OBJECTIVE To investigate the protective effect and mechanism of saikosaponin C （SSC） on acute liver injury （ALI） in mice induced by carbon tetrachloride （CCl4） based on serum metabolomics. METHODS Forty mice were divided into blank group （water）， model group （water）， positive control drug group （Biphenyl diester drop pills， 150 mg/kg）， and SSC low- and high-dose groups （2.5， 10 mg/kg） using the random number table method， with 8 mice in each group. They were given water/ relevant drugs， once a day， for 7 consecutive days. One hour after the last administration， all mice were intraperitoneally injected with 0.2% CCl4 olive oil to induce ALI model， except for the blank group. After 17 hours of the modeling， the liver index of mice was calculated. The levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， lactate dehydrogenase （LDH）， tumor necrosis factor-α （TNF-α）， interleukin-6 （IL-6）， and IL-1β in serum of mice were detected. The histopathological changes of liver tissue were observed. Meanwhile， the serum metabolomics of mice were analyzed by liquid chromatography-mass spectrometry. RESULTS Compared with the blank group， the levels of liver index， ALT， AST， LDH， TNF-α， IL-6， and IL-1β in the model group were significantly increased （P＜0.01）. Hepatocytes were edema， vacuolar degeneration， more necrosis， and a large number of inflammatory cells were infiltrated. Compared with the model group， liver index and serum index levels of mice were significantly decreased （P＜0.05 or P＜0.01）， accompanied by marked improvement in histopathological damage to the liver tissue. The metabolomics results showed that compared with the model group， there were 63 up-regulated and 256 down-regulated differential metabolites in the serum of mice in the SSC high-dose group， including prostaglandin B2， 20-hydroxy-leukotriene B4， 5- hydroxy-L-tryptophan， 7α -hydroxycholesterol， etc.； these metabolites were primarily involved in metabolic pathways such as arachidonic acid metabolism， 5-hydroxytryptamine synapse， primary bile acid biosynthesis. CONCLUSIONS SSC exerts a protective effect against CCl4-induced ALI by down-regulating the level of key metabolites such as prostaglandin B2 and 20-hydroxy-leukotriene B4， and then ruducing metabolic pathways such as arachidonic acid metabolism， 5- hydroxytryptamine synapse， and primary bile acid biosynthesis.

BACKGROUND: The primary limitation to kidney transplantation is organ shortage. Recent progress in gene editing and immunosuppressive regimens has made xenotransplantation with porcine organs a possibility. However, evidence in pig-to-human xenotransplantation remains scarce, and antibody-mediated rejection (AMR) is a major obstacle to clinical applications of xenotransplantation. METHODS: We conducted a kidney xenotransplantation in a brain-dead human recipient using a porcine kidney with five gene edits (5GE) on March 25, 2024 at Xijing Hospital, China. Clinical-grade immunosuppressive regimens were employed, and the observation period lasted 22 days. We collected and analyzed the xenograft function, ultrasound findings, sequential protocol biopsies, and immune surveillance of the recipient during the observation. RESULTS: The combination of 5GE in the porcine kidney and clinical-grade immunosuppressive regimens prevented hyperacute rejection. The xenograft kidney underwent delayed graft function in the first week, but urine output increased later and the single xenograft kidney maintained electrolyte and pH homeostasis from postoperative day (POD) 12 to 19. We observed AMR at 24 h post-transplantation, due to the presence of pre-existing anti-porcine antibodies and cytotoxicity before transplantation; this AMR persisted throughout the observation period. Plasma exchange and intravenous immunoglobulin treatment mitigated the AMR. We observed activation of latent porcine cytomegalovirus toward the end of the study, which might have contributed to coagulation disorder in the recipient. CONCLUSIONS 5GE and clinical-grade immunosuppressive regimens were sufficient to prevent hyperacute rejection during pig-to-human kidney xenotransplantation. Pre-existing anti-porcine antibodies predisposed the xenograft to AMR. Plasma exchange and intravenous immunoglobulin were safe and effective in the treatment of AMR after kidney xenotransplantation.
Transplantation, Heterologous/methods* ; Kidney Transplantation/methods* ; Heterografts/pathology* ; Immunoglobulins, Intravenous/administration & dosage* ; Graft Survival/immunology* ; Humans ; Animals ; Sus scrofa ; Graft Rejection/prevention & control* ; Kidney/pathology* ; Gene Editing ; Species Specificity ; Immunosuppression Therapy/methods* ; Plasma Exchange ; Brain Death ; Biopsy ; Male ; Aged

Cancer multidrug resistance (MDR) impairs the therapeutic efficacy of various chemotherapeutics. Novel approaches, particularly the development of MDR reversal agents, are critically needed to address this challenge. This study demonstrates that tenacissoside I (TI), a compound isolated from Marsdenia tenacissima (Roxb.) Wight et Arn, traditionally used in clinical practice as an ethnic medicine for cancer treatment, exhibits significant MDR reversal effects in ABCB1-mediated MDR cancer cells. TI reversed the resistance of SW620/AD300 and KBV200 cells to doxorubicin (DOX) and paclitaxel (PAC) by downregulating ABCB1 expression and reducing ABCB1 drug transport function. Mechanistically, protein arginine methyltransferase 1 (PRMT1), whose expression correlates with poor prognosis and shows positive association with both ABCB1 and EGFR expressions in tumor tissues, was differentially expressed in TI-treated SW620/AD300 cells. SW620/AD300 and KBV200 cells exhibited elevated levels of EGFR asymmetric dimethylarginine (aDMA) and enhanced PRMT1-EGFR interaction compared to their parental cells. Moreover, TI-induced PRMT1 downregulation impaired PRMT1-mediated aDMA of EGFR, PRMT1-EGFR interaction, and EGFR downstream signaling in SW620/AD300 and KBV200 cells. These effects were significantly reversed by PRMT1 overexpression. Additionally, TI demonstrated resistance reversal to PAC in xenograft models without detectable toxicities. This study establishes TI's MDR reversal effect in ABCB1-mediated MDR human cancer cells through inhibition of PRMT1-mediated aDMA of EGFR, suggesting TI's potential as an MDR modulator for improving chemotherapy outcomes.
Humans ; Protein-Arginine N-Methyltransferases/antagonists & inhibitors* ; Drug Resistance, Neoplasm/drug effects* ; ErbB Receptors/genetics* ; Animals ; Cell Line, Tumor ; Drug Resistance, Multiple/drug effects* ; Methylation/drug effects* ; Saponins/administration & dosage* ; Mice ; Mice, Nude ; Mice, Inbred BALB C ; ATP Binding Cassette Transporter, Subfamily B/genetics* ; Doxorubicin/pharmacology* ; Paclitaxel/pharmacology* ; Female ; Repressor Proteins

Objective:To explore the influence of static mechanical strain(SMS)on the osteoclastogenic gene expression of healthy periodontal ligment stem cells(HPDLSCs)and periodontitis periodontal ligment stem cells(PPDLSCs).Methods:HPDLSCs and PP-DLSCs were respectively isolated and cultured by low density in vitro.The expression of mesenchymal stem cell markers were detected by flow cytometry.Then,6％,8％,10％,12％and 14％SMS were respectively loaded to the HPDLSCs and PPDLSCs by Flexcell Tension Unit,and the expression of RANKL and C-fos was detected by real time RT-PCR.Results:Both HPDLSCs and PPDLSCs strongly expressed the mesenchymal stem cell markers STRO-1,CD146,CD90 and CD29,and higher expression of the above markers was found in HPDLSCs compared with PPDLSCs(P＜0.05).The expression of RANKL and C-fos in PPDLSCs was more obvious than that in HPDLSCs without SMS loading(P＜0.05).For HPDLSCs,the SMS of 14％induced significant up-regulation of RANKL and C-fos(P＜0.05),while no alteration was confirmed for the above osteoclastogenic genes when the SMS≤ 12％(P＞O.05).In addition,the expression of RANKL and C-fos was up-regulated significantly in PPDLSCs when the SMS≥ 10％(P＜0.05),and the expression of the above genes was not activated when the SMS ≤8％.Conclusion:HPDLSCs and PPDLSCs response differently to SMS,and ex-cessive SMS may lead to enhanced expression of osteoclastogenic genes in PPDLSCs.

Frailty is a complex age-related clinical condition characterized by a decline in physiological capacity across several organ systems, with a resultant increased susceptibility to stressors.The relationship between frailty assessment and elderly patients’ safety during perioperative period has received widespread attention.This paper reviews research progress of frailty assessment in elderly patients during perioperative period in the department of urology.

OBJECTIVE To investigate the damage effect and potential toxic mechanism of SARS-CoV-2 spike protein(S protein)on human neuroblastomacells(SH-SY5Y).METHODS SH-SY5Y were treated with S protein at concentrations of 25,50,75,and 100 mg·L-1 for 24 h.Cell viability of SH-SY5Y was detected using the CCK-8 assay.The cytotoxic lactate dehydrogenase(LDH)detection kit was used to measure the release rate of LDH,and the 5-ethynyl-2′-deoxyuridine(EdU)-488 cell prolifera-tion kit was used to assess cell proliferation.The ATP detection kit was used to measure intracellular ATP content.The JC-1 fluorescent probe method was employed to detect the mitochondrial membrane potential(MMP)of cells.Seahorse XF was used to measure mitochondrial respiratory and glycolytic capacity.RESULTS Compared with the cell control group,cell viability was significantly reduced in S protein 25,50,75 and 100 mg·L-1 groups(P<0.01),and the half-inhibition concentration(IC50)was 65.05 mg·L-1.The LDH release rate wassignificantly increased(P<0.01)and the proportion of EdU positive cellswas significantly reduced(P<0.01)in S protein 25,50,75 and 100 mg·L-1 groups.S protein signifi-cantly reduced intracellular ATP content(P<0.01)at the concentrations of 75 and 100 mg·L-1,while significantly reduced intracellular MMP(P<0.05,P<0.01)at the concentrations of 50 and 75 mg·L-1.S protein 50 mg·L-1 increased the maximum value of basal glycolysis levels and glycolytic capacity(P<0.05,P<0.01),and S protein 25 and 50 mg·L-1 increased the maximum value of respiration capacity(P<0.05,P<0.01).SH-SY5Y cell viability was positively correlated with the intracellular ATP content and the MMP level(r2=0.9209,P=0.001;r2=0.6170,P=0.0025),and negatively correlated with the maximum level of basal glycolysis and glycolytic capacity(r2=0.5194,P=0.0285;r2=0.6664,P=0.0073),and nega-tively correlated with ATP production capacity(r2=0.8204,P=0.0008).CONCLUSIONS protein decreases the viability of SH-SY5Y cells and inhibited cell proliferation.The mechanism may be closely related to the disorder of energy metabolism.

OBJECTIVE To investigate the damage effect and mechanisms of cyclophosphamide(CTX)and its active metabolite derivative 4-hydroperoxycyclophosphamide(4-HC)to human neuroblas-toma SH-SY5Y cells.METHODS SH-SY5Y cells were treated with CTX[0(cell control),0.01,0.1,1,5,10,20,40 and 80 mmol·L-1]and 4-HC[0(cell control),0.01,0.1,1,5,10,20,40 and 80 μmol·L-1]for 48 h.Cell confluence and morphology were observed by the IncuCyte ZOOM system.Cell viability was assessed by CCK-8 assay.Lactate dehydrogenase(LDH)release was measured by LDH assay kit.SH-SY5Y cells were treated with CTX(0,1,5,10 and 20 mmol·L-1)and 4-HC(0,1,5,10 and 20 μmol·L-1)for 48 h before cell proliferation was analyzed by 5-ethynyl-2′-deoxyuridine(EdU)staining assay.Immunofluorescence was employed to assess the levels of the DNA double-strand break marker γ-H2AX and to evaluate changes in mitochondrial membrane potential.SH-SY5Y cells were treated with CTX(0,1,5 and 10 mmol·L-1)and 4-HC(0,1,5 and 10 μmol·L-1)for 48 h,and the alterations in glycolysis and oxidative phosphorylation levels were analyzed using the Seahorse XFe96 Analyzer.RESULTS Compared with the cell control group,cell confluence and cell viability were significantly reduced in the CTX and 4-HC groups(P<0.01),and the half-maximal inhibitory concentrations(IC50)for CTX and 4-HC were 4.44 mmol·L-1 and 4.78 μmol·L-1,respectively.The release rate of LDH was signif-icantly increased while the percentage of EdU+cells was significantly reduced in the CTX and 4-HC groups(P<0.01).The percentage of γ-H2AX+cells was significantly increased and mitochondrial membrane potential significantly decreased in the CTX and 4-HC group(P<0.05).Treatment with CTX and 4-HC resulted in reduced levels of maximum glycolytic capacity,glycolytic reserve,maximal respi-ration,and ATP production(P<0.05).CONCLUSION CTX and 4-HC exert significant cytotoxic effects on SH-SY5Y cells by disrupting cell membrane structure,impeding cell proliferation,and reducing cell viability.The mechanisms underlying these effects may involve intracellular DNA damage,disturbance of energy metabolism and mitochondrial dysfunction.

Objective:To prepare a novel 68Ga-labeled bicyclic peptide targeting poliovirus receptor related protein 4 (PVRL4, Nectin-4), and evaluate its feasibility for breast cancer imaging via in vitro and in vivo experiments. Methods:A Biotin-modified bicyclic peptide targeting Nectin-4, Biotin-BMIC, was synthesized, and its targeting properties were preliminarily evaluated by in vitro cell staining experiments. BMIC was modified by 1, 4, 7-triazonane-1, 4-diacetic acid (NODA) and the labeling precursor NODA-BMIC was prepared. A potential PET probe targeting Nectin-4, 68Ga-NODA-BMIC was prepared by one-step labeling strategy. The imaging properties of the probe were investigated by in vivo microPET imaging and in vitro experiments in mice bearing breast tumors. Data were analyzed by independent-sample t test and repeated measures analysis of variance. Results:Fluorescence staining of the cells showed that the fluorescently labeled bicyclic peptide, Biotin-BMIC, was highly aggregated in Nectin-4 positive BT474 breast cancer cells compared to those in Nectin-4 negative MDA-MB-231 cells. The uncorrected yield of 68Ga-NODA-BMIC was (71.5±2.2)% and the radiochemical purity was greater than 95%. The specific activity was greater than 3 GBq/μmol. After incubation 10, 30, 60 and 120 min, higher radioactivity uptakes were found in BT474 breast cancer cells compared to those in MDA-MB-231 breast cancer cells respectively ( F=1 302.00, P<0.001). MicroPET imaging showed that the BT474 xenograft tumors were clearly visible with favorable contrast. A significant statistical difference in uptakes between BT474 and MDA-MB-231 xenograft tumor uptake at 10, 30, 60, and 120 min after probe injection respectively was existed ( F=1 826.00, P<0.001). At 60 min postinjection, the uptake value of BT474 tumors was (5.03±0.14) percentage activity of injection dose per gram of tissue (%ID/g), which was significantly higher than that of MDA-MB-231 tumors ((0.19±0.04) %ID/g; t=79.40, P<0.001). Meanwhile, the tumor-to-muscle ratios in the former were also greater than those in the latter ( F=222.00, P<0.001). At 60 min postinjection, the tumor-to-muscle ratio in the former was significantly higher than that in the latter (24.75±3.10 vs 1.30±0.15; t=14.31, P=0.002). The results were consistent with the immunohistochemistry staining. Conclusions:A novel bicyclic peptide PET probe targeting Nectin-4, 68Ga-NODA-BMIC, is easy to be synthesized and owns satisfactory labeling yield and radiological purity. The imaging performance is good and the target tissues could be visualized. It may play a unique role in the diagnosis and treatment of breast cancer.

Objective To investigate the effects of different approaches of periacetabular osteotomy(PAO)on traumatic stress,lower limb strength and prognosis of acetabular dysplasia(DDH).Methods Ninety-seven patients with DDH in our hospital from January 2021 to January 2022 were randomly divided into two groups.Among them,48 patients in the control group received conventional ilioinguinal approach PAO treatment,while 49 patients in the study group received modified ilioinguinal approach PAO treatment.The surgical and postoperative rehabilitation indexes,Mckay clinical efficacy,lower limb line of force,complications,traumatic stress factors before and after surgery[adrenalin(NE),cortisol(COR),angiotensin Ⅱ(AngⅡ)],hip imaging indexes[anterior CE Angle(ACEA),lateral CE Angle(LCEA),acetabular index(AI)]and changes were compared between the two groups Good Harris hip function Score(mHHS),hip outcome score Daily Living Ability Scale(HOS-ADL).Results The operation time of the study group and the control group were(128.64±18.73)min and(141.80±21.59)min respectively,the intraoperative blood loss were(472.95±35.18)ml and(495.68±40.26)ml respectively,the postoperative drainage were(242.39±32.74)ml and(305.81±39.56)ml respectively,and the hospital stay were(11.57±2.29)D and(12.86±2.41)d respectively,with significant differences between the two groups(P＜0.05);The excellent and good rate of McKay's clinical efficacy in the study group(95.92％)was higher than that in the control group(81.25％),and the differences were statistically significant(P＜0.05);Serum NE in the study group and control group at 1 d,3 d,and7 d postoperatively were(73.16±8.07)ng/L and(81.33±8.52)ng/L,(65.81±7.29)ng/L and(72.24±7.65)ng/L,(45.98±6.31)ng/L and(50.37±7.02)ng/L,respectively,and COR were(164.84±19.35)ng/L and(178.62±21.46)ng/L,(142.69±17.81)ng/L and(157.36±19.22)ng/L,(88.79±16.13)ng/L and(97.62±17.50)ng/L,respectively,and AngⅡ was(138.74±20.51)mmol/L and(150.19±21.36)mmol/L,(128.35±17.69)mmol/L and(137.18±19.24)mmol/L,and(119.82±17.41)mmol/L and(128.73±18.50)mmol/L,respectively,and the differences between the two groups were all statistically significance(P＜0.05);ACEA at 3,6,and 12 months postoperatively in the study and control groups were(29.71±4.81)° and(27.68±4.53)°,(29.80±4.75)° and(27.72±4.60)°,(29.64±4.79)° and(27.63±4.51)°,respectively,and LCEA was(33.79±6.12)° and(31.04±5.83)°,(33.82±6.10)° and(31.10±5.90)°,(33.75±6.08)° and(31.05±5.77)°,and AI was(6.15±1.86)° and(7.03±1.94)°,(6.08±1.82)° and(7.01±1.89)°,(6.12±1.84)° and(7.06±1.90)°,respectively.mHHS scores was(72.15±7.65)and(68.23±7.71),(76.51±7.52)and(72.19±7.94),(90.13±5.16)and(86.76±5.72),and HOS-ADL scores were(79.92±7.50)and(76.26±7.62)points,(80.85±7.42)and(77.13±7.66)points,(89.73±6.37)and(86.25±7.15)points,and the differences between the two groups were statistically significant(P＜0.05);the comparison of lower limb force lines and complications between the two groups showed no statistically significant differences(P＞0.05).Conclusion Modified ilioinguinal approach PAO in the treatment of DDH can optimize the operation,reduce traumatic stress factors,further improve the curative effect,improve the status of femoral head coverage and hip function,and improve the quality of life,with high safety.

Currently, the application rate of virtual reality technology in the fields of medical education and medical treatment is relatively low. In this study, based on the structure of the human stomach, virtual reality technology, sensing technology, big data technology, and cloud computing technology were integrated. A new form of medical education was established to include the online website platform for data storage and analysis and the offline VR glasses for the physical operation. Through the use of 3d Max technology, Unity3D technology, and C# language, we constructed a three-dimensional model of the human stomach to present the stomach in three dimensions. This enables the users to immerse in it to achieve true human-computer interactive learning. This study updates the concept of medical education, effectively improves the quality and efficiency of medical education, and facilitates the development of surgical programs and reduction in the risk of surgery, as well as provides experimental materials for scientific research. In the future, it can be further developed to include the three-dimensional structures of the circulatory system and other major systems in the human body.