ObjectiveTo investigate the effects and potential mechanisms of Tongmai Yangxin Pills （通脉养心丸） （TYPs） in preventing ischemia-reperfusion （I/R）-induced arrhythmia. MethodsSixty male SD rats were randomly assigned to sham operation group， model group， amiodarone group， low-dose and high-dose TYPs group， with 12 rats in each group. The sham operation group and the model group received 10 g/（kg·d） normal saline by gavage， the amiodarone group received 60 mg/（kg·d） amiodarone， and the low-dose and high-dose TMP groups received 1 g/（kg·d） and 2 g/（kg·d） TYPs solution respectively， for 21 days， administered twice daily. On the day after the last administration， the I/R model was established in the model and medication groups by ligation of the left anterior descending coronary artery with a cannula， while the sham operation group underwent the same procedure without ligation. Electrocardiogram recordings were continuously monitored throughout the modeling process. Heart rate was recorded at five time points， before ischemia （t-0）， 5-10 min after ischemia （t-1）， 10-15 min after ischemia （t-2）， 15-30 min after ischemia （t-3）， and during the first 2 min of reperfusion （t-4）； the incidence of arrhythmia including ventricular premature beats （VPB）， ventricular tachycardia （VT）， and ventricular fibrillation （VF） was recorded； arrhythmia scores were calculated. After 24 hours of reperfusion， left ventricular myocardial tissue was collected. Hematoxylin-eosin （HE） staining was performed to observe pathological changes. RT-PCR was used to detect the mRNA expression of microRNA-1 （miRNA-1）， microRNA-133a （miRNA-133a）， and potassium （K⁺） and calcium （Ca²⁺） ion channel-related genes including KCND2， KCNH2， KCNE2， KCNQ1， KCNE1， KCNJ2， CACNA1C， and CACNB1. Western blot analysis was used to measure protein levels of transient outward potassium current protein （Kv4.2）， rapidly activating delayed rectifier potassium current protein （HERG）， slowly activating delayed rectifier potassium current protein （KvLQT1）， inward rectifier potassium current protein （Kir2.1）， and L-type calcium channel protein （Cav1.2）. ResultsCompared with sham operation group， the model group showed diffuse myocardial hemorrhage， inflammatory cell infiltration， myocardial necrosis， nuclear pyknosis， vacuolar degeneration， and disrupted myocardial fibers； the model group also exhibited a decreased heart rate （t-1 to t-4）， increased arrhythmia scores， elevated miRNA-1 and miRNA-133a expression， and decreased mRNA expression of KCND2， KCNH2， KCNE2， KCNQ1， KCNE1， KCNJ2， CACNA1C， and CACNB1 in myocardial tissue； additionally， Kv4.2， HERG， KvLQT1， Kir2.1， and Cav1.2 protein levels significantly reduced （P＜0.01）. Compared to the model group， all medication-treated groups showed reduced myocardial damage， including less hemorrhage， reduced inflammatory infiltration， and improved myocardial structure， with the high-dose TYPs group exhibiting the most significant improvement； the amiodarone group and high-dose TYPs group showed a significant increase in heart rate （t-1 to t-4）， lower arrhythmia scores， reduced miRNA-1 and miRNA-133a expression； the high-dose TYPs group exhibited significantly increased mRNA expression levels of KCND2， KCNH2， KCNQ1， KCNJ2， and CACNA1C， as well as elevated protein levels of Kv4.2， HERG， KvLQT1， Kir2.1， and Cav1.2 （P＜0.05 or P＜0.01）. ConclusionTMPs can improve myocardial damage and reduce the incidence of ventricular arrhythmia in I/R rats. The underlying mechanism may be related to the downregulation of miRNA-1 and miRNA-133a gene expression， as well as the upregulation of K⁺ and Ca²⁺ channel-related genes and proteins.

Objective To systematically evaluate the efficacy and safety of Compound Danshen Dripping Pills combined with conventional western medicine in the treatment of coronary heart disease complicated with diabetes mellitus.Methods Wanfang database,CNKI,VIP,SinoMed,Cochrane Library,PubMed,Embase and Web of Science databases were searched to collect randomized controlled trials(RCTs)of Compound Danshen dripping pills combined with conventional western medicine in the treatment of coronary heart disease complicated with diabetes mellitus.The retrieval time is from the establishment of the database to May 2023.Two researchers independently completed literature screening,data collation and risk of bias evaluation.RevMan 5.4 software was used for Meta-analysis.Results Twenty-four RCTs with a total of 2 546 patients were included.Meta-analysis showed that the treatment of Compound Danshen Dripping Pills combined with conventional western medicine was better than that of conventional western medicine in improving the total effective rate of coronary heart disease complicated with diabetes mellitus[OR=4.93,95%CI(3.49,6.98),P＜0.000 01],plasma adiponectin[MD= 2.79,95%CI(2.30,3.27),P＜0.000 01],reducing fasting blood glucose[SMD=-1.06,95%CI(-1.24,-0.88),P＜0.000 01],postprandial two-hour blood glucose[MD=-1.53,95%CI(-1.71,-1.35),P＜0.000 01],glycosylated hemoglobin[MD=-1.56,95%CI(-2.01,-1.11),P＜0.000 01],homocysteine[MD=-8.47,95%CI(-8.89,-7.97),P＜0.000 01],low density lipoprotein[MD=-0.46,95%CI(-0.69,-0.24),P＜0.000 01],total cholesterol[MD=-0.45,95%CI(-1.11,-0.20),P=0.005],triglycerides[MD=-0.42,95%CI(-0.50,-0.34),P＜0.000 01],interleukin 6[SMD=-1.34,95%CI(-1.61,-1.07),P＜0.000 01],cardiovascular adverse events[OR=0.35,95%CI(0.19,0.64),P=0.000 6],incidence of adverse reactions[OR=0.45,95%CI(0.24,0.86),P=0.01].Conclusion Compound Danshen Dripping Pills combined with conventional western medicine in the treatment of coronary heart disease with diabetes mellitus is more effective than conventional western medicine treatment,and with better safety.However,due to the limit of the quality of the included studies and other factors,the above conclusions still need to be verified by more high-quality clinical trials.

Objective To explore the expression of autophagy-related gene 5(ATG5)in human malignant pleural mesothelioma(MPM)cells and tissues and analyze the correlation between ATG5expression and patient clinicopathological parameters and prognosis.Methods Real-time quantitative PCR and Western blotting were used to detect differences in ATG5 expression between normal human pleural mesothelial cells and MPM cells and between non-MPM pleural mesothelial tissues and MPM tissues.The Cancer Genome Atlas was used to analyze correlations between ATG5 expression and clinicopathological characteristics and prognosis of patients with MPM.A Cox proportional hazard model was constructed to analyze factors affecting the prognosis of patients with MPM.The Gene Expression Profiling Interactive Analysis database was used to evaluate the correlation between ATG5and MPM tumor markers and novel serum markers.Tumor Immune Estimation Resource was used to analyze correlations between ATG5 and MPM immune cell infiltration and key immune regulatory genes.Results ATG5 was highly expressed in MPM cells and tissues and positively correlated with tumor stage(P<0.05).High ATG5 expression indicated poor prognosis(P<0.05).ATG5expression was significantly associated with various MPM tumor markers(MTAP,SETD2,NF2,and FIB3)and novel serum markers(HMGB1,SMPR,THBS2,and KRAS)(P<0.05).ATG5was associated with immune cell infiltration in MPM(B cells,CD4+T cells,and macrophages)and expression of immune-related genes(CD28,CUL48B,CD166,and MMP14)(P<0.05).Conclusion ATG5 is upregulated in MPM and is associated with poor prognosis and immune cell infiltration.ATG5 could be a key biomarker for early screening,diagnosis,and prognostic assessment of MPM.

Femoral neck fracture (FNF) in the elderly patients is currently a major health challenge worldwide, with excessive consumption of medical resources, high incidence of complications as well as suboptimal outcome and prognosis. Hip joint arthroplasty (HJA) has been the mainstream treatment for FNF in the elderly, but the conventional surgical approaches and techniques are still confronted with a series of bottlenecks such as dislocation, limp and limb length discrepancy. In recent years, direct anterior approach (DAA) for HJA (DAA-HJA) has been a major new choice in the field of joint replacement, which achieves improved clinical effectiveness of HJA in the treatment of elderly FNF, due to the fact that DAA approach involves the neuromuscular interface and accords with the idea of soft tissue retention and enhanced recovery after surgery. However, there is still a lack of unified understanding of standard technique and procedure of DAA-HJA in the treatment of elderly FNF. Therefore, relevant experts from the Hip Joint Group of Chinese Orthopedics Association of Chinese Medical Association, Youth Arthrology Group of Orthopedic Committee of PLA, Orthopedic Committee of Chongqing Medical Association, Branch of Orthopedic Surgeons of Chongqing Medical Doctor Association and Sport Medicine Committee of Chongqing Medical Association were organized to formulate the " Chinese expert consensus on the technical standard of direct anterior hip arthroplasty for elderly femoral neck fracture ( version 2023)" based on evidence-based medicine. This consensus mainly proposed 13 recommendations covering indications, surgical plans, prosthesis selections, surgical techniques and processes, and postoperative management of DAA-HJA in elderly patients with FNF, aiming to promote standardized, systematic and patient-specific diagnosis and treatment to improve the functional prognosis of the patients.

Objective:To construct a psychological support training program for palliative care specialist nurses based on positive psychology theory and make a preliminary application.Methods:From August to September, 2020, 142 palliative nursing specialist nurses in Hunan Provincial Palliative Nursing Specialist Training Base were selected as the research objects. Based on positive psychology therapy, using literature review and analysis, semi-structured interviews, expert group meetings and other methods to construct the program, and the work stress, psychological resilience and subjective well-being scores of nurses before and after 8 weeks of psychological support training were compared.Results:The program included three dimensions: cognition, emotion and behavior. After the psychological support training for 8 weeks, the nursing staff′s work stress score 80.76 ± 20.43 was lower than that before the psychological support training 84.70 ± 19.88, the difference was statistically significant( t=6.59, P<0.05), and the psychological resilience score 64.40 ±14.26 was higher than that before the psychological support training 60.19 ±15.85, the difference was statistically significant ( t=-7.39, P<0.05), and the subjective well-being score 75.70 ± 7.70 was higher than that before the psychological support training 74.13 ± 5.98, the difference was statistically significant ( t=-3.52, P<0.05). Conclusions:The psychological support training program based on positive psychology theory can reduce the the work stress, improve psychological resilience and increase subjective well-being of palliative care specialist nurses.

Objective:To obtain purified protein antigen of guertu virus (GTV) nucleoprotein (NP) and establish a rapid and accurate enzyme-linked immunosorbent assay (ELISA) method for detection of GTV antibody.Methods:Codon optimized GTV NP encoding genes were synthesized, cloned into the pet32a (+) vector, and recombinant expression plasmids were constructed and transformed into BL21 (DE3). Recombinant protein (rNP) obtained from the optimized expression were purified over a Ni column and identified by SDS-PAGE and Western blot. The purified protein was used as the antigen to optimize the reaction conditions, and an indirect ELISA assay for GTV IgG antibody was developed and optimized, which was evaluated and initially applied.Results:The prokaryotic expression plasmid pet32a-NP was successfully constructed, the recombinant protein was highly expressed in E. coli in the form of inclusion bodies, the size was about 44 kD, and the results of Western blot indicated that the recombinant protein had good antigenicity with GTV positive serum. The optimized ELISA (GTV-rNP-iELISA) established in this study showed strong specificity, high sensitivity, and the coefficient of variation within and between batches is less than 10%, and has good repeatability; the detection results are consistent with the IFA detection results. Using the established ELISA method to detect 162 sheep sera from some regions of Xinjiang in 2017-2019, the total positive rate of antibodies was 39.8%.Conclusions:The GTV NP antibody detection ELISA method has good sensitivity, reproducibility, and specificity and has the potential to be a powerful tool for the diagnosis and serological investigation of GTV infection.

Objective:To obtain purified protein antigen of guertu virus (GTV) nucleoprotein (NP) and establish a rapid and accurate enzyme-linked immunosorbent assay (ELISA) method for detection of GTV antibody.Methods:Codon optimized GTV NP encoding genes were synthesized, cloned into the pet32a (+) vector, and recombinant expression plasmids were constructed and transformed into BL21 (DE3). Recombinant protein (rNP) obtained from the optimized expression were purified over a Ni column and identified by SDS-PAGE and Western blot. The purified protein was used as the antigen to optimize the reaction conditions, and an indirect ELISA assay for GTV IgG antibody was developed and optimized, which was evaluated and initially applied.Results:The prokaryotic expression plasmid pet32a-NP was successfully constructed, the recombinant protein was highly expressed in E. coli in the form of inclusion bodies, the size was about 44 kD, and the results of Western blot indicated that the recombinant protein had good antigenicity with GTV positive serum. The optimized ELISA (GTV-rNP-iELISA) established in this study showed strong specificity, high sensitivity, and the coefficient of variation within and between batches is less than 10%, and has good repeatability; the detection results are consistent with the IFA detection results. Using the established ELISA method to detect 162 sheep sera from some regions of Xinjiang in 2017-2019, the total positive rate of antibodies was 39.8%.Conclusions:The GTV NP antibody detection ELISA method has good sensitivity, reproducibility, and specificity and has the potential to be a powerful tool for the diagnosis and serological investigation of GTV infection.

Objective:To investigate the relevant factors on serum C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) which did not meet the 2011 Musculoskeletal Infection Society (MSIS) diagnostic criteria in patients with periprosthetic joint infection (PJI).Methods:During December 2011 to December 2019, a total of 328 patients with PJI were hospitalized for surgery or antibiotic administration, including 152 males and 166 females, aged 62.10±13.74 (range 24-87) years. All patients underwent CRP and ESR before the antibiotic administration or the revision surgery. PJI was diagnosed based on the 2011 MSIS diagnostic criteria. There were 172 knee PJIs (52.4%), 151 hip PJIs (46.0%), 4 elbow PJIs (1.2%) and 1 shoulder PJI (0.3%). Patients were classified according to Tsukayama type, pathogen and immune status. We, further, analyzed relevant factors on CRP and ESR levels in PJI patients.Results:There were 119 patients with CRP and ESR did not meet the MSIS diagnostic criteria, accounting for 36.3% (119/328). Furthermore, there was no significant difference in Tsukayama types among them (χ 2=7.224, P=0.065). In addition, the ratio was 46.4% in patients with negative culture results, which was higher than that in positive culture results (27.4%, χ 2=12.276, P<0.001). The ratio was 42.9% in patients with normal immune status (grade A), which was higher than that of immune grade B (30.6%) and of immune grade C (23.8%) (χ 2=6.586, P=0.037). Multivariate logistic regression analysis showed the negative association between positive culture results and immune grade B with nonconformity ( OR=0.420, P=0.001; OR=0.578, P=0.04). Conclusion:The serum tests level unmet the threshold in MSIS criteria usually present in PJI patients with normal immune status and negative culture results. Thus, we should utilize other methods for diagnosing PJI.

Objective To establish a mouse model of pneumonia with C57BL/6 and MyD88KO mice after infection with an isolated ST23 Klebsiella pneumonia (KP) strain, which was an epidemic strain and identified by multilocus sequence typing (MLST). Methods Fifty C57BL/6 mice were randomly di-vided into three groups:KP infection,control and immunosuppressive groups. Thirty MyD88KO mice were divided into KP infection and control groups. All mice in the KP infection groups were infected with 50 μl of ST23 KP strain through nasal dripping. Equal volume of PBS was used to set up the control groups. Mice in the immunosuppressive group were first injected with cyclophosphamide for three days and then infected with equal volume of ST23 KP strains through nasal dripping. Clinical signs and survival curves during KP infec-tion were monitored. Moreover,pulmonary bacterial loads and histopathological changes in the KP-infected mice were detected at different time points. Results ST23 KP-infected C57BL/6 mice showed inflammatory cell infiltration in lung tissues on the 10th day and remained alive on the 21st day. All ST23 KP-infected MyD88KO mice died on the 5th day with severe histopathological damage in lung tissues. C57BL/6 mice that pretreated with cyclophosphamide had similar symptoms with MyD88KO mice after infection and died on the 5th day. Some critical inflammatory mediators such as TNF-a,nitric oxide synthase 1 (NOS1) and NF-κBp65 were up-regulated in lung tissues of mice after KP infection. No inflammatory syndromes were found in the mice of PBS control groups. Conclusion This study suggests that the mouse model of pneumonia is successfully established with KP strain. It will help researchers to study the characteristics and pathogenesis of ST23 KP strain-induced pneumonia and to seek safe treatments in the future.

OBJECTIVE:To establish a method for the contents determination of psoralen,isopsoralen,psoralenoside and isop-soralenoside in Danzhi qing’e tablet. METHODS:HPLC performed on the column of Eclipse XDB-C18 with mobile phase of metha-nol-water(51∶49,V/V)(isocratic elution,for psoralen and isopsoralen)and acetonitrile-0.1% formic acid(12∶88,V/V)(isocratic elution,for psoralenoside and isopsoralenoside)at a flow rate of 1.0 ml/min,the detection wavelength was 246 nm,column tem-perature was 30℃,and injection volume was 10 μl. RESULTS:The linear range was 3.138-200.8 μg/ml for psoralen(r=0.999 9), 3.175-203.2μg/ml for isopsoralen(r=0.999 9),3.181-101.8μg/ml for psoralenoside(r=0.999 9)and 3.169-101.4μg/ml for isopso-ralenoside (r=0.999 9);RSDs of precision,stability and reproducibility tests were lower than 2%;limits of quantitation were 0.627 5 ng,0.635 0 ng,3.181 0 ng and 3.169 0 ng,the limits of detection were 0.251 0 ng,0.254 0 ng,1.273 0 ng and 1.268 0 ng;recoveries were 95.68%-102.80%(RSD=2.4%,n=6),95.91%-102.10%(RSD=2.3%,n=6),98.64%-99.13%(RSD=0.23%,n=6) and 100.20%-101.70%(RSD=0.69%,n=6),respectively. CONCLUSIONS:The method is simple and accurate, and can be used for the simultaneous determination of psoralen,isopsoralen,psoralenoside and isopsoralenoside in Danzhi qing’e tablet.