In the context of the development of transplant oncology, it is of great clinical significance to find a drug with both antitumor and immunosuppressive effects for liver transplantation patients with hepatocellular carcinoma (HCC). The antitumor effect of ginkgolic acid (GA) has been confirmed, and some studies suggest that GA may also have an immunosuppressive effect. The immunosuppressive effect of GA was evaluated by histopathology, T-cell subpopulation, and cytokine detection in rat liver transplantation and mouse cardiac transplantation models, and transcriptomic and metabolomic analysis was used to explore the underlying mechanism of the GA immunosuppressive effect. Metabolites, activation, and ferroptosis markers of CD8⁺ T cells were detected in vivo and in vitro. Based on rat liver transplantation and mouse cardiac transplantation models, the immunosuppressive effect of GA was first confirmed by histopathology, T-cell subpopulation, and cytokine detection. In the mouse cardiac transplantation model, transcriptomics combined with metabolomics demonstrated for the first time that GA inhibited lactate dehydrogenase A (LDHA) expression and pyruvate metabolism in CD8⁺ T cells. It was confirmed in vivo and in vitro that GA inhibited pyruvate metabolism of CD8⁺ T cells through LDHA, inhibiting their activation and inducing ferroptosis. Overexpression of LDHA partially reversed the effect of GA on the metabolism, activation, and ferroptosis of CD8⁺ T cells in vitro. GA mediates metabolic reprogramming through LDHA to inhibit the activation and induce ferroptosis of CD8⁺ T cells to exert an immunosuppressive effect, which lays an experimental foundation for the future clinical application of its immunosuppressive effect.

Objective To explore the feasibility of phase-contrast CT for three-dimensional visualization of in vitro intrahepatic vessels and necrotic foci in mouse model of acute liver injury(ALI).Methods Six clean grade male wild-type C57BL/6 mice were randomly divided into model group and control group(each n=3).The mice in model group were fed with overdose of acetaminophen to induce ALI,while those in control group were fed with the same amount of distilled water.After 24 h,the mice were all sacrificed,the livers were harvested and then fixed and dehydrated.Phase-contrast CT images of in vitro liver were acquired,sectional and three-dimensional images were reconstructed.The effect of phase-contrast CT for displaying the outline of liver and internal vessels and necrotic foci were observed,the maximum diameter and volume of necrotic foci were quantitatively analyzed,and the results of phase-contrast CT were compared with pathological findings.Results The original projection and sectional images of phase-contrast CT clearly showed the outline of in vitro liver and internal vasculatures.Necrotic foci within the liver were found in model group,but not in control group.The findings of phase-contrast CT corresponded accurately to those of pathology.Three-dimensional reconstruction images of phase-contrast CT clearly displayed intrahepatic portal vein system and hepatic vein system in both groups,and discontinuous punctate necrotic foci within the liver were found in model group,mainly distributed around hepatic vein,with a median maximum diameter of 18.50 μm and a median volume of 5 870.11 μm3,but was not observed in control group.Conclusion Phase-contrast CT could be used in three-dimensional visualization of intrahepatic vessels and necrotic foci of in vitro liver in mouse model of ALI.

Objective:To investigate the clinical characteristics of urea cycle disorder (UCD), the efficacy and safety of glyceryl phenylbutyrate (GPB) therapy in pediatric patients with UCD.Methods:This study was a retrospective, single-arm, multicenter clinical study. The clinical data of 20 pediatric patients with UCD who received GPB treatment at 9 hospitals nationwide between December 2021 and August 2024 were collected. The clinical manifestations, laboratory results, and molecular genetic characteristics were analyzed, ammonia levels and other laboratory results were evaluated pre-post GPB therapy by paired t-tests or Wilcoxon tests. Results:Among the 20 pediatric patients with UCD, there were 8 males and 12 females, and the onset age was 2.8 (1.4, 5.7) years. The ammonia levels were 174 (125, 342) μmol/L at first onset. The symptoms included vomiting in 6 cases, drowsiness in 5 cases, epilepsy in 5 cases, developmental delay in 5 cases, psychiatric and behavioral abnormalities in 3 cases, and lethargy in 1 case, and 18 cases exhibited abnormal liver function. Twenty cases included 6 UCD subtypes, with 11 cases being ornithine transcarbamylase deficiency. A total of 27 variants were identified, 11 (41%) of which were novel. The age of patients who began GPB therapy was 4.0 (1.5, 6.6) years. Ten cases stopped GPB after 4.2 (3.4, 5.3) months, with 4 patients undergoing liver transplantation and 6 discontinuing for financial reasons. The remaining ten patients continued GPB therapy for 11.6 (8.6, 14.0) months. The duration of GPB treatment was 6.0 (4.2, 12.3) months, at the final visit, the levels of ammonia, platelets and aspartate aminotransferase were lower compared to those of pre-treatment (all P<0.05). The serum albumin level was higher than that of pre-treatment ( P=0.016). Two patients suffered only one episode of acute hyperammonaemia, with ammonia levels of 232 and 141 μmol/L, respectively. Nine cases experienced adverse effects potentially related to GPB, decreased appetite in 6 cases, vomiting in 3 cases, abnormal skin oil odor in 2 cases, somnolence, fatigue and diarrhea each in 1 case, with symptoms improved within 6 (3, 10) days. Conclusions:UCD primarily manifests with neurological and gastrointestinal symptoms, and early diagnosis of UCD could be achieved through the analysis of ammonia. GPB may effectively reduce ammonia levels in UCD pediatric patients, with favorable safety and tolerability.

To screen and identify a chitosanase with high stability, we cloned the chitosanase gene from Bacillus atrophaeus with a high protease yield from the barren saline-alkali soil and expressed this gene in Escherichia coli. The expressed chitosanase of B. atrophaeus (BA-CSN) was purified by nickel-affinity column chromatography. The properties including optimal temperature, optimal pH, substrate specificity, and kinetic parameters of BA-CSN were characterized. The results showed that BA-CSN had the molecular weight of 31.13 kDa, the optimal temperature of 55 ℃, the optimal pH 5.5, and good stability at temperatures below 45 ℃ and pH 4.0-9.0. BA-CSN also had good stability within 4 h of pH 3.0 and 10.0, be activated by K⁺, Na⁺, Mn²⁺, Ca²⁺, Mg²⁺, and Co²⁺, (especially by Mn²⁺), and be inhibited by Fe³⁺, Cu²⁺, and Ag⁺. BA-CSN showcased the highest relative activity in the hydrolysis of colloidal chitosan, and it had good hydrolysis ability for colloidal chitin. Under the optimal catalytic conditions, BA-CSN demonstrated the Michaelis constant K_m and maximum reaction rate V_max of 9.94 mg/mL and 26.624 μmoL/(mL·min), respectively, for colloidal chitosan. In short, BA-CSN has strong tolerance to acids and alkali, possessing broad industrial application prospects.
Bacillus/genetics* ; Hydrogen-Ion Concentration ; Escherichia coli/metabolism* ; Glycoside Hydrolases/biosynthesis* ; Substrate Specificity ; Enzyme Stability ; Chitosan/metabolism* ; Temperature ; Kinetics ; Cloning, Molecular ; Bacterial Proteins/biosynthesis* ; Recombinant Proteins/genetics*

In the context of the development of transplant oncology,it is of great clinical significance to find a drug with both antitumor and immunosuppressive effects for liver transplantation patients with hepatocellular carcinoma(HCC).The antitumor effect of ginkgolic acid(GA)has been confirmed,and some studies suggest that GA may also have an immunosuppressive effect.The immunosuppressive effect of GA was evaluated by histopathology,T-cell subpopulation,and cytokine detection in rat liver transplantation and mouse cardiac transplantation models,and transcriptomic and metabolomic analysis was used to explore the underlying mechanism of the GA immunosuppressive effect.Metabolites,activation,and ferroptosis markers of CD8+T cells were detected in vivo and in vitro.Based on rat liver transplantation and mouse cardiac transplantation models,the immunosuppressive effect of GA was first confirmed by histopathology,T-cell subpopulation,and cytokine detection.In the mouse cardiac transplantation model,transcriptomics combined with metabolomics demonstrated for the first time that GA inhibited lactate dehydrogenase A(LDHA)expression and pyruvate metabolism in CD8+T cells.It was confirmed in vivo and in vitro that GA inhibited pyruvate metabolism of CD8+T cells through LDHA,inhibiting their activation and inducing ferroptosis.Over-expression of LDHA partially reversed the effect of GA on the metabolism,activation,and ferroptosis of CD8+T cells in vitro.GA mediates metabolic reprogramming through LDHA to inhibit the activation and induce ferroptosis of CD8+T cells to exert an immunosuppressive effect,which lays an experimental foundation for the future clinical application of its immunosuppressive effect.

Objective To mine and analyze signals of acute kidney injury(AKI)related to drugs,comprehensively summarize the potential risk drugs,and provide a reference for clinically safe medication.Methods The AKI reports from January 2004 to September 2023 in the US FDA Adverse Event Reporting System(FAERS)were retrieved.Disproportionality methods were used to explore the relationship between drugs and AKI,and demographic information,time to onset,and patient outcomes were analyzed.Results Out of 1 253 drugs,159 were identified as AKI signal drugs.Among these,there were 49 antimicrobial agents(30.82%),including 35 antibiotics and 14 antiviral agents;33 antineoplastic agents(20.75%);and 25 hypotensive agents(15.72%).Drug-related AKI occurred mostly in the elderly,and the male-to-female ratio was 124∶100.The median time to onset for AKI related to antibiotics was≤8 d,with the third quartile≤21 d.Rivaroxaban and aspirin had higher proportions of death reports,with 33.03%and 31.44%respectively.Conclusions A multitude of drugs pose a risk for acute kidney injury,necessitating caution in their clinical application and the implementation of monitoring of renal function.The elderly are a high-risk group for drug-related AKI,and there are more males than females.For antibiotics,the first 21 days are the key monitoring period.For drugs that require long-term use,regular monitoring is necessary.

Objective:To investigate the impact of different luteal phase support protocols on pregnancy outcomes in patients aged ≤35 years undergoing modified natural cycle frozen-thawed embryo transfer (mNC-FET).Methods:A retrospective cohort study was conducted to analyze 2 086 cycles of patients aged ≤35 years who received mNC-FET cycles in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2020. The cycles were divided into three groups based on luteal phase support protocols used. The patients received a combination of progesterone soft capsule and dydrogesterone in the group A (446 cycles), the patients received dydrogesterone in the group B (439 cycles), and the patients received a combination of progesterone vaginal sustained-release gel and dydrogesterone in the group C (1 201 cycles). The pregnancy and perinatal outcomes were compared between groups A and B, groups C and B after matching the baseline data in a ratio of 1∶1 using the propensity score matching (PSM). The effect of different luteal phase support on live birth rate was analyzed after adjusting for confounding factors affected by univariate and multivariate generalized estimating equation (GEE).Results:After PSM, there were no significant differences between groups A and B, groups C and B in human chorionic gonadotropin positive rate, clinical pregnancy rate, ectopic pregnancy rate, live birth rate in transplant cycle, incidence of low weight, macrosomia, premature delivery rate, pregnancy complication rate and incidence of birth defects (all P>0.05). GEE analysis showed that three different luteal phase support regimens were not associated with live birth rate. Conclusion:In the mNC-FET cycle, patients aged ≤35 years who chose dydrogesterone alone as luteal phase support drug, had no difference in live birth rate and perinatal outcome between progesterone soft capsules or progesterone vaginal sustained-release gel combined with dydrogesterone, but the outcome still needs to be confirmed by large sample prospective studies.

Objective:To investigate the impact of different luteal phase support protocols on pregnancy outcomes in patients aged ≤35 years undergoing modified natural cycle frozen-thawed embryo transfer (mNC-FET).Methods:A retrospective cohort study was conducted to analyze 2 086 cycles of patients aged ≤35 years who received mNC-FET cycles in Reproductive Health Hospital of the Third Affiliated Hospital of Zhengzhou University from January 2018 to December 2020. The cycles were divided into three groups based on luteal phase support protocols used. The patients received a combination of progesterone soft capsule and dydrogesterone in the group A (446 cycles), the patients received dydrogesterone in the group B (439 cycles), and the patients received a combination of progesterone vaginal sustained-release gel and dydrogesterone in the group C (1 201 cycles). The pregnancy and perinatal outcomes were compared between groups A and B, groups C and B after matching the baseline data in a ratio of 1∶1 using the propensity score matching (PSM). The effect of different luteal phase support on live birth rate was analyzed after adjusting for confounding factors affected by univariate and multivariate generalized estimating equation (GEE).Results:After PSM, there were no significant differences between groups A and B, groups C and B in human chorionic gonadotropin positive rate, clinical pregnancy rate, ectopic pregnancy rate, live birth rate in transplant cycle, incidence of low weight, macrosomia, premature delivery rate, pregnancy complication rate and incidence of birth defects (all P>0.05). GEE analysis showed that three different luteal phase support regimens were not associated with live birth rate. Conclusion:In the mNC-FET cycle, patients aged ≤35 years who chose dydrogesterone alone as luteal phase support drug, had no difference in live birth rate and perinatal outcome between progesterone soft capsules or progesterone vaginal sustained-release gel combined with dydrogesterone, but the outcome still needs to be confirmed by large sample prospective studies.

Objective To mine and analyze signals of acute kidney injury(AKI)related to drugs,comprehensively summarize the potential risk drugs,and provide a reference for clinically safe medication.Methods The AKI reports from January 2004 to September 2023 in the US FDA Adverse Event Reporting System(FAERS)were retrieved.Disproportionality methods were used to explore the relationship between drugs and AKI,and demographic information,time to onset,and patient outcomes were analyzed.Results Out of 1 253 drugs,159 were identified as AKI signal drugs.Among these,there were 49 antimicrobial agents(30.82%),including 35 antibiotics and 14 antiviral agents;33 antineoplastic agents(20.75%);and 25 hypotensive agents(15.72%).Drug-related AKI occurred mostly in the elderly,and the male-to-female ratio was 124∶100.The median time to onset for AKI related to antibiotics was≤8 d,with the third quartile≤21 d.Rivaroxaban and aspirin had higher proportions of death reports,with 33.03%and 31.44%respectively.Conclusions A multitude of drugs pose a risk for acute kidney injury,necessitating caution in their clinical application and the implementation of monitoring of renal function.The elderly are a high-risk group for drug-related AKI,and there are more males than females.For antibiotics,the first 21 days are the key monitoring period.For drugs that require long-term use,regular monitoring is necessary.

Objective To explore the feasibility of phase-contrast CT for three-dimensional visualization of in vitro intrahepatic vessels and necrotic foci in mouse model of acute liver injury(ALI).Methods Six clean grade male wild-type C57BL/6 mice were randomly divided into model group and control group(each n=3).The mice in model group were fed with overdose of acetaminophen to induce ALI,while those in control group were fed with the same amount of distilled water.After 24 h,the mice were all sacrificed,the livers were harvested and then fixed and dehydrated.Phase-contrast CT images of in vitro liver were acquired,sectional and three-dimensional images were reconstructed.The effect of phase-contrast CT for displaying the outline of liver and internal vessels and necrotic foci were observed,the maximum diameter and volume of necrotic foci were quantitatively analyzed,and the results of phase-contrast CT were compared with pathological findings.Results The original projection and sectional images of phase-contrast CT clearly showed the outline of in vitro liver and internal vasculatures.Necrotic foci within the liver were found in model group,but not in control group.The findings of phase-contrast CT corresponded accurately to those of pathology.Three-dimensional reconstruction images of phase-contrast CT clearly displayed intrahepatic portal vein system and hepatic vein system in both groups,and discontinuous punctate necrotic foci within the liver were found in model group,mainly distributed around hepatic vein,with a median maximum diameter of 18.50 μm and a median volume of 5 870.11 μm3,but was not observed in control group.Conclusion Phase-contrast CT could be used in three-dimensional visualization of intrahepatic vessels and necrotic foci of in vitro liver in mouse model of ALI.