Objective To investigate the effects of Jianpi Huatan Fang on lipid metabolism and expression levels of FOXO1 and PDK4 in rats with polycystic ovary syndrome(PCOS)and insulin resistance(IR).Methods Forty female rats were divided randomly into a blank control group(n=10)and a model group(n=30).A PCOS-IR rat model was established using a high-fat diet(8 weeks)combined with letrozole(added at weeks 4～8).Thirty successfully modeled rats were then divided randomly into a model control group,a Jianpi Huatan Fang group(11.07 g/kg),and a metformin group(0.2 g/kg)(n=10 rats per group).Drug intervention was given for 4 weeks.The general status,Traditional Chinese medicine syndrome score,and weight changes were observed in each group.Histopathological changes in the ovary were detected by hematoxylin and eosin staining.Fasting blood glucose and blood lipids were measured using a blood biochemical analyzer.Levels of sex hormones including follicle-stimulating hormone(FSH),luteinizing hormone(LH),testosterone(T),estradiol(E2),and insulin were determined by enzyme linked immunosorbent assay and changes in the expression of FOXO1 and PDK4 in the ovaries were determined by Western Blot and real-time fluorescence quantitative polymerase chain reaction.Results Rats in the model control group exhibited symptoms of spleen deficiency and phlegm-dampness and showed significant weight gain,sex hormone disorders,polycystic ovarian changes,and dysregulation of glucose and lipid metabolism,compared with rats in the blank control group(P＜0.01).In addition,weight increase was slower in rats in the Jianpi Huatan Fang group and the metformin group compared with that in the model control group,and follicle development was improved.Serum LH,T,LH/FSH,fasting blood glucose,insulin,homeostatic model assessment for IR,cholesterol,triglycerides,and low-density lipoprotein cholesterol decreased,estradiol and FSH increased,and ovarian FOXO1 and PDK4 mRNA and protein expression levels were down regulated in the Jianpi Huatan Fang group compared with the findings in the model control group(P＜0.05,P＜0.01).Serum high-density lipoprotein cholesterol content was also increased,but the difference was not significant(P＞0.05).Conclusions Jianpi Huatan Fang can effectively regulate sex hormone secretion,improve ovarian reproductive function,and regulate glucose and lipid metabolism in obese PCOS-IR rats,possibly via inhibiting the FOXO1/PDK4 pathway.

Objective To explore the value of coronary fat attenuation index(FAI)combined with laboratory indicators in predicting the risk of acute coronary syndrome(ACS)in patients with coronary heart disease(CHD).Methods A retrospective analysis was conducted on 454 patients who were diagnosed with CHD in Guangdong Provincial People's Hospital SCAD group(n=233)and an ACS group(n=221).Univariate and multivariate Logistic regression analyses were performed on the FAI values of the main coronary branches[right coronary artery(RCA),left anterior descending branch(LAD),left circumflex branch(LCX)],laboratory indicators,and clinical data,to identify independent risk factors for ACS in CHD patients.Receiver operating characteristic curves were constructed,and area under the curve(AUC)was calculated to evaluate the predictive performance of the independent risk factors and their combinations.Results LAD-FAI,RCA-FAI,and high-sensitivity C-reactive protein(hs-CRP)were independent influencing factors for ACS in CHD patients.The AUC for the prediction of ACS occurrence in CHD patients based on LAD-FAI,RCA-FAI,and elevated hs-CRP values alone were 0.568,0.703,and 0.749,respectively.When these three factors were analyzed in combination,the AUC was 0.815.Conclusion The combined analysis of LAD-FAI,RCA-FAI,and hs-CRP has good predictive performance for assessing the risk of ACS in CHD patients.

Objective To investigate the difference of electrocardiographic(ECG)features between total left main artery(LM)occlusion and subtotal occlusion,and analyze risk factors of in-hospital mortality.Methods A total of 94 patients with left main complete occlusion and 99 patients with subtotal occlusion were included.ECG characteristics,coronary angiography and other clinical data were compared,and factors of hospital death were analyzed.The receiver operating characteristics(ROC)curve was used to analyze the predictive value of ECG characteristics in hospital death risk in patients with LM occlusion.The relationship between ECG characteristics,shock and collateral circulation were analyzed in patients with LM occlusion.Results Compared with the subtotal occlusion group,patients with LM occlusion presented with more ST-segment elevation(STE)in Ⅰ,avL,V2-V5,more STE in avR and avL,more left anterior fascicular block+right bundle branch block,prolonged QRS duration,less STE in avR and less STE in avR+V1.The in-hospital mortality was 46.8%(44/94)in LM occlusion and 14.1%(14/99)in LM subtotal occlusion.STE in avR+avL predicted total LM occlusion with a specificity of 0.97,and left anterior branch+right bundle branch block predicted total LM occlusion with a specificity of 1.00.In patients with total LM occlusion,STE in Ⅰ,avL,V2-V5,prolongation of QRS duration,shock,no collateral circulation,STE in Ⅰ,avL,V2-V5 combined with left anterior fascicular block+right bundle branch block,and STE in Ⅰ,avL,V2-V5 combined with shock predicted in-hospital mortality,with the area under the curve of 0.716,0.619,0.766,0.688,0.572,0.785,respectively.The diagnostic specificity of STE in Ⅰ,avL,V2-V5 combined with shock was 0.82,and the sensitivity was 0.75.STE in Ⅰ,avL,V2-V5 combined with left anterior fascicular block+right bundle branch block predicted in-hospital death in LM occlusion with a specificity of 0.94.The proportion of shock was higher in patients with STE in Ⅰ,avL,V2-V5,left anterior fascicular block+right bundle branch block and collateral flow absence(P＜0.05).In patients with total occlusion,no collateral flow was observed in patients with STE in Ⅰ,avL,V2-V5.In patients with STE in avR(including avR+V1),82.4%of patients presented with right coronary collateral circulation supplying the left anterior descending coronary artery and left circumflex artery territory.In patients with STE in avR+avL,69.2%of patients presented with right coronary collateral circulation supplying left anterior descending coronary artery territory.Conclusion Total LM occlusion presents with different ECG features compared with subtotal occlusion.In LM total occlusion,the ECG features predict in-hospital mortality and are associated with different collateral circulation.

Objective To explore the value of CT-derived fractional flow reserve(CT-FFR)and pericoronary fat attenuation index(FAI)combined with clinical and coronary CT angiography(CCTA)characteristics for predicting major adverse cardiovascular events(MACE)after aortic valve replacement(AVR).Methods Data of 139 patients with aortic stenosis who underwent AVR were retrospectively analyzed.According to occurrence of MACE or not during follow-up,the patients were divided into MACE group and non-MACE group.Cox proportional hazard regression was used to analyze clinical and CCTA data,as well as CT-FFR and FAI to screen independent predictors of MACE after AVR,and nested models based on clinical data,CCTA characteristics,CT-FFR and right coronary artery(RCA)FAI were constructed.Receiver operating characteristic(ROC)curves were drawn,the area under the curve(AUC)and Harrell C index(C-index)were calculated to assess the diagnostic efficacy of each model,and their goodness of fit were evaluated.Results There were 22 cases in MACE group and 117 in non-MACE group.CT-FFR(HR=3.683)and RCA-FAI(HR=3.261)were both independent predictors of MACE in patients after AVR.The AUC of clinical model,modelclinical+CCTA,modelclinical+CCTA+CT-FFR and modelclinical+CCTA+CT-FFR+RCA-FAI was 0.636,0.730,0.758 and 0.817,and the C-index was 0.614,0.707,0.733 and 0.782,respectively.The predicted results of modelclinical+CCTA+CT-FFR+RCA-FAI were most consistent with actual results,with the best goodness of fit.Conclusion CT-FFR and RCA-FAI combined with clinical and CCTA characteristics could effectively predict MACE in patients after AVR.

Objective To observe the effects of Daizong Prescription on glycogen metabolism in adipose tissue of obese mice;To explore its regulatory mechanism in activating browning in the white adipose tissue.Methods A obesity model was established by feeding high-fat diet to C57BL/6J mice.The obese mice were divided into model group,metformin group(0.15 g/kg),and Daizong Prescription low-(0.20 g/kg)and high-dosage(0.40 g/kg)groups.Mice fed a standard diet were set as the normal group,with 12 mice in each group.Each medication group was given corresponding drugs by gavage for 6 consecutive weeks.Body mass and fasting blood glucose were monitored,serum triglycerides(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C),and low-density lipoprotein cholesterol(LDL-C)contents were measured.Brown adipose tissue from the interscapular region and white adipose tissue from the inguinal,perirenal and epididymal region were collected,the adipose tissue mass was measured,and the body fat coefficient was calculated.HE staining was performed to observe morphological changes in adipose tissue,PAS staining was used to observe glycogen distribution in adipose tissue,immunohistochemistry staining was performed to detect the expressions of Gys2,Ppp1r3c,and GSK-3β in inguinal white adipose tissue.Results Compared with the normal group,the body mass and fasting blood glucose in different time points of the model group significant increase(P<0.05,P<0.01),and serum TC and HDL-C contents significantly increased(P<0.01);the mass and body fat coefficient of white adipose tissue in inguinal,perirenal,and epididymis significantly increased(P<0.01),the cells in white adipose tissue in inguinal were hypertrophic and appeared as large vacuoles,with less glycogen accumulation,the expressions of Gys2 and Ppp1r3c significantly decreased(P<0.01).Compared with the model group,the mice in Daizong Prescription high-dosage group showed a significant decrease in body mass and fasting blood glucose at 4 and 6 weeks of administration(P<0.05,P<0.01),and the contents of serum TG,TC,HDL-C,and LDL-C were significantly decreased(P<0.01);the mass and body fat coefficient in white adipose tissue of perirenal and epididymal significantly decreased(P<0.05,P<0.01),and the mass of inguinal white adipose tissue significantly decreased(P<0.05),multiple irregularly shaped small vacuoles could be seen in inguinal white adipose tissue,accompanied by nuclear aggregation and increased glycogen accumulation,the expressions of Gys2 and Ppp1r3c significantly increased(P<0.01).There was no significant difference in the expression of GSK-3β inguinal white adipose tissue of mice among the groups.Conclusion Daizong Prescription can increase the activity of Gys2 by upregulating the expression of Ppp1r3c,promote glycogen synthesis,induce browning of adipose tissue,increase fat heat production,and improve obesity and related disorders of glycolipid metabolism.

ObjectiveTo investigate the possible mechanism of Huatan Sanjie Formula （化痰散结方， HSF） in treating Graves' disease （GD） from the perspective of thyroid angiogenesis. MethodsThirty-six BALB/c female mice were randomly divided into a normal control group （n=9） and a modeling group （n=27）. Mice in the modeling group were injected with 2.0×10⁹ PFU/ml of Ad-TSHR289 adenovirus into the tibialis anterior muscle to build GD model. Nine weeks after immunization， the successfully modeled mice were randomly divided into model group， methimazole （MMI） group and HSF group， with 9 mice in each group. The MMI group was given 5.2 mg/（kg·d） of methimazole tablets by gavage， while the HSF group was given HSF at a relative crude drug dosage of 7.02 g/（kg·d） by gavage. The normal control group and the model group were given 0.1 ml/10 g of pure water by gavage. All groups were administered intragastrically once a day for a total of 4 weeks. The levels of thyroxine （T4） and thyrotropin receptor autoantibodies （TRAb） in serum were detected by radioimmunoassay， while the pathological changes of the thyroid gland were assessed by HE staining. The vascular morphology of thyroid tissue was observed by CD34 immunohistochemical staining， and the microvessel density （MVD） was counted. The protein expression of vascular endothelial growth factor A （VEGFA） and vascular endothelial growth factor receptor 2 （VEGFR2） in thyroid was detected by Western-blot. ResultsCompared to those in the normal control group， the thyroid volume of the mice in the model group significantly increased with excessive congestion， and the pathology showed significant thyroid follicular hyperplasia， columnar and proliferated epithelial cells， and enlarged follicle size； serum T4 and TRAb significantly increased， as well as the count of thyroid MVD， and the protein expressions of thyroid VEGFA and VEGFR2 （P＜0.01）. Compared to those in the model group， the thyroid glands of the mice in the MMI group and the HSF group were significantly reduced， and the congestion was improved； pathology showed that thyroid follicular hyperplasia and epithelial cell proliferation were reduced， with smooth edges of the follicles and the significantly reduced inward protrusion； serum T4 and TRAb significantly decreased， as well as the thyroid MVD， thyroid VEGFA and VEGFR2 protein expressions （P＜0.05 or P＜0.01）. There was no significant difference in all indicators between the MMI group and the HSF group （P＞0.05）. ConclusionHSF may inhibit thyroid angiogenesis by down-regulating thyroid VEGFA/VEGFR2 signaling pathway， thereby improving goitre and hyperfunction in GD mice.

Objective:To investigate the protective effect of quercetin against LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, providing valuable insights into the use of quercetin as a virulence inhibitor for Pseudomonas aeruginosa infection treatment. Methods:This was an experimental study. The experimental strain was the standard strain. The LasB protein was obtained utilizing protein recombination technology, while the enzyme activity of LasB was assessed through both the Elastin Congo red assay and fluorescently labelled elastin assay. The LasB-induced THP-1 macrophage infection model was established, and quercetin was utilized for intervention. Cell viability was evaluated via CCK-8 assay, while cell morphology was observed under an inverted microscope. Apoptosis detection involved employing both TUNEL and Annexin V/PI staining. The mRNA expression and protein levels of inflammatory cytokines and COX-2 were determined by RT-qPCR and ELISA respectively. Intracellular ROS levels were quantified using the DCFH-DA fluorescent probe. One-way ANOVA was used for statistical analysis, and Tukey test was used for multiple comparisons. Results:The pLasB with a molecular weight of 33 000 and acceptable enzymatic activity (purity>90%), was successfully obtained. THP-1 macrophages treated with pLasB at a concentration of 100 μg/ml presented significantly decreased viability and integrity rate when compared with the normal control group. Additionally, pLasB promoted apoptosis, up-regulated the levels of inflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12, and TNF-α, increased intracellular ROS fluorescence intensity, and elevated COX-2 mRNA expression level. Furthermore, the viability of THP-1 macrophages was significantly enhanced under quercetin intervention at concentrations of 2.5 μmol/L, 5 μmol/L and 10 μmol/L. The apoptosis rate exhibited a significant reduction from 18.32%±0.17% to 13.17%±0.20%, 11.43%±0.06% and 7.74%±0.04%, respectively ( F=1 679, P<0.05). There was a notable down-regulation of pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-12 and TNF-α while the anti-inflammatory cytokine IL-10 showed a significant up-regulation. Both intracellular ROS fluorescence intensity ( F=86.92, P<0.05) and COX-2 level ( F=24.62, P<0.05) demonstrated a substantial decrease. Conclusion:Quercetin demonstrates significant efficacy in inhibiting LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, which highlights immense potential as a potent virulence inhibitor of Pseudomonas aeruginosa.

Objective:To investigate the protective effect of quercetin against LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, providing valuable insights into the use of quercetin as a virulence inhibitor for Pseudomonas aeruginosa infection treatment. Methods:This was an experimental study. The experimental strain was the standard strain. The LasB protein was obtained utilizing protein recombination technology, while the enzyme activity of LasB was assessed through both the Elastin Congo red assay and fluorescently labelled elastin assay. The LasB-induced THP-1 macrophage infection model was established, and quercetin was utilized for intervention. Cell viability was evaluated via CCK-8 assay, while cell morphology was observed under an inverted microscope. Apoptosis detection involved employing both TUNEL and Annexin V/PI staining. The mRNA expression and protein levels of inflammatory cytokines and COX-2 were determined by RT-qPCR and ELISA respectively. Intracellular ROS levels were quantified using the DCFH-DA fluorescent probe. One-way ANOVA was used for statistical analysis, and Tukey test was used for multiple comparisons. Results:The pLasB with a molecular weight of 33 000 and acceptable enzymatic activity (purity>90%), was successfully obtained. THP-1 macrophages treated with pLasB at a concentration of 100 μg/ml presented significantly decreased viability and integrity rate when compared with the normal control group. Additionally, pLasB promoted apoptosis, up-regulated the levels of inflammatory cytokines IL-1α, IL-1β, IL-6, IL-10, IL-12, and TNF-α, increased intracellular ROS fluorescence intensity, and elevated COX-2 mRNA expression level. Furthermore, the viability of THP-1 macrophages was significantly enhanced under quercetin intervention at concentrations of 2.5 μmol/L, 5 μmol/L and 10 μmol/L. The apoptosis rate exhibited a significant reduction from 18.32%±0.17% to 13.17%±0.20%, 11.43%±0.06% and 7.74%±0.04%, respectively ( F=1 679, P<0.05). There was a notable down-regulation of pro-inflammatory cytokines IL-1α, IL-1β, IL-6, IL-12 and TNF-α while the anti-inflammatory cytokine IL-10 showed a significant up-regulation. Both intracellular ROS fluorescence intensity ( F=86.92, P<0.05) and COX-2 level ( F=24.62, P<0.05) demonstrated a substantial decrease. Conclusion:Quercetin demonstrates significant efficacy in inhibiting LasB-induced apoptosis, inflammation, and oxidative stress in THP-1 macrophages, which highlights immense potential as a potent virulence inhibitor of Pseudomonas aeruginosa.

Objective:To investigate the effects of Jianpi Huatan Prescription on a rat model of polycystic ovary syndrome (PCOS) with insulin resistance (IR) induced by letrozole combined with high fat diet based on the liver kinase B1 (LKB1)/AMP activated protein kinase (AMPK) signaling pathway; To discuss its mechanism.Methods:A total of 40 SD female rats were divided into model group (30 rats) and blank group (10 rats). The model group rats were fed with high-fat feed from the first week of modeling for 8 consecutive weeks, and from the fourth week onwards, were given a daily gavage of letrozole solution for 5 consecutive weeks; the blank group was fed normally, and from the 4th week onwards, an equal amount of sodium carboxymethyl cellulose solution was administered. After the completion of modeling, the modeling group was divided into Jianpi Huatan Prescription group, Metformin group, and model group according to random number table method. The Jianpi Huatan Prescription group and the metformin group were given corresponding drug interventions, while the other two groups were simultaneously administered an equal amount of physiological saline for a total of 4 weeks. Ovarian morphological changes were observed using HE staining; intraperitoneal glucose tolerance test (IPGTT) was used to detect glucose tolerance in rats; Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estrogen (E 2), total testosterone (T), fasting blood glucose (FBG), and fasting insulin (FINS) in rats; Real time fluorescence quantitative PCR and Western blot techniques were used to detect changes in LKB1, AMPK mRNA and protein expression in the LKB1/AMPK signaling pathway of rat ovarian tissue. Results:Compared with the model group, the polycystic changes in the ovaries of rats in the Jianpi Huatan Prescription group were improved, and the growth rate of body weight decreased. The levels of FBG, FINS, HOMA-IR, and IPGTT decreased ( P<0.05 or P<0.01), while serum FSH and HDL-C levels were significantly up-regulated ( P<0.05 or P<0.01). The levels of T, LH, LH/FSH, TC, TG, and LDL-C decreased ( P<0.05 or P<0.01); The mRNA and protein expression levels of LKB1 and AMPK in ovarian tissue increased ( P<0.05). Conclusion:Jianpi Huatan Prescription can improve PCOS-IR in rats caused by the combination of letrozole and high-fat diet, regulate various levels of sex hormones, and improve glucose and lipid metabolism disorders by regulating the LKB1/AMPK signaling pathway, which has a repairing effect on polycystic ovary disease in rats.

Bone defects caused by different causes such as trauma, severe bone infection and other factors are common in clinic and difficult to treat. Usually, bone substitutes are required for repair. Current bone grafting materials used clinically include autologous bones, allogeneic bones, xenografts, and synthetic materials, etc. Other than autologous bones, the major hurdles of rest bone grafts have various degrees of poor biological activity and lack of active ingredients to provide osteogenic impetus. Bone marrow contains various components such as stem cells and bioactive factors, which are contributive to osteogenesis. In response, the technique of bone marrow enrichment, based on the efficient utilization of components within bone marrow, has been risen, aiming to extract osteogenic cells and factors from bone marrow of patients and incorporate them into 3D scaffolds for fabricating bone grafts with high osteoinductivity. However, the scientific guidance and application specification are lacked with regard to the clinical scope, approach, safety and effectiveness. In this context, under the organization of Chinese Orthopedic Association, the Expert consensus for the clinical application of autologous bone marrow enrichment technique for bone repair ( version 2023) is formulated based on the evidence-based medicine. The consensus covers the topics of the characteristics, range of application, safety and application notes of the technique of autologous bone marrow enrichment and proposes corresponding recommendations, hoping to provide better guidance for clinical practice of the technique.