ObjectiveTo investigate the mechanisms by which the combination of berberine （BBR） and baicalin （BAI） ameliorates type 2 diabetes mellitus （T2DM） due to internal accumulation of dampness-heat from the perspectives of gut microbiota and metabolomics. MethodsAntibiotics were used to induce pseudo-sterile mice. Thirty pseudo-sterile mice were randomized into a normal fecal microbiota transplantation group （n=10） and a T2DM （syndrome of internal accumulation of dampness-heat） fecal microbiota transplantation group （n=20）. The mice were then administrated with suspensions of fecal microbiota from healthy volunteers and a patient with T2DM due to internal accumulation of dampness-heat by gavage， respectively. Each mouse received 200 µL suspension every other day for a total of 15 times to reshape the gut microbiota. The T2DM model mice were then assigned into a model group （n=8） and a BBR-BAI group （n=11）. BBR was administrated at a dose of 200 mg·kg^-1， and BAI was administrated in a ratio of BBR-BAI 10∶1 based on preliminary research findings. The administration lasted for 8 consecutive weeks. Fasting blood glucose （FBG）， glycated hemoglobin （HbA1c）， insulin （INS）， triglycerides （TG）， total cholesterol （CHOL）， low-density lipoprotein cholesterol （LDL-C）， high-density lipoprotein cholesterol （HDL-C）， aspartate aminotransferase （AST）， and alanine aminotransferase （ALT） levels were measured to evaluate the effects of the BBR-BAI combination on glucose and lipid metabolism and liver function in T2DM mice. Hematoxylin-eosin staining was employed to observe pathological changes in the colon tissue. The expression of claudin-1， zonula occludens-1 （ZO-1）， and occludin in the colon tissue was determined by Western blot. Real-time quantitative polymerase chain reaction（Real-time PCR） was employed to assess the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the colon tissue. The fecal microbiota composition and differential metabolites were analyzed by 16S rRNA sequencing and ultra-high performance liquid chromatography-quadrupole-time of flight tandem mass spectrometry （UPLC-Q-TOF-MS）， respectively. ResultsThe BBR-BAI combination lowered the FBG， HbA1c， and INS levels （P<0.05， P<0.01） and alleviated insulin resistance （P<0.01） in T2DM mice. Additionally， BBR-BAI elevated the levels of ZO-1， occludin， and claudin-1 （P<0.05， P<0.01） and down-regulated the expression levels of TNF-α， IL-1β， and IL-6 in the colon （P<0.05， P<0.01）. The results of 16S rRNA sequencing showed that BBR-BAI increased the relative abundance of Ligilactobacillus， Phascolarctobacterium， and Akkermansia （P<0.05）， while significantly decreasing the relative abundance of Alistipes， Odoribacter， and Colidextribacter （P<0.05）. UPLC-Q-TOF-MS identified 28 differential metabolites， which were primarily involved in arachidonic acid metabolism and α-linolenic acid metabolism. ConclusionBBR-BAI can ameliorate T2DM due to internal accumulation of dampness-heat by modulating the relative abundance of various bacterial genera in the gut microbiota and the expression of fecal metabolites.

Objective:To explore the value of neutrophil extracellular traps (NETs) and interleukin (IL)-33 in the early diagnosis of contrast-induced acute kidney injury (CIAKI).Methods:It was a prospective cohort study. The clinical data of patients who underwent coronary angiography (CAG) in Sir Run Run Hospital, Nanjing Medical University from December 2022 to December 2023 were collected. The main indicators of NETs included myeloperoxidase (MPO), neutrophil elastase (NE), citrullinated histone H3 (H3Cit) and antimicrobial peptide LL-37 amide (LL-37). Serum samples were collected before CAG, and 2 hours and 12 hours after CAG, and the levels of MPO, NE, H3Cit, LL-37, IL-33 and neutrophil gelatinase-associated lipocalin (NGAL) were detected. The differences of clinical data between CIAKI group and non-CIAKI group were compared. Multivariate logistic regression model was applied to analyze the risk factors of CIAKI. The receiver- operating characteristic curve was used to evaluate the predictive performance of biomarkers. Spearman correlation analysis was used to analyze the correlations among those biomarkers.Results:A total of 280 eligible patients with CAG were included in this study, with age of (65±13) years and 203 males (72.5%). The incidence rate of CIAKI was 11.8% (33/280). Compared with non-CIAKI group, the proportions of diabetes ( χ2=5.302, P=0.021), preoperative positive urine protein ( χ2=6.871, P=0.009), taking beta-blockers ( χ2=4.580, P=0.032), diuretics ( χ2=21.987, P<0.001) and calcium channel blocker ( χ2=10.424, P=0.001), preoperative blood glucose ( Z=2.807, P=0.005), preoperative blood urea nitrogen ( Z=2.504, P=0.012), neutrophil at 24 hours after CAG ( Z=2.173, P=0.030), serum creatinine at 24 hours after CAG ( Z=4.000, P<0.001), and blood urea nitrogen at 24 hours after CAG ( Z=4.459, P<0.001) were higher, while the preoperative hemoglobin ( Z=-2.380, P=0.017) and serum albumin ( Z=-2.556, P=0.011) were lower in CIAKI group. Multivariate logistic regression analysis showed that increasing neutrophil at 24 hours after CAG ( OR=1.180，95% CI 1.037-1.341), diuretics ( OR=5.615，95% CI 2.294-13.745) and calcium channel blockers ( OR=3.141，95% CI 1.374-7.182) were independent influencing factors of CIAKI. There were statistically significant differences in the levels of serum NE, MPO, H3Cit, LL-37, NGAL and IL-33 among before CAG, 2 hours after CAG and 12 hours after CAG in the overall population, CIAKI group and non-CIAKI group (all P<0.05). In addition, the changes of IL-33 before CAG and 12 hours after CAG was positively correlated with the changes of MPO, NE, H3Cit, LL-37, NGAL, serum creatinine and blood urea nitrogen before CAG and 12 hours after CAG (all P<0.05). The levels of NE ( Z=3.435, P=0.001; Z=6.164， P<0.001), MPO ( Z=3.627, P<0.001; Z=4.729, P<0.001), H3Cit ( Z=5.174, P<0.001; Z=6.241, P<0.001), LL-37 ( Z=4.986, P<0.001; Z=6.346, P<0.001), NGAL ( Z=2.956, P=0.003; Z=4.263, P<0.001) and IL-33 ( Z=5.056, P<0.001; Z=6.240, P<0.001) in CIAKI group at 2 h and 12 h after CAG were significantly higher than those in non-CIAKI group. The receiver-operating characteristic curve indicated that the combined AUC of neutrophil 24 hours after CAG, diuretics and calcium channel blockers in predicting CIAKI was 0.791. NE ( AUC=0.701), MPO ( AUC=0.712), H3Cit ( AUC=0.777), LL-37 ( AUC=0.767) and IL-33 ( AUC=0.795) at 2 hours after CAG predicted CIAKI relatively well. NE ( AUC=0.865), MPO ( AUC=0.758), H3Cit ( AUC=0.834), LL-37 ( AUC=0.840) and IL-33 ( AUC=0.867) at 12 hours after CAG had better prediction effect for CIAKI. The AUC of NETs combined with IL-33 in predicting CIAKI at 2 hours and 12 hours after CAG was 0.874 and 0.956, respectively. Conclusions:CIAKI patients exhibit elevated levels of NETs and IL-33. Serum MPO, NE, H3Cit, LL-37 and IL-33 at 12 hours after CAG can predict the occurrence of CIAKI. The combination of NETs and IL-33 is more effective in predicting CIAKI.

Toxoplasma gondii is a single-celled parasite that infects nearly all warm-blooded animals, including humans (Montoya and Liesenfeld, 2004). It occurs worldwide and can persist for a lifetime in mammals. Humans get infected by eating undercooked meat of animals containing the tissue cysts of this parasite. In immune-competent individuals, T. gondii infection usually does not cause significant clinical symptoms, whereas in pregnant or immunocompromised individuals, T. gondii infection (toxoplasmosis) can cause more serious problems like abortion and even death (Dunn et al., 1999; Wang et al., 2017). A combination of pyrimethamine and sulfadiazine is usually used to treat toxoplasmosis, although it is generally inefficient and causes side effects (Alday and Doggett, 2017). Worse still, there is a lack of vaccines to prevent T. gondii infection in humans or animals.
Toxoplasma/enzymology* ; Animals ; Humans ; Toxoplasmosis ; Mitochondria/enzymology* ; Protozoan Proteins/metabolism*

To evaluate the immunogenicity and immunoprotective effects of a tRNA thiouridylase TgMnmA deletion strain of Toxoplasma gondii(T.gondii)on ICR mice,we constructed a mouse model immunized with RH△MnmA.Mice were immunized with 10 RH△MnmA tachyzoites by in-traperitoneal injection.After 30 d,indirect ELISA was used to detect the specific IgG antibody and its subtypes of immunized mice.Spleen lymphocyte suspension was prepared,and the splenic lym-phocyte subsets were analyzed by flow cytometry.Moreover,the relative expression level of cyto-kine mRNA was detected by real-time fluorescence quantitative PCR.After 30 d of immunization,mice were intraperitoneally inoculated with RH△ku80 tachyzoites.At 5 d post infection,the para-site load in the ascites,heart,liver and brain of mice was measured,and the survival of mice within 30 d after infection was observed and recorded.The results showed that compared with the control PBS group,RH△MnmA immunized group produced higher level of IgG and IgG2a antibodies,higher mRNA relative expression level of cytokines IL-2,IL-4,IL-6,IL-10,IL-12 and IFN-γ,and the number of CD4+and CD8a+in spleen lymphocytes also increased significantly.Mean-while,for the attack of RH△ku80 strain,the immune group can effectively reduce the parasite load in the ascites and some tissues,inhibit the reproduction of parasites In vivo,and significantly improve the survival rate of mice.The results of this study showed that the TgMnmA deletion strain of T.gondii can induce strong humoral and cellular immune responses in mice,and provide good immune protection against the infection of RHΔku80 strain,which has the potential to be-come a potentially promising live attenuated vaccine candidate against T.gondii.

Objective: To investigate the association between overweight, obesity, central obesity and hypertension, so as to provide the basis for formulating targeted hypertension prevention and control strategies. Methods: Permanent residents aged ≥18 years were selected in Wenzhou City, Zhejiang Province from June 2023 to August 2024 by a multistage cluster random sampling method. Data on demographic information, lifestyle, height, weight, waist circumference (WC), blood pressure, and blood biochemical indicators were collected through questionnaire surveys, physical examinations, and laboratory tests. The prevalence of hypertension was calculated and standardized using the data of the Sixth National Population Census in 2010. Body mass index (BMI) was calculated to determine overweight and obesity, while WC was used to identify central obesity. The association between overweight, obesity, central obesity and hypertension were analyzed using multivariable logistic regression models. Results: A total of 38 593 residents were surveyed, including 19 481 (50.48%) males and 19 112 (49.52%) females. The median age was 46.00 (interquartile range, 26.00) years. The rates of overweight, obesity, and central obesity were 32.74% (12 634 individuals), 10.27% (3 963 individuals), and 27.87% (10 755 individuals), respectively. There were 11 813 cases of hypertension, with a prevalence and standardized prevalence of 30.61% and 24.41%, respectively. Multivariable logistic regression analysis showed that after adjusting for demographic information, lifestyle, diabetes and dyslipidemia, the likelihood of hypertension in the overweight and obesity groups was 1.927 (95%CI: 1.815-2.045) times and 3.724 (95%CI: 3.404-4.073) times that of the normal BMI group, respectively. The likelihood of hypertension in the central obesity group was 2.346 (95%CI: 2.214-2.486) times that of the normal WC group. The likelihood of hypertension in the central obesity only, overweight only, overweight with central obesity, obesity only and obesity with central obesity groups was 1.586 (95%CI: 1.391-1.809), 1.704 (95%CI: 1.582-1.835), 2.433 (95%CI: 2.254-2.626), 1.768 (95%CI: 1.424-2.194), and 4.466 (95%CI: 4.053-4.921) times that of the normal BMI and WC group, respectively. Conclusions Overweight, obesity and central obesity were all associated with hypertension among adult residents. The highest likelihood of hypertension was observed among adult residents with both general obesity and central obesity.

To evaluate the immunogenicity and immunoprotective effects of a tRNA thiouridylase TgMnmA deletion strain of Toxoplasma gondii(T.gondii)on ICR mice,we constructed a mouse model immunized with RH△MnmA.Mice were immunized with 10 RH△MnmA tachyzoites by in-traperitoneal injection.After 30 d,indirect ELISA was used to detect the specific IgG antibody and its subtypes of immunized mice.Spleen lymphocyte suspension was prepared,and the splenic lym-phocyte subsets were analyzed by flow cytometry.Moreover,the relative expression level of cyto-kine mRNA was detected by real-time fluorescence quantitative PCR.After 30 d of immunization,mice were intraperitoneally inoculated with RH△ku80 tachyzoites.At 5 d post infection,the para-site load in the ascites,heart,liver and brain of mice was measured,and the survival of mice within 30 d after infection was observed and recorded.The results showed that compared with the control PBS group,RH△MnmA immunized group produced higher level of IgG and IgG2a antibodies,higher mRNA relative expression level of cytokines IL-2,IL-4,IL-6,IL-10,IL-12 and IFN-γ,and the number of CD4+and CD8a+in spleen lymphocytes also increased significantly.Mean-while,for the attack of RH△ku80 strain,the immune group can effectively reduce the parasite load in the ascites and some tissues,inhibit the reproduction of parasites In vivo,and significantly improve the survival rate of mice.The results of this study showed that the TgMnmA deletion strain of T.gondii can induce strong humoral and cellular immune responses in mice,and provide good immune protection against the infection of RHΔku80 strain,which has the potential to be-come a potentially promising live attenuated vaccine candidate against T.gondii.

Objective:To explore the value of neutrophil extracellular traps (NETs) and interleukin (IL)-33 in the early diagnosis of contrast-induced acute kidney injury (CIAKI).Methods:It was a prospective cohort study. The clinical data of patients who underwent coronary angiography (CAG) in Sir Run Run Hospital, Nanjing Medical University from December 2022 to December 2023 were collected. The main indicators of NETs included myeloperoxidase (MPO), neutrophil elastase (NE), citrullinated histone H3 (H3Cit) and antimicrobial peptide LL-37 amide (LL-37). Serum samples were collected before CAG, and 2 hours and 12 hours after CAG, and the levels of MPO, NE, H3Cit, LL-37, IL-33 and neutrophil gelatinase-associated lipocalin (NGAL) were detected. The differences of clinical data between CIAKI group and non-CIAKI group were compared. Multivariate logistic regression model was applied to analyze the risk factors of CIAKI. The receiver- operating characteristic curve was used to evaluate the predictive performance of biomarkers. Spearman correlation analysis was used to analyze the correlations among those biomarkers.Results:A total of 280 eligible patients with CAG were included in this study, with age of (65±13) years and 203 males (72.5%). The incidence rate of CIAKI was 11.8% (33/280). Compared with non-CIAKI group, the proportions of diabetes ( χ2=5.302, P=0.021), preoperative positive urine protein ( χ2=6.871, P=0.009), taking beta-blockers ( χ2=4.580, P=0.032), diuretics ( χ2=21.987, P<0.001) and calcium channel blocker ( χ2=10.424, P=0.001), preoperative blood glucose ( Z=2.807, P=0.005), preoperative blood urea nitrogen ( Z=2.504, P=0.012), neutrophil at 24 hours after CAG ( Z=2.173, P=0.030), serum creatinine at 24 hours after CAG ( Z=4.000, P<0.001), and blood urea nitrogen at 24 hours after CAG ( Z=4.459, P<0.001) were higher, while the preoperative hemoglobin ( Z=-2.380, P=0.017) and serum albumin ( Z=-2.556, P=0.011) were lower in CIAKI group. Multivariate logistic regression analysis showed that increasing neutrophil at 24 hours after CAG ( OR=1.180，95% CI 1.037-1.341), diuretics ( OR=5.615，95% CI 2.294-13.745) and calcium channel blockers ( OR=3.141，95% CI 1.374-7.182) were independent influencing factors of CIAKI. There were statistically significant differences in the levels of serum NE, MPO, H3Cit, LL-37, NGAL and IL-33 among before CAG, 2 hours after CAG and 12 hours after CAG in the overall population, CIAKI group and non-CIAKI group (all P<0.05). In addition, the changes of IL-33 before CAG and 12 hours after CAG was positively correlated with the changes of MPO, NE, H3Cit, LL-37, NGAL, serum creatinine and blood urea nitrogen before CAG and 12 hours after CAG (all P<0.05). The levels of NE ( Z=3.435, P=0.001; Z=6.164， P<0.001), MPO ( Z=3.627, P<0.001; Z=4.729, P<0.001), H3Cit ( Z=5.174, P<0.001; Z=6.241, P<0.001), LL-37 ( Z=4.986, P<0.001; Z=6.346, P<0.001), NGAL ( Z=2.956, P=0.003; Z=4.263, P<0.001) and IL-33 ( Z=5.056, P<0.001; Z=6.240, P<0.001) in CIAKI group at 2 h and 12 h after CAG were significantly higher than those in non-CIAKI group. The receiver-operating characteristic curve indicated that the combined AUC of neutrophil 24 hours after CAG, diuretics and calcium channel blockers in predicting CIAKI was 0.791. NE ( AUC=0.701), MPO ( AUC=0.712), H3Cit ( AUC=0.777), LL-37 ( AUC=0.767) and IL-33 ( AUC=0.795) at 2 hours after CAG predicted CIAKI relatively well. NE ( AUC=0.865), MPO ( AUC=0.758), H3Cit ( AUC=0.834), LL-37 ( AUC=0.840) and IL-33 ( AUC=0.867) at 12 hours after CAG had better prediction effect for CIAKI. The AUC of NETs combined with IL-33 in predicting CIAKI at 2 hours and 12 hours after CAG was 0.874 and 0.956, respectively. Conclusions:CIAKI patients exhibit elevated levels of NETs and IL-33. Serum MPO, NE, H3Cit, LL-37 and IL-33 at 12 hours after CAG can predict the occurrence of CIAKI. The combination of NETs and IL-33 is more effective in predicting CIAKI.

YE Tianshi possessed a comprehensive theoretical system and extensive therapeutic experience in the treatment of warm disease caused by latent pathogenic qi,all of which was recorded in his medical records.YE Tianshi elucidated the characteristics of the pathogenesis of the three types of warm disease caused by latent pathogenic qi,namely spring warmth,summer heat,and winter warmth,and explained the pathogenesis of latent pathogenic qi from the perspective of vital qi and pathogen,and pointed out that the weakness of vital qi and the pathogenic qi led to the concealment of pathogenic qi,and that the struggle between vital qi and pathogen led to the onset of latent pathogenic qi.In the treatment,YE Tianshi emphasized the importance of clearly identifying the level of qi and blood in the internal organs where the latent pathogenic qi is located,and focused on the syndrome differentiation of weifen,qifen,yingfen,and xuefen,and combined with the syndrome differentiation of zang-fu viscera,the principle and method of treatment and medication law of warm disease caused by latent pathogenic qi were formulated.Although warm disease caused by latent pathogenic qi is a heat syndrome,YE Tianshi also attached importance to the deficiency and excess of yang qi,pointed out that latent pathogenic qi had the possibility of transforming into cold syndrome,and discussed the rules of medication for cold latent pathogenic qi in the spleen,kidney,and eight extraordinary meridians.YE Tianshi's treatment of warm disease caused by latent pathogenic qi not only emphasizes the nourishment of yin,but also emphasizes clearing qi and blood,and the circulation of qi in order to clear and penetrate the evil qi;he also pays attention to the sanjiao transmission of the latent pathogenic evil.The study of the law of YE Tianshi's differentiation and treatment of warm disease caused by latent pathogenic qi may provide ideas for the clinical treatment of COVID-19,as well as diseases in various disciplines.

It is estimated that approximately seven million people worldwide are affected by inflammatory bowel disease(IBD),causing a huge burden on healthcare systems and society.In the occurrence,progression,and treatment of IBD,the intestinal microbiota and its key metabolic product,bile acids,play a crucial role.The intestinal microbiota not only participates in the biotransformation of bile acids,enriching the diversity of bile acids,but also regulates their synthesis and transport through the farnesoid X receptor(FXR).Meanwhile,bile acids contribute to regulating the structure and function of the intestinal microbiota by supporting microbial diversity,exerting direct toxicity,participating in indirect antimicrobial pathways,and influencing microbial metabolic capabilities.Furthermore,under normal physiological conditions,intestinal microbiota-derived bile acids facilitate the repair process of the intestinal epithelial barrier.They also promote the balance of the immune system by modulating the functions of various immune cells including helper T(Th)cells 17,regulatory T(Treg)cells,CD8+T cells and natural killer T(NKT)cells,thereby slowing down the development of IBD.This article focuses on exploring the role of intestinal microbiota and bile acids in the onset and progression of IBD,and investigating new effective treatment strategies by targeting intestinal microbiota and bile acids,such as bile acid receptor modulators,probiotics,prebiotics,fecal microbiota transplantation(FMT),and phage therapy.

AIM:To evaluate the bioequivalence of the two rivaroxaban tablets in Chinese healthy subjects.METHODS:Twenty-eight subjects under fasting status and twenty-eight subjects under fed status were enrolled in the study.This study was designed as a four period,fully repetitive,cross-over study.All subjects were administered test(T)and reference(R)rivaroxaban tablets(10 mg)un-der fasting and fed condition respectively.Liquid chromatography-tandem mass spectrometry was used to detect the concentrations of rivaroxaban in plasma.WinNonlin 7.0 was used to calculate the main pharmacokinetic parameters(PK)and to eval-uate the bioequivalence.RESULTS:In fasting group,the main pharmacokinetic parameters of T and R preparation were as follows:Cmax were(186.57±56.41)and(187.61±50.89)ng/mL;AUC0-t were(1 156.21±335.85)and(1 177.59±343.72)h·ng·mL-1;AUC0-∞ were(1 235.77±384.03)and(1223.53±392.10)ng·h·mL-1.The 90％confidential interval(CI)of the three main parameters were 90.81％-105.67％,92.83％-103.85％and 95.04％-107.13％.The upper limit of the 90％CI for the test-to-reference ratio of the within-subject of Cmax,AUC0-t and AUC0-∞ were 1.56,1.41 and 1.73.In fed group,the main pharmacokinetic parameters of T and R preparation were as follows:Cmax were(207.81±45.26)and(211.04±36.62)ng/mL;AUC0-t were(1 271.26±260.92)and(1 233.23±201.85)h·ng·mL-1;AUC0-∞ were(1 290.76±264.90)and(1251.68±203.73)ng·h·mL-1.The 90％CI of the three main parameters were 92.82％-102.28％,97.68％-106.68％and 97.71％-106.68％.The upper limit of the 90％CI for the test-to-reference ratio of the within-subject of Cmax,AUC0t and AUC0-∞were 1.76,1.47 and 1.47.CONCLUSION:The two preparations of rivaroxaban tablets were bioequiva-lent.