AIM:To investigate the therapeutic ef-fect of crocetin on diabetic lower limb ischemia and explore its underlying mechanism.METHODS:Male C57BL/6J mice aged 8 weeks were used to es-tablish a diabetic mouse model through intraperito-neal injection of streptozotocin(STZ).Following successful induction of diabetes,femoral artery li-gation(FAL)surgery was performed to create a model of diabetic lower limb ischemia.Fourteen days after STZ injection,the mice were treated with crocetin(3 mg/kg and 6 mg/kg)by gavage for 21 consecutive days.Post-FAL surgery,Doppler flowmetry was employed to assess blood flow in the mice of each group.Immunofluorescence stain-ing techniques were utilized to observe the expres-sion levels of platelet-endothelial cell adhesion molecule(CD31),α-smooth muscle actin(α-SMA),and inflammatory-related factors including tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),in-terleukin-6(IL-6),and transforming growth factor-β(TGF-β)in the gastrocnemius muscle of diabetic mice with lower limb ischemia.RESULTS:Com-pared to the normal group,the model group exhib-ited significantly elevated blood glucose levels and significantly reduced lower limb blood flow(P＜0.05).While CD31 and α-SMA expression showed no significant change,the expression levels of TNF-α,IL-1β,IL-6,and TGF-β were significantly in-creased(P＜0.05,P＜0.01,P＜0.05,P＜0.05).Com-pared to the model group,crocetin-treated groups showed no significant change in blood glucose lev-els but demonstrated significantly increased lower limb blood flow(P＜0.05).The high-dose crocetin group(6 mg/kg)significantly enhanced CD31 andα-SMA expression(P＜0.0001,P＜0.05)and signifi-cantly reduced the expression levels of IL-1β,IL-6,and TGF-β(P＜0.001,P＜0.05,P＜0.05).CONCLU-SION:Crocetin may exert its beneficial effects on diabetic lower limb ischemia through anti-inflam-matory and angiogenic mechanisms.

Objective:To explore the prospective association between physical activity level and mor-tality risk in Chinese adults with chronic obstructive pulmonary disease(COPD).Methods:Based on the China Kadoorie Biobank(CKB)who had COPD at the baseline survey,this study employed the Cox proportional hazards regression model to estimate the prospective associations between the overall physical activity,different intensities(low-level,moderate-to-vigorous-level),and types(occupational,non-occupational)of physical activity level and the risks of all-cause and cause-specific mortality,such as vascular diseases,cancer,and respiratory diseases.Based on the quintiles of physical activity level,par-ticipants were divided into five groups(Q1-Q5),with the lowest quintile group(Q1)as the reference group.Hazard ratio(HR)and 95％confidence interval(95％CI)were calculated for the remaining.In our study,we also performed sensitivity and subgroup analyses,including age,gender,self-rated health status,severity of COPD,etc.Results:Among 33 588 COPD patients at the baseline survey,8 314(22.2％)deaths were documented during an average follow-up of(11.1±3.1)years.Negative linear associations between the overall physical activity level and mortality risk from all-cause,vascular,and respiratory diseases were observed(P trend for linear correlation being＜0.001,0.002,＜0.001).Compared with the lowest quintile group of total physical activity(Q1),the hazard ratios(HR)and 95％confidence intervals(CI)for all-cause mortality,vascular disease mortality,and respiratory disease mortality in the highest quintile group(Q5)were 0.77(0.70,0.85),0.77(0.65,0.91),and 0.58(0.48,0.71),respectively.The low-level and moderate-to-vigorous-level physical activity were nega-tively associated with all-cause mortality in the COPD patients(P trend for linear correlation:0.002,＜0.001,respectively).Compared with the lowest quintile group of low-intensity and moderate-to-vigorous intensity physical activity(Q1),the HRs(95％CI)for all-cause mortality in the highest quintile group(Q5)were 0.89(0.82,0.97)and 0.79(0.72,0.87),respectively.The occupational and non-occupational physical activity were also found to have a linear inverse association with all-cause mortality risk among the COPD patients(P trend＜0.001 and 0.015,respectively).Compared with the lowest quintile group of occupational and non-occupational physical activity(Q1),the HR(95％CI)for all-cause mortality in the highest quintile group(Q5)were 0.69(0.61,0.78)and 0.91(0.84,0.98),respectively.The associations between overall physical activity and all-cause mortality risk were stronger for patients aged 60 and above,female,and who reported poor health status(P for interaction:0.028,0.012,0.010).The protective effect of total physical activity was also applicable to the COPD patients of varying severity.Conclusion:Physical activity could reduce the mortality risk in a dose-response relationship among COPD patients,regardless of its intensity and type,especially among indi-viduals aged 60 and above,females,and those with poor self-report health status.

Objective To establish the therapeutic effect prediction model of platelet transfusion in hematological patients,and receiver operating characteristic(ROC)curve and clinical cases are used to evaluate the clinical application value of the predic-tion model.Methods A total of 485 patients with hematological diseases who received platelet transfusion therapy in Kailuan General Hospital from January 2020 to December 2023 were selected,corrected count increment(CCI)was used to divide the patients into platelet transfusion effective group(n=340)and transfusion ineffective group(n=145).Multivariate Logistic regres-sion analysis was used to establish the prediction model of platelet infusion efficacy,and ROC curve was used to evaluate the application effect of the forcasting model.109 clinical cases were used to verify the practical application effect of the model,and the sensitivity,specificity and accuracy were calculated.Results Among 485 patients with hematological diseases,the incidence of ineffective platelet transfusion was 29.90%(145/485).Compated with the effective group,the ineffective group had more previous platelet transfusions was higher,and the difference was statistically significant(t=-4.435,P<0.05).In the ineffective group,there were more cases of hyperplenism,aplastic anemia and lymphoma,higher infection rate and higher positive rate of platelet antibody,and the differences were statistically significant(χ2=6.301～37.522,all P<0.05).Multivariate Logistic regres-sion analysis found that previous platelet infusion times,infection,leukemia,aplastic anemia and platelet antibodies were risk factors for ineffective platelet transfusion in patients with hematological diseases(Wald χ2=5.224～21.548,all P<0.05).Based on these risk factors,platelet infusion effect prediction models 1 and 2 were constructed.ROC curve was used to evaluate the application effect of the prediction model.The area under the curve(AUC),cut-offpoint,sensitivity and specificity of model 1 were 0.884,0.042,82.35%,88.89%.The AUC,cut-offpoint,corresponding sensitivity and specificity of prediction model 2 were 0.910,59.784,81.18%,94.44%,respectively.The Z values of model 1 and model 2 were 12.159 and 13.151,respectively.The prediction effect of model 2 was better than that of model 1.The actual application results showed that the sensitivity,specificity and accuracy of prediction model 1,2 were 85.71%,92.05%,90.89%and 90.48%,93.18%,92.66%,respectively.Conclusion The ineffective rate of platelet transfusion in hematological patients is relatively high.The prediction models 1 and 2 for platelet transfusion effectiveness have good results in predicting ineffective platelet transfusion,and prediction model 2 is better than pre-diction model 1,which can provide reliable basis for hematological patients on accurate platelet transfusion.

This study was aimed at establishing a method of ultra-fast quantitative PCR for Brucella detection.We used an exogenous recombinant plasmid as the internal reference and targeted the T4SS secretion system,an important Brucella viru-lence factor,to design specific primers and probes.The sensitivity,specificity,and repeatability of this method were evaluated,and a standard curve was constructed.The coincidence rate of detection findings with this method versus quantitative PCR was determined.This method markedly decreased the detection time to only 10 minutes.The standard curve demonstrated a good linear relationship(Y=-3.410 7x+38.357,R2=0.998 5)with a low minimum detection limit of 10 copies/μL.The method exhibited good specificity and did not specifically amplify several common clinical bacteria other than Brucella.The de-tection of three concentrations of positive plasmids yielded coefficients of variation(CVs)of 0.20％to 0.91％,thus demonstra-ting the method's excellent repeatability.Furthermore,140 clinical samples were analyzed concurrently with the fluorescence PCR method,which yielded a 100％compliance rate and consistent results.Our findings indicated that the Brucella ultra-fast quantitative PCR was ultrafast;had high sensitivity,high specificity,and good specificity;and can be used for the clinical de-tection of Brucella and emergency investigation of epidemics.Therefore,this method is valuable for the early diagnosis of Bru-cella.

Objective To establish the therapeutic effect prediction model of platelet transfusion in hematological patients,and receiver operating characteristic(ROC)curve and clinical cases are used to evaluate the clinical application value of the predic-tion model.Methods A total of 485 patients with hematological diseases who received platelet transfusion therapy in Kailuan General Hospital from January 2020 to December 2023 were selected,corrected count increment(CCI)was used to divide the patients into platelet transfusion effective group(n=340)and transfusion ineffective group(n=145).Multivariate Logistic regres-sion analysis was used to establish the prediction model of platelet infusion efficacy,and ROC curve was used to evaluate the application effect of the forcasting model.109 clinical cases were used to verify the practical application effect of the model,and the sensitivity,specificity and accuracy were calculated.Results Among 485 patients with hematological diseases,the incidence of ineffective platelet transfusion was 29.90%(145/485).Compated with the effective group,the ineffective group had more previous platelet transfusions was higher,and the difference was statistically significant(t=-4.435,P<0.05).In the ineffective group,there were more cases of hyperplenism,aplastic anemia and lymphoma,higher infection rate and higher positive rate of platelet antibody,and the differences were statistically significant(χ2=6.301～37.522,all P<0.05).Multivariate Logistic regres-sion analysis found that previous platelet infusion times,infection,leukemia,aplastic anemia and platelet antibodies were risk factors for ineffective platelet transfusion in patients with hematological diseases(Wald χ2=5.224～21.548,all P<0.05).Based on these risk factors,platelet infusion effect prediction models 1 and 2 were constructed.ROC curve was used to evaluate the application effect of the prediction model.The area under the curve(AUC),cut-offpoint,sensitivity and specificity of model 1 were 0.884,0.042,82.35%,88.89%.The AUC,cut-offpoint,corresponding sensitivity and specificity of prediction model 2 were 0.910,59.784,81.18%,94.44%,respectively.The Z values of model 1 and model 2 were 12.159 and 13.151,respectively.The prediction effect of model 2 was better than that of model 1.The actual application results showed that the sensitivity,specificity and accuracy of prediction model 1,2 were 85.71%,92.05%,90.89%and 90.48%,93.18%,92.66%,respectively.Conclusion The ineffective rate of platelet transfusion in hematological patients is relatively high.The prediction models 1 and 2 for platelet transfusion effectiveness have good results in predicting ineffective platelet transfusion,and prediction model 2 is better than pre-diction model 1,which can provide reliable basis for hematological patients on accurate platelet transfusion.

Objective:To describe the prevalence of preserved ratio impaired spirometry (PRISm) in participants from the China Kadoorie Biobank (CKB) and explore the influencing factors.Methods:The CKB project conducted the baseline survey, the first and the second resurvey in 2004-2008, 2008, and 2013-2014, respectively. Based on the lung function tests, the participants were categorized into three groups: regular, PRISm, and airflow obstruction. The prevalence of PRISm was reported by gender, age, and region at the baseline survey. The secular trend in the prevalence of PRISm was described during the three surveys. Finally, we used the multiple logistic regression model to examine the factors related to PRISm in the baseline survey.Results:After standardization for gender, age, and region according to the sixth national census data in 2010, the overall prevalence of PRISm and airflow obstruction among the 434 760 participants at baseline was 24.8% and 6.1%, respectively. The prevalence of PRISm was higher in rural (25.4%) than that in urban areas (24.3%). Of the 10 study regions, Gansu had the highest prevalence of PRISm (56.0%), while Henan had the lowest (15.4%). After standardization for gender, age, and region according to the baseline population, the prevalence of PRISm decreased from 24.9% at baseline to 15.7% in the second resurvey, and the prevalence of airflow obstruction increased from 5.9% to 21.4%. Unmarried status, current smoking, using solid fuels for cooking, low body weight, being overweight, obesity, and central obesity were associated with an increased risk of PRISm. In contrast, higher education attainments, increased household income, and maintaining a specific degree of physical activity were associated with a reduced risk of PRISm.Conclusions:The prevalence of PRISm was high in adults aged 40 years and above in China, and it varied by sociodemographic and lifestyle factors.

Objective To investigate the effect and mechanism of curculigoside(CUR)on hippocampal neuron injury in epileptic rats.Methods Forty-eight rats were selected,and the epileptic rat models were established by induction with 35 mg/kg pentylenetetrazol.The epileptic rats were randomly divided into the epilepsy group,the epilepsy+low-dose CUR group(the epilepsy+CUR-L group),the epilepsy+high-dose CUR group(the epilepsy+CUR-H group),and the epilepsy+CUR-H+PI3K activator 740Y-P group(the epilepsy+CUR-H+740Y-P group),with 12 rats in each group.Another 12 rats that were intraperitoneally injected with the same amount of normal saline were taken as the control group.Four weeks after administration,the behavioral changes of rats in each group were observed.Nissl staining was applied to determine the neuronal changes in hippocampal tissue.Immunohistochemistry was applied to detect the number of microglia in hippocampal tissue.ELISA was applied to detect the levels of interleukin-6(IL-6),IL-1β,and gamma-aminobutyric acid(GABA)in hippocampal tissue.TUNEL staining was applied to analyze the level of neuronal apoptosis in hippocampal tissue.Western blot was applied to detect the phosphorylation levels of PI3K,Akt and mTOR in hippocampal tissue.Results Compared with the control group,the Racine score and the frequency of epileptic seizures in rats,and the number of microglia,level of IL-6,level of IL-1β,the apoptosis rate of neurons,and levels of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR in hippocampal tissue in the epilepsy group were obviously increased(P<0.05),while the level of GABA in hippocampal tissue was obviously reduced(P<0.05),and the arrangement of neurons in hippocampal tissue was disordered,with neuronal loss.Compared with the epilepsy group,the Racine score,and the frequency of epileptic seizures in rats,and the number of microglia,levels of IL-6,levels of IL-1β,the apoptosis rates of neurons,and levels of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR in hippocampal tissue in the epilepsy+CUR-L group and the epilepsy+CUR-H group were significantly decreased(P<0.05),while the levels of GABA in hippocampus tissue were significantly increased(P<0.05),the loss and necrosis of neurons in hippocampus tissue were decreased,and the disorder of cell arrangement was improved.Compared with the epilepsy+CUR-H group,the Racine score and the frequency of epileptic seizures in rats,and the number of microglia,level of IL-6,level of IL-1β,the apoptosis rate of neurons,and levels of p-PI3K/PI3K,p-Akt/Akt,p-mTOR/mTOR in hippocampal tissue in the epilepsy+CUR-H+740Y-P group were significantly increased(P<0.05),while the level of GABA in hippocampus tissue was significantly decreased(P<0.05),the loss and necrosis of hippocampal neurons increased,and the cell arrangement was disordered.Conclusion CUR may reduce hippocampal neuron damage in epileptic rats by downregulating the PI3K/Akt/mTOR signaling pathway.

Objective:To investigate the specific mechanisms underlying the protective effect of interleukin (IL) -27 in the pathogenesis of psoriasis.Methods:Five skin tissue samples from healthy individuals and 6 lesional skin samples from psoriasis patients were collected, and IL-27 expression was determined by immunohistochemical staining. Il27ra gene knockout (KO) mice were constructed. Psoriasis-like mouse models were established with topical imiquimod in 5 wild-type (WT) mice and 6 KO mice. Mouse skin lesions were evaluated using the modified Psoriasis Area and Severity Index (mPASI), and lesional skin tissues were collected for hematoxylin and eosin (HE) staining to observe changes in epidermal thickness. Single-cell suspensions were prepared with skin lesions and skin-draining lymph nodes of 4 WT mice and 3 KO mice, and changes in immune cells (including T cells, γδ T cells, and neutrophils) were analyzed using flow cytometry. Additionally, skin-draining lymph node cells were isolated from 9 normal WT mice, and IL-17A expression was stimulated using a T-cell receptor agonist (CD3/28 activating antibodies, αCD3/28) or cytokines (IL-23 + IL-1β), followed by the addition of IL-27; peripheral blood mononuclear cells (PBMCs) were isolated from 6 psoriasis patients, and IL-17A expression was stimulated using the T-cell receptor agonist, followed by the addition of IL-27; the effect of IL-27 on IL-17A expression in T cells was analyzed using flow cytometry and enzyme-linked immunosorbent assay (ELISA). Measurement data were compared between two groups using the t test. Results:Immunohistochemical staining revealed a significant reduction in IL-27 expression in psoriatic lesions (mean fluorescence intensity: 9.85 ± 3.07) compared with the normal skin (19.45 ± 2.51, t = 5.60, P < 0.001). Animal experiments demonstrated that the KO mice exhibited significantly aggravated psoriasis-like skin inflammation (mPASI: 4.00 ± 0.89) and significantly increased epidermal thickness (115.50 ± 7.69 μm) compared with the WT mice (mPASI: 2.80 ± 0.84, t = 2.28, P = 0.049; epidermal thickness: 92.26 ± 8.76 μm, t = 4.70, P = 0.001) ; compared with the WT mice, the KO mice showed significantly increased proportions of T cells (11.22% ± 2.76% vs. 7.08% ± 0.85%) and dermal γδ T cells (4.78% ± 0.39% vs. 2.78% ± 0.49%) among live cells in the lesions ( t = 2.91, 2.75, respectively, both P < 0.05), as well as significantly increased proportions of Th17, IL-17 + γδ T, Th22, and IL-22 + γδ T cells in the skin-draining lymph nodes (all P < 0.05), but no significant difference in the proportion of neutrophils in the lesions (WT: 13.57% ± 8.36%, KO: 14.43% ± 9.13%; t = 0.13, P = 0.902). Experiments with different stimuli showed that IL-27 significantly suppressed T-cell receptor agonist-induced IL-17A expression in murine γδ T cells (αCD3/28 group: 1.00 ± 0.11, αCD3/28 + IL-27 group: 0.76 ± 0.13; t = 3.54, P = 0.004), while there was no significant difference in IL-17A expression between cells induced by IL-23 + IL-1β with the IL-27 co-culture and those without ( t = 1.34, P > 0.05). ELISA showed that IL-27 significantly reduced the IL-17A concentration in the culture supernatant of draining lymph node cells stimulated by the T-cell receptor agonist (αCD3/28 group: 1 535.00 ± 97.76 pg/ml, αCD3/28 + IL-27 group: 1 030.00 ± 287.90 pg/ml, t = 3.29, P = 0.031), but did not reduce the IL-17A concentration induced by IL-23 + IL-1β ( t = 0.09, P > 0.05). Flow cytometry indicated that IL-27 significantly inhibited the T-cell receptor agonist-induced IL-17A expression in T cells from psoriasis patients (αCD3/28 group: 4.28 ± 3.25, αCD3/28 + IL-27 group: 3.04 ± 2.65, t = 4.46, P = 0.007) . Conclusion:IL-27 appeared to play a protective role in psoriasis by suppressing IL-17A secretion from T cells.

Background: s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort. Results: In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM). Conclusions Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.

Background: s/Aims: Non-invasive models stratifying clinically significant portal hypertension (CSPH) are limited. Herein, we developed a new non-invasive model for predicting CSPH in patients with compensated cirrhosis and investigated whether carvedilol can prevent hepatic decompensation in patients with high-risk CSPH stratified using the new model. Methods: Non-invasive risk factors of CSPH were identified via systematic review and meta-analysis of studies involving patients with hepatic venous pressure gradient (HVPG). A new non-invasive model was validated for various performance aspects in three cohorts, i.e., a multicenter HVPG cohort, a follow-up cohort, and a carvediloltreating cohort. Results: In the meta-analysis with six studies (n=819), liver stiffness measurement and platelet count were identified as independent risk factors for CSPH and were used to develop the new “CSPH risk” model. In the HVPG cohort (n=151), the new model accurately predicted CSPH with cutoff values of 0 and –0.68 for ruling in and out CSPH, respectively. In the follow-up cohort (n=1,102), the cumulative incidences of decompensation events significantly differed using the cutoff values of <–0.68 (low-risk), –0.68 to 0 (medium-risk), and >0 (high-risk). In the carvediloltreated cohort, patients with high-risk CSPH treated with carvedilol (n=81) had lower rates of decompensation events than non-selective beta-blockers untreated patients with high-risk CSPH (n=613 before propensity score matching [PSM], n=162 after PSM). Conclusions Treatment with carvedilol significantly reduces the risk of hepatic decompensation in patients with high-risk CSPH stratified by the new model.