Objective: To investigate the expression of troponin T1(TNNT1) in clinical gastric cancer tissue samples and gastric cancer cell line HGC-27. Methods : The expression of TNNT1 protein in gastric cancer tissues was analyzed using the UALCAN and GEPIA databases to assess its differential expression. Western blot was used to detect the protein expression in clinical samples of gastric cancer tissue. The effects of TNNT1 protein on proliferation, invasion and migration of gastric cancer cell HGC-27 and relationship with clinicopathological features of patients were quantitatively detected and analyzed by MTT and Transwell assays. Results: The expression of TNNT1 protein was up-regulated in HGC-27 cells. The expression of TNNT1 protein was down-regulated(P<0.01) and the proliferation, invasion, migration of HGC-27 cells were inhibited(P<0.01) after transfection with sh-TNNT1. The expression of TNNT1 protein was down-regulated in gastric cancer tissues and correlated with TNM stage(P=0.049) and depth of tumor invasion(P=0.011); There was no significant correlation with patient age, tumor diameter, lymph node metastasis, and tumor differentiation. Conclusion Down-regulating of TNNT1 protein expression can inhibit the proliferation, invasion and migration of gastric cancer cells HGC-27.

Objective To investigate the effects of long non-coding RNA 00894(LINC00894)gene on prolifera-tion and metastasis of human gastric cancer cells,and to verify the regulatory relationship of LINC00894,miR-205-5p and ZEB1 in gastric cancer.Methods The expression level of LINC00894 in gastric cancer cell lines,normal gastric lines,clinical gastric cancer and normal gastric tissue samples were determined by RT-qPCR.Through fol-low-up,the relationship between the expression level of LINC00894 and the prognosis of gastric cancer patients was explored.LINC00894 knockdown cell lines and overexpression cell lines were constructed,and the knockdown and overexpression efficiency was detected by RT-qPCR.Cell proliferation and metastatic capacity were determined by CCK 8,clone formation and Transwell assays.Dual-luciferase reporter assays,RT-qPCR assays and Western blot assays were used to examine the targeted regulatory relationships of LINC00894,miR-205-5p and ZEB1.Results The expression of LINC00894 gene in gastric cancer tissues or cells was significantly higher than that in normal gas-tric tissues or cells,moreover,gastric cancer patients with high LINC00894 gene expression had a worse prognosis.The knockdown of LINC00894 inhibited the viability,clonogenesis,migration and invasion ability of gastric cancer cells,and conversely,the overexpression of LINC00894 obtained the opposite results.LINC00894 promoted ZEB1 expression by targeted downregulation of miR-205-5p expression.LINC00894 promoted the expression of ZEB1 by targeting miR-205-5p and down-regulating its expression.Conclusion LINC00894 serves as an oncogene in gastric cancer and may promote proliferation and metastasis of gastric cancer cells via regulating miR-205-5p/ZEB1 axis.

Objective Based on a novel type of cell death induced by disulfide stress,known as disulfidptosis,this study explores the role of long non-coding RNA(LncRNA)in gastric cancer and establishes a prognosis model re-lated to disulfidptosis,providing a new method for assessing the prognosis of gastric cancer treatment.Methods Transcriptomic data from gastric cancer and normal tissue samples were obtained from the public database TCGA,and disulfidptosis-related LncRNAs were selected through Pearson analysis and LASSO-Cox regression analysis.A relevant prognostic model for gastric cancer was constructed based on the above LncRNAs and validated by function-al enrichment analysis,tumour microenvironment and immune cell infiltration analysis,drug sensitivity analysis and quantitative reverse transcription PCR(RT-qPCR).Results In this study,400 disulfide death-associated LncR-NAs were identified and five of them were screened to construct a prognostic model for assessing the prognosis of gastric cancer patients.The models showed in validation that the survival of the high-risk score group was shorter than that of the low-risk score group(P＜0.05).In addition,the predictive ability of the prognostic model(AUC=0.725)was better than that based only on basic characteristics such as age and gender.The expression levels of disulfide death-associated LncRNAs differed between normal and gastric cancer tissues(P＜0.001).Conclusion The disulfidptosis-related LncRNA prognosis model developed in this study can effectively assess the prognosis of gastric cancer patients and the tumor microenvironment,providing potential targets and a theoretical basis for new immunotherapeutic strategies for gastric cancer.

Objective:To analyze the epidemiological and clinical characteristics of respiratory syncytial virus (RSV) infection in children in Jiangsu Province from 2014 to 2023.Methods:The acute respiratory infection cases in children aged 0-14 years were selected from outpatient/emergency or inpatient departments in 2 surveillance sentinel hospitals, respectively, in Nanjing, Suzhou and Taizhou of Jiangsu from 1 July 2014 to 31 December 2023, and RSV nucleic acid test was conducted and the intensity of the RSV infection was accessed by WHO influenza epidemiological threshold method, and case information and clinical data were collected. χ2 test was used to compare the differences between groups, and the Bonferroni method was used for pairwise comparisons between groups. Results:In 4 946 cases of acute respiratory infections, the RSV positive rate was 8.21% (406/4 946), and the age M( Q1, Q3) of the cases was 1 (0, 3) years. The RSV positive rate was 10.92% (258/2 362) during 2014-2019 and 6.06% (118/1 948) during 2019-2023, the difference was significant ( χ2=31.74, P<0.001). RSV infection mainly occurred from October to March during 2014-2019, with the incidence peak in December and moderate or higher intensity. The seasonality of RSV infection was not obvious during 2019-2023, with low intensity. The RSV positive rate was highest in children in age group 0- years (17.85%, 151/846), and the positive rate declined gradually with age ( χ2=184.51, P<0.001). The RSV positive rate was higher in inpatient cases (9.84%, 244/2 480) than in outpatient/emergency cases (6.57%, 162/2 466) ( χ2=17.54, P<0.001). In the 155 RSV infection cases with complete clinical data, the clinical symptoms mainly included cough (99.35%, 154/155), fever (55.48%, 86/155), and shortness of breath (45.16%, 70/155). In the cases aged <6 months, the proportion of those with fever was low, but the proportion of those with shortness of breath, transferred to intensive care units, and receiving oxygen therapy were higher (all P<0.05). Children aged <6 months and those with underlying diseases were more likely to have severe RSV infection (all P<0.05). Conclusions:RSV infection in children in Jiangsu Province showed seasonal prevalence in winter from 2014 to 2019. Since 2020, the seasonal characteristics of the epidemic have changed, the epidemic period has been dispersed and the epidemic intensity has decreased. Infants <1 year old were at high risk for RSV infection, and those <6 months old and with underlying diseases might have severe infection.

Objective:To analyze the effects of SARS-CoV-2 infection and vaccination on virus mutation.Methods:The whole genome sequencing sequences of 2 659 local SARS-CoV-2 specimens from Jiangsu Province in 2023 were selected for analysis, and relevant information such as demographic and clinical characteristics were collected, and the effects of infection and vaccination on the genome-wide mutation rate and S gene′s selective pressure of the virus were analyzed by univariate and multivariate linear regression models.Results:The average age of these infected patients was 55.0 (31.0, 74.0) years, 1 150 cases (43.2%) in the age group of ≥60 years, 1 367 cases (51.4%) were males, 2 044 cases (76.9%) had a history of COVID-19 vaccination, and 1 629 cases (61.3%) had the first-time infection. The clinical symptoms of the infected patients were mainly mild, with a total of 2434 cases (91.5%), and 29 cases (1.1%) with severe symptoms or more. The average substitution rate of SARS-CoV-2 was 9.69 (9.38, 9.98)×10 -4 subs/site/year, and the dN/dS value of the S gene was 6.08 (5.56, 8.66), which was significantly greater than that of 1 ( P<0.001), indicating positive selection. The result of univariate and multivariate linear regression model analysis showed that the SARS-CoV-2 substitution rate was higher in those with vaccination history and reinfection, aged 20-30 years, ≥60 years, and the SARS-CoV-2 substitution rate was lower in males with moderate clinical symptoms and severe disease and above. Those with a history of vaccination and reinfection, aged 50-60 years old, ≥60 years old have smaller S gene dN/dS. Conclusions:Under the immune pressure exerted by vaccination and infection, the genome-wide mutation of SARS-COV-2 accelerated, but the non-synonymous mutation rate of the S gene decreased. The mechanism causing these phenomena needs further study.

Clinicians need to focus on various points in the diagnosis and treatment of insomnia.This article prescribed the treatment protocol based on the unique features,such as insomnia in the elderly,women experiencing specific physiologi-cal periods,children insomnia,insomnia in sleep-breathing disorder patients,insomnia in patients with chronic liver and kidney dysfunction.It pro-vides some reference for clinicians while they make decision on diagnosis,differentiation and treat-ment methods.

Objective:To develop an integrated point-of-care testing (POCT) reagent for genotying respiratory syncytial virus (RSV) and evaluate its performance.Methods:Specific primers and probes were designed based on the conserved sequences of the genomes of RSV A and B as well as ON1 and BA9 genotypes. The PCR reaction system and conditions were optimized. The vitrification technology of reagents and multiplex detection platform were integrated to develop the RSV genotyping POCT reagent. The sensitivity, specificity, reproducibility, and clinical performance of the product were then evaluated.Results:The sensitivity of the developed integrated RSV genotyping POCT reagent reached 500 copies/ml. It exhibited good specificity with no cross-reaction with clinically similar pathogens. The coefficient of variation of Ct values for both inter-batch and intra-batch reproducibility was less than 5%, indicating good reproducibility. In testing 53 clinical samples, the detection results showed high consistency and concordance with the reference reagent, with a positive concordance rate of up to 98.11%.Conclusions:The developed integrated RSV genotyping POCT reagent incorporates nucleic acid extraction, purification, and detection into a single process, achieving a "sample in, result out" workflow. It is simple to operate and provides accurate, reliable, and stable detection results. This product can be used for the genotyping of RSV A and B in POCT, offering support for the prevention, control, and diagnosis of RSV.

Image 1.

OBJECTIVE To establish the HPLC fingerprint of Jianpi huayu decoction， and to determine the contents of 8 components. METHODS Thermo Hypersil Gold C18 column was used with mobile phase consisted of methanol-0.05% phosphoric acid aqueous solution （gradient elution） at the flow rate of 1.0 mL/min. The column temperature was 30 ℃， the injection volume was 5 μL. The detection wavelength of matrine was 211 nm， and the other components’ detection wavelength was 283 nm. The similarity evaluation of HPLC fingerprints for 10 batches of Jianpi huayu decoction was performed by using the Similarity Evaluation System of Chromatographic Fingerprint of Traditional Chinese Medicine （2012 edition）. The contents of chlorogenic acid， vanillic acid， p-coumaric acid， ferulic acid， hesperidin， quercetin， bergapten and matrine in the samples were determined by HPLC. RESULTS HPLC fingerprint of Jianpi huayu decoction was established. A total of 27 common peaks were identified， and 8 components were identified. The similarity between 10 batches of samples and the control map ranged from 0.942-0.999. RSDs of precision， repeatability and stability tests were less than 3% （n＝6）. The average recoveries of chlorogenic acid， vanillic acid， p-coumaric acid， ferulic acid， hesperidin， quercetin， bergapten and matrine were 99.48%， 101.32%， 101.18%， 100.79%， 101.12%， 99.19%， 99.81% and 102.46%， respectively； RSDs were 1.34%， 0.93%， 1.90%， 1.84%， 0.54%， 1.53%， 1.33% and 1.01%， respectively （n＝6）. The contents were 0.021-0.061， 0.025-0.034， 0.116-0.295， 0.006- 0.062， 0.014-0.053， 0.017-0.026， 0.014-0.027 and 14.05-24.11 mg/g， respectively. CONCLUSIONS The established fingerprint and content determination method can provide a reference for the quality control and subsequent preparation development for Jianpi huayu decoction.

Both cholinergic dysfunction and protein citrullination are the hallmarks of rheumatoid arthritis (RA), but the relationship between the two phenomena remains unclear. We explored whether and how cholinergic dysfunction accelerates protein citrullination and consequently drives the development of RA. Cholinergic function and protein citrullination levels in patients with RA and collagen-induced arthritis (CIA) mice were collected. In both neuron-macrophage coculture system and CIA mice, the effect of cholinergic dysfunction on protein citrullination and expression of peptidylarginine deiminases (PADs) was assessed by immunofluorescence. The key transcription factors for PAD4 expression were predicted and validated. Cholinergic dysfunction in the patients with RA and CIA mice negatively correlated with the degree of protein citrullination in synovial tissues. The cholinergic or alpha7 nicotinic acetylcholine receptor (α7nAChR) deactivation and activation resulted in the promotion and reduction of protein citrullination in vitro and in vivo, respectively. Especially, the activation deficiency of α7nAChR induced the earlier onset and aggravation of CIA. Furthermore, deactivation of α7nAChR increased the expression of PAD4 and specificity protein-3 (SP3) in vitro and in vivo. Our results suggest that cholinergic dysfunction-induced deficient α7nAChR activation, which induces the expression of SP3 and its downstream molecule PAD4, accelerating protein citrullination and the development of RA.