ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ （AngⅡ） and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment， AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h， and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 （CCK-8） to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of B-type natriuretic peptide （BNP） in cell culture supernatant， and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently， the experiment was divided into a blank group （20% blank serum）， a model group （20% blank serum + 5×10^-5 mol·L^-1 AngⅡ）， low-， medium-， and high-dose （5%， 10% and 20%） serum containing Zhenwutang groups， an autophagy inhibitor group （1×10^-4 mol·L^-1 3-MA）， and autophagy inducer group （1×10^-7 mol·L^-1 rapamycin）. The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe （Lyso Tracker） and mitochondrial probe （Mito Tracker） co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect Caspase-3， Caspase-9， B-cell lymphoma 2 （Bcl-2）， Bcl-2-related X protein （Bax）， and cytochrome C （Cyt C） in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase （Parkin）， phosphatase and tensin homolog （PTEN）-induced kinase 1 （PINK1）， and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3， cleaved Caspase-9， Bax， Bcl-2， and Cyt C in apoptosis-related pathways， phosphorylated ubiquitin ligase （p-Parkin）， phosphorylated PTEN-induced kinase 1 （p-PINK1）， p62， and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways， and the change of microtubule-associated protein 1 light chain 3 （LC3） Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10^-5 mol·L^-1， the cell activity was the lowest， and there was no cytotoxicity. At this concentration， the surface area of cardiomyocytes was significantly increased （P<0.01）， and the content of BNP in the supernatant of culture medium was significantly increased （P<0.05）. Therefore， AngⅡ with a concentration of 5×10^-5 mol·L^-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group， the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate （P<0.01） and significantly decreased mitochondrial membrane potential （P<0.01）. The results of immunofluorescence co-localization showed that compared with the blank group， the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group， the relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 in the model group was significantly up-regulated （P<0.01）， while the relative mRNA expression of Bcl-2， Parkin， and PINK1 was significantly down-regulated （P<0.01）. In addition， the relative protein expression of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 was significantly up-regulated （P<0.01）. The LC3Ⅱ/Ⅰ was significantly decreased， and the relative protein expression of Bcl-2， p-Parkin， p-PINK1， and Beclin1 was significantly down-regulated （P<0.01）. Compared with the model group， the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate （P<0.01）， and the decrease ratio of mitochondrial membrane potential is significantly lowered （P<0.01） in a dose-dependent manner. Additionally， both red and green fluorescence spots became more in these groups. In the 3-MA group， the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax， Caspase-3， Caspase-9， Cyt C， and p62 was significantly down-regulated （P<0.05， P<0.01）， while that of Bcl-2， Parkin， and PINK1 was significantly up-regulated （P<0.01）. In the serum containing Zhenwutang groups， the relative protein expression levels of Bax， cleaved Caspase-3， cleaved Caspase-9， Cyt C， and p62 were significantly down-regulated （P<0.05，P<0.01）. The LC3Ⅱ/Ⅰ was significantly increased， and the relative protein expression levels of Bcl-2， p-Parkin， p-PINK1， and Beclin1 were significantly up-regulated （P<0.01）. ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.

With the widespread use of antibiotics, drug-resistant bacterial infections have become a significant threat to human health. Finding new antibacterial strategies that can effectively control drug-resistant bacterial infections has become an urgent task. Unlike small molecule drugs that target bacterial proteins, antisense oligonucleotide (ASO) can target genes related to bacterial resistance, pathogenesis, growth, reproduction and biofilm formation. By regulating the expression of these genes, ASO can inhibit or kill bacteria, providing a novel approach for the development of antibacterial drugs. To overcome the challenge of delivering antisense oligonucleotide into bacterial cells, various drug delivery systems have been applied in this field, including cell-penetrating peptides, lipid nanoparticles and inorganic nanoparticles, which have injected new momentum into the development of antisense oligonucleotide in the antibacterial realm. This review summarizes the current development of small nucleic acid drugs, the antibacterial mechanisms, targets, sequences and delivery vectors of antisense oligonucleotide, providing a reference for the research and development of antisense oligonucleotide in the treatment of bacterial infections.

Objective: To create a prediction model for stroke risk among middle-aged and elderly populations, so as to provide a basis for early identification of high-risk population for stroke. Methods: From October to December 2023, residents aged ≥45 years in Chongchuan District, Nantong City, Jiangsu Province were selected using a multi-stage stratified random sampling method. The demographic information, life behavior, and chronic disease data were collected through a questionnaire survey. The standardized prevalence of stroke was calculated using data from the seventh National Population Census. The subjects were randomly divided into the training set and the internal validation set according to the ratio of 8∶2. The basic demographic information, life behavior, and chronic diseases of residents aged ≥45 years in Rugao City were collected from July to August 2023 as the external validation set. Predictive factors were selected using multivariable logistic regression model, and a nomogram for stroke among residents aged ≥45 years was established. The prediction effect was evaluated using the area under the curve (AUC) of the receiver operating characteristic (ROC), calibration curve, and Hosmer-Lemeshow goodness of fit test. Results: A total of 6 290 residents aged ≥45 years were included, including 2 975 males (47.30%) and 3 315 females (52.70%). The average age was (61.90±10.20) years. The prevalence of stroke was 3.80%, and the standardized prevalence was 3.36%. The multivariable logistic regression showed that age, smoking, hypertension, and hyperlipidemia were predictors of stroke risk among residents aged ≥45 years, and the prediction model was ln[p/(1-p)]=-4.619+0.046×age+0.383×smoking+0.887×hypertension+0.678×hyperlipidemia. The AUC values of the training set, internal validation set, and external validation set were 0.748, 0.755, and 0.738, respectively. The consistency indexes were 0.748, 0.755, and 0.738, respectively. The Hosmer-Lemeshow goodness of fit test showed a good fitting effect (P>0.05). Conclusion The prediction model based on age, smoking, hypertension, and hyperlipidemia has good discrimination and calibration, and can be used to predict the risk of stroke among middle-aged and elderly populations aged ≥45 years.

Objective To explore the mechanism of kaempferol in improving airway inflammation in chronic obstructive pulmonary disease(COPD)rats.Methods Twenty-for male SD rats were randomly divided into control group,model group(exposed to second-hand smoke for 36 weeks)and experimental group(intragastric administration of 20 mg·kg-1 kaempferol from the 7th day of exposure to secondhand smoke).Pulmonary function was measured by small animal pulmonary function test system,lung index was calculated by body weight and lung weight,inflammatory infiltration and fibrosis changes in lung tissue were observed histologically,the expression of interleukin-1(IL-1),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in bronchoalveolar lavage fluid was detected by enzyme linked immunosorbent assay(ELISA);the expression of superoxide dismutase(SOD)and malondialdehyde(MDA)was detected by the detection kit;and the protein expression of Toll-like receptor 4(TLR4)and nuclear factor-κB(NF-κB)p65 in lung tissue was detected by Western blotting.Results The peak expiratory flow in the control group,model group and experimental group were(19.48±1.61),(15.86±1.00)and(18.07±0.83)mL·s-1;valsalva maneuver were(231.22±10.26),(244.94±9.32)and(225.71±3.80)mL·min-1;lung compliance were(0.53±0.08),(0.29±0.02)and(0.48±0.03)mL·cm-1 H2O;total airway resistance were(0.19±0.12),(0.21±0.01)and(0.20±0.01)cmH2O·mL-1;running economy were(0.19±0.01),(0.26±0.03)and(0.21±0.01)mL·s-1;lung weight were(2.76±0.10),(2.94±0.18)and(2.29±0.26)g;body weight were(375.13±23.55),(243.00±26.75)and(325.38±23.80)g;lung index were 0.74±0.02,1.22±0.09 and 0.70±0.05;the expression levels of cytokines IL-1 in bronchoalveolar lavage fluid(BALF)were(32.39±3.37),(161.88±9.02)and(66.44±6.81)pg·mL-1;IL-6 were(75.07±8.87),(129.58±13.29)and(99.94±9.92)pg·mL-1;TNF-α were(441.76±9.92),(814.47±122.02)and(656.70±70.06)pg·mL-1;MDA expression were(135.73±6.20),(179.05±15.18)and(149.40±13.83)nmol·mL-1;SOD expression were(4 258.55±384.12),(1 232.24±237.08)and(2 134.33±197.99)U·mL-1;TLR4 protein expression level in lung tissue were 0.98±0.02,1.55±0.04,1.34±0.02;NF-κB p65 protein expression level were 0.98±0.02,1.61±0.03,1.09±0.03;the above indicators,control group compared with model group,experimental compared with and model group,the differences were all statistically significant(all P＜0.05).Conclusion Kaempferol ameliorates lung injury induced by airway inflammation in COPD rats by inhibiting TLR4/NF-κB signaling pathway.

Manganese superoxide dismutase catalyzes the dismutation of two molecules of superoxide radicals to one molecule of oxygen and one molecule of hydrogen peroxide. The oxidation of superoxide anion to oxygen by Mn³⁺SOD proceeds at a rate close to diffusion. The reduction of superoxide anion to hydrogen peroxide by Mn²⁺SOD can be progressed parallelly in either a fast or a slow cycle pathway. In the slow cycle pathway, Mn²⁺SOD forms a product inhibitory complex with superoxide anion, which is protonated and then slowly releases hydrogen peroxide out. In the fast cycle pathway, superoxide anion is directly converted into product hydrogen peroxide by Mn²⁺SOD, which facilitates the revival and turnover of the enzyme. We proposed for the first time that temperature is a key factor that regulates MnSOD into the slow- or fast-cycle catalytic pathway. Normally, the Mn²⁺ rest in the pent-coordinated state with four amino acid residues (His26, His74, His163 and Asp159) and one water (WAT1) in the active center of MnSOD. The sixth coordinate position on Mn (orange arrow) is open for water (WAT2, green) or O₂^• to coordinate. With the cold contraction in the active site as temperature decreases, WAT2 is closer to Mn, which may spatially interfere with the entrance of O₂^• into the inner sphere, and avoid O₂^•/Mn²⁺ coordination to reduce product inhibition. Low temperature compels the reaction into the faster outer sphere pathway, resulting in a higher gating ratio for the fast-cycle pathway. As the temperature increases in the physiological temperature range, the slow cycle becomes the mainstream of the whole catalytic reaction, so the increasing temperature in the physiological range inhibits the activity of the enzyme. The biphasic enzymatic kinetic properties of manganese superoxide dismutase can be rationalized by a temperature-dependent coordination model of the conserved active center of the enzyme. When the temperature decreases, a water molecule (or OH^-) is close to or even coordinates Mn, which can interfere with the formation of product inhibition. So, the enzymatic reaction occurs mainly in the fast cycle pathway at a lower temperature. Finally, we describe the several chemical modifications of the enzyme, indicating that manganese superoxide dismutase can be rapidly regulated in many patterns (allosteric regulation and chemical modification). These regulatory modulations can rapidly and directly change the activation of the enzyme, and then regulate the balance and fluxes of superoxide anion and hydrogen peroxide in cells. We try to provide a new theory to reveal the physiological role of manganese superoxide dismutase and reactive oxygen species.

italic>Lamiophlomis rotata is an important medicinal plant species endemic to the Tibetan Plateau, which is prone to strong climate change impacts on its habitable range due to the high sensitivity of the Tibetan Plateau to climate change. Accurate quantification of species vulnerability to climate change is essential for assessing species extinction risk and developing effective conservation strategies. Therefore, we carried out the α-shape analysis to determine the habitat of L. rotata. We then carried out the climate-niche factor analysis (CNFA) to assess the vulnerability of L. rotata to climate change based on five climate variables (i.e., mean diurnal range, temperature seasonality, mean temperature of warmest quarter, precipitation of driest month and precipitation of warmest quarter) in the context of two shared socioeconomic pathways (i.e., SSP126 and SSP585) and three global climate models (CMCC-ESM2: Centro Euro-Mediterraneo sui Cambiamenti Climatici-Earth System Model version 2; HadGEM3-GC31-LL: Hadley Global Environment Model version 3-Global Coupled configuration 3.1; IPSL-CM6A-LR: Institut Pierre Simon Laplace-Climate Model version 6) during two different periods (2041-2060 and 2081-2100). The vulnerability of L. rotata to climate change was calculated by integrating the sensitivity and exposure indices of L. rotata to five climate variables. The results showed that L. rotata had the highest vulnerability to the precipitation of warmest quarter. Its vulnerability within its habitat range generally showed a spatial pattern of high value in the southern region and low in the northern region, high in the western region and low in the eastern region. In general, the vulnerability of L. rotata under the SSP585 scenario was higher than that under the SSP126 scenario. The climate data of different global climate models have some influence on the results, while the resulted uncertainty can be reduced by data integration methods. As a result of climate change, the pressure on the survival of L. rotata in the future will be intensified in the low-altitude areas such as the Yarlung Zangbo River, Yigongzangbu River, Zayu River, and Jiaomuzu River, etc., while the highly weathered scree flats or stony alpine meadows in the high-altitude zones, such as the eastern Tanggula Mountain Range, the northern part of Hengduan Mountain Range, and the western part of the Qinling Mountains, may become its refuge. It is necessary to focus on and strengthen the protection and management of L. rotata resources in these vulnerble and critical areas.

Objective To analyze the epidemiological characteristics and related occupational diagnosis situations of occupational heat illness (OHI) diagnosed by Guangdong Province Hospital for Occupational Disease Prevention and Treatment (GDHOD) from 2011 to 2022. Methods A total of 203 patients who applied for OHI diagnosed in GDHOD from 2011 to 2022 were recruited as research subjects using the retrospective analysis method. Clinical data of OHI diagnosed cases was collected from "Occupational Diseases and Hazards Monitoring Information System" under "China Disease Prevention and Control Information System", and data of cases that diagnosed as non-OHI were collected from the occupational disease diagnosis file of GDHOD for retrospective analysis. Resultsi) The number of OHI diagnosed patients was 174 among 203 cases, with a rate of 85.7%. ii) The heat stroke accounted for 72.4% with a mortality rate of 7.5%, male patients accounted for 86.2% among the 174 OHI diagnosed cases. The median age of onset was 47 years, and patients aged 45-60 years accounting for 56.3%. The median working period of onset of illness was 80 days, with 20.1%, 40.8%, 53.4% and 70.1% of cases occurring within 1, 30, 90 and 365 days of working, respectively. The patients were concentrated in the Pearl River Delta region, accounting for 93.1%. The peak incidence of OHI occurred from May to September, accounted for 96.6%, with the highest incidence occurred in July. About 67.3% of cases occurred on days when the maximum temperature was ≥35.0 ℃. Cases occurred between 12:00 and 18:00 accounting for 74.1%, and cases occurred within 8 hours of work accounting for 75.2%. OHI of patients occurred during operations in non-heat-source workshops accounted for 43.7% of cases, while outdoor operations accounted for 39.7%. OHI patients in manufacturing and small private enterprises accounted for 55.2% and 62.6%, and the main occupations were workshop operators, loaders/unloaders, and sanitation workers, comprising 63.2% of cases. iii) Among the 29 cases diagnosed as non-OHI, accounting for 86.2% patients with high-temperature work history who did not meet clinical diagnostic criteria, most of them were diagnosed as OHI precursor. Conclusion OHI patients in Guangdong Province predominantly occur in summer and autumn, with heat stroke being the primary condition. Middle-aged males, workers in non-heat-source workshops and outdoor settings have higher risk of OHI. OHI cases are concentrated in specific region and enterprise. The OHI prevention should be enhanced on high-risk workers, who work in the Pearl River Delta region, manufacturing, and small private enterprises.

Objective To construct a radiomics nomogram combining clinical and a radiomics signature for distinguishing type Ⅱpapillary renal cell carcinoma(pRCC)from atypical clear cell renal cell carcinoma(ccRCC).Methods Clinical and CT data of patients with pathologically confirmed type Ⅱ pRCC(62 cases)and atypical ccRCC(56 cases)were analyzed.A random sample was divided into a training set(82 cases)and a test set(36 cases)in a ratio of 7∶3.Clinical factors were screened to construct clinical factor models.A total of 1 595 radiomics features of tumors were extracted from the corticomedullary phase CT images and based on the most effective features to construct a radiomics signature and calculate the radiomics score(Rad-score).A radiomics nomogram was constructed by combining the Rad-score and independent clinical factors.Receiver operating characteristic(ROC)curve was used to assess the clini-cal usefulness of the models.Decision curve analysis(DCA)was used to assess the difference between the models.Results The radiomics signature showed good discrimination in training set area under the curve(AUC)0.894[95％confidence interval(CI)0.834-0.947]and test set AUC 0.879(95％CI 0.774-0.963).The AUC of the clinical factors model in training set and test set were 0.725(95％CI 0.646-0.804)and 0.698(95％CI 0.567-0.819).The AUC of the radiomics nomogram in training set and test set were 0.901(95％CI 0.840-0.953)and 0.901(95％CI 0.809-0.975).DCA demonstrated the radiomics nomogram outmatched the clinical factors model and radiomics signature in the aspects of clinical usefulness.Conclusion Radiomics nomogram based on enhanced CT can provide good prediction of type Ⅱ pRCC and atypical ccRCC preoperatively,improve the diagnostic accuracy and provide guidance for future clinical treatment.

ObjectiveCongenital heart disease （CHD） is a common birth defect in children， with its incidence increasing annually. Because of their special health status， the vaccination situation for children with CHD has become a focal point of attention. This paper systematically reviews the current status of vaccination among children with CHD in China to identify existing issues in vaccination efforts for children with CHD， provide guidance for increasing vaccination rates among children with CHD， reduce the incidence of vaccine-preventable diseases， and provide ideas for subsequent research. MethodsLiterature on the vaccination of children with CHD in China was searched in both Chinese and English databases， including CNKI， Wanfang Data Knowledge Service Platform， VIP， Chinese Biomedical and Web of Science， PubMed， EmBase， and the Cochrane Library. Relevant literature was identified based on inclusion and exclusion criteria， and data extraction was carried out to summarize the research results. ResultsA total of 15 studies were included. Three articles focusing on expert consensus （guideline recommendations） for vaccination of children with CHD in China all agreed that vaccination is both feasible and necessary for children with CHD. Systematic analysis found that the overall timely vaccination rate for children with CHD was between 34% and 50%， with a notable delay in vaccination. However， after professional outpatient assessment， the vast majority of children with CHD were advised to be vaccinated according to the national recommended schedule， with only about 2% of them being advised to delay vaccination. The proportion of children with CHD who reported suspected adverse events following immunization （AEFI） was low and not significantly different from that of healthy/control children. ConclusionThe safety of vaccinations for children with CHD in China is relatively high， but the total timely vaccination rate is currently low. In order to improve the coverage and timeliness of vaccination in children with CHD， it is recommended that relevant departments provide vaccination policy support. There is a lack of research on the effectiveness of vaccination in children with CHD， so further studies are urgently needed to further improve the vaccination strategy for children with CHD in China.

BACKGROUND:Tyrosine kinase inhibitors,as well as other types of small-molecule cancer drugs,can cause severe cardiotoxicity. OBJECTIVE:To perform a heart safety re-evaluation by observing the effects of antitumor drugs on isolated heart electrocardiograph,cardiac action potential and associated ion channels and cytotoxicity. METHODS:Extracorporeal cardiac perfusion was given to the isolated rabbit heart using Langendorff perfusion:Sunitinib(0.3,3,10 μmol/L),Crizotinib(0.3,1,3 μmol/L),and Doxorubicin(1,30 μmol/L)were perfused sequentially for 120 minutes to record electrocardiograph and left ventricular pressure.A blank control group was set for comparison.Manual patch clamp was used to record the effects of Crizotinib,Sunitinib,Doxorubicin on hERG,Cav1.2,Nav1.5 channel currents and action potential in human induced pluripotent stem cell derived cardiomyocytes.Adenosine triphosphate level in human induced pluripotent stem cell derived cardiomyocytes was detected by CellTiter-Glo luminescent cell viability assay. RESULTS AND CONCLUSION:Isolated rabbit heart using Langendorff perfusion:Compared with the blank ontrol group,Sunitinib and Crizotinib at≥3 μmol/L decreased heart rate(P<0.01)and prolonged QT/QTc interval(P<0.01),and reduced left ventricular pressure to different extents.Manual patch clamp recording:Compared with the blank control group,Sunitinib and Crizotinib at 3 μmol/L inhibited the activities of hERG,Nav1.5 and Cav1.2 channels and significantly prolonged the duration of action potential(P<0.01).According to the analysis of the test article,the difference between the labeled concentration and the measured concentration of the recovered solution was not significant.Cell viability assays:Compared with the blank control group,adenosine triphosphate content in human induced pluripotent stem cell derived cardiomyocytes significantly decreased after treatment with Sunitinib(IC50=4.64 μmol/L),Doxorubicin(IC50=4.21 μmol/L)and Crizotinib(IC50=2.87 μmol/L),indicating that cell viability significantly decreased(P<0.01).To conclude,this study successfully established an early cardiac safety evaluation method for antitumor drugs,which provides good support and help for the subsequent development of antitumor drugs.