[摘 要] 目的：探讨绿茶单体表没食子儿茶素没食子酸酯（EGCG）对大鼠肝癌发生过程中肠道菌群结构变化的影响。方法：建立二乙基亚硝胺（DEN）诱导的肝癌大鼠模型。将26只SD大鼠随机分成3组，分别为正常对照组、肝癌组和EGCG干预组。从实验开始第1天起，EGCG干预组每日给予EGCG（40 mg/kg）灌胃，正常对照组和肝癌组给予等量生理盐水灌胃，1次/日，持续至第20周。灌胃结束后，采集大鼠粪便样本，提取DNA，进行高通量16S rRNA V3-V4区测序；处死大鼠，取肝，观察肿瘤形成情况，并计算肝癌发生率。对测序数据进行生物信息学分析：经质控、聚类获得操作分类单元（OTU）表，据此计算α多样性指数（包括Observed species、Chao1、Shannon和Simpson指数），并进行β多样性分析。同时，对物种进行分类学注释，比较各组间菌群组成与丰度差异。结果： EGCG干预组大鼠（8只）肝脏肿瘤形成率明显低于肝癌组（10只）（50% vs 100%， P = 0.023），正常对照组（8只）大鼠无肿瘤发生。在肠道菌群方面，肝癌组操作分类单元（OTU）数量远低于正常对照组（P <0.001），而EGCG干预组OTU数量总体上高于肝癌组（P = 0.021）。α多样性分析显示，肝癌组Shannon指数低于正常对照组（P < 0.05）；此外，与肝癌组相比，EGCG干预组的Observed species指数、Chao1指数、Shannon指数和Simpson指数均显著提高（P < 0.05）。β多样性分析及主坐标分析（PCoA）表明，三组肠道菌群结构存在显著分离（PERMANOVA， R² = 0.3918， P = 0.001），其中EGCG干预组群落结构介于肝癌组与正常对照组之间，并更接近于正常对照组。肝癌组大鼠较正常大鼠肠道菌群中链球菌等潜在致病菌富集，丁酸产生相关菌（如丁酸球菌属、瘤胃球菌属等）丰度显著降低（P < 0.05）。相比之下，在EGCG干预肝癌发生过程中，大鼠肠道菌群结构相对稳定。厚壁菌门/拟杆菌门比值较肝癌组显著提高（P < 0.05），益生菌（如双歧杆菌属、乳杆菌属等）和丁酸产生相关菌（如丁酸球菌属）富集。结论：EGCG干预可降低DEN诱导的大鼠肝癌发生率，并有助于稳定肠道菌群结构，其作用可能与增加菌群多样性、促进益生菌及丁酸产生菌富集、恢复菌群平衡有关。

BACKGROUND:Psoralen has a strong anti-osteoporotic activity and may have a restorative effect on chemotherapy-induced osteoporosis. OBJECTIVE:To explore the restorative effect of psoralen on the osteogenic differentiation of bone marrow mesenchymal stem cells in mice inhibited by cyclophosphamide and its mechanism. METHODS:C57BL/6 mouse bone marrow mesenchymal stem cells were isolated and cultured.Effect of psoralen on viability of bone marrow mesenchymal stem cells was detected by MTT assay.Osteogenic induction combined with alkaline phosphatase staining was used to determine the optimal dose of psoralen to restore the osteogenic differentiation of bone marrow mesenchymal stem cells inhibited by cyclophosphamide.The mRNA expression levels of Runx2,alkaline phosphatase,Osteocalcin,osteoprotegerin,and Wnt/β-catenin signaling pathway-related genes Wnt1,Wnt4,Wnt10b,β-catenin,and c-MYC were measured by RT-qPCR at different time points under the intervention with psoralen.The protein expression of osteogenic specific transcription factor Runx2 and Wnt/β-catenin signaling pathway related genes Active β-catenin,DKK1,c-MYC,and Cyclin D1 was determined by western blot assay at different time points under the intervention with psoralen. RESULTS AND CONCLUSION:(1)There was no significant effect of different concentrations of psoralen on the viability of bone marrow mesenchymal stem cells.The best recovery of the inhibition of osteogenic differentiation of bone marrow mesenchymal stem cells caused by cyclophosphamide was under the intervention of psoralen at a concentration of 200 μmol/L.(2)Psoralen reversed the reduction in osteogenic differentiation marker genes Runx2,alkaline phosphatase,Osteocalcin and osteoprotegerin mRNA expression and Runx2 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(3)Psoralen reversed the decrease in Wnt/β-catenin pathway-related genes Wnt4,β-catenin,c-MYC mRNA and Active β-catenin,c-MYC,and Cyclin D1 protein expression and the increase in DKK1 protein expression in bone marrow mesenchymal stem cells caused by cyclophosphamide conditioned medium.(4)The results showed that cyclophosphamide inhibited osteogenic differentiation of bone marrow mesenchymal stem cells in mice,and psoralen had a restorative effect on it.The best intervention effect was achieved at a concentration of 200 μmol/L psoralen,and this protective effect might be related to the activation of Wnt4/β-catenin signaling pathway by psoralen.

Objective:To explore the effect of necroptosis inhibitor necrosulfonamide (NSA) on traumatic brain injury (TBI) mouse model and BV2 cell pyroptosis model and their mechanisms.Methods:(1) In vivo experiments: 50 mice were randomly divided into sham-operated group, TBI group, TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group, and TBI+10 mg/kg NSA group, with 10 mice in each group. TBI model was established using a modified Feeney's weight-drop method; 4 h after modeling, 90% corn oil, 1 mg/kg NSA, 5 mg/kg NSA, or 10 mg/kg NSA was administered into the mice, respectively. Mice in the sham-operated group only had circular bone window opened without being subjected to impact. At 48 hours after modeling, neurological function was evaluated by modified neurological function score (mNSS), serum lactate dehydrogenase (LDH) content was detected by LDH detection kit, contents of interleukin (IL)-18, IL-1β and tumor necrosis factor-α (TNF-α) in the brain tissues were detected by enzyme-linked immunosorbent assay (ELISA), and expressions and localizations of ionized calcium binding adaptor molecule 1 (IBA-1), cysteinyl aspartate specific proteinase-1 (Caspase-1) p20 and gasdermin D (GSDMD) in the injured parietal cortex were detected by double immunofluorescent staining. (2) In vitro experiments: BV2 cells were divided into control group, lipopolysaccharide (LPS)+adenosine triphosphate (ATP)+dimethyl sulfoxide (DMSO) group, LPS+ATP+5 μmol/L NSA group, LPS+ATP+10 μmol/L NSA group, and LPS+ATP+15 μmol/L NSA group. Cells in the latter 4 groups were induced by LPS+ATP to establish BV2 cell pyroptosis model, and incubated with 2 μL DMSO, 5 μmol/L NSA, 10 μmol/L NSA, and 15 μmol/L NSA for 1 hour, respectively; cells in the control group were cultured conventionally. Contents of LDH, IL-1β, IL-18, and TNF-α in the cell culture supernatant were detected by ELISA; pyroptosis was detected by calcein acetoxymethyl ester (CAM)/propidium iodide (PI) double staining; protein expressions of nucleotide binding domain-like receptor protein 3 (NLRP3), Caspase-1 p20, GSDMD, and N-terminal fragment of GSDMD (GSDMD-N) were detected by Western blotting. Results:(1) Compared with the TBI group, the TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group and TBI+10 mg/kg NSA group had decreased mNSS score and serum LDH content, decreased IL-1β and IL-18 contents in the brain tissues and number of Caspase-1 p20 + cells in the injured parietal cortex, successively, with significant differences ( P<0.05). Compared with the TBI group ([287.80±12.26] cells/mm 2), the TBI+1 mg/kg NSA group, TBI+5 mg/kg NSA group, and TBI+10 mg/kg NSA group had decreased number of Iba-1 +GSDMD + cells in the injured parietal cortex ([213.70±11.87] cells/mm 2, [205.30±9.15] cells/mm 2, [131.70±13.69] cells/mm 2),successively, with significant differences ( P<0.05). Compared with the TBI group, the TBI+5 mg/kg NSA group and TBI+10 mg/kg NSA group had significantly decreased number of Iba-1 + cells in the injured parietal cortex, and the TBI+10 mg/kg NSA group had significantly decreased TNF-α content in the brain tissues and number of GSDMD + cells in the injured parietal cortex ( P<0.05). Compared with the TBI group ([247.20±9.88] cells/mm 2), the TBI+10 mg/kg NSA group had significantly decreased number of Iba-1 +Caspase-1 p20 + cells in the injured parietal cortex ([181.70±9.37] cells/mm 2, P<0.05). (2) Compared with the LPS+ATP+DMSO group, the LPS+ATP+5 μmol/L NSA group, LPS+ATP+10 μmol/L NSA group, and LPS+ATP+15 μmol/L NSA group had decreased IL-18 content in the supernatant, successively, with significant differences ( P<0.05); and compared with the LPS+ATP+DMSO group, the LPS+ATP+10 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased contents of LDH, IL-1β, and TNF-α in the supernatant and ratio of PI +/CAM + cell counts ( P<0.05). Compared with the LPS+ATP+DMSO group (2.62±0.50), the LPS+ATP+10 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased Caspase-1 p20 protein expression (1.36±0.14, 1.32±0.07, P<0.05). Compared with the LPS+ATP+DMSO group (5.00±1.67), the LPS+ATP+5 μmol/L NSA group and LPS+ATP+15 μmol/L NSA group had significantly decreased GSDMD protein expression (1.42±0.26, 1.68±0.32, P<0.05). Compared with the LPS+ATP+DMSO group (2.28±0.24), the LPS+ATP+15 μmol/L NSA group had significantly decreased GSDMD-N protein expression (1.23±0.08, P<0.05). Conclusion:NSA can inhibit microglial pyroptosis after TBI by inhibiting the Caspase-1 p20/GSDMD pathway, thereby playing a neuroprotective role.

Purpose This study aims to analyze the clinical,pathological,and molecular genetic characteristics of nodal follicular helper T-cell lymphoma,follicular type(nTFHL-F).Methods 7 cases of nTFHL-F were re-viewed.Clinical data were collected,tissue morphology was summarized,and immunohistochemical staining and mo-lecular testing were performed.Results The median age of patients was 62 years with a male-to-female ratio of 6:1.The initial symptoms included neck lymphadenectasis in 6 cases and abdominal discomfort in one.Six cases were in ad-vanced stages,while 1 case was in the localized stage.The tumors exhibited a vague,irregular follicular nodular pat-tern,without significant polymorphic inflammatory background or high endothelial vascular proliferation.Five cases showed a progressive transformation resembling germinal center pattern,and two cases exhibited a follicular lymphoma-like growth pattern.Tumor cells presented three distinct morphologies:centrocyte-like appearance,monocytoid B cell-like appearance,and atypical cells with abundant,transparent cytoplasm.Tumor cells expressed at least three follicu-lar T-cell markers.Testing for ITK::SYK gene fusion was negative in all cases(0/7).Next generation sequencing i-dentified mutations in TET2 gene in two cases(2/2),the RHOA gene in one case(1/2),and VAV1 gene in one case(1/2).The follow-up duration ranged from 2 to 64 months,with three deaths(3/7),and a median survival time of 37 months.Conclusion nTFHL-F predominantly occurs in middle-aged to elderly males,presenting with advanced clinical stages,and has a poor prognosis.nTFHL-F is closely associated with nodal follicular helper T-cell lymphoma,angioimmunoblastic type,and mya coexist within the same individual.

OBJECTIVE To explore the effects of meropenem exposure and degradation levels on clinical efficacy in patients with purulent meningitis （PM）. METHODS A total of 131 PM patients treated with meropenem at the Affiliated Suzhou Hospital of Nanjing Medical University from January 2022 to June 2025 were prospectively included. Relevant data were collected and divided into a cured group （91 cases） and a non-cured group （40 cases） based on the efficacy. High-performance liquid chromatography-tandem mass spectrometry was used to determine the concentration of meropenem and its open-loop metabolites. Risk factors that affect efficacy were screened， and their predictive power and correlation were evaluated by univariate analysis， and multivariate Logistic regression analysis， receiver operating characteristic （ROC） curves， and correlation analysis. RESULTS Univariate analysis showed that serum creatinine， creatinine clearance rate， minimum inhibitory concentration of meropenem ≥16 μg/mL， cerebrospinal fluid red blood cell count， cerebrospinal fluid white blood cell count， cerebrospinal fluid glucose content， blood trough concentration， blood open-loop metabolite concentration/trough concentration ratio， and intrathecal injection were all correlated with efficacy （P＜0.05）. The results of multiple Logistic regression analysis showed that serum creatinine blood open-loop metabolite concentration/trough concentration ratio， intrathecal injection， and cerebrospinal fluid glucose content were influencing factors for suboptimal anti-infective ltt efficacy （P＜0.05）. ROC curve analysis showed that when the blood open-loop metabolite concentration/trough concentration ratio was greater than 2.854 （AUC＝0.647）， serum creatinine was less than 59.5 μmol/L （AUC＝0.647）， and cerebrospinal fluid glucose content was less than 3.37 mmol/L （AUC＝0.709）， the risk of treatment failure significantly increased （P＜0.05）. Correlation analysis showed that the blood trough concentration of meropenem was positively correlated with the concentration of its open-loop metabolites （R 2＝0.134 5， P＜0.000 1）. CONCLUSIONS Insufficient exposure level and rapid degradation of meropenem are key mechanisms affecting the anti-infective efficacy of PM. Elevated blood open-loop metabolite concentration/ trough concentration ratio， low serum creatinine level， lack of intrathecal injection， and low cerebrospinal fluid glucose content are independent risk factors for poor efficacy.

Objective:To investigate the role of Golgi protein 73(GP73)in the diagnosis of primary biliary cholangitis(PBC)and its association with disease progression and therapeutic efficacy monitoring.Methods:Serum samples were collected from 70 PBC pa-tients,36 patients with liver diseases other than autoimmune liver disease(non-AILD group),and 40 healthy controls(HC group),and ELISA was used to measure the serum level of GP73.For the inpatients with PBC,serum samples were collected before and after treat-ment to measure GP73.Results:There was a significant difference in the distribution of serum GP73 concentration between the PBC group,the non-AILD group,and the HC group(P<0.001),and the receiver operating characteristic(ROC)curve showed that GP73 had an area under the ROC curve of 0.839 in the diagnosis of PBC.Serum GP73 level was positively correlated with aspartate amino-transferase(AST)(r=0.337,P=0.009),alkaline phosphatase(ALP)(r=0.380,P=0.003),total bilirubin(r=0.330,P=0.010),and direct bilirubin(r=0.371,P=0.004),while it was negatively correlated with prothrombin activity(r=-0.329,P=0.036)and cholinesterase(r=-0.518,P<0.001).The PBC patients with liver cirrhosis had a significantly higher serum GP73 level than those without liver cirrhosis(P=0.002).There was no significant difference in GP73 content between the patients with positive anti-mitochondrial antibodies-M2,anti-BCOADC-E2PDC-E2 OGDC-E2 antibodies,and anti-SPl00 antibodies and those with negative antibodies.The PBC patients had significant reductions in the serum levels of AST,ALP,gamma-glutamyl transpeptidase,and GP73 after liver-protecting treatment and improvement in cholestasis(P<0.05).Conclusion:GP73 plays an important role in the diagnosis,disease progression,and efficacy monitoring of PBC and is expected to become a potential disease marker for PBC.

Objective:To explore the funding status and research hotspots of the National Natural Science Foundation of China (NSFC) for projects in the field of TCM for the prevention and treatment of heart failure.Methods:The research projects of TCM in the prevention and treatment of heart failure funded were retrieved from NSFC Big Data Knowledge Management Service Platform from January 1, 2010 to December 31, 2023. Excel 2022 software was used to analyze the data of the number of funded projects, the amount of funding, the supporting units and the regions. VOSviewer 1.6.20 software was used to analyze the co-occurrence of keywords with frequency≥3.Results:A total of 202 research projects were funded with a total funding amount of 89.8 million RMB, and the number and amount of projects showed a fluctuating upward trend. The funding categories were mainly general programs and youth science foundation projects. There were 42 supporting units involved, and TCM universities and colleges were the main recipients of funding. Regional distribution was uneven, mainly in Beijing, Shanghai, Guangdong and other regions. There were 17 secondary discipline codes involved, and the discipline categories were concentrated in Internal Medicine of TCM (H3108), Clinical Basis of Integrative Medicine (H3302) and Cardiovascular Pharmacology of Chinese Medicine (H3209). Therapeutic modalities covered compounding, monomers, proprietary Chinese medicines, and drug pairs, and research hotspots included aspects of myocardial energy metabolism remodeling, mitochondrial autophagy, macrophage polarization, endoplasmic reticulum stress, calcium homeostasis, ferroptosis, exosomes, micro-RNAs, and so on.Conclusion:NSFC has provided strong support to the research in the field of TCM for heart failure. In the future, the supporting units should pay attention to the interdisciplinary integration and development to promote the vigorous development of TCM.

Objective:To explore the correlation between smoking behavior of employees in heavy industry enterprises and risk factors for cardiovascular and cerebrovascular diseases.Methods:A cross-sectional study design was adopted. A questionnaire survey and physical examination data collection were conducted on employees of a heavy industry enterprise in Shaanxi Province using a cluster sampling method from March to April 2024. A total of 2 209 cases met the inclusion criteria. According to the smoking index (SI), participants were divided into a non-smoking group (SI=0, 1 316 cases), a low smoking group (SI400, 656 cases), and a high smoking group (SI≥400, 237 cases). According to the fagerstrom test for nicotine dependence (FTND) score, participants were divided into a non-dependence group (FTND=0, 1 316 cases), a mild dependence group (FTND=1-3, 623 cases), a moderate dependence group (FTND=4-6, 204 cases), and a severe dependence group (FTND≥7, 66 cases). Spearman rank correlation, univariate, and multivariate logistic regression analyses were used to compare differences in blood pressure, fasting blood glucose, total cholesterol, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, triglycerides, uric acid, homocysteine, and body mass index among the groups.Results:As the SI and FTND scores increased, blood pressure, low-density lipoprotein cholesterol, triglycerides, uric acid, and homocysteine all showed an increasing trend, while high-density lipoprotein cholesterol showed a decreasing trend (all P0.05). The systolic blood pressure, diastolic blood pressure, fasting blood glucose, low density lipoprotein cholesterol (LDL-C), triglycerides, serum uric acid, and homocysteine in the high smoking group were higher than those in the non-smoking group [(124.93±16.55) vs (122.32±16.62) mmHg (1 mmHg=0.133 kPa), (80.86±11.10) vs (76.18±11.63) mmHg, 5.58 vs 5.29 mmol/L, (3.13±0.65) vs (2.95±0.56) mmol/L, 1.70 vs 1.09 mmol/L, (336.80±69.94) vs (299.00±86.43) μmol/L, 14.20 vs 11.76 μmol/L, all P0.001], the high-smoking group had lower HDL cholesterol than the non-smoking group [(1.34±0.30) vs (1.39±0.27) mmol/L, P0.001], the systolic blood pressure, diastolic blood pressure, fasting blood glucose, total cholesterol, low-density lipoprotein cholesterol, triglycerides, serum uric acid, and homocysteine in the severely dependent group were higher than those in the non-dependent group [(132.09±16.69 )vs (122.32±16.62) mmHg, (81.32±12.97) vs (76.18±11.63) mmHg, 5.61 vs 5.29 mmol/L, (4.98±0.91 )vs (4.70±0.88) mmol/L, (3.19±0.62) vs (2.95±0.56) mmol/L, 1.87 vs 1.09 mmol/L, (328.95±75.21) vs (299.03±86.43) μmol/L, 14.38 vs 11.76 μmol/L, all P0.05], and the high density lipoprotein cholesterol (HDL-C) in the severely dependent group was lower than that in the non-dependent group [(1.32±0.30) vs (1.39±0.27) mmol/L, P0.001]. Multivariate analysis showed that SI and degree of tobacco dependence (DTD), were significantly correlated with fasting blood glucose, triglycerides, and LDL cholesterol, but the effect size of SI was too small ( OR=1.001, 95% CI: 1.000-1.001, P=0.002, OR=1.001, 95% CI: 1.001-1.002, P0.001, OR=1.001, 95% CI: 1.000-1.001, P=0.032). Conclusions:Smoking behavior among employees in heavy industry enterprises is significantly correlated with risk factors for cardiovascular and cerebrovascular diseases. The degree of tobacco dependence is closely related to the risk of abnormal fasting blood glucose, low-density lipoprotein cholesterol, and triglycerides.

Objective:To analyze the clinical features and genetic sequencing results of a patient with Kallmann syndrome(KS) carrying digenic mutations who initially presented with osteoporosis, and to enhance awareness of this disease phenotype.Methods:Clinical data were collected, and peripheral blood DNA was extracted for whole-exome sequencing. Relevant literature was reviewed to summarize phenotypes associated with digenic/oligogenic variants involving CHD7.Results:The patient exhibited back pain, delayed development of secondary sexual characteristics, and hyposmia. Laboratory tests revealed reduced sex hormones and gonadotropin levels, while pituitary imaging was unremarkable. Bone mineral density imaging confirmed osteoporosis, and thoracolumbar X-rays showed multiple vertebral compression fractures. Genetic analysis identified a heterozygous splice-site mutation in CHD7(c.2698-1G>T) and a heterozygous missense mutation in WDR11(c.439G>A: p.D147N). According to ACMG criteria, the CHD7 mutation was classified as pathogenic, while the WDR11 variant was defined as a variant of uncertain significance(VUS). Literature review indicated that 40% of KS patients with digenic/oligogenic variants involving CHD7 presented with hearing or ocular abnormalities.Conclusion:This study reports a novel CHD7 mutation and a previously undescribed digenic combination of CHD7 and WDR11 variants in a KS patient. CHD7 variants may be implicated in auditory or ocular involvement in KS cases with digenic/oligogenic inheritances. KS patients may also manifest skeletal abnormalities in addition to hypogonadotropic hypogonadism. Tailored management of sex hormones and osteoporosis therapies across life stages is essential for optimizing bone health in KS.

Purpose This study aims to analyze the clinical,pathological,and molecular genetic characteristics of nodal follicular helper T-cell lymphoma,follicular type(nTFHL-F).Methods 7 cases of nTFHL-F were re-viewed.Clinical data were collected,tissue morphology was summarized,and immunohistochemical staining and mo-lecular testing were performed.Results The median age of patients was 62 years with a male-to-female ratio of 6:1.The initial symptoms included neck lymphadenectasis in 6 cases and abdominal discomfort in one.Six cases were in ad-vanced stages,while 1 case was in the localized stage.The tumors exhibited a vague,irregular follicular nodular pat-tern,without significant polymorphic inflammatory background or high endothelial vascular proliferation.Five cases showed a progressive transformation resembling germinal center pattern,and two cases exhibited a follicular lymphoma-like growth pattern.Tumor cells presented three distinct morphologies:centrocyte-like appearance,monocytoid B cell-like appearance,and atypical cells with abundant,transparent cytoplasm.Tumor cells expressed at least three follicu-lar T-cell markers.Testing for ITK::SYK gene fusion was negative in all cases(0/7).Next generation sequencing i-dentified mutations in TET2 gene in two cases(2/2),the RHOA gene in one case(1/2),and VAV1 gene in one case(1/2).The follow-up duration ranged from 2 to 64 months,with three deaths(3/7),and a median survival time of 37 months.Conclusion nTFHL-F predominantly occurs in middle-aged to elderly males,presenting with advanced clinical stages,and has a poor prognosis.nTFHL-F is closely associated with nodal follicular helper T-cell lymphoma,angioimmunoblastic type,and mya coexist within the same individual.