Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.

Aim To investigate the regulatory mecha-nism of butin on the proliferation,migration,cycle blockage and pyroptosis related inflammatory factors in human fibroblast-like synoviocytes of rheumatoid arthri-tis(HFLS-RA).Methods Cell proliferation,migra-tion and invasion were studied using cell migration and invasion assays.Cell cycle was detected by flow cytom-etry,and the expression of the pyroptosis-associated in-flammatory factors IL-1β,IL-18,caspase-1 and caspase-3 was detected by ELISA,RT-qPCR and West-ern blot.Results Migration and invasion experiments showed that the cell proliferation rate of the butin group was lower than that of the blank control group(P＜0.05).Cell cycle analysis demonstrated that in the G0/G1 phase,the DNA expression was elevated in the medium and high-dose groups of butin(P＜0.05),while in the G2 and S phases,the DNA expression was reduced in the medium and high-dose groups of butin(P＜0.05).The results of ELISA,RT-qPCR and Western blot assay revealed that the expression of IL-1β,IL-1 8,caspase-1,and caspase-3 decreased in the butin group compared with the IL-1β+caspase-3 in-hibitor group(P＜0.05).Conclusions Butin inhib-its HFLS-RA proliferation by inhibiting the synthesis of inflammatory vesicles by caspase-1 in the pyroptosis pathway,thereby reducing the production and release of inflammatory factors such as IL-1β and IL-18 down-stream of the pathway,and also inhibits HFLS-RA pro-liferation by exerting a significant blocking effect in the G1 phase,which may be one of the potential mecha-nisms of butin in the treatment of RA.

Aim To investigate the impact of Cinobu-facini on the proliferation,invasion,and apoptosis of HepG2 cells and the underlying mechanism.Methods The proliferation of HepG2 cells was assessed using the CCK-8 method following treatment with Cinobufaci-ni.The invasion capability of HepG2 cells was evalua-ted through Transwell assay after exposure to Cinobufa-cini.The apoptosis rates of HepG2 cells post Cinobufa-cini intervention were measured using flow cytometry,and the expression levels of VEGF in the culture medi-um of HepG2 cells were determined using enzyme-linked immunoassay.Furthermore,qRT-PCR and Western blot analyses were conducted to assess the im-pact of Cinobufacini on mRNA and protein expression levels related to the CXCL5/FOXD1/VEGF pathway.The interaction between CXCL5 and FOXD1 was inves-tigated via co-immunoprecipitation.Results Cinobufa-cini treatment led to a gradual decrease in HepG2 cell viability in a dose-dependent manner compared to the control group(P＜0.05).Moreover,Cinobufacini sig-nificantly suppressed HepG2 cell invasion(P＜0.05)while enhancing cell apoptosis(P＜0.05).Notably,Cinobufacini exhibited inhibitory effects on the CX-CL5/FOXD1/VEGF pathway,as evidenced by re-duced expression of related mRNA and proteins(P＜0.05).FOXD1 was identified as the binding site of CXCL5.Overexpression of CXCL5 resulted in in-creased proliferation and VEGF secretion by HepG2 cells(P＜0.05),and increased expression of FOXD1 and VEGF(P＜0.05).However,Cinobufacini inter-vention effectively inhibited liver cancer cell prolifera-tion and invasion(P＜0.05),promoted apoptosis(P＜0.05),reduced VEGF secretion by HepG2 cells(P＜0.05),and downregulated the expression of CXCL5 and FOXD1 in HepG2 cells(P＜0.05);but com-pared with the unexpressed group of Cinobufacini,its ability to inhibit cell activity was weakened(P＜0.05),and its ability to inhibit the expression of CX-CL5,FOXD1,and VEGF was weakened(P＜0.05).Conclusion Cinobufacini may inhibit HepG2 cell pro-liferation and invasion and promote HepG2 cell apopto-sis by regulating the CXCL5/FOXD1/VEGF pathway.

Objective To investigate the effectiveness of real-time tracking and virtual reality technology(RTVI)used to assist the intraoperative alignment of the trauma orthopaedic surgery robot for the treatment of femoral neck fractures and its impact on the treatment outcome.Methods A retrospective analysis was conducted on 60 patients with femoral neck fractures treated with trauma orthopedic robotic surgery from September 2020 to September 2022.Patients were divided into two groups according to whether RTVI technology was used during surgery to assist robotic surgery.There were 28 patients in the RTVI group(12 males and 16 females),with an average age of(46.2±9.3)years old ranging from 28 to 60 years old.There were 32 patients in the simple Tianji surgical robot group,including 15 males and 17 females,aged(48.2±7.8)years old ranging from 32 to 58.The number of registered fluoroscopy,operation time,total number of intraoperative fluoroscopy,intraoperative blood loss,and hospitalization time of the two groups of patients were observed and recorded.All patients received regular follow-up after surgery,and hip X-rays were routinely reviewed to record Garden alignment index,fracture healing time,postoperative complications,and Harris score.Results All 60 patients were followed up.The RTVI group was followed up for 9 to 16 months with an average of(13.0±l.2)months,and the Tianji surgical robot group alone was followed up for 10 to 14 months with an average of(12.0±1.3)months.During the follow-up period,the femoral neck fractures of both groups of patients healed well,and no complications such as internal fixation loosening and incision infection occurred.The number of registered fluoroscopy,operation time,and number of intraoperative fluoroscopy of patients in the RTVI group were significantly better than those in the simple Tianji surgical robot group(P＜0.01).There was no statistically significant difference in intraoperative blood loss,hospital stay,Garden alignment index,fracture healing time,and hip Harris score between two groups(P＞0.05).Conclusion Although RTVI technology assisted by the surgical robot for femoral neck fracture surgery has little impact on its postoperative outcome,it can effectively reduce the operating time,the number of intraoperative X-ray projections,and the risk of intraopera-tive radiation exposure to patients.It also shortened the learning curve of the operator and better reflected the precision and ef-ficiency of the trauma orthopaedic surgery robot.

Objective:To assess the clinical efficacy and safety of ultrasound-guided brachial plexus block combined with superficial cervical plexus block during the internal fixation of clavicle fractures.Methods:A prospective randomized controlled study was conducted on 100 patients suffering from clavicular fractures who underwent open reduction and internal fixation at Yiwu Central Hospital between June 2019 and March 2022. Using a random number table method, the patients were divided into two groups: a conventional group ( n = 50) and an ultrasound-guided group ( n = 50). The conventional group received blind brachial plexus block combined with superficial cervical plexus block, while the ultrasound-guided group received brachial plexus block combined with superficial cervical plexus block under ultrasound guidance. The anesthesia indices of the patients were recorded, intraoperative vital signs were monitored, the anesthesia effect was evaluated, and adverse reactions to anesthesia were observed. Results:There were no significant differences in general data between the two groups, including age, sex, and ASA grading ( P > 0.05). The duration of anesthesia operation and the onset time of nerve block in the ultrasound-guided group [(5.70 ± 0.79) minutes and (7.28 ± 0.90) minutes] were significantly shorter compared with the conventional group [(12.18 ± 1.03) minutes and (14.55 ± 1.17) minutes, t = 18.92, 21.63, both P < 0.001]. However, the duration of nerve block in the ultrasound-guided group was (417.84 ± 13.88) minutes, which was significantly longer than that in the conventional group [(361.47 ± 12.19) minutes, t = 16.37, P < 0.001]. Additionally, the mean arterial pressure and heart rate in the ultrasound-guided group [(103.36 ± 2.22) mmHg (1 mmHg = 0.133 kPa), (103.02 ± 2.26) mmHg, (76.66 ± 4.51) beats/minute and (75.94 ± 4.34) beats/minute] were significantly lower compared with the conventional group [(103.36 ± 2.22) mmHg, (103.02 ± 2.26) mmHg, (76.66 ± 4.51) beats/minute, and (75.94 ± 4.34) beats/minute, t = 6.83, 5.52, 6.27, 4.96, all P < 0.001]. Furthermore, the effective rate of anesthesia in the ultrasound-guided group was 94.00% (47/50), which was significantly higher than that in the conventional group [76.00% (38/50), χ2 = 6.35, P < 0.05]. The overall incidence of adverse reactions to anesthesia in the ultrasound-guided group was 4.00% (2/50), which was significantly lower than that in the conventional group [20.00% (10/50), χ2 = 6.06, P < 0.05]. Conclusion:Ultrasound-guided brachial plexus block combined with superficial cervical plexus block for internal fixation of clavicular fractures can effectively shorten the onset time of anesthesia, minimize fluctuations in patients' intraoperative vital signs, improve the efficiency of anesthesia, and reduce the risk of adverse reactions.

AIM:To compare the risk of hypergly-caemia with PCSK9 inhibitors versus statins,based on U.S.Food and Drug Administration Adverse Events Reporting System(FAERS).METHODS:The hyperglycaemia reports induced by"alirocumab","evolocumab","atorvastatin"and"rosuvastatin"were utilized as the first suspected drugs from the database of FAERS from 2016 to the third quarter of 2023.The report odd ratio(ROR)method was employed.RESULTS:Based on the FAERS database,the ROR(95％CI)for hyperglycaemia due to Ali-rocumab versus Atorvastatin and Rosuvastatin were 0.628(0.545,0.724)and 0.307(0.263,0.357),the ROR(95％CI)for hyperglycaemia due to Evoloc-umab were 0.817(0.750,0.889)and 0.399(0.361,0.441),all generated no adverse reaction signals.The ROR(95％CI)for hyperglycaemia due to Ali-rocumab and Evolocumab versus all other drugs in FAERS were 1.488(1.315,1.682)and 1.934(1.845,2.027),all generated adverse reaction signals,re-spectively.CONCLUSION:Based on the FAERS data-base,PCSK9 inhibitors have a lower risk of hyper-glycemia than statins and deserve clinical attention.

AIM To study the chemical constituents from the branches and leaves of Toona ciliata Roem.var.pubescens(Franch.)Hand-Mazz.and their antitumor activities.METHODS The compounds were isolated and purified by silica gel,RP-18 reverse phase silica gel and semi-preparative HPLC,the structures of compounds were identified by physicochemical properties and spectral data.The antitumor activities were determined by MTT method.RESULTS Fifteen compounds were isolated and identified as toonaolide D(1),toonaciliatin E(2),bourjotinolone A(3),(21R,23R)-epoxy-21α-ethoxy-24S,25-dihydroxyapotirucalla-7-en-3-one(4),(Z)-toonasterone C(5),(E)-toonasterone(6),3-epi-dyscusin C(7),(Z)-aglawone(8),(E)-volkendousin(9),8(14),15-isopimaradiene-2α,3α,19-triol(10),(-)-loliolide(11),cyclohexenone(12),pubinernoid A(13),quercetin-3-O-(4″-methoxy)-α-L-rahmnopyranosyl(14),5-hydroxymethyl-2-furancarboxaldehyde(15).The IC50 values of compounds 3 and 4 on K562 cells were 54.2 and 47.3 μmol/L,respectively,and the IC50 values on HEL cells were 47.3 and 61.1 μmol/L,respectively.CONCLUTION Compounds 4,7,10 and 11 are isolated from Toona genus for the first time,and compounds 2,15 are first isolated from this plant.Compounds 3 and 4 show weak antitumor activities.

Objective:To explore the impact of the hospital health literacy environment on patients′ postoperative pain self-management behaviors, aiming to provide insights for hospitals to implement the Comprehensive Pain Management Pilot Work Program in hospitals and to promote self-health management among patients with other diseases or symptoms. Methods:From November to December 2023, a convenience sampling method was used to select postoperative patients from three grade A tertiary general hospitals in Zhejiang Province for an on-site questionnaire survey. The Chinese version of brief health literacy screen (BHLS), short-form health literacy environment scale (SF-HLES) and postoperative pain self-management behavior questionnaire (PPSMB) were used as survey tools to investigate the health literacy level of patients, the health literacy environment of the hospital, and the postoperative pain management behaviors of patients. Two-way ANOVA was used to compare the impact of different dimensions of the hospital health literacy environment on postoperative pain management behaviors among patients with different levels of health literacy. Multiple linear regression analysis was used to explore the relationship between the hospital health literacy environment, individual health literacy, and patients′ postoperative pain self-management behaviors, and to discuss the impact of individual health literacy on patients′ postoperative pain self-management behaviors under different hospital health literacy environments.Results:341 valid questionnaires were collected. The average score of the hospitals′ SF-HLES was (73.62±19.54) points. The average score of the patients′ BHLS was (9.65±2.88) points. The average score of the patients′ PPSMB was (25.99±6.35) points. Two-way ANOVA results showed that the interaction between individual health literacy and the clinical dimension ( F=5.463, P=0.020) and structural dimension ( F=6.470, P=0.011) of the hospital health literacy environment had a statistically significant impact on patients′ postoperative pain self-management behaviors, while the interaction with the interpersonal dimension ( F=0, P=0.984) had no statistically significant impact on pain self-management behaviors. Simple effect analysis indicated that only in the high health literacy environment of the clinical and structural dimensions did the difference in pain self-management behaviors between patients with good health literacy and those with limited health literacy had statistical significance ( P<0.001). Multiple linear regression analysis results showed that for each 1-point increase in the patients′ BHLS score, their PPSMB score increased by 3.74 points ( β1=0.832, P<0.001); for each 1-point increase in the hospital′s SF-HLES score, the patients′ PPSMB score could increase by 0.198 points ( β2=0.610, P<0.001). In a low health literacy environment, individual health literacy did not affect pain self-management behaviors ( P>0.05); however, in a high health literacy environment, for each 1-point increase in the patients′ BHLS score, their PPSMB score correspondingly increased by 4.037 points ( β4=0.317, P<0.001). Conclusions:The positive impact of individual health literacy on pain self-management is contingent upon a high-quality hospital health literacy environment. This suggests that optimizing the hospital health literacy environment is a necessary precondition for implementing the relevant content of the Comprehensive Pain Management Pilot Work Program and can provide a reference for promote self-health management among patients with pain and other diseases or symptoms.

Objective: First-line bevacizumab plus carboplatin and paclitaxel (CP) is approved for stage III/IV ovarian cancer treatment following initial surgical resection, based on global phase III GOG-0218 and ICON7 trials. This study evaluated the efficacy and safety of bevacizumab + CP as first-line ovarian cancer therapy in Chinese patients. Methods: Patients with newly diagnosed, International Federation of Gynecology and Obstetrics (FIGO) stage III/IV epithelial ovarian, fallopian tube, or primary peritoneal cancer post-primary surgery were randomized 1:1 to receive 6 cycles of CP with bevacizumab/ placebo, followed by bevacizumab/placebo maintenance until unacceptable toxicity or disease progression. Primary endpoint was investigator-assessed progression-free survival (PFS). Stratification factors were FIGO stage and debulking status (stage III optimally debulked vs stage III suboptimally debulked vs stage IV) and Eastern Cooperative Oncology Group performance status (0 vs 1 or 2). Results: Of randomized patients, 51 received bevacizumab + CP and 49 received placebo + CP. Median PFS was 22.6 months with bevacizumab + CP (95% confidence interval [CI]=18.6, not estimable) and 12.3 months (95% CI=9.5, 15.0) with placebo + CP (stratified hazard ratio=0.30; 95% CI=0.17, 0.53). Treatment-related grade 3/4 adverse events occurred in 46 of 49 (94%) patients receiving bevacizumab + CP, and 34 of 50 (68%) receiving placebo + CP. Conclusion Bevacizumab + CP showed clinically meaningful improvement in PFS vs placebo + CP, consistent with GOG-0218 results. Safety data were aligned with the known bevacizumab safety profile. These results support first-line bevacizumab + CP therapy in Chinese patients with ovarian cancer.

Objective To analyze the relationship between expression level of protein O-fucosyltransferase 1(POFUT1)and tumor immune infiltration level and prognosis of patients,and explore the value of POFUT1 in tumor immunotherapy.Methods Based on the pan-cancer data,R software was used to analyze the changes in POFUT1 expression levels of various tumor tissues and their correlation with the risk ratio of various tumor patients,and screen for tumor types related to patient prognosis and POFUT1 expression.Through the String database,a protein interaction network was constructed,POFUT1-associated genes were screened,and functional enrichment analysis was performed.The estimate software package was used to analyze the correlation between POFUT1 expression and immune infiltration score in selected tumor tissues.The TIMER database was used to analyze the correlation between the expression level of POFUT1 and various levels of immune cell infiltration,and the correlation between the infiltration level of immune cells and the prognosis of patients.Results The expression levels of POFUT1 were significantly high in 14 types of tumors(P<0.05),and were associated with poor prognosis of low-grade gliomas,lung adenocarcinoma,and thyroid cancer(Hazard Ratio>1,P<0.05).POFUT1 and its associated genes were highly involved in Notch signaling pathway,lymphocyte activation and immune regulation processes.The expression level of POFUT1 was positively correlated with the infiltration degree of B cells,CD8+T cells,CD4+T cells,macrophages,neutrophils and dendritic cells in low-grade gliomas,as well as positively associated with the infiltration level of neutrophils in LUAD,and B cells,CD4+ T cells and macrophages in thyroid cancer(|r|>0.2,P<0.05).The high infiltration levels of the above 6 types of immune cells were correlated with the poor prognosis of patients with low-grade glioma(Hazard Ratio>1,P<0.05).Conclusion POFUT1 was highly involved in the process of immune regulation,affecting the immune infiltration level of some tumors and further influencing patient prognosis.It may have the potential to become a target for tumor immunotherapy.