Objective:To investigate the role and underlying mechanisms of methyltransferase (Mettl) 3 in the process of angiotensin Ⅱ (Ang Ⅱ)-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis.Methods:C57BL/6J mice were used, in cell experiments, mouse renal pericytes were isolated and cultured using magnetic bead sorting. These pericytes were then induced to transdifferentiate into myofibroblasts with 1×10 6 mmol/L Ang Ⅱ, which was the Ang Ⅱ group, while pericytes cultured in normal conditions served as the control group. Successful transdifferentiation was verified by immunofluorescence staining, Western blotting, and real-time reverse transcription PCR (RT-qPCR) for α-smooth muscle actin (α-SMA). The levels of m6A modifications and related enzymes (Mettl3, Mettl14), Wilms tumor 1-associated protein (WTAP), fat mass and obesity protein (FTO), ALKBH5, YTHDF1, YTHDF2, YTHDC1, YTHDC2, YTHDC3 were assessed by Dot blot, RT-qPCR and Western blot. Mettl3 expression was inhibited in cells using lentivirus-mediated Mettl3-shRNA transfection, creating sh-Mettl3 and Ang Ⅱ+sh-Mettl3 groups, while lentivirus empty vector transfection served as the negative control (Ang Ⅱ+sh-NC group). The impact of Ang Ⅱ on pericyte transdifferentiation was observed, and the expression of downstream phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway proteins, including PI3K, AKT, phosphorylated AKT at serine 473 (p-AKT (S473)), and phosphorylated AKT at threonine 308 (p-AKT (T308)), were examined. PI3K gene transcription was inhibited by co-culturing cells with actinomycin D, and the half-life of PI3K mRNA was calculated by measuring residual PI3K mRNA expression over different co-culture time. The reversibility of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was assessed by adding the AKT activator SC79 to the Ang Ⅱ+sh-Mettl3 group. In animal experiments, mice were divided into these groups: sham group (administered 0.9% sterile saline), Ang Ⅱ group (infused with Ang Ⅱ solution), sh-Mettl3 group (injected with Mettl3 shRNA lentivirus solution), Ang Ⅱ+sh-Mettl3 group (infused with Ang Ⅱ solution and injected with Mettl3 shRNA lentivirus solution), and Ang Ⅱ+sh-Mettl3+SC79 group (administered Ang Ⅱ solution and Mettl3 shRNA lentivirus, with an additional injection of SC79). Each group consisted of six subject mice. Blood pressure was measured using the tail-cuff method before and after surgery, and serum creatinine, urea, and urinary albumin levels were determined 4 weeks post-surgery. Kidney tissues were collected at 28 days and stained using hematoxylin-eosin (HE) and Masson′s trichrome to assess the extent of renal fibrosis. Results:Primary renal pericytes were successfully obtained by magnetic bead sorting, and intervened with 1×10 6 mmol/L Ang Ⅱ for 48 hours to induce pericyte-to-myofibroblast transdifferentiation. Dot blot results indicated higher m6A modification levels in the Ang Ⅱ group compared to the control group ( P<0.05). RT-qPCR and Western blot results showed upregulation of Mettl3 mRNA and protein levels in the Ang Ⅱ group compared to the control group (both P<0.05). In the Ang Ⅱ+sh-Mettl3 group, Mettl3 protein expression was lower than that in the Ang Ⅱ group, with reduced expression levels of α-SMA, vimentin, desmin, fibroblast agonist protein (FAPa) and type Ⅰ collagen (all P<0.05). Compared to the control group, PI3K mRNA expression level was elevated in the Ang Ⅱ group, along with increased p-AKT (S473) and p-AKT (T308) expressions. In the Ang Ⅱ+sh-Mettl3 group, PI3K mRNA expression and p-AKT (S473) and p-AKT (T308) levels were decreased (all P<0.05). The half-life of PI3K mRNA was shorter in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ+sh-NC group (2.34 h vs. 3.42 h). The ameliorative effect of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was reversible by SC79. Animal experiments showed higher blood pressure, serum creatinine, urea, and 24-hour urinary protein levels, and a larger fibrosis area in the Ang Ⅱ group compared to the sham group (all P<0.05). The fibrosis area was smaller in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ group ( P<0.05), but increased again upon addition of SC79. Conclusion:Mettl3-mediated RNA m6A epigenetic regulation is involved in Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis, potentially by affecting PI3K stability and regulating the PI3K/AKT signaling pathway.

Background::Carotid intima-media thickness (IMT) and diameter, stiffness, and wave reflections, are independent and important clinical biomarkers and risk predictors for cardiovascular diseases. The purpose of the present study was to establish nationwide reference values of carotid properties for healthy Chinese adults and to explore potential clinical determinants.Methods::A total of 3053 healthy Han Chinese adults (1922 women) aged 18-79 years were enrolled at 28 collaborating tertiary centers throughout China between April 2021 and July 2022. The real-time tracking of common carotid artery walls was achieved by the radio frequency (RF) ultrasound system. The IMT, diameter, compliance coefficient, β stiffness, local pulse wave velocity (PWV), local systolic blood pressure, augmented pressure (AP), and augmentation index (AIx) were then automatically measured and reported. Data were stratified by age groups and sex. The relationships between age and carotid property parameters were analyzed by Jonckheere-Terpstra test and simple linear regressions. The major clinical determinants of carotid properties were identified by Pearson’s correlation, multiple linear regression, and analyses of covariance.Results::All the parameters of carotid properties demonstrated significantly age-related trajectories. Women showed thinner IMT, smaller carotid diameter, larger AP, and AIx than men. The β stiffness and PWV were significantly higher in men than women before forties, but the differences reversed after that. The increase rate of carotid IMT (5.5 μm/year in women and 5.8 μm/year in men) and diameter (0.03 mm/year in both men and women) were similar between men and women. For the stiffness and wave reflections, women showed significantly larger age-related variations than men as demonstrated by steeper regression slopes (all P for age by sex interaction <0.05). The blood pressures, body mass index (BMI), and triglyceride levels were identified as major clinical determinants of carotid properties with adjustment of age and sex. Conclusions::The age- and sex-specific reference values of carotid properties measured by RF ultrasound for healthy Chinese adults were established. The blood pressures, BMI, and triglyceride levels should be considered for clinical application of corresponding reference values.

Objective:To construct an intelligent medical emergency information platform for the 19th Hangzhou Asian Games events and evaluate its practical application.Methods:Addressing the medical support demands for Asian Games, based on the " Emergency medical service system for large-scale social activities" developed by Zhejiang Laboratory, a collaborative team comprising Hangzhou Emergency Center, the Second Affiliated Hospital of Zhejiang University,Zhejiang Lab and College of Biomedical Engineering and Instrument Science of ZhejiangUniversity developed an intelligent medical emergency information platform for the Asian Games. This system was implemented during the 19th Hangzhou Asian Games, collecting basic information, initial diagnosis locations, prognoses, severity grading, and occurrences of infectious diseases for all patients treated at support locations from September 16th to October 8th, 2023. The Modified Early Warning Score (MEWS) was employed for assessing patient conditions, and a symptom monitoring system was used to alert the occurrence of infectious diseases. Post-event, a satisfaction evaluation (on a 5-point scale) regarding the system's application was conducted.Results:The successful construction of the intelligent medical emergency information platform comprised a data dashboard display platform, a comprehensive command system on the management center, and a mobile platform covering on-site medical support, ambulances, and designated hospitals. ①A total of 11 068 patients were treated using this system, including 2 979 athletes, 3 429 staff members, 1 421 volunteers,1 051 spectators, and 2 188 others. The proportion of patients seeking treatment due to illnesses was significantly higher than those due to injuries. Among injury-related cases, athletes constituted 41.37%, while staff accounted for 32.5% of illness-related cases. ②The area under the ROC curve for MEWS regarding hospitalization rates was 0.81 for disease patients and 0.60 for injury patients. ③ There were 211 reported cases of infectious diseases during the event, constituting 1.91% of all treated cases.④ Satisfaction ratings: User-friendliness scored 4.47, completeness of functions scored 4.28, data security scored 4.38, and data accuracy scored 4.49. Compared to traditional paper-based reporting methods, 98.11% of users found the system more convenient, 92.45% were satisfied overall, and the recommendation rating was 4.36.Conclusions:The study findings confirm the critical role played by the information platform in enhancing comprehensive, accurate, timely, secure medical support information dissemination for the Asian Games and in early warning and control of infectious diseases.

Purpose To explore the association of tumor budding in preoperative biopsies with the clinicopathological characteristics of colorectal cancer(CRC),and its effects on the prognosis.Methods A total of 144 preoperative biopsies and corresponding radical resection specimens of colorectal ade-nocarcinoma were collected.Tumor budding was evaluated by HE staining and EnVision immunohistochemical staining,to an-alyze the association of tumor budding with the clinicopathologi-cal characteristics and prognosis in biopsies.Results The de-tection rate of intra-tumoral budding(ITB)in biopsy specimens was 39.6%(57/144),and peritumoural budding(PTB)in surgical specimens was 64.6%(93/144);ITB was significantly correlated with PTB(r =0.510,P<0.05).The proportion of patients with lymph node metastasis,pTNM stage,differentiated degree,vascular invasion and nerve recidivism in the tumor bud-ding group of biopsies was significantly higher than that in no tumor budding group(P<0.05).Kaplan-Meier survival analy-sis showed that ITB significantly affected the overall survival and progression free survival of CRC(P<0.05).Conclusion The presence of ITB in biopsies tissue may be an important indicator of invasive behavior and poor prognosis in CRC.

Osteochondral lesion of talus (OLT) is a foot and ankle disease characterized by ankle pain, which may impact the joint function and life quality. If managed improperly, it may lead to a further ankle arthritis, severely compromising the prognosis. The therapeutic effect of conservative treatment for OLT is still uncertain. Surgery is still the main treatment modality for OLT with various techniques. However, the optimized surgical technique is still inconclusive, furthermore, regeneration and repair of cartilage after debridement is also a great challenge for the treatment of OLT. Platelet-rich plasma (PRP) with good repair effect on cartilage injury is gradually applied in the treatment of OLT. However, there still lacks the unified understanding of the technique and specification of PRP for the treatment of OLT. Therefore, National Orthopedics Center of Shanghai Sixth People′s Hospital allied Foot Ankle Basic Research & Orthopedics Group, Chinese Association of Orthopedic Surgeons; Foot and Ankle Committee of Chinese Association of Sports Medicine Physicians; and Foot and Ankle Group of Orthopedic Specialized Branch of Shanghai Medical Association to organize related experts to formulate the Expert consensus on platelet- rich plasma treatment for osteochondral lesion of talus ( version2023). Fifteen recommendations were put forward upon PRP preparation, indications, contraindications and treatment methods of PRP for OLT, so as to standardize the PRP treatment for OLT.

Objective:To analyze the early volume characteristics of patients with severe cardiogenic shock treated with veno-arterial extracorporeal membrane oxygenation (VA-ECMO) and the relationship between their early volume and the prognosis.Methods:This study reviewed patients of Emergency Medical University , treated with VA-ECMO and screened the patients with severe cardiogenic shock and VA-ECMO running more than 72 h for further study. The basic condition of the patients was recorded, and the fluid balance in the first 72 h was analyzed. The patients were grouped according to their fluid balance in the first 72 h. The gender, age, survival rate, continuous renal replacement therapy (CRRT) rate, intra-aortic balloon pump (IABP) rate, and invasive mechanical ventilation rate were compared between the two groups, and the relative risk to the prognosis was calculated. The prognosis was compared between the two groups. Results:Totally 77 patients with severe cardiogenic shock were enrolled. Forty-one cases survived, with an overall survival rate of 53.2%. The volume balance at 48-72 h and the total volume balance at the first 72 h were different between the survival and dead groups. Compared with the positive balance group, patients in the negative balance group were less likely to receive CRRT or invasive mechanical ventilation during the first 72 h. Patients in the negative balance group during the first 72 h had a better survival rate, and their relative risk of survival was 1.81 (95% confidence interval: 1.101, 2.985). However, there was no significant difference in survival rate according to every 24 h fluid balance.Conclusions:Patients with severe cardiogenic shock treated with VA-ECMO who had negative total volume balance during the first 72 h are more likely to survive and less likely to require CRRT or invasive mechanical ventilation.

Objective:To establish rules for medical order review in parenteral nutrition, assist pharmacists in the efficient real-time pre-review of parenteral nutrition medical orders, and promote the standardized use of parenteral nutrition.Methods:Professional clinical nutrition pharmacists, prescription reviewing pharmacists, clinicians, and nutritionists jointly contributed to the formulation of custom prescription review rules for parenteral nutrition. Utilizing the prescription review system of our hospital, pharmacists reviewed and intervened parenteral nutrition medical orders in real time.Result:After the implementation of review rules, all of the medical orders for parenteral nutrition have been pre-reviewed in our hospital, and the related indicators that are out of normal range and the cases of single bottle infusion show significant reduction in frequency.Conclusion:The formulation and implementation of specific prescription review rules for parenteral nutrition can effectively assist pharmacists in real-time review of parenteral nutrition medical orders, and can solve the problems of inappropriate medical orders and single-bottle infusion.

Betacoronavirus ; isolation & purification ; pathogenicity ; Bile Acids and Salts ; metabolism ; Bile Ducts, Intrahepatic ; pathology ; virology ; Cell Culture Techniques ; Coronavirus Infections ; complications ; pathology ; Cytokine Release Syndrome ; etiology ; physiopathology ; Cytopathogenic Effect, Viral ; Epithelial Cells ; enzymology ; pathology ; virology ; Humans ; Hyperbilirubinemia ; etiology ; Liver ; pathology ; Organoids ; pathology ; virology ; Pandemics ; Peptidyl-Dipeptidase A ; analysis ; Pneumonia, Viral ; complications ; pathology ; Receptors, Virus ; analysis ; Serine Endopeptidases ; analysis ; Viral Load

Objective:To explore the predictive factors of renal replacement therapy (RRT) in extracorporeal membrane oxygenation (ECMO) patients.Methods:The clinical data of 68 ECMO patients treated at Emergence Department of Jiangsu Provincial Hospital from January 2015 to December 2018 were retrospectively analyzed. Vasoactive-inotropic score (VIS) was used to assess the usage of vasoactive-inotropic drugs on day 1, 2 and 3 of ECMO (24 h VIS, 48 h VIS, and 72 h VIS). According to received RRT or not, patients were divided into the RRT group and non-RRT group. Age, gender, weight, VIS, presence of cardiac arrest before ECMO, ECMO mode, and ECMO treatment time were compared. Logistic regression analysis was used to identify predictive factors for RRT in ECMO patients.Results:Of the enrolled patients, 73.5% of ECMO patients received RRT. The mean age, 24 h VIS, ECMO failure and mortality of the RRT group were significantly higher than those of the non-RRT group ( P <0.05). The use of RRT was 87.8% in elderly ECMO patients (> cutoff age of 38.5 years). According to the cutoff value of 24 h VIS (33.75), ECMO patients were divided into the high VIS group and low VIS group. The rates of RRT and mortality were both exceeded 90% in the high VIS group, which was significantly higher than that of the low VIS group ( P <0.05). Logistic regression analysis showed that age ( OR=1.223) and 24 h VIS ( OR=1.033) were predictive factors of RRT in ECMO patients ( P <0.05). Conclusions:Age and 24 h VIS show the predictive value for RRT in ECMO patients.

Objective:To explore the effects of different doses of Ethephon on testes of male pups.Methods:Thirty-two 45-day-old healthy female Sprague-Dawley (SD) rats were randomly divided into the control group and the low dose, middle dose and high dose Ethephon groups by the random figure table.The female rats in the low dose, middle dose and high dose Ethephon groups were given 200, 400, and 800 mg/kg Ethephon solution, respectively.The control group was treated with 9 g/L saline.After the birth of the offspring, the mother rats were not administrated with any medications, and the male offspring rats were given Ethephon solution instead.Twelve offspring male rats were randomly selected from each group and killed at the age of 0, 14 and 28 days after birth.Fresh testicular tissues were stained with hematoxylin eosin (HE), and the morphological changes of testicular tissues were observed under light microscope.The apoptotic cells were labeled by terminal dexynucleotidyl transferase (TdT) mediated dUTP nick end labeling (TUNEL) method and the apoptosis index (AI) of spermatogenic cells was detected by fluorescence microscope.Results:(1) Compared with the newborn rats in the middle dose group, low dose group and control group, se-miniferous tubules in the newborn rats of the high dose group were slightly thicker, and seminiferous cells were arranged slightly in disorder.The AI of the newborn rats in high dose group was significantly higher than that in the control group (1.00±0.06 vs.0.41±0.03, P<0.01). The AI of the newborn rats in the middle dose group was not significantly different from that in the control group and the low dose group ( P>0.05). (2) The seminiferous tubules of the 14-day-old rats in the low dose, middle dose and high dose Ethephon groups were significantly thicker and arranged more loosely than those in the control group.Compared with the control group, there were very few seminiferous cells, which were arranged disorderly in the low dose, middle dose and high dose Ethephon groups.The AI of the 14-day-old rats in the low dose, middle dose and high dose Ethephon groups was (2.13±0.10), (2.18±0.10) and (3.90±0.23), respectively, which were significantly higher than that in the control group (1.00±0.02) ( F=2 508.36, P<0.01). There was no significant difference in the AI between the middle dose and low dose groups ( P>0.05). (3) Compared with the control group, the seminiferous tubules of the 28-day-old rats in the low dose, middle dose and high dose groups were significantly thicker and arranged much more loosely, and spermatogenic cells were even less and arranged in a severely disordered way.The AI of 28-day-old rats in the low dose group (5.52±0.13), the middle dose group (9.44±0.07) and the high dose group (14.56±0.27) was significantly higher than that in the control group (3.11±0.13) ( F=10 784.69, P<0.01). Conclusions:Ethephon can thicken the seminiferous tubules of newborn and young rats, cause the germ cells to arrange disorderly, promote the apoptosis of spermatogenic cells and reduce the ability of spermatogenesis.Moreover, a longer exposure of the rats to a higher concentration of Ethephon will result in more serious damage to testicular tissues.