OBJECTIVES: To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Buyang Huanwu Decoction (BYHWT) on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients (FLS-RA) cultured under a hypoxic condition. METHODS: Forty normal Wistar rats were randomized into two groups (n=20) for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera. FLS-RA were cultured in routine condition or exposed to hypoxia (10% O₂) for 24 h wigh subsequent treatment with IL-1β, followed by treatment with diluted BYHWT-medicated serum (5%, 10% and 20%) or control serum. AnnexinV-APC/7-AAD double staining and T-AOC kit were used for detecting apoptosis and total antioxidant capacity of the cells, and the changes in ROS, ATP level, mitochondrial membrane potential and Ca²⁺ homeostasis were analyzed. The changes in mRNA and protein expressions of BNIP3, PI3K and AKT and mRNA expressions of LC3, Beclin-1 and P62 were detected using RT-qPCR and Western blotting. RESULTS: Treatment with BYHWT-medicated serum dose-dependently lowered apoptosis rate of IL-1β-induced FLS-RA with hypoxic exposure. The treatment significantly decreased T-AOC concentration, increased ROS production, autophagosome formation and ATPase levels, and lowered mitochondrial membrane potential and Ca²⁺ level in the cells. In IL-1β-induced FLS-RA with hypoxic exposure, treatment with BYHWT-medicated serum significantly increased BNIP3 protein expression, decreased the protein expressions of PI3K and AKT, increased the mRNA expressions of BNIP3 and P62, and lowered the mRNA expressions of PI3K, AKT, LC3 and Beclin-1 without significantly affecting Beclin-1 protein expression. The cells treated with 5% and 10% BYHWT-medicated serum showed no significant changes in LC3 expression. CONCLUSIONS BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.
Drugs, Chinese Herbal/pharmacology* ; Arthritis, Rheumatoid/pathology* ; Animals ; Signal Transduction/drug effects* ; Proto-Oncogene Proteins c-akt/metabolism* ; Autophagy/drug effects* ; Humans ; Fibroblasts/cytology* ; Rats, Wistar ; Membrane Proteins/metabolism* ; Rats ; Phosphatidylinositol 3-Kinases/metabolism* ; Mitochondria/metabolism* ; Cells, Cultured ; Proto-Oncogene Proteins/metabolism* ; Apoptosis/drug effects* ; Cell Hypoxia ; Synovial Membrane/cytology* ; Male ; Mitochondrial Proteins

Objective To investigate the role of the BNIP3-PI3K/Akt signaling pathway in mediating the inhibitory effect of Buyang Huanwu Decoction(BYHWT)on mitochondrial autophagy in human synovial fibroblasts from rheumatoid arthritis patients(FLS-RA)cultured under a hypoxic condition.Methods Forty normal Wistar rats were randomized into two groups(n=20)for daily gavage of BYHWT or distilled water for 7 days to prepare BYHWT-medicated or control sera.FLS-RA were cultured in routine condition or exposed to hypoxia(10％O2)for 24 h wigh subsequent treatment with IL-1β,followed by treatment with diluted BYHWT-medicated serum(5％,10％and 20％)or control serum.AnnexinV-APC/7-AAD double staining and T-AOC kit were used for detecting apoptosis and total antioxidant capacity of the cells,and the changes in ROS,ATP level,mitochondrial membrane potential and Ca2+homeostasis were analyzed.The changes in mRNA and protein expressions of BNIP3,PI3K and AKT and mRNA expressions of LC3,Beclin-1 and P62 were detected using RT-qPCR and Western blotting.Results Treatment with BYHWT-medicated serum dose-dependently lowered apoptosis rate of IL-1β-induced FLS-RA with hypoxic exposure.The treatment significantly decreased T-AOC concentration,increased ROS production,autophagosome formation and ATPase levels,and lowered mitochondrial membrane potential and Ca2+level in the cells.In IL-1β-induced FLS-RA with hypoxic exposure,treatment with BYHWT-medicated serum significantly increased BNIP3 protein expression,decreased the protein expressions of PI3K and AKT,increased the mRNA expressions of BNIP3 and P62,and lowered the mRNA expressions of PI3K,AKT,LC3 and Beclin-1 without significantly affecting Beclin-1 protein expression.The cells treated with 5％and 10％BYHWT-medicated serum showed no significant changes in LC3 expression.Conclusion BYHWT inhibits mitochondrial autophagy in IL-1β-induced FLS-RA with hypoxic exposure possibly by inhibiting BNIP3-mediated PI3K/AKT signaling pathway.

Objective To establish,evaluate and validate the Nomogram prediction model of radiation pneumonitis(RP)in intensity modulated radiotherapy(IMRT)with TCM syndrome elements.Methods 257 patients with locally advanved non-small cell lung cancer receiving IMRT were analyzed retrospectively.The total population was randomly divided into a training set and a validation set by 7:3.A prediction model was established by Lasso-Logistic regression analysis,and then visualized by Nomogram to evaluate and validate the model.Results Independent risk factors included in the prediction model included tumor stage(OR=6.576;P=0.003),position(OR=2.935;P=0.016),MLD of the affected lung(OR=1.001;P<0.001)and Yin deficiency(OR=3.861;P=0.003).Based on the above factors,the prediction model was constructed and visualized.The C-index of the training set and the validation set were 0.865 and 0.867,respectively.The calibration curves of the two sets had a good fit and had certain clinical usefulness.Conclusion Based on the clinical elements of Chinese and Western medicine,tumor stage,position,lung MLD and Yin deficiency,the model can accurately predict the occurrence of≥grade 2 RP,and provide a reference for clinical screening of high-risk patients and further improvement of treatment plan.

Objective To establish,evaluate and validate the Nomogram prediction model of radiation pneumonitis(RP)in intensity modulated radiotherapy(IMRT)with TCM syndrome elements.Methods 257 patients with locally advanved non-small cell lung cancer receiving IMRT were analyzed retrospectively.The total population was randomly divided into a training set and a validation set by 7:3.A prediction model was established by Lasso-Logistic regression analysis,and then visualized by Nomogram to evaluate and validate the model.Results Independent risk factors included in the prediction model included tumor stage(OR=6.576;P=0.003),position(OR=2.935;P=0.016),MLD of the affected lung(OR=1.001;P<0.001)and Yin deficiency(OR=3.861;P=0.003).Based on the above factors,the prediction model was constructed and visualized.The C-index of the training set and the validation set were 0.865 and 0.867,respectively.The calibration curves of the two sets had a good fit and had certain clinical usefulness.Conclusion Based on the clinical elements of Chinese and Western medicine,tumor stage,position,lung MLD and Yin deficiency,the model can accurately predict the occurrence of≥grade 2 RP,and provide a reference for clinical screening of high-risk patients and further improvement of treatment plan.

Objective To evaluate the effects of Baihe Gujin Decoction on long-term survival in patients with non-small cell lung cancer(NSCLC)receiving radiotherapy(RT)in the real world.Methods Retrospective data from patients with NSCLC receiving RT in the Longhua Hospital Affiliated to Shanghai University of Traditional Chinese Medicine(TCM)and data from patients in the Surveillance,Epidemiology and End Results(SEER)database were collected and grouped as the TCM+conventional Western medicine(CWM)cohort and CWM cohort,respectively.Propensity score matching(PSM)was used to balance confounders between groups.Kaplan-Meier curve and Cox regression model were used to estimate the effects of Baihe Gujin Decoction on survival outcomes of patients receiving RT.Results Totally 190 samples with a balance between groups were obtained using PSM.Overall survival(OS)analysis showed that the TCM+CWM cohort had significantly longer OS than the CWM cohort(P=0.004).Subgroup analysis showed that the Baihe Gujin Decoction was more advantageous in the characteristic population of age≥65 years(P=0.027),male(P=0.010),T3-T4 stage(P<0.001),N2-N3 stage(P=0.026),M1 stage(P<0.001),not receiving chemotherapy(P<0.001),and not undergoing surgery(P<0.001).Multivariate Cox regression confirmed that treatment group(P=0.003),T(P=0.003)and M stages(P=0.002)may be the independent prognostic factors for OS of NSCLC patients treated with RT.Conclusion Baihe Gujin Decoction probably can improve the long-term prognosis in NSCLC patients treatment with RT.

Public hospitals play a dominant role in providing medical services.Meanwhile,they are also critical for un-dertaking missions to handle health emergencies.This paper analyzes the necessity,current situation,and existing weaknesses of the health emergency capacity of public hospitals.It also conducts a comparative study of the emergency response mechanisms of foreign medical institutions.This paper aims to explore a well-developed design for health emergency in public hospitals that is suitable for China's medical conditions and to provide a feasible model for promoting high-quality health emergency management.

Objective:This study was conducted to investigate the independent risk factors for predicting the occurrence of acute Stanford type A aortic dissection(TAAD), and to construct a nomogram model for predicting the occurrence of TAAD.Methods:The clinical data of patients meeting the diagnostic criteria for TAAD admitted to Tianjin Chest Hospital from June 2016 to December 2021 and healthy people examined by the physical examination center of Tianjin Chest Hospital during the same period were retrospectively collected, and the independent risk factors for TAAD were predicted by propensity matching analysis. Univariate and multivariate Logistic regression were used to analyze the variables with statistical differences, and a nomogram model was constructed to predict the occurrence of TAAD disease according to the screened risk factors. Results:A total of 148 patients in the TAAD group and 5 690 patients in the control group were collected. After bias matching analysis, 148 pairs were successfully matched. Multivariate Logistic regression analysis was performed on the matching results. The results showed that hypertension(HBP), diabetes mellitus(T2DM), Lp(a), very low density lipoprotein(VLDL) and apolipoprotein A1/B(ApoA1/B) were independent risk factors for the development of TAAD. HBP, Lp(a) and ApoA1/B were pathogenic factors( OR 7.267, 1.010 and 2.199, P<0.05, respectively), while T2DM and VLDL were protective factors( OR 0.173 and 0.139, P<0.05). Based on the independent risk factors obtained by multi-factor Logistic regression analysis, a nomogram model of TAAD incidence was constructed. The area under ROC curve( AUC) for predicting the onset of TAAD was 81.6%(95% CI: 0.766-0.863), and the internal calibration curve was close to the standard curve. Conclusion:This model has a good degree of differentiation and calibration, which is helpful for clinicians to guide healthy people to prevent the occurrence of TAAD and provide a theoretical basis for the prevention of TAAD.

Objective　To perform the pathogenic and genomics analyses on isolates of Streptococcus suis (Ss) from three human infections in Shenzhen, aiming to provide a basis for the prevention and control of Ss outbreaks. Methods　The suspected bacterial strains from three blood plate cultures of three critically ill patients in three hospitals were subjected to biochemical identification, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and real-time fluorescent PCR identification, resulting in the identification of three strains positive for Streptococcus suis serotype 2(SS2). Pure positive cultures were taken for an antimicrobial susceptibility test and extracted nucleic acids for whole-genome sequencing and analysis. The whole-genome sequencing and analysis included species identification, antibiotic resistance genes alignment, multilocus sequence typing (MLST), virulence genes alignment, and coregene-based phylogenetic tree analysis. Results　The blood agar isolates from three patients were all identified as Ss, the VITEK 2 identified them as SS2, and MALDI-TOF-MS identified them as Ss. Real-time PCR results for the universal gene gdh and serotype 2 cps2 gene of Ss were both positive. The antimicrobial susceptibility test results showed that all three strains were resistant to erythromycin and clindamycin, with variable sensitivity to tetracycline. Whole-genome sequencing results showed that all three strains were identified as Ss, including one ST7 strain and two ST1 strains. The virulence gene prediction results based on the VFDB database showed that all three strains were positive for mrp, sly, and cps, indicating high virulence gene characteristics. The analysis of the phylogenetic tree based on coregene showed that the three strains were in different evolutionary branches, with two ST1 strains having a closer evolutionary distance. Conclusions　The pathogens responsible for these three critically ill patients were SS2, and all three strains were resistant to erythromycin and clindamycin. Genetically, they all carried virulence genes that are found in highly virulent strains, while showed differences in MLST typing and phylogenetic tree analysis, indicating the presence of different genotypes of high pathogenicity SS2 in Shenzhen area and had caused sporadic cases, which requires high attention.

Objective To study the effects of Guizhi Shaoyao Zhimu Decoction on factors related to bone destruction and bone protection in rats with collagen-induced arthritis(CIA)based on osteopro-tegerin(OPG)/receptor activator of NF-κB ligand(RANKL)/receptor activator of NF-κB(RANK)signaling pathway.Methods According to the body weight,60 female Wistar rats were randomly di-vided into the following six groups:the normal group,the model group,the Triperygium wilfordii mul-tiglucoside group(0.01 g/kg),the Guizhi Shaoyao Zhimu Decoction low-dose group(8.6 g/kg),the Guizhi Shaoyao Zhimu Decoction medium-dose group(17.2 g/kg),and the Guizhi Shaoyao Zhimu Decoction high-dose group(34.4 g/kg)(n=10 rats per group).The rats in all groups except for the normal group were given 100 μg bovine type Ⅱ collagen on the 1st and 8th days to establish the CIA model,and was injected into the left foot sole and tail root of the rats.After the successful modeling,the rats were treated by gavage for 4 weeks.The general state,body weight,and arthritis index(AI)score of rats were recorded,and the contents of RANKL and OPG in rat serum were determined by enzyme-linked immunosorbent assay.The mRNA and protein expressions of RANKL,RANK,and OPG in the ankle joint were determined through real-time PCR and Western blotting,respectively.Results Com-pared with the normal group,the general state of the model group was poor,the toe swelling was obvious,the AI score was increased,the serum RANKL content was increased,the serum OPG content was de-creased,and the mRNA and protein expressions of RANKL and RANK in the ankle joint were increased(P＜0.05).Compared with the model group,the degree of toe swelling and the AI score of rats in the Guizhi Shaoyao Zhimu Decoction low-,medium-,and high-dose groups were decreased,the serum RANKL content was decreased,the serum OPG content was increased,the mRNA and protein expres-sions of RANKL and RANK in the ankle joint were decreased,the mRNA and protein expressions of OPG were increased,and the RANKL/OPG ratio of the ankle joint was decreased(P＜0.05).Conclusion Guizhi Shaoyao Zhimu Decoction can improve the destruction of joint bone in CIA rats,and its mecha-nism of action may be related to reducing RANKL level,reducing RANKL/OPG ratio,and regulating bone balance.

ObjectiveTo explore the possible mechanism of Tongfeng Decoction （痛风汤） for preventing and treating acute gouty arthritis. MethodsSixty Wistar male rats were divided into normal group， model group， colchicine group and low-， medium- and high-dose Tongfeng Decoction groups according to the random number table， 10 rats in each group. Tongfeng Decoction of 11.34， 22.68 and 45.36 g/kg were given by gavage to low-， medium- and high-dose Tongfeng Decoction groups， colchicine 3.15×10^-4 g/（kg·d） to the colchicine group， and normal saline 10 ml/（kg·d） to the normal group and model group respectively for 7 consecutive days. After 1 hour of gavage on day 5， rats in all groups except the normal group were modelled as acute gouty arthritis in the ankle joint of the right hind limb with the modified Coderre's method； rats in the normal group were injected with 0.2 ml of normal saline at the same location. The swelling degree of the ankle joint was measured before modelling and after 6 h， 12 h， 24 h and 48 h of modelling， respectively. HE staining was used to observe the histopathological and morphological changes in the synovial tissue of the ankle joint； immunoblotting and RT-qPCR were used to detect NOD-like receptor protein 3 （NLRP3）， apoptosis-associated speck-like protein （ASC）， cysteine aspartate protease 1 （Caspase-1）， and interleukin 1β （IL-1β） protein and mRNA expression levels in ankle synovial tissues， respectively； immunohistochemistry was used to detect the positive expression of Caspase-1 and IL-1β in ankle synovial tissues； ELISA was used to detect the tumour necrosis factor alpha （TNF-α）， IL-1β， and interleukin in serum； immunofluorescence staining was used to observe the formation of neutrophil extracellular traps （NETs） in the synovial tissues of the ankle joints. ResultsCompared with the normal group， rats in the model group showed elevated joint swelling at all time points， significantly increased inflammatory cell infiltration and the number of synovial cell layers in the synovial tissues of the ankle joints， elevated NLRP3， Caspase-1， ASC and IL-1β protein and mRNA expression， elevated positive expression of Caspase-1 and IL-1β， increased formation of NETs， and elevation of TNF-α， IL-6， and IL-1β in serum （P＜0.05）. Compared with the model group， there was an improvement in synovial cell proliferation and inflammatory cell infiltration of the ankle joint in the colchicine group， high- and medium-dose Tongfeng Decoction groups. In the colchicine group and the high-， medium- and low-dose Tongfeng Decoction groups， the degree of joint swelling， positive expression of Caspase-1， IL-1β and formation of NETs in the synovial tissue of the ankle joint， and TNF-α， IL-6 and IL-1β in serum reduced 24 h and 48 h after modelling； in colchicine group and high-dose Tongfeng Decoction group， NLRP3， ASC， Caspase-1， IL-1β protein and mRNA expression in the synovial tissues of ankle joints all reduced （P＜0.05）. The colchicine group and high-dose Tongfeng Decoction group were superior to the low-dose Tongfeng Decoction group in reducing ASC， Caspase-1， IL-1β protein and mRNA expression in the ankle synovial tissues， the positive expression of Caspase-1， as well as TNF-α， IL-6， and IL-1β level in serum （P＜0.05）. ConclusionTongfeng Decoction showed effectiveness for the prevention and treatment of acute gouty arthritis， and its mechanism may be related to the inhibition of the assembly and activation of NLRP3 inflammatory vesicles in ankle synovial tissues， the inhibition of the release of inflammatory factors， and the formation of NETs.