1.Microbiological characterization of Staphylococcus epidermidis with hemolytic phenotype
Guiyun LENG ; Wei CHEN ; Chenghao WANG ; Jie YAO ; Chuanping CHEN ; Wei TANG
Acta Universitatis Medicinalis Anhui 2026;61(1):60-66
ObjectiveTo explore the microbiological characteristics of Staphylococcus epidermidis with hemolytic phenotype (SEHP). MethodsHemolytic phenotype was detected using the three-point inoculation method, involving a total of 5 strains of SEHP and 5 strains of Staphylococcus epidermidis with non-hemolytic phenotype (SENHP) . Bacterial species were identified using the Microflex LT MALDI-TOF mass spectrometer, and a phylogenetic tree was constructed through 16S rRNA sequence alignment. Growth curves were monitored through the microcultivation assay. Biofilm formation ability was assessed by microplate crystal violet staining. Red blood cell toxicity was detected using the microplate method. Antimicrobial susceptibility testing of SEHP and SENHP against commonly used antibiotics was performed using a VITEK 2 GP639 test kit. Antagonistic effects of SEHP and SENHP against Staphylococcus aureus and Corynebacterium striatum were evaluated by the Oxford cup inhibition assay. ResultsCompared with SENHP, SEHP exhibited a marked decrease in growth rate during the late logarithmic phase, accompanied by significant hemolytic toxicity. Additionally, it showed lower resistance rates to levofloxacin and moxifloxacin, and could antagonize Staphylococcus aureus and Corynebacterium striatum. ConclusionThe microbiological characteristics of SEHP differ from those of SENHP in that SEHP demonstrates antagonistic effects against S. aureus and C. striatum.
2.Research Progress on the Role of Programmed Cell Death in Flap Ischemia-Reperfusion Injury
Jiwei ZHANG ; Jie ZHANG ; Xinshan WANG ; Xingzhang YAO ; Zhenxing JIANG ; Zhijun HE ; Tao LIU ; Jianliang LI ; Hui YAO ; Jie AN ; Qiuyue ZHAO ; Xiaotao WEI ; M Rayan GHAZI
Medical Journal of Peking Union Medical College Hospital 2026;17(3):851-861
Flap transplantation is a critical surgical strategy for the reconstruction of tissue defects caused by trauma, tumor resection, and congenital malformations, and its survival rate directly determines surgical efficacy and patient prognosis. Following transplantation, flaps inevitably undergo ischemia-reperfusion (I/R) injury, during which oxidative stress, inflammatory responses, and metabolic disturbances are intricately intertwined, ultimately leading to cellular injury and tissue necrosis. Recent studies have demonstrated that multiple forms of programmed cell death—including apoptosis, pyroptosis, ferroptosis, necroptosis, and PANoptosis—play central roles in flap I/R injury. The extensive crosstalk and molecular interactions among these pathways form a highly complex cell death network. Specifically, apoptosis is mediated by the imbalance of Bcl-2 family proteins and the activation of cysteine-dependent aspartate-specific protease (caspase) cascades; pyroptosis is driven by the NLRP3-caspase-1-GSDMD axis, resulting in membrane pore formation and the release of pro-inflammatory cytokines; ferroptosis is characterized by iron-dependent lipid peroxidation and dysfunction of glutathione peroxidase 4 (GPX4); necroptosis is triggered by the receptor-interacting serine/threonine-protein kinase 1 (RIPK1)-RIPK3-MLKL signaling complex, leading to membrane rupture; and PANoptosis represents an integrated form of inflammatory cell death that coordinates multiple death pathways. Importantly, these forms of programmed cell death are not independent but are interconnected through extensive signaling crosstalk. Key regulatory molecules, including caspase-8, reactive oxygen species (ROS), nuclear factor-κB (NF-κB), and nuclear factor erythroid 2-related factor 2 (Nrf2), collectively modulate the dynamic balance among these pathways. Therefore, the multidimensional interplay and spatiotemporal dynamics of programmed cell death constitute a fundamental pathological basis of flap I/R injury. This review systematically summarizes the latest advances in the mechanisms and interactions of various programmed cell death pathways in flap I/R injury, aiming to elucidate the underlying regulatory network. These insights may provide novel theoretical foundations for optimizing flap protection strategies, improving flap survival, and promoting tissue repair.
3.Mechanism of active ingredient compatibility of Dimocarpus longan Lour. leaves in improving glucose and lipid metabolism disorders in type 2 diabetic mellitus rats
Yanli LIANG ; Shijia AN ; Fengsheng LI ; Jiani MAI ; Anqi HUO ; Jiali WEI ; Zejuan ZHANG ; Shuyan QIN ; Wenqing HUANG ; Jie LIANG
China Pharmacy 2026;37(13):1697-1703
OBJECTIVE To explore the mechanism of the active ingredient compatibility(quercetin, quercitrin and kaempferol at a mass ratio of 2∶9∶3)of Dimocarpus longan Lour. leaves(abbreviated as CDL) on ameliorating glucose and lipid metabolism disorders in type 2 diabetes mellitus (T2DM) rats. METHODS SD rats were randomly divided into blank control group, model group, metformin hydrochloride group (100 mg/kg), and CDL high-, medium- and low-dose groups (280, 140, 75 mg/kg), with 10 rats in each group. Rats in the blank control group were fed with standard chow, while rats in the other groups were given high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin to establish the T2DM rat model. After successful modeling, rats in each administration group were given corresponding drug solution, and rats in the blank control group and model group were intragastrically administered with equal volume of pure water, once a day, for consecutive 4 weeks. Fasting blood glucose (FBG) was detected at fixed time every week. The curves of oral glucose tolerance test (OGTT) and intraperitoneal insulin tolerance test (IPITT) were plotted, and the area under curve (AUC) was calculated. The pancreatic islet function indexes [fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI)],blood lipid indexes [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C)] and hepatic glycogen content were determined. The pathological morphological changes of liver and pancreatic tissues were observed. The protein and mRNA expression levels of molecules related to phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in liver tissues were detected. RESULTS Compared with the blank control group, the FBG, AUC of IPITT curve, AUC of OGTT curve, HOMA-IR, the levels of FINS, TC, TG and LDL-C, as well as the protein and mRNA expression of phosphatase and tensin homolog, forkhead box protein O1 and glycogen synthase kinase-3β in liver tissues were significantly increased in the model group ( P <0.05). ISI, the levels of HDL-C and hepatic glycogen content, along with the protein and mRNA expression of PI3K, insulin receptor substrate-1, Akt and protein expression of phosphorylated Akt in liver tissues were markedly decreased ( P <0.05). In model group rats, the arrangement of hepatocytes was irregular, the overall structure of pancreatic lobules was disordered, and a large number of inflammatory cell infiltration was observed. Compared with the model group, most of the above quantitative indexes were significantly reversed in the CDL high-dose group ( P <0.05), and the pathological lesions of liver and pancreas were obviously alleviated. CONCLUSIONS CDL can regulate glucose and lipid metabolism disorders, elevate insulin sensitivity and relieve insulin resistance in T2DM rats. Its mechanism may be related to the activation of the PI3K/Akt signaling pathway.
4.Mechanism of active ingredient compatibility of Dimocarpus longan Lour. leaves in improving glucose and lipid metabolism disorders in type 2 diabetic mellitus rats
Yanli LIANG ; Shijia AN ; Fengsheng LI ; Jiani MAI ; Anqi HUO ; Jiali WEI ; Zejuan ZHANG ; Shuyan QIN ; Wenqing HUANG ; Jie LIANG
China Pharmacy 2026;37(13):1697-1703
OBJECTIVE To explore the mechanism of the active ingredient compatibility(quercetin, quercitrin and kaempferol at a mass ratio of 2∶9∶3)of Dimocarpus longan Lour. leaves(abbreviated as CDL) on ameliorating glucose and lipid metabolism disorders in type 2 diabetes mellitus (T2DM) rats. METHODS SD rats were randomly divided into blank control group, model group, metformin hydrochloride group (100 mg/kg), and CDL high-, medium- and low-dose groups (280, 140, 75 mg/kg), with 10 rats in each group. Rats in the blank control group were fed with standard chow, while rats in the other groups were given high-sugar and high-fat diet combined with intraperitoneal injection of streptozotocin to establish the T2DM rat model. After successful modeling, rats in each administration group were given corresponding drug solution, and rats in the blank control group and model group were intragastrically administered with equal volume of pure water, once a day, for consecutive 4 weeks. Fasting blood glucose (FBG) was detected at fixed time every week. The curves of oral glucose tolerance test (OGTT) and intraperitoneal insulin tolerance test (IPITT) were plotted, and the area under curve (AUC) was calculated. The pancreatic islet function indexes [fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), insulin sensitivity index (ISI)],blood lipid indexes [total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C)] and hepatic glycogen content were determined. The pathological morphological changes of liver and pancreatic tissues were observed. The protein and mRNA expression levels of molecules related to phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway in liver tissues were detected. RESULTS Compared with the blank control group, the FBG, AUC of IPITT curve, AUC of OGTT curve, HOMA-IR, the levels of FINS, TC, TG and LDL-C, as well as the protein and mRNA expression of phosphatase and tensin homolog, forkhead box protein O1 and glycogen synthase kinase-3β in liver tissues were significantly increased in the model group ( P <0.05). ISI, the levels of HDL-C and hepatic glycogen content, along with the protein and mRNA expression of PI3K, insulin receptor substrate-1, Akt and protein expression of phosphorylated Akt in liver tissues were markedly decreased ( P <0.05). In model group rats, the arrangement of hepatocytes was irregular, the overall structure of pancreatic lobules was disordered, and a large number of inflammatory cell infiltration was observed. Compared with the model group, most of the above quantitative indexes were significantly reversed in the CDL high-dose group ( P <0.05), and the pathological lesions of liver and pancreas were obviously alleviated. CONCLUSIONS CDL can regulate glucose and lipid metabolism disorders, elevate insulin sensitivity and relieve insulin resistance in T2DM rats. Its mechanism may be related to the activation of the PI3K/Akt signaling pathway.
5.Research progress of nano drug delivery system based on metal-polyphenol network for the diagnosis and treatment of inflammatory diseases
Meng-jie ZHAO ; Xia-li ZHU ; Yi-jing LI ; Zi-ang WANG ; Yun-long ZHAO ; Gao-jian WEI ; Yu CHEN ; Sheng-nan HUANG
Acta Pharmaceutica Sinica 2025;60(2):323-336
Inflammatory diseases (IDs) are a general term of diseases characterized by chronic inflammation as the primary pathogenetic mechanism, which seriously affect the quality of patient′s life and cause significant social and medical burden. Current drugs for IDs include nonsteroidal anti-inflammatory drugs, corticosteroids, immunomodulators, biologics, and antioxidants, but these drugs may cause gastrointestinal side effects, induce or worsen infections, and cause non-response or intolerance. Given the outstanding performance of metal polyphenol network (MPN) in the fields of drug delivery, biomedical imaging, and catalytic therapy, its application in the diagnosis and treatment of IDs has attracted much attention and significant progress has been made. In this paper, we first provide an overview of the types of IDs and their generating mechanisms, then sort out and summarize the different forms of MPN in recent years, and finally discuss in detail the characteristics of MPN and their latest research progress in the diagnosis and treatment of IDs. This research may provide useful references for scientific research and clinical practice in the related fields.
6.The value of dual-energy CT Bone Marrow Edema in quantitative evaluation of rib fracture evolution timing
Fei YANG ; Yongjie YAO ; Jie LI ; Fan LI ; Meixia YANG ; Wenxin WEI
Chinese Journal of Forensic Medicine 2025;40(2):188-193
Objective To explore the value of dual-energy CT Bone Marrow Edema in quantitatively evaluating the timing of rib fracture evolution.Methods Sixty patients with thoracic trauma were scanned by dual-energy CT.Using CT Bone Marrow Edema technique,bone marrow CT values were standardized and quantified in both the Bone Marrow Edema area at the rib fracture site and in normal areas 1 cm on both sides of the fracture.The increments of standardized CT values for Bone Marrow Edema and VNCa were obtained for three stages of healing.The numerical variables were statistically described,and both the standardized CT value increments and VNCa standardized CT value increments were compared between the three stages and between each pair of stages.Diagnostic efficacy for stages with significant differences was evaluated using the area under the receiver operating characteristic(ROC)curve(AUC),and Cut-offvalues were calculated.Results There were significant differences in standardized CT value increments and VNCa standardized CT value increments across the three stages of Bone Marrow Edema(H=10.788,p=0.005;F=115.787,p=0.000).The standardized CT value increment of Bone Marrow Edema showed significant differences between the cartilaginous callus stage(fibrous callus stage)and the bony callus-remodeling stage(H=54.958,p=0.003),while the other pairwise comparisons showed no statistical significance(H=-25.603,p=0.183;H=29.354,p=0.113,respectively).However,the VNCa standardized CT value increments showed statistical significance across all three pairwise comparisons(all p=0.000).The ROC curve for differentiating the cartilaginous callus stage(fibrous callus stage)from the bony callus-remodeling stage using Bone Marrow Edema standardized CT value increment had an AUC of 0.652,with a Cut-offvalue of 81.575 Hu.The ROC curve for distinguishing the hematoma inflammation stage from the cartilaginous callus stage(fibrous callus stage)using VNCa standardized CT value increment had an AUC of 0.668,with a Cut-offvalue of 55.700 Hu.The ROC curve for differentiating the cartilaginous callus stage(fibrous callus stage)from the bony callus-remodeling stage using VNCa standardized CT value increment had an AUC of 0.905,with a Cut-offvalue of 37.625 Hu.Conclusion Dual-energy CT Bone Marrow Edema can quantitatively evaluate the timing of rib fracture evolution,and the differences in standardized CT value increments at different stages can provide a theoretical basis for forensic identification of fractures at different time periods.The Cut-offvalues of standardized CT value increments can,to some extent,predict the time stage of a fracture,providing quantitative evidence for forensic experts in the identification of rib fractures.
7.Predictive value of CTP parameters combined with serum PAC-1 level and HAT score for hemorrhagic transformation after intravenous thrombolysis in acute cerebral infarction
Zhixi CHEN ; Wei YAO ; Wei LIU ; Jie WANG ; Xingning ZHOU
International Journal of Laboratory Medicine 2025;46(20):2446-2451
Objective To investigate the predictive value of CT perfusion imaging(CTP)parameters com-bined with platelet activator combined-1(PAC-1)level and hemorrhage after thrombolysis(HAT)score for hemorrhagic transformation(HT)after intravenous thrombolysis in patients with acute cerebral infarction(ACI).Methods A total of 162 patients with ACI who were treated with intravenous thrombolysis in Cheng-du 363 Hospital from January 2022 to April 2023 were selected.All patients underwent head CT examination(including plain scan and CTP)before intravenous thrombolytic therapy,and serum PAC-1 level and HAT score were measured at admission.According to whether HT occurred after intravenous thrombolysis,the pa-tients were divided into HT group(HT group)and non-HT group(non-HT group).The differences of CTP parameters,serum PAC-1 level and HAT score between the two groups were compared.Multivariate Logistic regression was used to analyze the independent risk factors of HT after intravenous thrombolysis in ACI pa-tients.The predictive value of CTP parameters combined with serum PAC-1 and HAT scores alone and in combination for HT after intravenous thrombolysis in ACI patients was evaluated by drawing the receiver op-erating characteristic(ROC)curve and calculating the area under the curve(AUC).Results The cerebral blood flow and blood volume in the HT group were significantly lower than those in the non-HT group(P<0.05),while the serum PAC-1 level,peak time,mean transit time,flow extraction product,and HAT score were significantly higher than those in the non-HT group(P<0.05).Multivariate Logistic regression analysis showed that CTP integration parameters which were composed of cerebral blood flow,cerebral blood volume,peak time,mean transit time and flow extraction product,serum PAC-1 level and HAT score were independ-ent risk factors for HT after intravenous thrombolysis in ACI patients(P<0.05).ROC curve analysis showed that CTP integration parameters,serum PAC-1 level and HAT score all had certain predictive value for HT in ACI patients after intravenous thrombolysis,and their AUC and 95%CI were 0.766(95%CI:0.605-0.915),0.702(95%CI:0.451-0.955)and 0.741(95%CI:0.551-0.923),respectively.The com-bined application of CTP integration parameters,serum PAC-1 levels,and HAT scores significantly improved the predictive performance,and its AUC was 0.856(95%CI:0.750-0.951),which was higher than those of each index alone(Zcombination-CTP=2.177,P=0.032,Zcombination-PAC-1=2.917,P=0.004,Zcombination-HAT=2.537,P=0.013).Conclusion The combination of CTP integration parameters which are composed of cerebral blood flow,cerebral blood volume,peak time,mean transit time and flow extraction product,and serum PAC-1 level and HAT score has high predictive value for HT after intravenous thrombolysis in ACI patients,and has high clinical application potential.
8.Complete cfb gene deletion in Streptococcus agalactiae: molecular mechanism of qPCR false negatives
Xiu Wang ; Guiyun Leng ; Yunsi Yang ; Wei Tang ; Qiang Zhou ; Jie Yao
Acta Universitatis Medicinalis Anhui 2025;60(9):1624-1630
Objective :
To investigate the causes of false negative results in the detection of Streptococcus agalactiae using fluorescent quantitative PCR(qPCR) targeting the CAMP factor gene(cfb),and to perform a comprehensive analysis of the associated molecular mechanisms.
Methods:
A total of 76 vaginal secretion samples were evaluated using both qPCR based on cfb gene and bacterial culture methods. Four suspicious strains exhibiting negative qPCR results but positive culture findings were identified using matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF MS),latex agglutination antigen detection,and the CAMP test. Whole genome analysis was conducted utilizing the MGI DNBSEQ-T7 and Nanopore-PromethION 48 sequencing platforms. Phylogenetic and circular evolutionary trees were constructed using the 16S rRNA gene for strain verification. Multilocus sequence typing(MLST) was conducted,and cfb sequences were aligned and analyzed based on spliced sequences and original data. Specific primers targeting the cfb gene were designed for full-length amplification,followed by verification through agarose gel electrophoresis.
Results:
The four strains identified as suspicious were classified as S. agalactiae through MALDI-TOF MS,antigen detection,and 16S rRNA gene analysis,with MLST typing indicating ST-862. Phenotypic analysis revealed a negative CAMP test. Whole genome sequence alignment failed to detect the cfb gene or any homologous sequences,and molecular testing confirmed the absence of cfb gene PCR amplification products,thereby confirming its complete deletion.
Conclusion
This deletion is identified as the molecular mechanism responsible for the false negative qPCR detection of S. agalactiae when targeting this specific gene.It is recommended that the qPCR detection targeting a single cfb gene has limitation,and this may impact clinical diagnosis and treatment decisions. This limitation warrants carefulconsideration.
9.Hemolytic phenotype analysis of Staphylococcus aureus clinical isolates
Ju Gao ; Shengnan Weng ; Guiyun Leng ; Xin Li ; Jie Yao ; Qiang Zhou ; Wei Tang
Acta Universitatis Medicinalis Anhui 2025;60(7):1312-1319
Objective:
To investigate the hemolytic phenotypes of Staphylococcus aureus clinical isolates.
Methods:
The hemolytic phenotypes of 105 Staphylococcus aureus isolates were analyzed and summarized using the three-point inoculation method.Real-time fluorescence quantitative PCR was used to measure the mRNA expression levels of four hemolysin genes (hla,hlb,hlc,and hld) ; The VITEK 2 GP639 antimicrobial susceptibility card was used to detect resistance to commonly used antibiotics ; DNA gel electrophoresis was performed to determine the prevalence of the mecA,sea,tst,and pvl genes ; The microtiter plate crystal violet staining method was used to assess biofilm formation ability ; The CCK-8 assay was used to evaluate cytotoxicity against macrophages.
Results:
Seven hemo- lytic phenotypes were identified among the Staphylococcus aureus clinical isolates. Differences were found among Staphylococcus aureus clinical isolates with different hemolytic phenotypes in terms of mRNA expression levels of he- molysin genes,antibiotic resistance,virulence gene prevalence,biofilm formation ability,and cytotoxicity to mouse macrophages (P <0. 05 ) .
Conclusion
Staphylococcus aureus clinical isolates exhibit diverse hemolytic pheno- types,which should be a focus across multiple dimensions,including microbiological testing,clinical treatment, and nosocomial infection prevention and control.
10.Divergent activation patterns of BRS3 revealed by two Chinese herb-derived agonists.
Jie LI ; Changyao LI ; Qingtong ZHOU ; Wei HAN ; Mingzhu FANG ; Youwei XU ; Yiting MAI ; Yao ZHANG ; Jiahua CUI ; H Eric XU ; Yan ZHANG ; Wanchao YIN ; Ming-Wei WANG
Acta Pharmaceutica Sinica B 2025;15(10):5231-5243
Bombesin receptor subtype-3 (BRS3) is an orphan G protein-coupled receptor (GPCR) that plays critical roles in energy homeostasis, glucose metabolism, and insulin secretion. Recent structural studies have elucidated BRS3 signaling mechanisms using synthetic ligands, including BA1 and MK-5046. However, the molecular basis of BRS3 activation by bioactive natural compounds and their derivatives, particularly those derived from traditional Chinese medicine, remains unclear. Here, we present high-resolution cryogenic electron microscopy (cryo-EM) structures of the human BRS3-Gq complex in both unliganded and active states bound by two herb-derived compounds (DSO-5a and oridonin), at resolutions of 2.9, 2.8, and 2.9 Å, respectively. These structures display distinct ligand recognition patterns between DSO-5a and oridonin. Although both compounds bind to the orthosteric pocket, they differentially engage the interaction network of BRS3, as demonstrated by mutagenesis studies assessing calcium mobilization and inositol phosphate 1 (IP1) accumulation. These findings enhance our understanding of BRS3 activation and provide valuable insights into the development of small-molecule BRS3 modulators with therapeutic potential.


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