Humans ; Amyloidosis ; Amyloid ; Cardiomyopathies

Objective:To analyze the molecular evolutional characteristics of the hemagglutinin and neuraminidase genes of influenza A (H3N2) viruses isolated in Yancheng from 2022 to 2024.Methods:The throat swab specimens of influenza-like illness ( ILI) from sentinel surveillance hospital and outbreak sites were detected using the method of real time Rt-qPCR. The influenza A(H3N2) viruses were isolated using MDCK cells culture method from April 2022 to Marh 2024. The strains isolated from 2022 to 2024 were selected randomly and their sequences of the HA1 and NA genes were amplified through one step RT-PCR method and the PCR products were sequenced.The nucleotide and amino acid site variations and evolutionary characteristics of the genes were analyzed using relevant bioinformatics software. The mutations of genes and nucleic acid locus were analyzed and the evolutional trees were generated using bioinformatics software.Results:A total of 5 020 samples were collected between April 2022 and March 2024, the positive detection rate of influenza virus nucleic acid was 18.59%(933/5 020).The winter and spring influenza peaks were obvious in the two monitoring seasons from April 2022 to March 2024. Among them, the summer influenza peak was obvious in the monitoring season from April 2022 to March 2023, and the H3N2 subtype influenza virus was the dominant epidemic strain in the two monitoring seasons. Genetic evolution tree displayed: the clustering relationships of the respective branches of HA1 and NA genes of 32 strains isolated in Yancheng were basically the same.The HA1 and NA genes of 24 strains isolated from 2023-2024 in Yancheng and the 2022-2024 Northern Hemisphere vaccine strain A/Darwin/9/2021 (H3N2) were located in the 3C.2a1b2a.2a.3a.1 evolutionary lineage, while the 8 strains isolated in the 2022 in Yancheng and the 2021-2022 Northern Hemisphere vaccine strain A/Cambodia/e0826360/2020 (H3N2) were located in the 3C.2a1b.2a.1a evolutionary lineage.The 6 strains (A/JSTH/11735/2023, A/JSTH/11788/2023, A/JSTH/11974/2023, A/JSYD/353/2023, A/JSYD/354/2023, A/JSTH/138/2023) all exhibited variations in the F79L, N122D, P239S, and K276E amino acid sites, which were present in both sporadic and outbreak strains. Because the strains of the antigen epitopes, receptor binding sites and glycosylation sites in the HA1 genes had a certain degree of variations in Yancheng in the 2022-2024 year, the immunogenicity matching between the 24 strains isolated in the 2023-2024 and the Northern Hemisphere vaccine strain A/Darwin/9/2021 was good, while the immunogenicity matching between the 8 strains isolated in the 2022 and the Northern Hemisphere vaccine strain A/Cambodia/e0826360/2022 was good; 32 strains isolated from 2022 to 2024 had no mutations in catalytic residues and drug resistant sites of NA genes.Conclusion:These result indicated that the HA1 and NA genes of influenza A/H3N2 viruses circulated in Yancheng city from 2022 to 2024 are changed gradually.The accumulation of these mutations would result in antigenic drift of influenza A(H3N2) viruses and increase the mismatching of the recommended vaccine strain.Compared with the vaccine strain A/Darwin/9/2021(H3N2), the strains isolated in the 2022 had substantially result in antigenic drift on the whole.The influenza A(H3N2) viruses surveillance should be strengthened to find the new mutant of virus in time.

Objective To investigate the current status and awareness of pharmaceutical services in hospitals in Guizhou province and to provide a reference for exploring and carrying out pharmaceutical service fees.Methods The questionnaire was designed by the"wjx.cn"website.Three kinds of questionnaires were designed for pharmacists,doctors,nurses,and patients as the research objects,with corresponding differences in some questions,and promoted on WeChat,Dingxiangyuan,and other network platforms.Results A total of 655 questionnaires were collected,and 639 valid questionnaires were recovered,with an effective recovery rate of 97.56%.324 pharmacists(50.70%),82 doctors and nurses(12.83%),233 patients(36.46%)were surveyed.The average approval score of these three groups of respondents on pharmaceutical service fees was 4.67,4.23,and 4.22,respectively(full score:5).Conclusions Overall,pharmacists'professional services have received support from medical staff and patients.However,patients'pharmaceutical service projects currently focus on dispensing services.The recognition of pharmacists'work and the public's awareness of pharmaceutical services can be improved by enhancing the professional ability of pharmacists,strengthening publicity and guidance,and exploring"Internet+pharmaceutical services",etc.,to promote the sustainable development of pharmaceutical services.

Objectives:To discuss the inhibitory effect of Lactobacillus reuteri on the replication of rotavirus(RV)strain SA11 in vivo and in vitro,and to clarify its effect on the expression of related immune factors.Methods:For in vitro experiments,Lactobacillus reuteri was cultured and identified,and the standard curve and growth curve were plotted to screen the optimal time and concentration for Lactobacillus reuteri cultivation.The cells were infected with Lactobacillus reuteri at the concentrations of 5×108,10×108,50×108,100×108,200×108,and 500×108 CFU·mL-1,and the surival rates of Caco-2 cells were detected by trypan blue staining method.Various concentrations of Lactobacillus reuteri were co-incubated with RV in vitro and applied to the Caco-2 cells.The cells were divided into negative control group(NC group),positive control group(PC group),and 107,108,109,and 1010 CFU·mL-1 Lactobacillus reuteri groups.Immunofluorescence focus method was used to detect the viral titers in the Caco-2 cells after treated with Lactobacillus reuteri and real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the copy numbers of RV VP6 gene in the Caco-2 cells after treated with various concentrations of Lactobacillus reuteri.In in vivo experiments,25 litters of SPF suckling mice were divided into control group,RV group(infected with SA11 strain),Ab-NC group(treated with antibiotic to deplete gut microbiota),Ab-RV group(depleting gut microbiota and then infected with SA11 strain),and Ab-Lac-RV group(depleting gut microbiota,treated with Lactobacillus reuteri,and then infected with SA11 strain).The fecal samples were collected on days 2,4,6,8,and 10 gavage,colon tissue sample were collected on day 4 of and RT-qPCR method was used to detect the copy numbers of RV VP6 gene in feces and the mRNA expression levels of interleukin(IL)-1β,IL-8,IL-10,interferon-γ(IFN-γ),and tumor necrosis factor-α(TNF-α)in colon tissue of the suckling mice in vartious groups.Results:The Lactobacillus reuteri grew well,with round,smooth,and milky white convex colonies and neat edges.After Gram staining,the bacteria appeared purple,irregular,and square-shaped rods.16SrDNA sequencing showed 99%sequence homology,indicating successful activation of Lactobacillus reuteri.The number of live Lactobacillus reuteri was linearly related to the absorbance(A)value,and the standard curve for regression analysis was Y=0.437 5X+0.000 6,R2=0.999 4.During the 0-2 h cultivation period,the bacteria were at the logarithmic growth phase with slow growth;from 2-14 h,the bacteria grew rapidly and stabilized at 14-16 h,reaching the growth rate peak at 16 h,after which they entered the decline phase.Infection with Lactobacillus reuteri at concentrations of 5×108,10×108,50×108,100×108,and 200×108 CFU·mL-1 resulted in the survival rates of Caco-2 cells were all>90%,so these concentrations were selected for the further experiments.Compared with PC group,the copy numbers of RV VP6 gene in the Caco-2 cells in 5×108,10×108,50×108,100×108,and 200×108 CFU·mL-1 Lactobacillus reuteri groups were significantly decreased(P<0.01).Compared with PC group,the viral titers in the Caco-2 cells in 107,108,109,and 1010 CFU·mL-1 Lactobacillus reuteri groups were significantly decreased(P<0.01).Compared with control group,the numbers of gut microbiota colonies in Ab-NC,Ab-RV,and Ab-Lac-RV groups were significantly decreased,indicating successful depletion of gut microbiota in the suckling mice.On days 2 and 4 after gavage,the RV VP6 gene copy number in the feces in Ab-RV group was significantly lower than that in RV group(P<0.05).On days 4,6,8,and 10 after gavage,the RV VP6 gene copy number in the feces in Ab-Lac-RV group was significantly lower than that in Ab-RV group(P<0.05).Compared with control group,the expression levels of IL-1β,IL-10,IFN-γ,and TNF-α mRNA in colon tissue in Ab-RV and Ab-Lac-RV groups were significantly increased(P<0.05 or P<0.01),while the expression level of IL-8 mRNA was significantly decreased(P<0.05),and the expression level of IL-10 mRNA in colon tissue in Ab-LAC-RV group was significantly increased(P<0.01).Conclusion:Lactobacillus reuteri may inhibit the RV replication by upregulating the expressions of IL-1β,IL-10,IFN-γ,and TNF-α mRNA and downregulating the expression of IL-8 mRNA.

Objective:To accurately ascertain the whole genome sequencing and the composition of open reading frames (ORFs) of non-replicating Tiantan strain of vaccinia virus (NTV) using next-generation long-read sequencing technology.Methods:NTV, obtained from our laboratory stock, was amplified and purified on chicken embryo fibroblast cells(CEFs), and the full-length genomic nucleic acid of NTV was extracted. The PacBio HiFi sequencing platform was utilized for de novo assembly to obtain the complete genomic sequence of NTV. Using a homology annotation strategy, we identified its ORF composition and compared it with known non-replicating vaccinia virus strains. Results:The total length of NTV′s genome was 171 729 bp, with a GC content of 33%. Its unique inverted terminal repeat (ITR) region comprised hairpin structures, two tandem repeat regions, and three non-repeat regions. NTV contained 166 ORFs, with major differences observed in the ITR and its surrounding regions when compared to MVA-BN and NYVAC. These three strains shared a common set of 138 ORFs. NTV encoded six unique ORFs related to virus evasion of host antiviral response.Conclusions:This study accurately determines the whole genome sequencing and ORFs composition of NTV, and reveals its similarities and differences with other replication-deficient vaccinia virus strains, which pave a way for the development and application of the next generation of monkeypox vaccines and novel viral vectors.

Objective:To innovatively applicate and evaluate the efficacy of simultaneous off-pump coronary artery bypass grafting (OPCABG), modified pulmonary vein isolation and left atrial ablation treating coronary heart disease combined with atrial fibrillation.Methods:From January 2021 to August 2023, a retrospective analysis was conducted on the clinical data of 76 patients who underwent OPCABG, modified pulmonary vein isolation and left atrial ablation in our department. There were 57 males and 19 females, aged(63.42 ± 8.25)years, with a median duration of 26 months of atrial fibrillation. Follow up was conducted for 1 year after surgery, and 24-hour dynamic electrocardiogram was rechecked to observe the recurrence of atrial fibrillation.Results:All patients successfully completed the surgery without any conversion to extracorporeal circulation, and no perioperative death. On the first day after surgery, there was a conversion to sinus or junctional rhythm in 69(90.79%) cases. The median postoperative hospitalization time was 7 days, with no postoperative cerebral infarction or complications of thoracotomy hemostasis. All patients recovered and were discharged, with a sinus rhythm maintained at 67(88.16%) upon discharge. Compared with preoperation, there were 56 (73.68%) patients with sinus rhythm after 1 year of postoperative follow-up( P<0.05). Conclusion:The simultaneous OPCABG, modified pulmonary vein isolation and left atrial ablation treating coronary heart disease with atrial fibrillation was safe, feasible, and effective. The mid- and long-term efficacy needs to be further confirmed through multicenter and large-scale studies.

BACKGROUND: The purpose of this study was to investigate the specific effects of signal transducer and activator of transcription 4 (STAT4)-induced long intergenic nonprotein coding RNA 1278 (LINC01278) on the growth of non-small cell lung cancer (NSCLC) cells involved in the microRNA (miR)-877-5p/activated transcription factor 4 (ATF4) axis. METHODS: NSCLC tumor tissue and adjacent normal tissue were collected. Human normal lung epithelial cell BEAS-2B and human NSCLC cell lines (H1299, H1975, A549, H2228) were collected. The expression levels of STAT4, LINC01278, miR-877-5p, and ATF4 were detected. A549 cells were screened for subsequent experiments. The proliferation ability of cells was detected by colony formation experiment. Cell apoptosis was tested by flow cytometry. Scratch test and transwell assay were used to detect the migration and invasion ability of cells. Biological function of LINC01278 in NSCLC was confirmed by xenograft experiments. RESULTS: Low expression miR-877-5p and high expression of STAT4, LINC01278 and ATF4 were detected in NSCLC.Silenced LINC01278 in A549 cell depressed cell proliferation, migration and invasion, but facilitated cell apoptosis.LINC01278 was positively correlated with STAT4 and could directly bind to miR-877-5p. Upregulating miR-877-5p suppressed NSCLC cell progression, while downregulating miR-877-5p had the opposite effect. Upregulating miR-877-5p abrogated the effects of silenced LINC01278 on NSCLC cell progression. MiR-877-5p targeted ATF4. ATF4 upregulation could partly restore the carcinogenic effect of LINC01278 in vitro and in vivo. CONCLUSION Our data supports that STAT4-induced upregulation of LINC01278 promotes NSCLC progression by modulating the miR-877-5p/ATF4 axis, suggesting a novel direction for NSCLC treatment.

Objective:To evaluate the prognosis of off-pump coronary artery bypass grafting combined(OPCABG) with coronary endarterectomy(CE) treating the diffuse coronary artery disease.Methods:From January 2012 to December 2014, the clinical data of 2 496 OPCABG patients in our department were retrospectively analyzed, and they were divided into OPCABG group and OPCABG+ CE group. After 1∶1 matching via the propensity score matching method, the perioperative prognosis, long-term survival and adverse cardiovascular and cerebrovascular events(MACCE) were compared between the two groups.Results:A total of 238 pairs of patients were included after propensity score matching. The incidence of postoperative AMI in the OPCABG+ CE group was significantly higher than that in the OPCABG group(5.04% vs. 1.68%, P=0.042). With an average follow-up of 7.3 years, there was no significant difference in the cumulative survival rate(92.44% vs. 88.65%, P=0.159) and long-term MACCE(10.92% vs. 15.13%, P=0.173) between the two groups. Compared with the OPCABG group, the recurrence of angina pectoris(CCS grade Ⅲ-Ⅳ) in the OPCABG+ CE group increased significantly(20.16% vs. 12.60%, P=0.026). Conclusion:The risk of early AMI and long-term angina recurrence after OPCABG+ CE is significantly increased, but the long-term survival and MACCE of OPCABG+ CE and OPCABG are comparable.

Objective:To develop a recombinase-aided amplification (RAA)-clustered regularly interspaced short palindromic repeats(CRISPR)/Cas12a-based nucleic acid assay for monkeypox virus with high specificity and sensitivity.Methods:RAA primers and CRISPR RNA (crRNA) were designed based on the known conserved regions of the monkeypox virus gene and synthesized, and specific crRNAs were screened using fluorescence detection. The sensitivity and specificity of the detection system were evaluated.Results:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was developed with a sensitivity of 2.5 copies/reaction and high specificity without cross-reactivity with ectromelia virus and vaccinia virus.Conclusions:An RAA-CRISPR/Cas12a-based nucleic acid assay for monkeypox virus was established, which would provide a powerful tool for efficient, rapid and specific detection of monkeypox virus.

Objective:Based on targeted amplicon technology combined with high-throughput sequencing technology and bioinformatic analysis technology, to understand the characteristics of the whole genome of the monkeypox virus and its variation, and to construct a method for the analysis of monkeypox virus variation and molecular traceability of the case in Qinghai province, and to provide technical support for the prevention and control of monkeypox epidemic in the future.Methods:The extracted viral DNA was used as a template, and the genome of monkeypox virus was specifically amplified by Ion AmpliSeq Monkeypox Panel with the number of amplicons 1 609 and the length of 125 bp-275 bp, and the sequencing library was constructed by Ion AmpliSeq Library Kit Plus, and sequenced by Ion Torrent GeneStudio S5. The sequencing library was constructed by Ion AmpliSeq Library Kit Plus, and the monkeypox virus genome was sequenced using Ion Torrent GeneStudio S5 sequencer. Monkeypox virus was analyzed for genomic profiling and mutation site analysis using the online analysis tool Nextclade. The genomic sequence of the case virus in this study was compared with some sequences in the GIASID monkeypox virus database and a phylogenetic tree was constructed to analyze the potential origin of the case virus.Results:The Ct values of monkeypox virus genes in the rash swab and oropharyngeal swab samples were 32.13 and 36.91, respectively. The rash swab sample had a reads number match of 99.99% and a genome coverage of 99.45% after whole-genome sequencing of monkeypox virus, and the sequences belonged to the IIb (West African branch) B. 1.3 type. The analysis of nucleotide mutation sites and phylogenetic tree showed that the sequences were in the same branch with four monkeypox virus genome sequences recently submitted by China and Japan in the GISAID monkeypox virus database, and had the closest evolutionary relationship with the sequence EPI_ISL_18059184 (sampled on 2023-07-03) submitted by Yunnan, China, which shared 82 single-nucleotide mutation sites, among which the sequence from Yunnan was only present in all of the shared 82 single-nucleotide mutation sites. The sequence in this study has 2 additional nucleotide mutation sites on top of the shared 82 single nucleotide mutation sites. The sequence submitted by Japan, EPI_ISL_17692269 (sampled on 2023-04-28), is more closely related in evolution, sharing 78 single nucleotide mutation sites, with 7 single nucleotide mutation site differences, and the Japanese sequence shares 78 single nucleotide mutation sites. The Japanese sequence shared 78 mutation sites with one additional nucleotide mutation site (G57786A), while the present sequence had six additional nucleotide mutation sites (G13563A, C21062T, G101241A, C142797T, G152866A, T169721A).Conclusions:The whole genome sequence of monkeypox virus of 197 084 bp was successfully obtained from a sample with low viral load, and the average. We constructed a method for sequencing and analyzing the whole genome of monkeypox virus.