ObjectiveTo investigate the effect of interfering with long noncoding RNA （LncRNA） small nucleolar RNA host gene16 （SNHG16） on improving angiogenesis of ectopic endometrial stromal cells （EScs） in adenomyosis （AM） by targeting upregulation of miRNA （miR）-141-3p and inhibition of high mobility group box-1 protein （HMGB1）. MethodsThe expression levels of SNHG16， miR-141-3p and HMGB1 mRNA in endometrial tissues of 52 patients with adenomyosis （AM group） and 52 patients who needed hysterectomy due to cervical cancer or ovarian cancer （control group） were detected by quantitative reverse transcription polymerase chain reaction （qRT-PCR）. Y14 cells were divided into small hairpin RNA （shRNA） NC group， shRNA SNHG16 group， shRNA SNHG16+miR-141-3p inhibitor （inhibitor） group， shRNA SNHG16+inhibitor NC group and blank group. The targeting relationship between miR-141-3p and SNHG16 as well as HMGB1 was verified. The expression levels of SNHG16， miR-141-3p and HMGB1 mRNA in Y14 cells were detected by qRT-PCR. CCK-8 and Transwell assay were used to detect cell proliferation， invasion and migration. Microvascular density （MVD） was determined by immunofluorescence. The expressions of hypoxia-inducing factor α （HIF-1α）， cyclooxygenase-2 （Cox-2）， vascular endothelial growth factor （VEGF） and HMGB1 were detected by Western blot. ResultsThe expression of SNHG16 and HMGB1 mRNA in AM group was higher than that in control group， but the expression of miR-141-3p was lower than that in control group （P0.05）. The expression of SNHG16， HMGB1 mRNA， proliferation rate， migration， invasion number， MVD， expression of VEGF， HIF-1 α， Cox-2， and HMGB1 proteins in the shRNA SNHG16 group were lower than those in the blank group and shRNA NC group， while the expression of miR-141-3p was higher than that in the blank group and shRNA NC group （P0.05）. Inhibition of miR-141-3p reversed the improvement of EScs angiogenesis by interfering with SNHG16. ConclusionInterference with LncRNA SNHG16 improves EScs angiogenesis and inhibits proliferation， migration， and invasion of EScs by targeting upregulation of miR-141-3p and inhibition of HMGB1.

Objective To compare the effect of different intensity of neuromuscular training(NMT)on muscle strength and knee joint function of patients after anterior cruciate ligament reconstruction(ACLR).Methods From January,2023 to January,2024,60 ACLR patients in Changhai Hospital were selected,and they received the same intensity of NMT from one to eight weeks after surgery.Eight weeks after surgery,they were randomly divided into low intensity group(n=30)and high intensity group(n=30),and then they received different inten-sities of NMT from nine to 16 weeks after surgery,each training session lasted one hour,with three sessions per week,totaly 48 sessions.The Lysholm score,knee flexor and extensor muscle strength and muscle endurance-were compared at eight weeks and 16 weeks after surgery.Results After group training,the Lysholm score significantly increased in both groups(|t|>13.739,P<0.001),and was higher in the high intensity group than in the low intensity group(t=-2.574,P<0.05);in the high intensity group,the relative peak torque and endurance of the extensor and flexor muscles improved at angular velocities of 60°/s,120°/s and 180 °/s(|t|>2.320,P<0.05);in the low intensity group,the flexor peak torque improved at all the three angular velocities(t>2.177,P<0.05),the extensor peak torque improved at angular velocities of 60°/s and 180°/s(|t|>1.715,P<0.05),and the extensor endurance improved at angular velocity of 60°/s(t=-2.293,P<0.05).However,there was no significant difference in the relative peak torque and endurance of the extensor and flexor muscles at all the three angular velocities(P>0.05).Conclusion Both high and low intensity NMT could improve the muscle strength,muscle endurance and knee joint func-tion.Maybe,high intensity is superior to low intensity.Further verification is still needed.

Aim To explore the mechanism of rhein in the treatment of IgA nephropathy in rats by impro-ving spleen immune function based on mTORC1 path-way.Methods Thirty-two SD rats were randomly di-vided into control group,model group(IgAN),rhein group(RH)and rapamycin group.The deposition of IgA in mesangial region and pathological changes of spleen and kidney were observed,and the ratio of Th/Ts in spleen was determined by immunohistochemical method.The protein expressions of p-mTOR,p-p70S6k,TGF-β1 and TNF-α in spleen were detected by Western blot.Results Compared with the control group,IgAN group significantly increased urinary sludge red blood cell count,24 hour urinary total pro-tein,serum creatinine,blood urea nitrogen,serum IgA,Th/Ts,p-mtor,p-P70S6K and spleen TNF-αprotein expression levels(P＜0.01),and spleen TGF-β1 protein expression(P＜0.05).IgA deposi-tion in mesangial area of the kidney increased signifi-cantly,and pathological changes were observed in spleen and kidney.Compared with IgAN group,urine sludge red blood cell count,serum IgA,Th/Ts,p-mTOR,p-P70S6K and TNF-α protein expression lev-els in RH and RAPA groups were significantly reduced(P＜0.01).24 hour urinary total protein,blood urea nitrogen,serum creatinine and spleen TGF-β1 protein decreased(P＜0.05),the deposition of IgA in the mesangial region of the kidney decreased,and the pathological changes of spleen and kidney were allevia-ted.There was no significant difference between RH group and RAPA group.Conclusions RH can allevi-ate IgA nephropathy by inhibiting the activity of m-TORC1 pathway in the spleen of IgA nephropathy rats,thereby inhibiting the differentiation of splenic lympho-cytes and reducing the secretion of IgA.

Objective This study examines the factors influencing oral frailty in the elderly community,develops a risk prediction model,and validates its efficacy,so as to provide references for identifying and preventing oral weakness in the elderly.Methods 556 elderly individuals from 4 communities were selected by convenience sampling from June to August 2024 in Zigong City Sichuan Province.They were randomly divided into a training group(383 cases)and a validation group(165 cases).Data were collected by a general information questionnaire,Social Frailty Scale,Geriatric Depression Scale,and the Oral Frailty Index-8 screening tool.Logistic regression was used to determine the influencing factors,and R software was used to establish a nomogram model for predicting the risk of oral frailty.Bootstrap method and the validation group were used for internally validation of the model.Calibration curve was used to evaluate the prediction performance of the model.Results 548 valid questionnaires were collected.The final model variables included whether the age ≥80 years,wearing removable dentures,reduced frequency of going out,brushing teeth less than twice a day,frequent dry mouth,increased difficulty in eating hard foods,and choking.The area under the receiver operating characteristic curve of the training group was 0.95(95％CI:0.93～0.97),and the best cutoff value was 0.687.The model achieved an accuracy of 87％,sensitivity of 91％,specificity of 85％,positive predictive value of 0.75,and negative predictive value of 0.95.The Hosmer-Lemeshow fitting test show that x2=3.036,P=0.932,indicating a good model fit.Conclusion The oral frailty prediction model demonstrated a good discrimination,calibration,and clinical utility,which can provide a scientific basis for the prevention and early screening of oral frailty in the elderly.

Objective To explore the relationship between triglyceride-glucose(TyG)index and hypertension in patients with cerebral hemorrhage.Methods A total of 1718 patients with cere-bral hemorrhage admitted to our hospital from January,2013 to May,2023 were enrolled in this study.According to the TyG index quartile,437 cases were assigned into Q1 group(≤8.375),424 cases into Q2 group(TyG index 8.376～8.737),429 cases into Q3 group(TyG index 8.738～9.087),and 428 cases into Q4 group(≥9.088).The general clinical data were compared in the four groups.Logistic regression analysis was used to study the correlation between TyG index and hy-pertension.Results There were significant differences among the four groups in terms of age,hy-pertension,diabetes,SBP,DBP,TC,TG,HDL-C,LDL-C,FBG,glycated hemoglobin and TyG in-dex(P＜0.05,P＜0.01).Logistic regression analysis showed that when the TyG index was a con-tinuous variable,it was significantly correlated with the risk of hypertension(OR=1.999,95％CI:1.393-2.869,P=0.001).When the index was used as a categorical variable,with Q1as a ref-erence,TyG index in Q3 and Q4 was associated with an increase in OR of hypertension(OR=1.869,95％CI:1.220-2.865,P=0.004;OR=1.844,95％CI:1.125-3.020,P=0.015).After ad-justing cofounders,the association of TyG index and risk of hypertension was stronger in the fe-males(OR=2.618,95％CI:1.312-5.221,P=0.006)than the males(OR=1.783,95％CI:1.151-2.761,P=0.010),and in the patients ≥65 years old(OR=3.277,95％CI:1.600-6.741,P=0.001)than those＜65 years old(OR=1.782,95％CI:1.076-2.949,P=0.025).Conclusion TyG index is closely associated with hypertension in patients with cerebral hemorrhage,especially in women and elderly.

Aim To investigate the protective effect of Gualou Guizhi granules(GLGZG)on neuronal injury induced by LPS-activated microglia based on Notch signaling pathway.Methods LPS-activated microglia were co-cultured with neurons to construct neuron inju-ry models,and the cells were divided into the control group,model group,Notch inhibitor(DAPT)group,GLGZG(50,100,200 mg·L-1)group,DAPT+100 mg·L-1GLGZG group.After intervention,the activity of HT22 cells was detected by CCK-8 method,and rel-ative mRNA expression was detected by real-time PCR.The relative protein expression was detected by Western blot.Results Compared with the model group,after GLGZG intervention,the cell activity was significantly improved,GLGZG decreased IL-6,IL-12,Bax,Notch 1,caspase-3,Delta-1,NICD,RBPSUH,HES1 expression,and increased Bcl-2 expression(P＜0.05).Compared with the model group,the NICD,RBPSUH and HES1 mRNA and protein expressions significantly decreased after DAPT treatment(P＜0.05),and there was no superposition effect with GLG-ZG.Conclusion GLGZG may play a neuroprotective role by inhibiting inflammatory factors and apoptosis,and inhibiting Notch signaling pathway.

Aim To identify the underlying target of bullatine A(BA)against rheumatoid arthritis(RA)u-sing limited proteolysis-small molecule mapping(LiP-SMap)drug target proteomics and to provide a scientif-ic basis for clinical application of Aconiti brachypodi Radix in the treatment of RA.Methods LiP-SMap drug target proteomics was employed to perform bioin-formatics analysis for comparing and validating the dif-ferential protein expression after BA intervention.A collagen-induced arthritis(CIA)model was estab-lished in DBA/1 mice using bovine type Ⅱ collagen.The mice were then divided into the CIA model group,methotrexate-positive control group(MTX group),and BA groups(10 mg·kg-1 and 20 mg·kg-1)based on their clinical scores.After drug intervention,the thera-peutic efficacy against RA was assessed by joint index scores and foot thickness measurements.Histopatholog-ical changes in the arthritic joints of CIA mice were e-valuated using hematoxylin and eosin(HE)staining.Enzyme-linked immunosorbent assay(ELISA)was employed to detect inflammatory cytokines interleukin-17(IL-17)and total IgG and IgG3 anti-collagen-spe-cific antibodies levels from the serum of CIA mice.Flow cytometry was used to detect the expression levels of intracellular Th17 cells(IL-17+CD4+T cells)and Th1 cells(IFN-γ+CD4+T cells).Fluorescent quanti-tative PCR was performed to detect the expression of genes related to differential proteins.Results The proteomic analysis identified Serpinb1a as a protein with strong binding affinity to BA,and KEGG enrich-ment analysis indicated IL-17 signaling pathway was a crucial pathway of BA in against RA.BA treatment significantly reduced clinical scores and foot thickness,improved local arthritis symptoms in CIA mice,and al-leviated inflammatory cell infiltration into arthritic joints(P＜0.05).Differential protein validation re-sults showed that BA had strong affinity with Serpinb1a(-5.92 kJ·mol-1)and downregulated the expres-sion of Serpinb1a mRNA.Furthermore,the administra-tion of BA markedly reduced serum IL-17 A levels from CIA mice,inhibited the expression of intracellular IL-17 A and IFN-γ cytokines in splenic CD4+T cells(P＜0.05),and significantly downregulated the transcrip-tional expression of IL-17F(P＜0.05).Conclusion BA exhibits therapeutic effects on collagen-induced arthritis,and its mechanism of action may involve the regulation of Serpinb1a and the IL-17 signaling path-way.

AIM:This study aims to investigate the therapeutic impact of bone marrow mesenchymal stem cell(MSC)-derived exosomes(MSC-Exos)on septic lung injury by targeting Toll-like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway.METHODS:MSC-Exos were isolated from bone marrow MSCs using ultracentrifugation.The exosomes were characterized by transmission electron microscopy(TEM),nanoparticle tracking analysis(NTA),and Western blot.A septic lung injury model was established via cecal ligation and puncture(CLP).MSC-Exos were adminis-tered intraperitoneally 3 h after CLP.Blood and lung tissue samples were collected at 6,12,24,and 72 h after CLP.Con-centrations of interleukin-10(IL-10),IL-6,IL-1β,and tumor necrosis factor-α(TNF-α)were measured by ELISA.The lung tissue damage in mice was assessed by HE staining.Expression levels of TLR4 and NF-κB were detected by RT-qP-CR and Western blot.RESULTS:The results of TEM revealed typical round and oval exosome-like structures of MSC-Exos.Western blot showed positive expression of CD63,TSG101 and HSP-70.The results of NTA indicated that the parti-cle size of MSC-Exos was(107.5±16.6)nm.Septic lung injury began to manifest 12 h after CLP and progressively worsened,peaking at 72 h after CLP.At 6 h after CLP,the level of TNF-α was markedly elevated compared to the control group,reaching its peak at 72 h after CLP.At 12 h after CLP,the levels of IL-1β,IL-10,and IL-6 were elevated com-pared to the control group,and they reached their maximum point 72 h post-CLP.The MSC-Exos intervention group ex-hibited significantly reduced lung injury compared to the CLP group,as indicated by the lung pathology scoring.In the MSC-Exos intervention group,concentrations of IL-10,IL-1β,IL-6,and TNF-α peaked at 12 h after CLP,gradually de-clined,and returned to baseline levels by 72 h.TLR4 and NF-κB expression levels were significantly increased at 6 h af-ter CLP in the CLP group,peaking at 72 h.In contrast,MSC-Exos treatment normalized TLR4 and NF-κB expression levels at 72 h after CLP.CONCLUSION:Early intervention with bone marrow MSC-Exos significantly alleviated septic lung injury by reducing the inflammatory cytokine storm,likely through downregulation of the TLR4/NF-κB signaling path-way.

Nucleic acid detection technology has been widely applied in fields such as pathogen detection due to its characteristics of rapidity,sensitivity,and specificity.With the numerous nucleic acid markers related to diseases,the demand for multiplex nucleic acid detection is gradually increasing.Multiplex polymerase chain reaction(PCR)can simultaneously amplify multiple targets,but there are problems such as easy contamination when opening the tube during the analysis process after amplification and high technical requirements.With the continuous advancement of detection technology,a series of simple,re-liable single-tube multiplex PCR detection technologies that do not require opening the tube have emerged successively.A common technology is the single-closed tube multiplex PCR detection method based on fluorescent probes,which mainly uses different fluorescent labels to distinguish multiple targets.Com-bined with different specific enzymatic digestion reactions,it can achieve multiplex detection of rare tumor mutations and single nucleotide-specific genotyping.In addition,the monochromatic melting curve analysis method based on differences in melting temperatures enables parallel detection of multiple targets within a single fluorescence channel.When performed within multiple fluorescence channels,it is called the multicolor melting curve analysis method,which can increase the number of detected targets to doz-ens,greatly breaking through the limitation of the number of fluorescence channels on the multiplexity of detection.At the same time,the fluorescence coding method using different combinations of fluorescent labels also provides new ideas for single-closed tube multiplex PCR detection.These include encoding the sequence of signals generated by different fluorescent labels corresponding to the same target,using a combination of two fluorescent labels to identify specific targets,and controlling the amplitude of fluores-cent signals of different targets,all of which can also improve the multiplexity of detection.This article summarizes and prospects the research progress of single-closed tube multiplex PCR detection technology in recent years from multiple dimensions such as principles,applications,and the advantages and disad-vantages of the methods,providing valuable references for subsequent scientific research exploration and application.

Objective:To establish a portable gastrointestinal endoscopy telemedicine platform that enables the real-time transmission of endoscopy audio and video data to the consulting physicians through 5G communication technology, and to evaluate the effectiveness of this platform.Methods:From October 10 to October 20, 2023, 30 cases (16 cases from Sanya, Hainan Province, and 14 cases from Zhanjiang, Guangdong Province) from primary health institutions were selected for portable gastrointestinal endoscopy telemedicine consultations with a tertiary hospital in Shanghai. 5G network transmission parameters were recorded, and the lesion detection rate of the examination side and the consultation side, as well as the quality of the videos and images were compared.Results:The 5G network transmission parameters for telemedicine from Sanya and Zhanjiang to Shanghai included download network speeds of 168.3 Mbps and 160.4 Mbps, upload network speeds of 75.1 Mbps and 72.4 Mbps, and delays of 28 ms and 25 ms, respectively. There was no significant difference in the lesion detection rate [43.3% (13/30) VS 36.7% (11/30), χ2=0.792, P=0.598] between the physicians on the examination side and the consultation side. The endoscopic video quality (2.92±0.59 points VS 2.88±0.64 points, t=0.302, P=0.763) and picture quality (3.10±0.57 points VS 3.05±0.53 points, t=0.424, P=0.622) were compared by two senior endoscopists between the examination side and the consultation side, indicating no significant difference. Conclusion:The utilization of portable gastrointestinal endoscopy for telemedicine based on 5G network technology is effective in enabling telemedicine consultations and providing operational guidance. This approach enhances the procedural skills and diagnostic capabilities of physicians in primary healthcare settings, facilitating the equitable distribution of high-quality medical resources.