Andrographolide sulfonate (AS) is a sulfonated derivative of andrographolide extracted from Andrographis paniculata (Burm.f.) Nees, and has been approved for several decades in China. The present study aimed to investigate the novel therapeutic application and possible mechanisms of AS in the treatment of rheumatoid arthritis. Results indicated that administration of AS by injection or gavage significantly reduced the paw swelling, improved body weights, and attenuated pathological changes in joints of rats with adjuvant-induced arthritis. Additionally, the levels of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and IL-1β in the serum and ankle joints were reduced. Bioinformatics analysis, along with the spleen index and measurements of IL-17 and IL-10 levels, suggested a potential relationship between AS and Th17 cells under arthritic conditions. In vitro, AS was shown to block Th17 cell differentiation, as evidenced by the reduced percentages of CD4⁺ IL-17A⁺ T cells and decreased expression levels of RORγt, IL-17A, IL-17F, IL-21, and IL-22, without affecting the cell viability and apoptosis. This effect was attributed to the limited glycolysis, as indicated by metabolomics analysis, reduced glucose uptake, and pH measurements. Further investigation revealed that AS might bind to hexokinase2 (HK2) to down-regulate the protein levels of HK2 but not glyceraldehyde-3-phosphate dehydrogenase (GAPDH) or pyruvate kinase M2 (PKM2), and overexpression of HK2 reversed the inhibition of AS on Th17 cell differentiation. Furthermore, AS impaired the activation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) signals in vivo and in vitro, which was abolished by the addition of lactate. In conclusion, AS significantly improved adjuvant-induced arthritis (AIA) in rats by inhibiting glycolysis-mediated activation of PI3K/AKT to restrain Th17 cell differentiation.
Animals ; Th17 Cells/immunology* ; Diterpenes/pharmacology* ; Arthritis, Rheumatoid/metabolism* ; Proto-Oncogene Proteins c-akt/immunology* ; Glycolysis/drug effects* ; Cell Differentiation/drug effects* ; Phosphatidylinositol 3-Kinases/genetics* ; Rats ; Male ; Rats, Sprague-Dawley ; Humans ; Andrographis paniculata/chemistry* ; Arthritis, Experimental/drug therapy* ; Interleukin-17/immunology* ; Signal Transduction/drug effects*

OBJECTIVE To explore the risk factors for polymyxin-and carbapenem-resistant Klebsiella pneumoni-ae(PR-CRKP)infection and establish the prediction model.METHODS The clinical data were retrospectively col-lected from the patients with CRKP infection who were treated in the First Affiliated Hospital of Xi'an Jiaotong University from Jan.2023 to Mar.2024.The enrolled patients were divided into the CRKP group and the PR-CRKP group according to the result of drug susceptibility testing for polymyxin B.Totally 203 patients who were treated from Jan.2023 to Dec.2023 were assigned as the modeling group,and 91 patients who were treated from Jan.2024 to Mar.2024 were assigned as the validation group.Multivariate logistic regression analysis was per-formed for the risk factors for PR-CRKP infection,nomogram was built up for prediction of PR-CRKP infection by R software,and the predictive efficacy of the model was evaluated by means of receiver operating characteristic(ROC)curves.RESULTS The result of univariate analysis showed that the proportions of patients who received fi-berobronchoscopy,endotracheal intubation/tracheotomy,were complicated with other carbapenem-resistant or-ganisms(CROs)infections,bloodstream infections,were treated in intensive care unit(ICU)and had the history of exposure to polymyxins and carbapenems 3 months before the admission were higher in the PR-CRKP group than in the CRKP group(P＜0.05);the length of hospital stay and duration of use of polymyxins and carbapene-ms were longer in the PR-CRKP group than in the CRKP group(P＜0.05).Multivariate logistic regression analy-sis indicated that complication with other CROs infections,history of exposure to polymyxins and carbapenems 3 months before the admission,duration of use of polymyxins and ICU stay were the risk factors for the PR-CRKP infection(P＜0.05).The area under the curve(AUC)of the predictive model was 0.898 in the modeling group,with the sensitivity 80.33％,the specificity 84.51％.ROC curve analysis showed that the AUC was 0.901 in the validation group,with the sensitivity 75.00％,the specificity 92.73％.CONCLUSION The prediction model that is established based on the result of multivariate analysis has high value in prediction of PR-CRKP infection.

Objective:To evaluate the effect of Bu-Zhong-Yi-Qi-Tang (BZYQT) combined with low molecular weight heparin (LMWH) on lower limb deep vein thrombosis (DVT) after fracture fixation in aged rabbits.Methods:Thirty-two elderly New Zealand rabbits of either sex, aged 36 months, weighing 3.5-4.0 kg, were divided into 4 groups ( n=8 each) using the random number table method: control group (C group), fracture fixation group (F group), low molecular weight heparin group (L group) and BZYQT combined with LMWH group (BL group). A model of fixation surgery for right femoral shaft fracture was established in anesthetized rabbits in F, L and BL groups. Starting from postoperative day 1, BL group received BZYQT at 3.6 ml/kg via gavage, F and L groups received normal saline at 3.6 ml/kg via gavage, L and BL groups received subcutaneous injection of LMWH at 200 IU/kg, and all the administrations were performed daily at 08: 00 for 7 consecutive days. On postoperative day 7, ultrasound examination was performed to assess lower limb DVT. Blood samples were collected from the marginal ear vein for determination of the serum concentrations of cortisol (COR), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and endothelin-1 (ET-1) by enzyme-linked immunosorbent assay. Femoral vein specimens were obtained under anesthesia for microscopic examination of the pathological changes of the femoral vein lumen and endothelial tissues (by HE staining) and of the morphology of vascular endothelial cells (by the transmission electron microscopy). Results:Compared with C group, the concentrations of serum COR, TNF-α, IL-6 and ET-1 were significantly increased in F, L and BL groups, and the incidence of lower limb DVT within 7 days after surgery was significantly increased in F and L groups ( P<0.05). Compared with F group, the incidence of lower limb DVT within 7 days after surgery was significantly decreased in L and BL groups, and the concentrations of serum COR, TNF-α, IL-6 and ET-1 were significantly decreased ( P<0.05), and the pathological damage was significantly reduced in BL group. Compared with L group, the incidence of lower limb DVT and concentration of IL-6 were significantly decreased ( P<0.05), and pathological damage was significantly alleviated in BL group. Conclusions:BZYQT combined with LMWH can prevent the occurrence of lower limb DVT after fracture fixation in aged rabbits.

Objective:To evaluate the effect of Bu-Zhong-Yi-Qi-Tang (BZYQT) combined with low molecular weight heparin (LMWH) on lower limb deep vein thrombosis (DVT) after fracture fixation in aged rabbits.Methods:Thirty-two elderly New Zealand rabbits of either sex, aged 36 months, weighing 3.5-4.0 kg, were divided into 4 groups ( n=8 each) using the random number table method: control group (C group), fracture fixation group (F group), low molecular weight heparin group (L group) and BZYQT combined with LMWH group (BL group). A model of fixation surgery for right femoral shaft fracture was established in anesthetized rabbits in F, L and BL groups. Starting from postoperative day 1, BL group received BZYQT at 3.6 ml/kg via gavage, F and L groups received normal saline at 3.6 ml/kg via gavage, L and BL groups received subcutaneous injection of LMWH at 200 IU/kg, and all the administrations were performed daily at 08: 00 for 7 consecutive days. On postoperative day 7, ultrasound examination was performed to assess lower limb DVT. Blood samples were collected from the marginal ear vein for determination of the serum concentrations of cortisol (COR), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and endothelin-1 (ET-1) by enzyme-linked immunosorbent assay. Femoral vein specimens were obtained under anesthesia for microscopic examination of the pathological changes of the femoral vein lumen and endothelial tissues (by HE staining) and of the morphology of vascular endothelial cells (by the transmission electron microscopy). Results:Compared with C group, the concentrations of serum COR, TNF-α, IL-6 and ET-1 were significantly increased in F, L and BL groups, and the incidence of lower limb DVT within 7 days after surgery was significantly increased in F and L groups ( P<0.05). Compared with F group, the incidence of lower limb DVT within 7 days after surgery was significantly decreased in L and BL groups, and the concentrations of serum COR, TNF-α, IL-6 and ET-1 were significantly decreased ( P<0.05), and the pathological damage was significantly reduced in BL group. Compared with L group, the incidence of lower limb DVT and concentration of IL-6 were significantly decreased ( P<0.05), and pathological damage was significantly alleviated in BL group. Conclusions:BZYQT combined with LMWH can prevent the occurrence of lower limb DVT after fracture fixation in aged rabbits.

OBJECTIVE To investigate the risk factors for carbapenem-resistant gram-negative bacteria(GNB)bloodstream infection(BSI)in patients with hematological malignancies(HMs)and their prognosis,and to devel-op a nomogram prediction model.METHODS A total of 316 patients with HMs and GNB-BSI admitted to the First Affiliated Hospital of Xi'an Jiaotong University from Jan.2017 to Dec.2022 were selected as the training set,and 106 patients admitted from Jan.2023 to Oct.2024 were selected as the validation set.Variables were selected by lasso regression and multifactor logistic regression,and a nomogram model was constructed.The prediction model was internally validated by the receiver operating characteristic(ROC)curve,calibration curve and decision curve analysis(DCA),respectively.RESULTS Granulocytopenia for ≥7 days(OR=14.525),use of cephalosporins/β-lactamase inhibitors within 30 days before BSI(OR=3.510),exposure history of carbapenem antibacterial drug(OR=4.840)and albumin＜30 g/L(OR=2.697)were risk factors for CR-GNB BSI in patients with HMs(P＜0.05).Septic shock(OR=6.934),central venous catheterization(OR=5.586),inappropriate empirical antibac-terial drug therapy(OR=4.744),CR-GNB infection(OR=2.916)and albumin＜30 g/L(OR=3.324)were risk factors for 30-day mortality in patients with HMs and GNB-BSI(P＜0.05).Based on these indicators,two nomogram models were constructed.The areas under the ROC curve(AUC)for the internal validation set were 0.775 and 0.849,respectively.The calibration curves demonstrated high predictive performance for the pre-diction models(P=0.998 and 0.660,respectively),and DCA showed high clinical application value for both models.CONCLUSION The nomogram prediction model constructed in this study based on multifactor analy-sis not only demonstrates good predictive value but also exhibits significant clinical efficacy,aiding in the early i-dentification of high-risk patients for targeted therapy.

OBJECTIVE To investigate the risk factors for carbapenem-resistant gram-negative bacteria(GNB)bloodstream infection(BSI)in patients with hematological malignancies(HMs)and their prognosis,and to devel-op a nomogram prediction model.METHODS A total of 316 patients with HMs and GNB-BSI admitted to the First Affiliated Hospital of Xi'an Jiaotong University from Jan.2017 to Dec.2022 were selected as the training set,and 106 patients admitted from Jan.2023 to Oct.2024 were selected as the validation set.Variables were selected by lasso regression and multifactor logistic regression,and a nomogram model was constructed.The prediction model was internally validated by the receiver operating characteristic(ROC)curve,calibration curve and decision curve analysis(DCA),respectively.RESULTS Granulocytopenia for ≥7 days(OR=14.525),use of cephalosporins/β-lactamase inhibitors within 30 days before BSI(OR=3.510),exposure history of carbapenem antibacterial drug(OR=4.840)and albumin＜30 g/L(OR=2.697)were risk factors for CR-GNB BSI in patients with HMs(P＜0.05).Septic shock(OR=6.934),central venous catheterization(OR=5.586),inappropriate empirical antibac-terial drug therapy(OR=4.744),CR-GNB infection(OR=2.916)and albumin＜30 g/L(OR=3.324)were risk factors for 30-day mortality in patients with HMs and GNB-BSI(P＜0.05).Based on these indicators,two nomogram models were constructed.The areas under the ROC curve(AUC)for the internal validation set were 0.775 and 0.849,respectively.The calibration curves demonstrated high predictive performance for the pre-diction models(P=0.998 and 0.660,respectively),and DCA showed high clinical application value for both models.CONCLUSION The nomogram prediction model constructed in this study based on multifactor analy-sis not only demonstrates good predictive value but also exhibits significant clinical efficacy,aiding in the early i-dentification of high-risk patients for targeted therapy.

OBJECTIVE To explore the risk factors for polymyxin-and carbapenem-resistant Klebsiella pneumoni-ae(PR-CRKP)infection and establish the prediction model.METHODS The clinical data were retrospectively col-lected from the patients with CRKP infection who were treated in the First Affiliated Hospital of Xi'an Jiaotong University from Jan.2023 to Mar.2024.The enrolled patients were divided into the CRKP group and the PR-CRKP group according to the result of drug susceptibility testing for polymyxin B.Totally 203 patients who were treated from Jan.2023 to Dec.2023 were assigned as the modeling group,and 91 patients who were treated from Jan.2024 to Mar.2024 were assigned as the validation group.Multivariate logistic regression analysis was per-formed for the risk factors for PR-CRKP infection,nomogram was built up for prediction of PR-CRKP infection by R software,and the predictive efficacy of the model was evaluated by means of receiver operating characteristic(ROC)curves.RESULTS The result of univariate analysis showed that the proportions of patients who received fi-berobronchoscopy,endotracheal intubation/tracheotomy,were complicated with other carbapenem-resistant or-ganisms(CROs)infections,bloodstream infections,were treated in intensive care unit(ICU)and had the history of exposure to polymyxins and carbapenems 3 months before the admission were higher in the PR-CRKP group than in the CRKP group(P＜0.05);the length of hospital stay and duration of use of polymyxins and carbapene-ms were longer in the PR-CRKP group than in the CRKP group(P＜0.05).Multivariate logistic regression analy-sis indicated that complication with other CROs infections,history of exposure to polymyxins and carbapenems 3 months before the admission,duration of use of polymyxins and ICU stay were the risk factors for the PR-CRKP infection(P＜0.05).The area under the curve(AUC)of the predictive model was 0.898 in the modeling group,with the sensitivity 80.33％,the specificity 84.51％.ROC curve analysis showed that the AUC was 0.901 in the validation group,with the sensitivity 75.00％,the specificity 92.73％.CONCLUSION The prediction model that is established based on the result of multivariate analysis has high value in prediction of PR-CRKP infection.

Objective To explore the effect of a formative assessment of mini clinical evaluation exercise(Mini-CEX)and direct ob-servation of procedure skills(DOPS)on clinical internship teaching in periodontology.Methods Fifty-six undergraduates,who were interning in the Department of Periodontics,were selected and randomly divided into the control group and the teaching reform group.The traditional summative evaluation teaching mode was adopted in the control group,while the formative evaluation teaching methods of Mini-CEX and DOPS were used in the teaching reform group.The application effect was evaluated through the examination scores of two groups of students before and after the clinical practice,the Mini-CEX and DOPS scores of students in the teaching reform group before and after clinical practice,and a questionnaire survey.Results There was no significant difference in the scores of pre-clinical practice examination between the two groups(P＞0.05).However,after the clinical practice,the scores of case report and clinical skills of the students in the teaching reform group were significantly higher than those in the control group(P＜0.05).In addition,after the clinical practice,the scores of each item in the Mini CEX and DOPS of teaching reform group were significantly improved compared with those before clinical practice(P＜0.05).The students and teachers were both satisfied with the formative assessment and its effec-tiveness.Conclusion The formative evaluation system of Mini-CEX and DOPS can effectively improve the quality of clinical internship teaching in periodontology,which has been widely recognized by students and teachers and is worthy of promoting and application.

Objective To observe the effect of Shenfu injection on lung injury caused by hemor-rhagic shock(HS)in rats and explore the related potential mechanism.Methods Thirty-six SPF healthy male SD rats,aged 16-17 weeks,weighing 400-600 g,were randomly divided into three groups:sham op-eration group(group SH),HS group(group HS),and Shenfu injection group(group SF),12 rats in each group.In group SH,only the right femoral vein and femoral artery were separated after anesthesia,and ve-nous catheterization was not performed.HS model was established in groups SF and HS.In group HS,liquid resuscitation was performed through an intravenous catheter,and the resuscitation fluid consisted of the auto-blood lost and the compound sodium chloride injection equivalent to 1.5 times the blood loss and 10 ml/kg normal saline.In group SF,the resuscitation fluid consisted of the lost autoblood and the compound sodium chloride injection equivalent to 1.5 times the blood loss and Shenfu injection 10 ml/kg.The whole perfusion time was about 60 minutes.Six rats in the three groups were randomly anesthetized 24 and 48 hours after op-eration.The wet/dry weight ratio(W/D)of lung tissues was detected.The concentrations of interleukin-6(IL-6),IL-17,IL-10,and transforming growth factor-β(TGF-β)were detected by ELISA,the mRNA ex-pression of retinoic acid-related orphan nuclear receptor γt(RORγt),transcription factor forkhead box pro-tein 3(Foxp3),and hypoxia-inducible factor-1α(HIF-1α)in lung tissues were detected by PCR.The pro-tein contents of RORγt,Foxp3,HIF-1α,aquaporin 1(AQP1),and AQP5 in lung tissue were detected by Western blot.Pathological changesunder HE staining light microscope and lung injury scores were observed.Results Compared with 24 hours after operation,W/D,the concentrations of IL-6 and IL-17,mRNA ex-pression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concentrations of IL-10,and TGF-β,Foxp3 mRNA expression and protein content,and AQP1 protein content were significantly increased in group SF 48 hours after operation(P＜0.05).Compared with group SH,W/D,the concentrations of IL-6,IL-17,IL-10,and TGF-β,mRNA expression and protein content of RORγt,Foxp 3,and HIF-1α,and lung injury score were significantly increased(P＜0.05),AQP1 and AQP5 protein contents were significantly decreased in groups HS and SF 24 and 48 hours after operation(P＜0.05),and alveolar structure was damaged under light microscope and alveolar interstitium was filled with a large amount of edematous fluid,during which a large number of inflammatory cells infiltra-ted.Compared with group HS,W/D,the concentrations of IL-6 and IL-17,mRNA expression and protein content of RORγt and HIF-1α,and lung injury score were significantly decreased(P＜0.05),the concen-trations of IL-10 and TGF-β,Foxp3 mRNA expression and protein content,AQP1 and AQP5 protein con-tents were significantly increased in group SF 24 and 48 hours after surgery(P＜0.05),and the alveolar structure was improved under light microscope,and edema was reduced,and the number of inflammatory cells was reduced.Conclusion Shenfu injection can regulate the balance between pro-inflammatory factors IL-6 and IL-17,and anti-inflammatory factors IL-10 and TGF-β,increase the protein content of AQP1 and AQP5 in lung tissue,and decrease the W/D and injury score in lung tissue,thus alleviating lung injury in HS rats.The mechanism may be related to the regulation of HIF-1α-RORγt/Foxp3 balance.

ObjectiveTo establish a quantitative analysis multi-components by single marker method （QAMS） for five main components （aucubin， geniposidic acid， chlorogenic acid， asperuloside and rutin） in Eucommiae Folium， to verify its feasibility and applicability in the determination of Eucommiae Folium， so as to provide a scientific basis for the development of quality standard of this herb. MethodHigh performance liquid chromatography was performed on a Welch Boltmatetm™ C₁₈ column （4.6 mm×100 mm， 2.7 μm） with methanol （A）-0.2% phosphoric acid aqueous solution （B） as the mobile phase for gradient elution （0-8 min， 3%A； 8-10 min， 3%-11%A； 10-26 min， 11%A； 26-27 min， 11%-25%A； 27-60 min， 25%-32%A）， the column temperature was set at 30 ℃， the flow rate was 0.6 mL·min^-1， the detection wavelengths were at 210 nm and 254 nm. Chlorogenic acid was used as an internal reference to establish the relative correction factors （f） between it and the other four components， and the contents of the five components in 14 batches of Eucommiae Folium were determined by QAMS and external standard method （ESM）， respectively. ResultThe f values of chlorogenic acid to aucubin， geniposidic acid， asperuloside and rutin were 3.13， 1.45， 2.64 and 0.56， respectively. Repeatability was good under different experimental conditions， relative standard deviation （RSD） was <5.0%. The contents of aucubin， geniposidic acid， chlorogenic acid， asperuloside and rutin in 14 batches of Eucommiae Folium were 1.340-28.975， 0.252-36.086， 10.016-27.443， 1.396-8.646， 0.533-1.766 mg·g^-1， respectively. There were no significant difference between content results of QAMS and that of ESM （RSD<5.0%）. ConclusionQAMS established with chlorogenic acid as the internal reference can be used to determine the contents of five components in Eucommiae Folium， and this method is simple and accurate. After comprehensive evaluation， the quality standard of Eucommiae Folium in subsequent editions of Chinese Pharmacopoeia is suggested that three main active components， chlorogenic acid， aucubin and geniposidic acid， are selected as quality markers， and their content limits are recommended not less than 1.5%， 1.0% and 1.0%， respectively. This quality standard draft can avoid the potential quality risk due to poor specificity and low content limit of the index component （chlorogenic acid） in the previous editions of Chinese Pharmacopoeia.