OBJECTIVES: Bladder cancer is a common malignancy with high incidence and poor prognosis. N⁶-methyladenosine (m⁶A) modification is widely involved in diverse physiological processes, among which the m⁶A recognition protein YTH N⁶-methyladenosine RNA binding protein F2 (YTHDF2) plays a crucial role in bladder cancer progression. This study aims to elucidate the molecular mechanism by which O-linked N-acetylglucosamine (O-GlcNAc) modification of YTHDF2 regulates its downstream target, period circadian regulator 1 (PER1), thereby promoting bladder cancer cell proliferation. METHODS: Expression of YTHDF2 in bladder cancer was predicted using The Cancer Genome Atlas (TCGA). Twenty paired bladder cancer and adjacent normal tissues were collected at the clinical level. Normal bladder epithelial cells (SV-HUC-1) and bladder cancer cell lines (T24, 5637, EJ-1, SW780, BIU-87) were examined by quantitative real-time PCR (RT-qPCR), Western blotting, and immunohistochemistry for expression of YTHDF2, PER1, and proliferation-related proteins [proliferating cell nuclear antigen (PCNA), minichromosome maintenance complex component 2 (MCM2), Cyclin D1]. YTHDF2 was silenced in 5637 and SW780 cells, and cell proliferation was assessed by Cell Counting Kit-8 (CCK-8), colony formation, and EdU assays. Bioinformatics was used to predict glycosylation sites of YTHDF2, and immunoprecipitation (IP) was performed to detect O-GlcNAc modification levels of YTHDF2 in tissues and cells. Bladder cancer cells were treated with DMSO, OSMI-1 (O-GlcNAc inhibitor), or Thiamet G (O-GlcNAc activator), followed by cycloheximide (CHX), to assess YTHDF2 ubiquitination by IP. YTHDF2 knockdown and Thiamet G treatment were further used to evaluate PER1 mRNA stability, PER1 m⁶A modification, and cell proliferation. TCGA was used to predict PER1 expression in tissues; SRAMP predicted potential PER1 m⁶A sites. Methylated RNA immunoprecipitation (MeRIP) assays measured PER1 m⁶A modification. Finally, the effects of knocking down YTHDF2 and PER1 on 5637 and SW780 cell proliferation were assessed. RESULTS: YTHDF2 expression was significantly upregulated in bladder cancer tissues compared with adjacent tissues (mRNA: 2.5-fold; protein: 2-fold), which O-GlcNAc modification levels increased 3.5-fold (P<0.001). YTHDF2 was upregulated in bladder cancer cell lines, and its knockdown suppressed cell viability (P<0.001), downregulated PCNA, MCM2, and CyclinD1 (all P<0.05), reduced colony numbers 3-fold (P<0.01), and inhibited proliferation. YTHDF2 exhibited elevated O-GlcNAc modification in cancer cells. OSMI-1 reduced YTHDF2 protein stability (P<0.01) and enhanced ubiquitination, while Thiamet G exerted opposite effects (P<0.001). Thiamet G reversed the proliferation-suppressive effects of YTHDF2 knockdown, promoting cell proliferation (P<0.01) and upregulating PCNA, MCM2, and CyclinD1 (all P<0.05). Mechanistically, YTHDF2 targeted PER1 via m⁶A recognition, promoting PER1 mRNA degradation. Rescue experiments showed that PER1 knockdown reversed the inhibitory effect of YTHDF2 knockdown on cell proliferation, upregulated PCNA, MCM2, and Cyclin D1 (all P<0.05), and promoted bladder cancer cell proliferation (P<0.001). CONCLUSIONS O-GlcNAc modification YTHDF2 promotes bladder cancer development by downregulating the tumor suppressor gene PER1 through m⁶A-mediated post-transcriptional regulation.
Humans ; Urinary Bladder Neoplasms/metabolism* ; RNA-Binding Proteins/genetics* ; Cell Proliferation ; Cell Line, Tumor ; Disease Progression ; Acetylglucosamine/metabolism* ; Adenosine/metabolism* ; Gene Expression Regulation, Neoplastic ; Genes, Tumor Suppressor

Sustained inflammatory responses are closely related to various severe diseases, and inhibiting the excessive activation of inflammasomes and pyroptosis has significant implications for clinical treatment. Natural products have garnered considerable concern for the treatment of inflammation. Huanglian-Wumei decoction (HLWMD) is a classic prescription used for treating inflammatory diseases, but the necessity of their combination and the exact underlying anti-inflammatory mechanism have not yet been elucidated. Inspired by the supramolecular self-assembly strategy and natural drug compatibility theory, we successfully obtained berberine (BBR)-chlorogenic acid (CGA) supramolecular (BCS), which is an herbal pair from HLWMD. Using a series of characterization methods, we confirmed the self-assembly mechanism of BCS. BBR and CGA were self-assembled and stacked into amphiphilic spherical supramolecules in a 2:1 molar ratio, driven by electrostatic interactions, hydrophobic interactions, and π-π stacking; the hydrophilic fragments of CGA were outside, and the hydrophobic fragments of BBR were inside. This stacking pattern significantly improved the anti-inflammatory performance of BCS compared with that of single free molecules. Compared with free molecules, BCS significantly attenuated the release of multiple inflammatory mediators and lipopolysaccharide (LPS)-induced pyroptosis. Its anti-inflammatory mechanism is closely related to the inhibition of intracellular nuclear factor-kappaB (NF-κB) p65 phosphorylation and the noncanonical pyroptosis signalling pathway mediated by caspase-11.

Objective To explore the value of serum soluble triggering receptor expressed on myeloid cell-1(sTREM-1),interleukin-17(IL-17),and soluble Fas ligand(sFasL)in evaluating severity and prognosis of neonatal infection.Methods From November 2022 to November 2024,a total of 105 neonates with infections who visited the Department of Pediatrics in the hospital were labeled as infection group,and 105 healthy neo-nates were labeled as control group.Based on the severity of neonatal infection,the affected children were as-signed into extremely critical group,critical group,and non critical group.Based on prognosis,the affected children were grouped into adverse group and good group.Enzyme linked immunosorbent assay(ELISA)was used to measured serum sTREM-1,IL-17,and sFasL.Receiver operating characteristic(ROC)curve was used to evaluate the predictive value of serum sTREM-1,IL-17,and sFasL for prognosis.Results The serum levels of sTREM-1,IL-17,and sFasL in the infection group were significantly higher than those in the control group(P＜0.05).The serum sTREM-1,IL-17,and sFasL decreased in serum of extremely critical group,critical group,and non critical group in sequence(P＜0.05).Spearman correlation showed that serum sTREM-1,IL-17,and sFasL levels were positively correlated with disease severity(r=0.601,0.592,0.523,P＜0.05).The proportion of cesarean section,serum levels of sTREM-1,IL-17,and sFasL in the poor group were higher than those in the good group(P＜0.05),and the Apgar score was lower than that in the good group(P＜0.05).Logistic regression revealed that cesarean section,sTREM-1,IL-17,and sFasL were all factors affecting the adverse prognosis of neonatal infections(P＜0.05).ROC curve showed that the area under the curve(AUC)of serum sTREM-1,IL-17,sFasL,and their combined detection in predicting adverse prognosis of neonatal in-fections was 0.781,0.788,0.768,and 0.879,respectively.The AUC of joint prediction was greater than those of individual prediction(Zcombination-sTREM-1=2.238,P=0.025,Zcombination-IL-17=2.014,P=0.044,Zcombination-sFasL=2.248,P=0.025).Conclusion Serum sTREM-1,IL-17,and sFasL levels in neonates with infections are man-ifestly elevated,and they are closely related to the severity and prognosis.

BACKGROUND:Swimming is an important non-pharmacological treatment for knee osteoarthritis,which can effectively alleviate the disease.However,the effect and mechanism of swimming on senile knee osteoarthritis are still unclear. OBJECTIVE:To investigate the effect of swimming exercise on the articular cartilage of aged mice with knee osteoarthritis. METHODS:Six 3-month-old male C57BL/6 mice were selected as the young group,and twelve 18-month-old male C57BL/6 mice were randomized into the aged group and the swimming group,with six mice in each group.Mice in the swimming group received adaptive swimming for 1 week and formal swimming for 8 weeks.After the intervention,stride length analysis and sampling were performed in each group.The total number of leukocytes and lymphocytes in peripheral blood was detected by blood routine examinations.The morphology of the articular cartilage was observed by hematoxylin-eosin and safranin O-fast green staining.Chondrocyte counts and the modified Mankin's score were used to evaluate the degree of articular cartilage damage.The protein and mRNA expressions of type Ⅱ collagen,aggrecan and matrix metalloproteinase 13 in articular cartilage were detected by immunohistochemical staining and RT-qPCR. RESULTS AND CONCLUSION:Compared with the young group,the mice in the aged group showed significantly decreased stride length(P＜0.05),significantly increased numbers of peripheral leukocytes and lymphocytes(P＜0.05),significantly decreased count of chondrocytes(P＜0.05),significantly increased modified Mankin's score(P＜0.05),significantly decreased protein and mRNA expression of type Ⅱ collagen and aggreca(P＜0.05),and significantly increased matrix metalloproteinase 13 expression(P＜0.05).Moreover,hematoxylin-eosin and safranin O-fast green staining showed the uneven surface of the articular cartilage,abnormal chondrocytes,and proteoglycan loss in the aged group.Compared with the aged group,swimming exercise significantly improved the stride length of mice(P＜0.05),decreased the count of peripheral blood lymphocytes(P＜0.05),increased the count of chondrocytes(P＜0.05),decreased the modified Mankin's score(P＜0.05),increased the protein and mRNA expression of type Ⅱ collagen and aggrecan(P＜0.05),and decreased the expression of matrix metalloproteinase 13(P＜0.05).Hematoxylin-eosin and safranin O-fast green staining showed that the articular surface of mice in the swimming group was smooth,chondrocytes were normal,and proteoglycan loss was less.All these findings indicate that swimming exercise can reduce the number of inflammatory cells in the blood of aged mice,improve articular chondrocytes,matrix composition and cartilage tissue morphology;thus,it has a protective effect on the cartilage of aged mice with knee osteoarthritis.

Objective:To compare the clinical efficacy of fluorescent laparoscopy versus conventional laparoscopy for living-donor hepatectomy and explore the application value of fluorescent cholangiography in pediatric living donor liver transplantation (LT) using left lateral lobe.Methods:From October 2016 to August 2023, retrospective analysis was conducted for the relevant clinical data from 302 recipients of laparoscopic left lateral lobe donor hepatectomy. There were 90 cases in conventional laparoscopy group and 212 cases in fluorescent laparoscopy group. After propensity score matching (PSM), 88 patients were enrolled into each group. The perioperative data such as operative duration, donor liver thermal ischemic time, number of graft bile duct openings, liver function parameters and complication rates at Day 1/3 postoperation of donors and recipients were compared between two groups. SPSS 27.0 statistical software was utilized for analysis. Independent sample t, Mann-Whitney U and chisquare tests were utilized for examining the inter-group differences. Results:In fluorescent laparoscopy group, the proportion of grafts with a single open left hepatic duct was 84.1 %(74/88) versus 70.5% (62/88) in conventional laparoscopy group with statistically significant difference ( P=0.031). No statistically significant inter-group differences existed in postoperative liver function recovery or short-term postoperative complications ( P>0.05 ). No statistically significant inter-group differences existed in short-term postoperative liver function recovery or hospitalization length ( P> 0.05). In terms of biliary complications, fluorescent laparoscopy group had no biliary complications while the incidence of biliary complications was 3.4% (3/88) in conventional laparoscopy group. However, inter-group difference was statistically insignificant ( P>0.05 ) . Conclusions:Living-donor LT using left lateral lobe under fluorescent laparoscopy may enhance the surgical safety for both donors and recipients.

Objective:To explore the clinical efficacy of immunoadsorption in highly sensitized kidney transplant (KT) candidates.Methods:From September 2019 to April 2023, the relevant clinical data were retrospectively reviewed for 26 highly sensitized KT recipients. Protein A immunoadsorption desensitization therapy was offered after KT. The effect of immunosorbent on reducing anti-human leukocyte antigen (HLA) antibodies was summarized. And operative success rate and postoperative complication incidence were calculated.Results:The mean number of treatment session was (10.76±5.53). The highest level of HLA-Ⅰ antibody mean fluorescence intensity (MFI) dropped from (17 921±4 442) to (7 333±6 434) with a decline of 59% and HLA-Ⅱ antibody MFI decreased from (21 135±5 245) to (10 989±7 627) with a decline of 48%. The differences were statistically significant (both P<0.001). All kidneys were harvested from cadavers. The complications were acute antibody mediated rejection (7 cases), perioperative pulmonary infection (3 cases) and myelosuppression (2 cases). The average follow-up period was (30.8±12.6) month. The graft survival rate was 88.5% (23/26) and the recipient survival rate 100% (26/26) . Conclusions:Immunoadsorption therapy can effectively reduce HLA antibody in highly sensitized KT candidates, thereby increasing the probability of successful KT. In terms of safety, immunosorbent therapy may boost the potential risks of infection and myelosuppression. It requires heightened attention.

Objective　To investigate the current status and attrition among HIV-infected persons receiving antiretroviral therapy (ART), and to analyze factors affecting attrition in Hainan. Methods　In this study, HIV-infected patients who started ART treatment in Hainan Province from 2005 to 2022 were selected from the antiviral treatment submodule of China Disease Prevention and Control Information System.According to the inclusion and exclusion criteria,a total of 4 286 HIV-infected persons were receiving . A Cox proportional hazards regression model was used to analyze factors affecting attrition. Results　Among the 4 286 study subjects, 3 718 were males (86.7%), with a sex ratio of 6.55∶1. Unmarried individuals accounted for 58.4%, and the average age was (39.68±13.17) years. Transmission through homosexual contact accounted for 49.8%, and 84.3% were in WHO clinical stage I. Treatment regimens containing Efavirenz (EFV) accounted for 71.7%. During a follow-up of 19 677.44 person-years, the overall attrition rate was 0.80 per 100 person-years, with the first-year post-ART initiation attrition rate being 21.10 per 100 person-years. The results of Cox regression analysis showed that the time of treatment initiation in 2016-2022 (AHR=2.40, 95%CI: 1.40-4.10), and the last HIV viral load (VL) 20-<1 000 copies/mL (AHR=3.69, 95%CI: 2.08-6.54), the last HIV-1 VL≥1 000 copies/mL (AHR=15.98, 95%CI: 9.46-27.01), and no last HIV-1 VL test (AHR=92.90, 95%CI: 57.68-149.62), the time interval from diagnosis to treatment for 1-12 months (AHR=1.62, 95%CI: 1.12-2.36), and an interval longer than 12 months (AHR=1.68, 95%CI: 1.07-2.62) were the main factors that increased the risk of attrition. Treatment regimens containing Lopinavir/ritonavir (Lpv/r) (AHR=0.34, 95%CI:0.18-0.66) and treatment regimens containing integrase strand transfer inhibitors (INSTIs) (AHR=0.24, 95%CI: 0.09-0.58) were the factors that reduced the risk of attrition after antiretroviral therapy. Conclusions　The attrition of ART in HIV/AIDS patients in Hainan Province is related to a longer interval from diagnosis to treatment, treatment plan, and abnormal HIV viral load test results. Case-based measures should be taken to address factors influencing antiretroviral treatment attrition, while improving the timeliness of antiviral treatment and treatment management service quality to further improve the efficacy of antiviral treatment.

Objective To investigate the antibacterial performance and biocompatibility of silver nanoparticles-coated root canal nickel titanium instruments(AgNPs-NiTi).Methods AgNPs-NiTi was prepared using pulse electrochemical deposition.The morphol-ogy of AgNPs-NiTi was observed using field emission scanning electron microscopy(FE-SEM),and the elemental composition and con-tent were analyzed using X-ray diffraction(XRD)and energy dispersive spectroscopy(EDS).The mechanical properties of AgNPs-NiTi were tested.After Co-culturing AgNPs-NiTi with E.faecalis,the antibacterial effect was detected by colony-forming units method.By constructing an in vitro model of E.faecalis biofilm in the root canal of teeth,the antibacterial effect of AgNPs-NiTi was observed using FE-SEM and live/dead bacterial staining.In addition,AgNPs-NiTi was co-cultured with Raw 264.7 cells,and its cytotoxicity was de-tected by CCK-8.Results The pulse electrochemical deposition was used to construct a silver nanoparticle(AgNPs)coating on NiTi instruments with no significant change in the mechanical properties.AgNPs-NiTi significantly inhibited the proliferation of E.faecalis and damaged E.faecalis biofilm in the root canal.AgNPs-NiTi had no significant influence on the proliferation of Raw264.7 cells and had no cytotoxicity.Conclusion The mechanical properties of AgNPs-NiTi are similar to those of nickel titanium instruments.AgNPs-NiTi inhibits E.faecalis proliferation with good biocompatibility.

With advancements in radiology,endoscopic techniques,surgical treatments,cell biology and molecular biology,the un-derstanding of temporomandibular disorders(TMD)has increased.The temporomandibular joint(TMJ)is a complex structure comprising both soft and hard tissues.Within the TMJ,the temporomandibular disc is a soft tissue structure that connects the mandible to the skull,providing cushioning and stability during joint movement.Different imaging techniques have their own advantages and limi-tations in the diagnosis and treatment of TMD.Therefore,using image registration technology to assess the condition and position of the articular disc provides new research perspectives for evaluating TMD,which may contribute to the diagnosis and treatment.This article reviews the latest advancements in TMJ imaging,explores the applications of various image registration techniques,particularly in the context of TMD diagnosis and treatment,and discusses future prospects.Combining the research results of some scholars at home and a-broad with the author’s clinical experience,the article aims to provide valuable insights for clinicians.

The basic concept and related background of multivariate statistical process control(MSPC)were intro-duced.The basic process of MSPC implementation and the common statistics and control charts were described.The important role of MSPC in the pharmaceutical production process was emphasized,especially in the field of tra-ditional Chinese medicine,providing reference for the selection of quality control methods in drug production process.The advantages and disadvantages of MSPC were summarized,and the development trend of MSPC in drug production and quality control was prospected.