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 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.

Extracellular polymeric substances(EPS)constitutes crucial elements within bacterial biofilms,facili-tating accelerated antimicrobial resistance and conferring defense against the host's immune cells.Developing precise and effective antibiofilm approaches and strategies,tailored to the specific charac-teristics of EPS composition,can offer valuable insights for the creation of novel antimicrobial drugs.This,in turn,holds the potential to mitigate the alarming issue of bacterial drug resistance.Current analysis of EPS compositions relies heavily on colorimetric approaches with a significant bias,which is likely due to the selection of a standard compound and the cross-interference of various EPS compounds.Considering the pivotal role of EPS in biofilm functionality,it is imperative for EPS research to delve deeper into the analysis of intricate compositions,moving beyond the current focus on polymeric materials.This ne-cessitates a shift from heavy reliance on colorimetric analytic methods to more comprehensive and nuanced analytical approaches.In this study,we have provided a comprehensive summary of existing analytical methods utilized in the characterization of EPS compositions.Additionally,novel strategies aimed at targeting EPS to enhance biofilm penetration were explored,with a specific focus on high-lighting the limitations associated with colorimetric methods.Furthermore,we have outlined the challenges faced in identifying additional components of EPS and propose a prospective research plan to address these challenges.This review has the potential to guide future researchers in the search for novel compounds capable of suppressing EPS,thereby inhibiting biofilm formation.This insight opens up a new avenue for exploration within this research domain.

Objective:To investigate the characteristics of gut microbiota in patients with type 2 diabetes (T2DM) complicated by nonalcoholic fatty liver disease (NAFLD).Methods:A total of 74 patients first diagnosed with T2DM at the Endocrinology Department of Peking Union Medical College Hospital from April 2021 to October 2023 were included. Among them, 28 patients had concurrent NAFLD while 46 patients did not. Additionally, 51 healthy controls were matched (HC group). Clinical laboratory parameters were collected, and 16S rRNA sequencing with fecal samples was conducted to compare the differences in gut microbiota across the groups.Results:Compared to the group with T2DM, patients with concurrent T2DM and NAFLD were younger, had higher level of insulin resistance as assessed by the homeostatic model assessment of insulin resistance, higher body mass index (BMI), and higher triglyceride levels. There was no difference in α-diversity across the three groups ( P>0.05), while there was a significant difference in β-diversity ( P=0.03). The Eubacterium coprostanoligenes, Fusicatenibacter, Parasutterella and Tyzzerella 3 were enriched in the group with concurrent T2DM and NAFLD as shown by the relative abundance, while the relative abundance of Flavonifractor was decreased in this group. Tyzzerella 3 abundance was positively correlated with triglyceride and albumin levels and negatively correlated with high-density lipoprotein cholesterol (HDL-C) levels. Conclusion:Patients with T2DM complicated by NAFLD exhibit dysbiosis in gut microbiota composition and specific genera abundance, with Flavonifractor identified as a potential protective factor for T2DM complicated by NAFLD.

Objective:To explore the effect of early intermittent enteral nutrition (EN) in critically ill patients after cardiac surgery.Methods:We developed an early EN feeding regimen for critically ill patients after cardiac surgery. A prospective analysis was conducted on 178 critically ill patients admitted to the Cardiothoracic Surgery Intensive Care Unit of a tertiary hospital in Shanghai from May 2022 to May 2023. They were divided into the control group ( n=88) and the observation group ( n=90) using a random number table method. The control group received routine EN feeding, while the observation group received early intermittent EN feeding. Comparison between groups was performed in terms of the tolerability and interruption of EN administration, nutrition related outcome indicators, and prognosis indicators. Results:The observation group had significantly lower rates of abdominal distension, diarrhea, nausea and vomiting (or reflux), and EN interruption, as well as shorter duration of EN interruption, compared to the control group ( P<0.05). After 7 days of early EN, 87.8% of patients in the observation group met the calorie goal, which was significantly higher than that of 22.5% in the observation group ( P<0.05). The nutritional related indicators (serum prealbumin, hemoglobin, actual intake of nutritional fluids) were also better in the observation group patients compared to the control group ( P<0.05). The duration of mechanical ventilation and length of ICU stay in the observation group were significantly shorter than those in the control group, with statistical significance ( P<0.05). There was no significant difference in the length of hospital stay between the two groups. Conclusion:Early EN program can improve the symptoms of feeding intolerance in patients, reduce the occurrence and duration of feeding interruption, increase the rate of calorie goal fulfillment, improve nutritional status,and promote patient recovery and outcome in critically ill patients after cardiac surgery.

Objective:To explore the actor-partner interdependence between depressive symptoms and social participation, comorbidity in older couples in China, so as to provide new directions for depression intervention in older adults.Methods:A total of 4 346 older adults (2 173 older couples) were included using data from the 2020 China Health and Retirement Longitudinal Study (CHARLS). Spearman correlation was used to analyze the correlation between social participation, comorbidities, and depressive symptoms among older couples in China. AMOS software was used to establish an actor-partner interdependence model of depressive symptoms and social participation, comorbidities in older couples in China, and to analyze the actor-partner effects of social participation and comorbidities on depressive symptoms in older adults and their spouses.Results:The detection rate of depressive symptoms among 4 346 older adults was 38.66% (1 680/4 346), and the depressive symptom score of wives was higher than that of husbands ( Z=15.445, P<0.01). The analysis of the actor-partner interdependence model showed that the social participation and comorbidity of wives and husbands had a actor effect on their own depressive symptoms ( β=-0.097, P<0.01; β=0.275, P<0.01; β=-0.070, P<0.01; β=0.282, P<0.01), and the social participation of wives and husbands had a partner effect on their spouse's depressive symptoms ( β=-0.054, P<0.01; β=-0.054, P<0.01), and the comorbidity of husband had a partner effect on his wife's depressive symptoms ( β=0.057, P<0.01) . Conclusions:The dualistic interaction effects of actor-partner exist in the depressive symptoms, social participation, and comorbidities of older couples in China. Medical and nursing staff should pay attention to the dual effects of "patient-family caregiver" when intervening in depression and chronic diseases, and focus on the depressive symptoms of older women.

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.

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.