale (BBS), the Modified Barthel Index (MBI), Brunnstrom staging, the TecnoBody balance assessment system, and surface electromyography.Results:Compared with the sham stimulation group, the BBS score of the rTMS group was significantly higher after 2 weeks of treatment, and both the FMA-LE and BBS scores were significantly higher after 3 weeks. The average Brunnstrom stage in the sham group had increased significantly after 3 weeks, but in the rTMS group it had increased after 2 weeks. By 3 weeks there were no significant differences between the two groups. In terms of movement control, the average motion ellipse area in the rTMS group was significantly smaller than among the sham group after 2 weeks, and after 3 weeks the average motion trajectory length was significantly shorter than in the sham group. The average RMS of the rectus femoris in the rTMS group was significantly higher than the sham group′s average after 3 weeks of treatment, indicating improved muscle activation.Conclusions:High-frequency rTMS using a conical coil can effectively improve the lower limb motor function and balance ability of stroke survivors, demonstrating promising clinical application potential.

Objective:To investigate the related factors of cognitive impairment in middle-aged and old-aged patients with type 2 diabetes mellitus(T2DM).Methods:A total of 970 patients with T2DM(585 middle-aged group and 385 old-aged group)were selected from residents of a large community in Beijing from September to December 2018.The Mini-Mental State Examination(MMSE)was used to assess the cognitive func-tion.Multivariate logistic regression was used to analyze the related factors.Results:The detection rates of cognitive impairment were 12.0％and 13.5％in middle-aged and old-aged patients with T2DM,respectively.Among mid-dle-aged patients with T2DM,work(OR=0.22,95％CI:0.03-0.77)and education at the junior college or un-dergraduate level and above(OR=0.18,95％CI:0.04-0.55)were protective factors for cognitive impair-ment.Myocardial infarction(OR=4.13,95％CI:1.26-13.63)was a risk factor for cognitive impairment.Among old-aged patients with T2DM,drinking tea 1-2 times a week(OR=0.11,95％CI:0.01-0.58)and education at the junior college or undergraduate level and above(OR=0.19,95％CI:0.05-0.54)were protective factors for cognitive impairment.Stroke(OR=3.64,95％CI:1.55-8.39)and good sleep self-assessment(OR=2.75,95％CI:1.13-7.35)were risk factors for cognitive impairment.Conclusion:Cognitive impairment in middle-aged pa-tients with T2DM is related to work,education level and myocardial infarction,and cognitive impairment in old-aged patients with T2DM is related to lifestyle,education level and stroke.

Objective:To document any effect of repeated high-frequency repetitive transcranial magnetic stimulation (rTMS) using double-cone coils on the lower limb motor function of stroke survivors.Methods:A total of 40 stroke survivors were randomly divided into an rTMS group and a sham stimulation group, each of 20. The rTMS group received rTMS at 10Hz with a double-cone coil, while a coil that produced sound but no magnetic stimulation was used with the sham group. The treatments were administered daily, five times a week, for three weeks. Before as well as after 1, 2 and 3 weeks of treatment, lower limb motor function, balance, and the root mean square (RMS) and median frequency (MF) of the rectus femoris and tibialis anterior muscles were evaluated using the Fugl-Meyer lower extremity assessment (FMA-LE), the Berg Balance Scale (BBS), the Modified Barthel Index (MBI), Brunnstrom staging, the TecnoBody balance assessment system, and surface electromyography.Results:Compared with the sham stimulation group, the BBS score of the rTMS group was significantly higher after 2 weeks of treatment, and both the FMA-LE and BBS scores were significantly higher after 3 weeks. The average Brunnstrom stage in the sham group had increased significantly after 3 weeks, but in the rTMS group it had increased after 2 weeks. By 3 weeks there were no significant differences between the two groups. In terms of movement control, the average motion ellipse area in the rTMS group was significantly smaller than among the sham group after 2 weeks, and after 3 weeks the average motion trajectory length was significantly shorter than in the sham group. The average RMS of the rectus femoris in the rTMS group was significantly higher than the sham group′s average after 3 weeks of treatment, indicating improved muscle activation.Conclusions:High-frequency rTMS using a conical coil can effectively improve the lower limb motor function and balance ability of stroke survivors, demonstrating promising clinical application potential.

A 38-year-old male patient with ankylosing spondylitis received subcutaneous injection of adalimumab 40 mg once every 2 weeks. After 21 months of medication, the patient developed fever, fatigue, swelling, and pain in the right neck lymph node and throat. Laboratory tests showed that the tuberculin test was strong positive, mycobacterium tuberculosis γ-interferon release test was 1 911.98 ng/L, and erythrocyte sedimentation rate was 27 mm/1 h. The biopsy of right neck lymph node showed granulomatous inflammation of the lymph node. The patient was diagnosed with cervical lymph node tuberculosis, which was considered to be related to adalimumab. The drug was stopped and anti-tuberculosis treatments were given. The next day, the patient′s temperature returned to normal. After 5 days, the swelling and pain of cervical lymph nodes and throat, and the fatigue were relieved gradually. After 45 days, the above symptoms in the patient disappeared.