BACKGROUND: Pre-transplant exposure to immune checkpoint inhibitors (ICIs) significantly increases the risk of allograft rejection after liver transplantation (LT); however, whether ICI-related rejection leads to increased graft loss remains controversial. Therefore, this study aimed to investigate the association between ICI-related allograft rejection and perioperative graft loss. METHODS: This was a retrospective analysis of adult liver transplant recipients with early biopsy-proven T-cell-mediated rejection (TCMR) at Liver Transplantation Center of Sun Yat-sen Memorial Hospital from June 2019 to September 2024. The pathological features, clinical characteristics, and perioperative graft survival were analyzed. RESULTS: Twenty-eight patients who underwent early TCMR between June 2019 and September 2024 were included. Based on pre-LT ICI exposure, recipients were categorized into ICI-related TCMR (irTCMR, n = 12) and conventional TCMR (cTCMR, n = 16) groups. Recipients with irTCMR had a higher median Banff rejection activity index (RAI) (6 vs . 5, P = 0.012) and more aggressive tissue damage and inflammation. Recipients with irTCMR showed higher proportion of treatment resistance, achieving a complete resolution rate of only 8/12 compared to 16/16 for cTCMR. Graft loss occurred in 5/12 of irTCMR recipients within 90 days after LT, with no graft loss in cTCMRs recipients. Cox analysis demonstrated that irTCMR with an ICI washout period of <30 days was an independent risk factor for perioperative graft loss (hazard ratio [HR], 6.540; 95% confidence interval [CI], 1.067-40.067, P = 0.042). CONCLUSION IrTCMR is associated with severe pathological features, increased resistance to treatment, and higher graft loss in adult liver transplant recipients.
Humans ; Liver Transplantation/adverse effects* ; Male ; Female ; Middle Aged ; Retrospective Studies ; Graft Rejection/immunology* ; Immune Checkpoint Inhibitors/therapeutic use* ; Adult ; T-Lymphocytes/drug effects* ; Graft Survival/immunology* ; Aged

This study aimed to explore the molecular mechanism of rubioncolin C(RuC) in inhibiting gastric cancer(GC). AGS and MGC803 cell lines were selected as cellular models. After treating the cells with RuC at different concentrations, the effects of RuC on the proliferation ability of GC cells were assessed using the CCK-8 method, real-time cellular analysis(RTCA), and colony formation assays. Transmission electron microscopy was used to observe subcellular structural changes. Immunofluorescence was applied to detect LC3 fluorescent foci. Acridine orange staining was used to evaluate the state of intracellular lysosomes. Western blot was employed to detect the expression of autophagy-related proteins LC3Ⅱ, P62, and lysosomal cathepsin D(CTSD). The SuperPred online tool was used to predict the target proteins that bound to RuC, and molecular docking analysis was conducted to identify the interaction sites between RuC and CTSD. The drug affinity responsive target stability(DARTS) assay was performed to detect the direct binding interaction between RuC and CTSD. The results showed that RuC significantly inhibited the proliferation and colony formation of GC cells at low concentrations, with 24-hour half-maximal inhibitory concentrations(IC_(50)) of 3.422 and 2.697 μmol·L~(-1) for AGS and MGC803 cells, respectively. After 24 hours of treatment with RuC at concentrations of 1, 2, and 3 μmol·L~(-1), the colony formation rates for AGS cells were 61.0%±1.5%, 28.0%±0.5%, and 18.2%±0.5%, respectively, while the rates for MGC803 cells were 56.0%±0.5%, 23.3%±1.0%, and 11.8%±1.0%, all of which were significantly reduced. Transmission electron microscopy revealed that RuC promoted an increase in autophagosome formation in GC cells. Immunofluorescence detection showed that LC3 fluorescent foci of GC cells increased with the increase in RuC dose. RuC up-regulated the expression of autophagy-related proteins LC3Ⅱ and P62 in GC cells. Acridine orange staining indicated that RuC altered the acidic environment of lysosomes. SuperPred online prediction identified CTSD as a potential target protein of RuC. Western blot analysis revealed that RuC induced the up-regulation of the inactive precursor of CTSD in GC cells. CTSD activity assays indicated that RuC reduced the activity of CTSD. Molecular docking simulations found that RuC bound to the substrate-binding region of CTSD, forming hydrogen bonds with the Tyr205 and Asp231 residues. Microscale thermophoresis and DARTS assays further confirmed that RuC directly bound to CTSD. In summary, RuC inhibits lysosomal activity by targeting and down-regulating the expression of CTSD, thereby inducing autophagosome accumulation in GC cells.
Humans ; Stomach Neoplasms/enzymology* ; Cathepsin D/chemistry* ; Cell Line, Tumor ; Molecular Docking Simulation ; Cell Proliferation/drug effects* ; Autophagosomes/metabolism* ; Autophagy/drug effects*

OBJECTIVE: To explore the inhibitory effects of Nardostachys Jatamansi DC. volatile oil (NJVO) on psychological factors substance P (SP)/cortisol (CORT)-induced hyperpigmentation. METHODS: The model of psychologically-induced hyperpigmentation of B16F10 cells was created using SP (10 nmol/L) + CORT (10 µmol/L) for 72 h. The levels of melanin content, tyrosinase (TYR) activity using NaOH lysis and L-dihydroxyphenylalanine (L-DOPA) oxidation methods were assessed, respectively. The effect of NJVO on SP/CORT-induced normal human skin tissue pigmentation was detected by Masson staining. Protein expressions of tyrosinase-related protein 1 (TRP-1), tyrosinase-relative protein 2 (DCT), and microphthalmia-associated transcription factor were determined using Western blot. The melanosome number, maturation, and melanosomal structure changes were detected through transmission electron microscopy and immunofluorescence experiments. In vivo, zebrafish pigment content was evaluated in SP/CORT-induced zebrafish hyperpigmentation model. RESULTS: NJVO significantly reduced the melanin content (P<0.01) and inhibited tyrosinase activity (P<0.01), the pigmentation of the normal skin tissue in the NJVO group was significantly lower than that in the SP/CORT group (P<0.05). And NJVO considerably downregulated expressions of melanogenesis-related proteins (TYR, TRP-1, DCT) in cells (P<0.01). In addition, the number of melanosomes was decreased and the dentrites formation of B16F10 cells was inhibited after NJVO treatment (P<0.01). In vivo, NJVO significantly reduced the pigment content in the zebrafish body (P<0.01). CONCLUSION NJVO effectively reversed SP/CORT-induced hyperpigmentation by suppressing the activity and expression of TYR and TRPs and inhibiting melanosome maturation in mouse B16F10 melanoma cells.
Animals ; Hyperpigmentation/psychology* ; Zebrafish ; Oils, Volatile/therapeutic use* ; Melanins/metabolism* ; Humans ; Monophenol Monooxygenase/metabolism* ; Mice ; Nardostachys/chemistry* ; Substance P ; Hydrocortisone ; Skin Pigmentation/drug effects* ; Cell Line, Tumor ; Melanosomes/ultrastructure* ; Microphthalmia-Associated Transcription Factor/metabolism* ; Melanoma, Experimental ; Oxidoreductases/metabolism* ; Intramolecular Oxidoreductases/metabolism*

Background and aims:Despite growing evidence linking pretransplant exposure to immune checkpoint inhibitors(ICIs)to increased allograft rejection risk after liver transplantation(LT),a lack of comparative studies to definitively establish the correlation between ICI exposure and adverse short-term outcomes after LT exists.This study aimed to analyze the impact of preoperative ICI exposure on short-term post-LT prognosis and allograft rejection risk.Methods:This retrospective cohort study included 121 recipients who underwent LT for hepatocellular carcinoma(HCC)between June 2019 and March 2023.The recipients were categorized into ICI(n=35)and non-ICI(n=86)exposure groups based on pretransplant ICI exposure.Demographics,clinical characteristics,and short-term outcomes were compared between the cohorts.Kaplan-Meier analysis evaluated the impact of ICI exposure on graft survival.Univariate and multivariate logistic regression models assessed the impact of patient characteristics on allograft rejection.Results:Recipients with or without ICI exposure exhibited comparable demographic baseline charac-teristics.The incidences of early allograft dysfunction and biliary and vascular complications were similar between both groups.Post-transplant infection incidence was 37.1％and 20.9％in the ICI and non-ICI groups,respectively(P=0.064).Allograft rejection rates were significantly higher in the ICI group than in the non-ICI group(22.9％vs.5.8％,P=0.015).The ICI group exhibited a higher 90-day post-transplant mortality rate than that of the non-ICI group(14.3％vs.2.3％,P=0.034).Logistic regression analyses demonstrated that allograft rejection independently correlated with 90-day post-transplant mortality,with ICI exposure being an independent risk factor for allograft rejection.In recipients with ICI exposure,a shorter interval between ICIs and LT(washout period)was significantly associated with a higher allograft rejection risk,with the optimal washout period identified as 21 days for predicting 90-day rejection-free survival(P=0.0001).Moreover,in recipients with allograft rejection,the peripheral CD4+/CD8+T cell ratio was much lower in the ICI group than in the non-ICI group.Conclusions:Pretransplant ICI exposure was an independent risk factor for allograft rejection and was significantly associated with 90-day post-transplant mortality after LT for HCC.A ≤21-day washout period was significantly associated with allograft rejection.Future multicenter studies with larger cohorts and prospective designs are essential to validate these findings,confirm causality,and establish standardized clinical guidelines for ICI use before transplantation.Trail registration:ClinicalTrials.gov NCT05913583.

AIM:This study is aimed to investigate the impact of 3,5,6,7,8,3',4'-heptamethoxyflavone(HMF)on the proliferation,invasion,and migration of human colorectal cancer(CRC)cell lines(SW480 and HCT116)and preliminarily explore the underlying molecular mechanisms.METHODS:Human colorectal cancer cells(SW480 and HCT116)cultured in vitro were subjected to various concentrations of HMF(0,12.5,25 and 50 μmol/L)for 48 h.Proliferation levels were assessed using the CCK-8 assay,invasion abilities were examined via the Transwell assay,migra-tion rates were measured using the scratch assay,and oxidative stress levels were determined by the DCF-DA reactive oxy-genation assay.The mRNA expression levels of heme oxygenase-1(HO-1)mRNA and NAD(P)H:quinone oxidoreduc-tase-1(NQO-1)were quantified using RT-qPCR.RESULTS:Treatment with varying concentrations of HMF resulted in a significant reduction in the proliferative capacity of SW480 and HCT116 cancer cells,as was indicated by CCK-8 experi-ments(P<0.05).Transwell assays demonstrated a pronounced attenuation in the invasive potential of SW480 and HCT116 following HMF treatment(P<0.05).Scratch assays highlighted a notable constraint on the migratory capabilities of SW480 and HCT116 after HMF treatment(P<0.05).DCF-DA staining revealed a substantial increase in reactive oxy-gen species(ROS)levels within SW480 and HCT116 cells after HMF treatment(P<0.05).Furthermore,RT-qPCR ex-periments elucidated that HMF markedly suppressed the mRNA expression of antioxidant genes HO-1 and NQO-1.CON-CLUSION:HMF induces oxidative stress response in SW480 and HCT116 cells,consequently inhibiting their prolifera-tion,invasion and migration.

Objective To investigate the prognosis of patients with hepatitis B virus(HBV)-related intrahepatic cholangiocarcinoma(ICC)whose HBV DNA was negative before surgical.Methods A retrospective analysis was conducted on the clinical data of 97 ICC patients who underwent surgery resection at the Fifth Medical Center of Chinese PLA General Hospital between October 2010 and January 2017.All patients were divided into HBV-related ICC(HBV-ICC)group(n=62)and non-HBV-related ICC(Con-ICC)group(n=35).HBV-ICC group included 34 patients with HBV core antigen positive(HBcAb+)and HBV surface antigen positive(HBsAg+),and 28 patients with HBcAb positive and HBsAg negative.Kaplan-Meier analysis was used to plot survival curves and compare the overall survival(OS)and postoperative recurrence-free survival(RFS)among patients in Con-ICC,ICC patients with HBsAg+/HBcAb+,and ICC patients with HBsAg-/HBcAb+.Univariate and multivariate Cox proportional hazard models were used to analyze independent influencing factor for OS,RFS and early postoperative recurrence among gender,age,pathogenic factor,liver cirrhosis,Child-Pugh grade,carbohydrate antigen 19-9(CA199),alpha-fetoprotein(AFP),glutamine transferase(GGT),alkaline phosphatase(ALP),total bilirubin(TBil),direct bilirubin(DBil),American Joint Committee on Cancer(AJCC)stage,tumor size,tumor number,tumor differentiation,microvascular invasion,lymph node metastasis,hepatectomy procedure,cholecystectomy,and follow-up treatment.Results Of the 97 patients,the median age was 56 years,and 79(81.4%)of them were male.The median follow-up time was 92.2 months.Eighty-eight(90.7%)patients presented with tumor recurrence and 73(75.3%)died.In multivariate analyses,HBV-ICC and CA199>37 kU/L were independent predictors of OS(HR=0.45,95%CI 0.26-0.77,P=0.003;HR=2.10,95%CI 1.24-3.57,P=0.006),RFS(HR=0.43,95%CI 0.27-0.68,P<0.001;HR=1.78,95%CI 1.12-2.81,P=0.014),and postoperative early recurrence(HR=0.42,95%CI 0.26-0.70,P=0.001;HR=2.02,95%CI 1.20-3.39,P=0.008).AJCC stage Ⅲ was an independent risk factor for postoperative RFS(HR=1.81,95%CI 1.04-3.14,P=0.037).Multiple tumor lesions was an independent risk factor for postoperative RFS and early recurrence(HR=1.73,95%CI 1.07-2.77,P=0.024;HR=1.90,95%CI 1.12-3.24,P=0.017).There was no statistically significant difference in OS,RFS,and early recurrence between HBV-ICC patients with HBsAg-/HBcAb+and Con-ICC patients(P<0.05),whereas HBsAg+/HBcAb+was a significant factor affecting postoperative OS(HR=0.32,95%CI 0.16-0.62,P=0.001),RFS(HR=0.32,95%CI 0.18-0.55,P<0.001),and early recurrence(HR=0.29,95%CI 0.15-0.54,P<0.001)in ICC patients.Conclusions The prognosis of HBV-ICC patients with preoperative HBV-DNA-is better than that of Con-ICC patients.The prognosis of HBV-ICC patients with HBcAb+/HBsAg-is worse than that of HBV-ICC patients with HBcAb+/HBsAg+,but similar to Con-ICC patients.Therefore,the postoperative stratified management of HBV-ICC patients should be emphasized.

Purpose: This cross-sectional cohort-comparison observational study investigated the value of high-frame-rate vector flow (V Flow) imaging for evaluating differences in carotid plaque shape and biomechanical parameters in patients with mild stenosis according to a recent history of ipsilateral ischemic stroke. Methods: The present study included 352 patients from February 2023 to October 2023, who were categorized as symptomatic or asymptomatic based on a history of recent ischemic stroke and ipsilateral ischemic lesions detected on head computed tomography or magnetic resonance imaging. A Mindray Resona R9 system was used for B-mode ultrasonography and V Flow imaging. The upstream and downstream surfaces of the plaques were examined at the carotid bifurcation for wall shear stress (WSS), oscillatory shear index (OSI), and turbulence index, which performed peri-plaque biomechanical condition. Multivariable logistic regression models were used to determine associations between plaque shape, V Flow parameters, and ischemic stroke. Results: Symptomatic patients exhibited higher WSS values for the upstream and downstream surfaces of carotid plaque, as well as higher OSI and turbulence index values for the downstream surface. Type Ⅲ plaques and higher WSS and OSI values for the downstream surface of the plaque were significantly associated with ischemic stroke. Type Ⅲ plaques were more prevalent in symptomatic patients and demonstrated much higher WSS and OSI values for the downstream plaque surface in both groups. Conclusion High-frame-rate V Flow imaging could assess peri-plaque biomechanical forces and may provide effective imaging biomarkers for early prediction of ischemic stroke in patients with mild stenosis.

Purpose: This study aimed to determine whether micro-flow imaging (MFI) offers diagnostic performance comparable to that of contrast-enhanced ultrasonography (CEUS) in detecting segmental congestion among patients undergoing living donor liver transplantation (LDLT). Methods: Data from 63 patients who underwent LDLT between May and December 2022 were retrospectively analyzed. MFI and CEUS data collected on the first postoperative day were quantified. Segmental congestion was assessed based on imaging findings and laboratory data, including liver enzymes and total bilirubin levels. The reference standard was a postoperative contrast-enhanced computed tomography scan performed within 2 weeks of surgery. Additionally, a subgroup analysis examined patients who underwent reconstruction of the middle hepatic vein territory. Results: The sensitivity and specificity of MFI were 73.9% and 67.5%, respectively. In comparison, CEUS demonstrated a sensitivity of 78.3% and a specificity of 75.0%. These findings suggest comparable diagnostic performance, with no significant differences in sensitivity (P=0.655) or specificity (P=0.257) between the two modalities. Additionally, early postoperative laboratory values did not show significant differences between patients with and without congestion. The subgroup analysis also indicated similar diagnostic performance between MFI and CEUS. Conclusion MFI without contrast enhancement yielded results comparable to those of CEUS in detecting segmental congestion after LDLT. Therefore, MFI may be considered a viable alternative to CEUS.

The global increase in livestock production has correspondingly intensified farm odors due to harmful bacteria, reduced immunity, and disease progression. In this study, we treated feces with Biomagic-Enzyme complex for 4 months to understand the relationship between farm odor, immunity against common viral diseases, immune cytokines, and changes in the microbiota. A gas meter (MultiRAE) was used to measure ammonia (NH3) and hydrogen sulfide (H2S) while odor intensity and offensiveness were characterized by the non-objective scaling method. A complete blood count was performed and plasma was obtained after blood centrifugation at 3,000 rpm for 20 minutes. The cytokine profile was evaluated using commercial kits. Microbial DNA was extracted and purified from fecal samples to analyze the microbiota. Microbial DNA and viral RNA/DNA were obtained from fecal samples and amplified to determine the expression of transmissible gastroenteritis virus (TGEV), porcine reproductive and respiratory syndrome (PRRS), and porcine circovirus type 2 (PCV2). Our results indicated that Biomagic reduced odor nuisance by decreasing ammonia levels, resulting in faint and fairly offensive odor intensity. After the enzyme treatment, Escherichia coli populations significantly reduced across all 3 farms. In contrast, beneficial Lactobacillus spp. levels remained stable, indicating the enzyme selectively targeted harmful bacteria while preserving beneficial ones. The beneficial Lachnospiraceae, Spirochaetaceae, and Bacteroidaceae were found to be higher in the third month of treatment. TGEV was not detected, while PRRS and non-pathogenic PCV2 showed a positive infection rate. In conclusion, Biomagic reduced ammonia, prevented viral infection from pig farms, and improved gut-beneficial bacteria and microbiota.

The correspondence between the five zang organs and the five phases exhibited variations in the previous and the current literature， which is related to the understanding and interpretation of the historical construction of the basic theories of traditional Chinese medicine， and is an important issue that has been debated for a long time in the history of academia but has not been resolved. Through combing and analyzing the relevant literature handed down during the pre-Qin and Han dynasties， which includes Master LYU's Spring and Autumn Annals （《吕氏春秋》）， The Book of Rites： Monthly Ordinances （《月令》）， Huainanzi （《淮南子》）， Records of the Grand Historian： Biography of BIAN Que and CANG Gong （《史记·扁鹊仓公列传》）， The Inner Canon of Yellow Emperor （《黄帝内经》）， as well as excavated literature like the Warring States bamboo slips Tang Zai Chi Men （《汤在啻门》） collected by Tsinghua University， Taichan Shu （《胎产书》） unearthed from the Mawangdui Han Dynasty Tombs， Tianhui Medical Manuscripts （《天回医简》） unearthed from the Han Dynasty Tombs of Lao Guan Mountain， Chengdu， this paper finds that the current five phases are newly proposed by previous medical practitioners on the basis of the objective understanding of the physiology and pathology of the five zang organs， which is a unique contribution of traditional Chinese medicine to the traditional Chinese culture. The lacquered acupuncture figurine unearthed from the Han Dynasty Tombs of Lao Guan Mountain， Tianhui Town， Chengdu， Sichuan Province， is engraved on its back with the inscriptions of "heart， lungs， liver， stomach， and kidneys"， which refer to the back-Shu points of the five zang organs， and the order follows the sequence of “fire - metal - wood - earth - water”， which is the order of the five phases being restricted. It is believed that this is a reflection of the idea of “heart is the head of the five organs” in the early medical scriptures， which can be used as a basis for judging the chronological order of the relevant chapters of the modern version of The Inner Canon of Yellow Emperor. This order is also seen in the Tianhui Medical Manuscripts unearthed from the same tombs as the acupuncture figurine and is preserved in many chapters of the modern version of The Inner Canon of Yellow Emperor.