Median arcuate ligament compression syndrome (MALS) presents with atypical clinical manifestations. MALS is rarely reported in pediatric patients but is recognized as an independent risk factor for postoperative hepatic artery thrombosis in liver transplant recipients. We report a case of a pediatric liver transplant recipient with hepatolenticular degeneration, cirrhosis, and acute liver failure. Despite undergoing artificial liver support therapy, the patient showed no significant improvement in liver function and subsequently underwent liver transplantation. Intraoperatively, weak arterial pulsation and the absence of a pulsatile waveform in the hepatic artery anastomosis raised suspicion of MALS. The condition was successfully managed by releasing and transecting the median arcuate ligament, along with ligation of the splenic and left gastric arteries. To optimize transplantation outcomes, meticulous preoperative imaging assessment, particularly focusing on characteristic findings in CT angiography, is essential. Additionally, individualized surgical planning and intraoperative adjustments based on ultrasound monitoring and arterial pulsation assessments are critical for ensuring successful transplantation.

This study assessed the role and mechanism of the recombinant Bacillus Calmette-Gue?rin vaccine(rBCG)with c-di-AMP adjuvant in regulating metabolism and immunity in epididymal white adipose(eWAT)in mice.Male C57BL/6 mice were intravenously immunized with BCG and rBCG,and their body weights were monitored.eWAT was isolated from the mice,and the stromal vascular fractions(SVFs)cell number was counted with a hemocytometer.Sections of mouse adipose tissue were prepared,and the size,number,and morphology of eWAT adipocytes and crown-like structure(CLS)formation were compared under a microscope after HE staining.The transcription levels of lipid metabolism-associated factors,cytokines and aging-associated genes in each group were determined with qRT-PCR.The body weights of mice gradually increased after immunization with BCG and rBCG.The proportions of eWAT increased,and the SVFs cell number decreased,in rBCG immunized mice.HE staining indicated that BCG immunization promoted hyperplasia,whereas rBCG immunization promoted hypertrophy of eWAT adipocytes;moreover,both BCG and rBCG immunization induced CLS formation in eWAT.The qRT-PCR results indicated that rBCG immunization inhibited the expression of genes associated with lipolysis and energy expenditure in eWAT.BCG immunization had little effect on cytokine transcription,whereas rBCG significantly induced the transcription of IFN-γ and IL-1Ra,and inhibited that of IL-15 and IL-2,but did not induce the expression of aging-associated genes.Thus,rBCG immunization induced eWAT adipocyte hypertrophy,which was associated with the inhibition of eWAT lipolysis and the regulation of cytokine expression.

This study was aimed at developing an in vitro fluorescent substrate assay for the activity of leucyl aminopeptid-ase(LAP)from Echinococcus multilocularis and comparing it with the chemical chromogenic substrate enzyme activity assay.Through the establishment of reaction conditions for the fluorescent substrate-based in vitro enzyme activity assay,we com-pared the differences between the fluorescent substrate L-Leucine-7-amido-4-methylocoumarin(Leu-AMC)and the chemical chromogenic substrate L-Leucine-4-nitroanilide(Leu-pNA)through molecular docking,inhibition rates,and precision measures.Molecular docking revealed that the fluorescent substrate Leu-AMC had higher affinity for the protein than the chemical chromogenic substrate Leu-pNA.Through analysis of the effects of varying reaction conditions on fluorescence intensi-ty,we optimized the fluorescent substrate enzyme activity assay to demonstrate favorable performance at a reaction temperature of 37℃,a pH of 9.0,a protein concentration of 800 nmol/L,and a reaction duration of 60 minutes.Leu-AMC exhibited significant and distinct responses at a 5 μmol/L substrate concentration,under varying substrate conditions.The fluo-rescent substrate assay demonstrated more significant intergroup differences than the chemical chromogenic substrate assay when various inhibitors were added.This study established a fluorescence-based enzyme activity assay for leucyl aminopeptidase from Echinococcus multilocularis by using Leu-AMC as the substrate;this method demonstrated a more significant intergroup difference and sensitivity than the chemical chromogenic substrate assay.

Objective To analyze the effect of sodium cantharidinate and vitamin B6 injection(SCV)on four human hepatocellular carcinoma(HCC)cell lines(SMMC-7721,Bel-7402,Huh7,and HepG2)and explore its mechanism.Methods Normal hepatic cell line L02 was treated with SCV at concentrations of 0 μmol/L(control),0.5,1,2,4,8,16,and 32 μmol/L,and the cytotoxicity of SCV on L02 cells was detected using CCK-8 assay.Human HCC cell lines(SMMC-7721,Bel-7402,Huh7,and HepG2)were cultured.SCV-untreated control group(0 μmol/L)and 2,4,and 8 μmol/L SCV-treated groups were set up.CCK-8 assay,plate cloning formation assay,Transwell assay,wound healing assay,and flow cytometry were used to detect the effects of SCV on the growth and proliferation capacity,colony formation ability,invasion and migration capabilities,cell cycle,and apoptosis of the four hepatocellular carcinoma cell lines,respectively.Western blotting was performed to detect the expression levels of apoptosis-related proteins,including nuclear factor kappa-B subunit p65(p65),B-cell lymphoma 2(Bcl-2),and Caspase-3,and to preliminarily explore the underlying mechanism.Results The CCK-8 assay showed that SCV at 0.5,1,2,4,and 8 μmol/L had no significant cytotoxic effect on L02 cells compared with untreated control group,so 2,4,and 8 μmol/L SCV were selected for subsequent experiments.Compared with the untreated control group(0 μmol/L),SCV at different concentrations(2,4,and 8 μmol/L)significantly inhibited the proliferation of the four HCC cell lines(P<0.001).The plate cloning formation assay showed that SCV at different concentrations(2,4,and 8 μmol/L)significantly reduced the colony formation ability of the four HCC cell lines(P<0.05 or P<0.01 or P<0.001).In addition,Transwell and wound healing assays revealed that SCV at different concentrations(2,4,and 8 μmol/L)significantly inhibited the invasion and migration of HCC cells(P<0.05 or P<0.01 or P<0.001).In the above results,the inhibitory effect of SCV was concentration-dependent.Flow cytometry analysis indicated that SCV arrested cells in the G2/M phase(P<0.05 or P<0.01 or P<0.001)and significantly promoted cell apoptosis(P<0.05 or P<0.01 or P<0.001).Western blotting showed that SCV significantly down-regulated the expression of p65(P<0.05 or P<0.01)and Bcl-2(P<0.05),and up-regulated the expression of Caspase-3(P<0.05 or P<0.01).Conclusions SCV can significantly inhibit the proliferation,colony formation,invasion,and migration of multiple human HCC cell lines and arrest the cell cycle.SCV may inhibit the expression of p65 and Bcl-2,thereby lifting their inhibitory effect on the apoptotic pathway and activating Caspase-3 to promote apoptosis.

Objective To evaluate the efficacy of a combined immunosuppression regimen in modulating rejection in genetically modified pig-to-macaque xenogeneic heart transplantation.Methods Two xenogeneic heart transplantation models were constructed using genetically modified pigs and macaques.Dynamic monitoring of recipient peripheral blood immune parameters and observation of graft pathological changes were performed.Results Regimen 1,featuring B-cell depletion,T-cell inhibition,and C3 complement suppression,reduced lymphocyte levels but failed to control acute humoral rejection and macrophage infiltration.Regimen 2,adding C5 complement inhibition and interleukin-6 inhibition to Regimen 1,more effectively lowered lymphocyte levels,inhibited acute humoral rejection and complement activation,and decreased antibody deposition.However,a late-phase cytokine storm and residual T cells emerged.Conclusions Regimen 2 reduces the hyperacute and acute rejection risks through multi-target intervention.Yet,it requires balancing medication complexity and safety.This indicates the need to optimize cellular immune regulation and adjust the plan through dynamic multidimensional monitoring.

This study assessed the role and mechanism of the recombinant Bacillus Calmette-Gue?rin vaccine(rBCG)with c-di-AMP adjuvant in regulating metabolism and immunity in epididymal white adipose(eWAT)in mice.Male C57BL/6 mice were intravenously immunized with BCG and rBCG,and their body weights were monitored.eWAT was isolated from the mice,and the stromal vascular fractions(SVFs)cell number was counted with a hemocytometer.Sections of mouse adipose tissue were prepared,and the size,number,and morphology of eWAT adipocytes and crown-like structure(CLS)formation were compared under a microscope after HE staining.The transcription levels of lipid metabolism-associated factors,cytokines and aging-associated genes in each group were determined with qRT-PCR.The body weights of mice gradually increased after immunization with BCG and rBCG.The proportions of eWAT increased,and the SVFs cell number decreased,in rBCG immunized mice.HE staining indicated that BCG immunization promoted hyperplasia,whereas rBCG immunization promoted hypertrophy of eWAT adipocytes;moreover,both BCG and rBCG immunization induced CLS formation in eWAT.The qRT-PCR results indicated that rBCG immunization inhibited the expression of genes associated with lipolysis and energy expenditure in eWAT.BCG immunization had little effect on cytokine transcription,whereas rBCG significantly induced the transcription of IFN-γ and IL-1Ra,and inhibited that of IL-15 and IL-2,but did not induce the expression of aging-associated genes.Thus,rBCG immunization induced eWAT adipocyte hypertrophy,which was associated with the inhibition of eWAT lipolysis and the regulation of cytokine expression.

Objective To clarify the effect of preoperative oral carbohydrate on overweight or obese patients undergoing laparoscopic hysterectomy in Trendelenburg position.Methods A total of 72 overweight or obese patients who underwent laparoscopic hysterectomy in Trendelenburg position at Yancheng Third People's Hospital from September 2022 to August 2024 were selected,and randomly divided into the control group and the observation group,with 36 cases in each group.The control group received routine restriction of fasting and fluid starting from 22:00 on the day before surgery,while the observation group was given an additional 300 mL of carbohydrate orally 2 hours before anesthesia on the basis of the control group.The preoperative ultrasound results of gastric antrum,Perlas classification,full stomach status,visual analogue scale(VAS)scores for thirst and hunger,awakening time,tracheal extubation time,intraoperative hypothermia,gastro-intestinal function recovery time,and adverse reactions were compared between the two groups.Results Before anesthesia,there was no statistically significant difference in the cross-sectional area of the gastric antrum(CSA)in the right lateral position,gastric volume(GV),or the ratio of gastric volume to body weight(GV/W)between the two groups(P>0.05).There was also no statistically significant difference in the Perlas classification of gastric antrum ultrasound or the incidence of full stomach between the two groups(P>0.05).Immediately before anesthesia,the VAS scores for thirst and hunger in the observation group were significantly lower than those in the control group(P<0.05).The awakening time,tracheal extubation time,and incidence of intraoperative hypothermia in the observation group were significantly shorter/lower than those in the control group(P<0.05).The observation group had significantly shorter time of first exhaust and defecation after surgery,as well as lower incidence of postoperative bloating,compared with the control group(P<0.05).Conclusion For overweight or obese patients undergoing laparoscopic hysterectomy in the Trendelenburg position,oral intake of 300 mL carbohydrate 2 hours before anesthesia does not increase the risk of regurgitation or aspiration,with safety and reliability.It not only can improve comfort of patients and promote post-anesthesia awakening,but also can reduce the incidences of intraoperative hypothermia and gastrointestinal adverse reactions,which is conducive to accelerating postoperative recovery.

This study was aimed at developing an in vitro fluorescent substrate assay for the activity of leucyl aminopeptid-ase(LAP)from Echinococcus multilocularis and comparing it with the chemical chromogenic substrate enzyme activity assay.Through the establishment of reaction conditions for the fluorescent substrate-based in vitro enzyme activity assay,we com-pared the differences between the fluorescent substrate L-Leucine-7-amido-4-methylocoumarin(Leu-AMC)and the chemical chromogenic substrate L-Leucine-4-nitroanilide(Leu-pNA)through molecular docking,inhibition rates,and precision measures.Molecular docking revealed that the fluorescent substrate Leu-AMC had higher affinity for the protein than the chemical chromogenic substrate Leu-pNA.Through analysis of the effects of varying reaction conditions on fluorescence intensi-ty,we optimized the fluorescent substrate enzyme activity assay to demonstrate favorable performance at a reaction temperature of 37℃,a pH of 9.0,a protein concentration of 800 nmol/L,and a reaction duration of 60 minutes.Leu-AMC exhibited significant and distinct responses at a 5 μmol/L substrate concentration,under varying substrate conditions.The fluo-rescent substrate assay demonstrated more significant intergroup differences than the chemical chromogenic substrate assay when various inhibitors were added.This study established a fluorescence-based enzyme activity assay for leucyl aminopeptidase from Echinococcus multilocularis by using Leu-AMC as the substrate;this method demonstrated a more significant intergroup difference and sensitivity than the chemical chromogenic substrate assay.

Objective To clarify the effect of preoperative oral carbohydrate on overweight or obese patients undergoing laparoscopic hysterectomy in Trendelenburg position.Methods A total of 72 overweight or obese patients who underwent laparoscopic hysterectomy in Trendelenburg position at Yancheng Third People's Hospital from September 2022 to August 2024 were selected,and randomly divided into the control group and the observation group,with 36 cases in each group.The control group received routine restriction of fasting and fluid starting from 22:00 on the day before surgery,while the observation group was given an additional 300 mL of carbohydrate orally 2 hours before anesthesia on the basis of the control group.The preoperative ultrasound results of gastric antrum,Perlas classification,full stomach status,visual analogue scale(VAS)scores for thirst and hunger,awakening time,tracheal extubation time,intraoperative hypothermia,gastro-intestinal function recovery time,and adverse reactions were compared between the two groups.Results Before anesthesia,there was no statistically significant difference in the cross-sectional area of the gastric antrum(CSA)in the right lateral position,gastric volume(GV),or the ratio of gastric volume to body weight(GV/W)between the two groups(P>0.05).There was also no statistically significant difference in the Perlas classification of gastric antrum ultrasound or the incidence of full stomach between the two groups(P>0.05).Immediately before anesthesia,the VAS scores for thirst and hunger in the observation group were significantly lower than those in the control group(P<0.05).The awakening time,tracheal extubation time,and incidence of intraoperative hypothermia in the observation group were significantly shorter/lower than those in the control group(P<0.05).The observation group had significantly shorter time of first exhaust and defecation after surgery,as well as lower incidence of postoperative bloating,compared with the control group(P<0.05).Conclusion For overweight or obese patients undergoing laparoscopic hysterectomy in the Trendelenburg position,oral intake of 300 mL carbohydrate 2 hours before anesthesia does not increase the risk of regurgitation or aspiration,with safety and reliability.It not only can improve comfort of patients and promote post-anesthesia awakening,but also can reduce the incidences of intraoperative hypothermia and gastrointestinal adverse reactions,which is conducive to accelerating postoperative recovery.