ObjectiveTo systematically evaluate the content differences of 4 components in Leonuri Herba granules， reveal the quality fluctuation patterns of products from the same and different manufacturers， providing scientific basis for the optimization of production process and quality control. MethodsHigh performance liquid chromatography-diode array detector-charged aerosol detector（HPLC-DAD-CAD） was employed to determine the contents of 4 components（syringic acid， leonurine hydrochloride， ferulic acid， and stachydrine hydrochloride） in samples from 19 manufacturers（53 batches， 159 boxes）. Additionally， fingerprint profiles were constructed， and the fingerprint dissimilarity（P_S） and relative standard deviation（RSD） of different samples from the same manufacturer were calculated. A principal component analysis（PCA） model was established with P_S and the RSD values of the 4 components as variables to classify the manufacturers. Finally， samples from 5 manufacturers（M1-M5） covering three consistency groups were selected to calculate three quality consistency parameters， namely intra-batch consistency（P_A）， inter-batch consistency（P_B）， and P_S. Then， PCA was performed with P_A， P_B， and P_S of these 5 manufacturers as variables. ResultsThe average total content of the 4 index components per bag across the 19 manufacturers ranged from 41.10 mg to 97.54 mg. Among them， the content of stachydrine hydrochloride（a pharmacopoeial quality control component） was 32.46-72.70 mg per bag， all meeting the requirements of the 2025 edition of the Pharmacopoeia of the People's Republic of China， with RSD of 1.7%-17.1%. The content ranges of the other 3 components were as follows：syringic acid of 1.43-41.92 mg per bag， leonurine hydrochloride of 0.67-11.85 mg per bag， and ferulic acid of 0.11-3.81 mg per bag. Notably， leonurine hydrochloride exhibited the most significant content fluctuation among samples from the same manufacturer（RSD of 4.8%-59.2%）. PCA results showed that the 19 manufacturers could be classified into 3 categories. Samples from 8 manufacturers（M2， M6， M7， M8， M10， M15， M17， M18） demonstrated relatively high consistency， five manufacturers（M3， M9， M12， M13， M14） showed moderate consistency， six manufacturers（M1， M4， M5， M11， M16， M19） exhibited low consistency. The two methods yielded consistent classification results for the 5 representative manufacturers， verifying the reliability of the proposed method. Among these， manufacturer M2 showed the best quality consistency and the highest total content of indicator components among M1-M5. ConclusionThe HPLC-DAD-CAD multi-detector hyphenation technology established in this study enables the accurate detection of 4 components in Leonuri Herba granules. Significant differences in the total content of these four components are observed among products from 19 manufacturers. The application of 2 consistency evaluation methods combined with PCA can effectively classify their consistency into 3 categories， and the classification results of the 2 methods are highly consistent. This study provides scientific basis for the process optimization and quality standard improvement of Leonuri Herba granules.

ObjectiveTo systematically evaluate the content differences of 4 components in Leonuri Herba granules， reveal the quality fluctuation patterns of products from the same and different manufacturers， providing scientific basis for the optimization of production process and quality control. MethodsHigh performance liquid chromatography-diode array detector-charged aerosol detector（HPLC-DAD-CAD） was employed to determine the contents of 4 components（syringic acid， leonurine hydrochloride， ferulic acid， and stachydrine hydrochloride） in samples from 19 manufacturers（53 batches， 159 boxes）. Additionally， fingerprint profiles were constructed， and the fingerprint dissimilarity（P_S） and relative standard deviation（RSD） of different samples from the same manufacturer were calculated. A principal component analysis（PCA） model was established with P_S and the RSD values of the 4 components as variables to classify the manufacturers. Finally， samples from 5 manufacturers（M1-M5） covering three consistency groups were selected to calculate three quality consistency parameters， namely intra-batch consistency（P_A）， inter-batch consistency（P_B）， and P_S. Then， PCA was performed with P_A， P_B， and P_S of these 5 manufacturers as variables. ResultsThe average total content of the 4 index components per bag across the 19 manufacturers ranged from 41.10 mg to 97.54 mg. Among them， the content of stachydrine hydrochloride（a pharmacopoeial quality control component） was 32.46-72.70 mg per bag， all meeting the requirements of the 2025 edition of the Pharmacopoeia of the People's Republic of China， with RSD of 1.7%-17.1%. The content ranges of the other 3 components were as follows：syringic acid of 1.43-41.92 mg per bag， leonurine hydrochloride of 0.67-11.85 mg per bag， and ferulic acid of 0.11-3.81 mg per bag. Notably， leonurine hydrochloride exhibited the most significant content fluctuation among samples from the same manufacturer（RSD of 4.8%-59.2%）. PCA results showed that the 19 manufacturers could be classified into 3 categories. Samples from 8 manufacturers（M2， M6， M7， M8， M10， M15， M17， M18） demonstrated relatively high consistency， five manufacturers（M3， M9， M12， M13， M14） showed moderate consistency， six manufacturers（M1， M4， M5， M11， M16， M19） exhibited low consistency. The two methods yielded consistent classification results for the 5 representative manufacturers， verifying the reliability of the proposed method. Among these， manufacturer M2 showed the best quality consistency and the highest total content of indicator components among M1-M5. ConclusionThe HPLC-DAD-CAD multi-detector hyphenation technology established in this study enables the accurate detection of 4 components in Leonuri Herba granules. Significant differences in the total content of these four components are observed among products from 19 manufacturers. The application of 2 consistency evaluation methods combined with PCA can effectively classify their consistency into 3 categories， and the classification results of the 2 methods are highly consistent. This study provides scientific basis for the process optimization and quality standard improvement of Leonuri Herba granules.

ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9（CRISPR/Cas9）gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity， which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system， which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana， endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase （LUC） reporter gene， and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements， upstream sequence element （USE） and TATA-box， which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3， AaU6-1， and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter， with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua， providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.

Bovine rotavirus(BRV)is an important pathogen causing diarrhea in calves.To understand the genomic charac-teristics and genetic variations in bovine rotavirus,and to further enrich data on the biological characteristics of rotavirus,we aimed to amplify 11 gene segments of the isolated and cultured G6P[1]bovine rotavirus BLL strain,perform whole genome se-quencing,and analyze the molecular characteristics.MEGA7.0 and DNAMAN software were used for homology and typing a-nalysis,and the whole genome phylogenetic tree was constructed to analyze genetic evolution relationships.The complete geno-type of the BLL strain was G6-P[1]-I2-R2-C2-M2-A3-N2-T6-E2-H3.Phylogenetic analysis of the VP7 and VP4 genes of the BLL strain showed that the VP7 gene had the highest homology with RVA/Cow-wt/HB01/China/2021,and the VP4 gene of the BLL strain was in the same branch as RVA/Human-tc/ISR/Ro8059/1995.From the sequence alignment of VP8*amino acids,the sialic acid domain of the BLL strain was found to be similar to that in other P[1]strains,but different from those in other types of strains,except for residue 189,which was the same as that in Ro8059 but different from that in other strains.The results suggested that the BLL strain might potentially infect humans.Therefore,continued monitoring and study of the biological characteristics of this strain are necessary to provide more information and evidence supporting further research on the cross-species transmission of group A rotavirus in China.

Gastric cancer remains one of the most prevalent and lethal malignancies of the digestive tract worldwide,underscoring the urgent need for more effective targeted therapeutic strategies.Poly(ADP-ri-bose)polymerase(PARP)inhibitors have demonstrated remarkable efficacy in tumors with homologous recombination repair(HRR)deficiency;however,their clinical application in gastric cancer remains limited.Clinical evidence suggests that patients harboring Helicobacter pylori infection in combination with HRR gene mutations exhibit a significantly elevated risk of developing gastric cancer,thereby supporting the potential benefit of PARP inhibition in this setting.In this study,a kinase inhibitor library was screened in combination with the PARP inhibitor olaparib in gastric cancer cells.And we identify the cy-clin-dependent kinase 8/19(CDK8/19)inhibitor Senexin A as a compound that synergistically enhances the cytotoxic effect of PARP inhibition(P<0.05).Phenotypic validation using CCK-8 and colony for-mation assays demonstrated that the combination treatment significantly suppressed cellular proliferation and clonogenic potential compared to either monotherapy(P<0.0001).Mechanistically,alkaline comet assays revealed a significant increase in DNA damage in the combination treatment group relative to either single-agent group(P<0.0001),suggesting that the synergistic effect results from the exacerbation of DNA damage via impaired DNA repair mechanisms.In addition,treatment with CDK8/19 inhibitors a-lone markedly increased the formation of γH2AX and 53BP1 foci in irradiated gastric cancer cells(P<0.0001),indicating inhibition of DNA damage repair pathways.Transcriptome sequencing further re-vealed that CDK8/19 inhibition impacts critical cellular pathways,including DNA repair,cell cycle reg-ulation,and RNA splicing.Co-immunoprecipitation assays confirmed that inhibition of CDK8/19 kinase activity significantly reduces the phosphorylation level of PARP1,suggesting a potential regulatory inter-action.Immunohistochemical analysis of tumor and adjacent non-tumor tissues from gastric cancer pa-tients demonstrated that CDK8 is significantly overexpressed in tumor tissues,supporting its potential as both a prognostic biomarker and a therapeutic target.Collectively,this study elucidates a mechanistic ba-sis by which CDK8/19 inhibition enhances the sensitivity of gastric cancer cells to PARP inhibitors.These findings provide a strong rationale for the combined use of CDK8/19 and PARP inhibitors as a tar-geted therapeutic strategy and offer promising translational implications for advancing personalized medi-cine in gastric cancer treatment.

Objective:To study the anti-inflammatory and protective effects of electroacupuncture on rats with cardiopulmo-nary resuscitation based on cholinergic anti-inflammatory pathway(CAP).Methods:Sixty male SD rats were randomly divided into sham operation group(Sham group),cardiopulmonary resuscitation group,electroacupuncture group,antagonist of α7-nicotinic ace-tylcholine receptor(α7nAChR)α Bungarus toxin group(αBGT group,1 μg/kg)and agonist of α7nAChR 3-(2,4-dimethoxybenzyli-dene)anabaseine(GTS-21)group(5 mg/kg),with 12 rats in each group.Model of cardiac arrest was established by inducing ventric-ular fibrillation in rats with percutaneous epicardial electrical stimulation,and routine resuscitation was performed,the defibrillation times,cardiopulmonary resuscitation duration and 72 h survival rate of rats in each group were observed and recorded.Neurological damage of rats in each group was scored(NDS score).Levels of inflammatory factors TNF-α,IL-1β and IL-6 in serum,heart,lung and brain of rats were measured by ELISA.HE staining was applied to evaluate the degree of tissue damage of heart,lung and brain tis-sues of rats in each group.Expressions of α7nAChR,NF-κB/MAPKs signal pathway related proteins in heart,lung and brain of rats in each group were detected by Western blot.Results:Compared with Sham group,the number of defibrillations,duration of cardiopul-monary resuscitation,NDS score,levels of TNF-α,IL-1β and IL-6 in serum,heart,lung and brain tissues,and NF-κB p65 protein in heart,lung and brain tissues in cardiopulmonary resuscitation group increased greatly(P<0.05),the 72 h survival rate,phosphory-lation levels of α7nAChR protein,ERK1/2,JNK and p38MAPK in heart,lung and brain tissues decreased greatly after restoration of spontaneous circulation(P<0.05),cells and neurons in the CA1 region of heart,lung and hippocampus were severely damaged.Com-pared with cardiopulmonary resuscitation group,change trend of relevant indicators in electroacupuncture group was opposite to the above,damage of cells and neurons in rat heart,lung and brain tissue was reduced(P<0.05).αBGT reduced the anti-inflammatory effect of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).GTS-21 enhanced the anti-inflammatory protection of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).Conclusion:Electroacupuncture has anti-inflammatory and pro-tective effects on cardiopulmonary resuscitation rats based on CAP,and NF-κB/MAPKs participate in α7nAChR mediated CAP.

With the latest development of information and communication technology, mobile applications for health care have attracted much attention to help cancer patients better manage their diseases. Digital therapeutics is an evidence-based intervention driven by high-quality software programs that prevent, manage, or treat medical disorders or diseases. This paper summarizes the origin, concept and characteristics of digital therapeutics and the application form, effect, challenge and opportunity of symptom management in cancer patients, in order to provide reference for the scientific and efficient application of Digital therapeutics to cancer patients.

Objective:To study the anti-inflammatory and protective effects of electroacupuncture on rats with cardiopulmo-nary resuscitation based on cholinergic anti-inflammatory pathway(CAP).Methods:Sixty male SD rats were randomly divided into sham operation group(Sham group),cardiopulmonary resuscitation group,electroacupuncture group,antagonist of α7-nicotinic ace-tylcholine receptor(α7nAChR)α Bungarus toxin group(αBGT group,1 μg/kg)and agonist of α7nAChR 3-(2,4-dimethoxybenzyli-dene)anabaseine(GTS-21)group(5 mg/kg),with 12 rats in each group.Model of cardiac arrest was established by inducing ventric-ular fibrillation in rats with percutaneous epicardial electrical stimulation,and routine resuscitation was performed,the defibrillation times,cardiopulmonary resuscitation duration and 72 h survival rate of rats in each group were observed and recorded.Neurological damage of rats in each group was scored(NDS score).Levels of inflammatory factors TNF-α,IL-1β and IL-6 in serum,heart,lung and brain of rats were measured by ELISA.HE staining was applied to evaluate the degree of tissue damage of heart,lung and brain tis-sues of rats in each group.Expressions of α7nAChR,NF-κB/MAPKs signal pathway related proteins in heart,lung and brain of rats in each group were detected by Western blot.Results:Compared with Sham group,the number of defibrillations,duration of cardiopul-monary resuscitation,NDS score,levels of TNF-α,IL-1β and IL-6 in serum,heart,lung and brain tissues,and NF-κB p65 protein in heart,lung and brain tissues in cardiopulmonary resuscitation group increased greatly(P<0.05),the 72 h survival rate,phosphory-lation levels of α7nAChR protein,ERK1/2,JNK and p38MAPK in heart,lung and brain tissues decreased greatly after restoration of spontaneous circulation(P<0.05),cells and neurons in the CA1 region of heart,lung and hippocampus were severely damaged.Com-pared with cardiopulmonary resuscitation group,change trend of relevant indicators in electroacupuncture group was opposite to the above,damage of cells and neurons in rat heart,lung and brain tissue was reduced(P<0.05).αBGT reduced the anti-inflammatory effect of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).GTS-21 enhanced the anti-inflammatory protection of electroacupuncture on cardiopulmonary resuscitation rats(P<0.05).Conclusion:Electroacupuncture has anti-inflammatory and pro-tective effects on cardiopulmonary resuscitation rats based on CAP,and NF-κB/MAPKs participate in α7nAChR mediated CAP.

Gastric cancer remains one of the most prevalent and lethal malignancies of the digestive tract worldwide,underscoring the urgent need for more effective targeted therapeutic strategies.Poly(ADP-ri-bose)polymerase(PARP)inhibitors have demonstrated remarkable efficacy in tumors with homologous recombination repair(HRR)deficiency;however,their clinical application in gastric cancer remains limited.Clinical evidence suggests that patients harboring Helicobacter pylori infection in combination with HRR gene mutations exhibit a significantly elevated risk of developing gastric cancer,thereby supporting the potential benefit of PARP inhibition in this setting.In this study,a kinase inhibitor library was screened in combination with the PARP inhibitor olaparib in gastric cancer cells.And we identify the cy-clin-dependent kinase 8/19(CDK8/19)inhibitor Senexin A as a compound that synergistically enhances the cytotoxic effect of PARP inhibition(P<0.05).Phenotypic validation using CCK-8 and colony for-mation assays demonstrated that the combination treatment significantly suppressed cellular proliferation and clonogenic potential compared to either monotherapy(P<0.0001).Mechanistically,alkaline comet assays revealed a significant increase in DNA damage in the combination treatment group relative to either single-agent group(P<0.0001),suggesting that the synergistic effect results from the exacerbation of DNA damage via impaired DNA repair mechanisms.In addition,treatment with CDK8/19 inhibitors a-lone markedly increased the formation of γH2AX and 53BP1 foci in irradiated gastric cancer cells(P<0.0001),indicating inhibition of DNA damage repair pathways.Transcriptome sequencing further re-vealed that CDK8/19 inhibition impacts critical cellular pathways,including DNA repair,cell cycle reg-ulation,and RNA splicing.Co-immunoprecipitation assays confirmed that inhibition of CDK8/19 kinase activity significantly reduces the phosphorylation level of PARP1,suggesting a potential regulatory inter-action.Immunohistochemical analysis of tumor and adjacent non-tumor tissues from gastric cancer pa-tients demonstrated that CDK8 is significantly overexpressed in tumor tissues,supporting its potential as both a prognostic biomarker and a therapeutic target.Collectively,this study elucidates a mechanistic ba-sis by which CDK8/19 inhibition enhances the sensitivity of gastric cancer cells to PARP inhibitors.These findings provide a strong rationale for the combined use of CDK8/19 and PARP inhibitors as a tar-geted therapeutic strategy and offer promising translational implications for advancing personalized medi-cine in gastric cancer treatment.

Objective:To explore the correlation between serum ferritin(SF)levels and the efficacy of platelet transfusion in patients with malignant hematological diseases.Methods:Patients with malignant hematological diseases who received repeated transfusions of apheresis platelets in Department of Hematology of Aerospace Center Hospital in 2023 were selected.The platelet corrected count increment(CCI)was used to evaluate the efficacy of platelet transfusion.The correlations between sex,age,disease type,transplantation history,red blood cell transfusion history,and SF level and the efficacy of platelet transfusion were analyzed.Results:A total of 87 patients were included,with a cumulative 326 person-times platelet transfusions.As suggested by one-way analysis of variance,compared with the patients in the age groups of 24-45 years old and 46-66 years old,the patients in the age group of 2-23 years old had a better efficacy of platelet transfusion(P=0.004,P=0.004).There was no significant difference in the efficacy of platelet transfusion between the patients in the age group of 24-45 years old and those in the age group of 46-66 years old(P=0.876).Compared with the patients who had a history of red blood cell transfusion within 3 days,the patients without a history of red blood cell transfusion within 3 days had a better efficacy of platelet transfusion(P＜0.001).Compared with the groups with SF levels of 1.44-2.78 ng/L and＞2.78 ng/L,the group with SF levels＜1.44 ng/L had a better efficacy of platelet transfusion(P=0.028,P＜0.001).Compared with the group with SF levels＞2.78 ng/L,the group with SF levels of 1.44-2.78 ng/L had a better efficacy of platelet transfusion(P=0.001).After adjusting for age and the history of red blood cell transfusion,the transfusion efficacy of the group with SF levels＜1.44 ng/L was better than that of the groups with SF levels of 1.44-2.78 ng/L and＞2.78 ng/L(P=0.021,P＜0.001);Compared with the group with SF levels＞2.78 ng/L,the group with SF levels of 1.44-2.78 ng/L had a better efficacy of platelet transfusion(P=0.001).Both univariate and multivariate linear regression models showed that SF levels were negatively correlated with the efficacy of platelet transfusion(P＜0.001).Conclusion:There is a negative correlation between SF levels and the efficacy of platelet transfusion in patients with malignant hematological diseases.Detection of SF levels may provide guidance for predicting the efficacy of platelet transfusion.