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Chaihu and Longgu Muli Decoction has demonstrated significant efficacy in the treatment of tachyarrhythmia accompanied by anxiety and depression. However, there is a lack of standardized guidelines for its clinical application. In this study, the Chaihu and Longgu Muli Decoction was investigated through extensive research on ancient and modern literature, as well as a collection of clinical medical records. The basic information, medication details, and diagnostic information from medical records, personal experience literature, and clinical cases in the treatment of tachyarrhythmia accompanied by anxiety were extracted and analyzed to preliminarily identify the prescription characteristics and syndrome patterns. Subsequently, the Delphi method was employed to construct an item pool based on the data obtained in the first step. An expert questionnaire was prepared to collect scores and revision opinions from experts regarding these items. After statistical analysis and group discussions, a second round of questionnaires was formed by screening out certain items. This process was repeated until a final item set for the treatment of tachyarrhythmia accompanied by anxiety with Chaihu and Longgu Muli Decoction was determined. These findings provided guidance for clinical prescription practices. By extracting 71 syndromes and signs, as well as 33 tongue and pulse characteristics, the main syndrome features included palpitations, chest tightness, irritability, etc., which were basically consistent with the ancient syndromes. Through frequency analysis and group discussions, 71 items were screened out. After screening, modification, and primary and secondary division, 11 main diagnostic items and 10 secondary diagnostic items were determined. On this basis, the research team believes that Chaihu and Longgu Muli Decoction is mainly indicated for the following syndromes in the treatment of tachyarrhythmia accompanied by anxiety(palpitations, poor sleep, bitter taste, dry mouth, irritability/easily angered/anxiety/fearfulness/easily startled, red tongue with greasy yellow coating, rapid pulse, high work/life pressure, tachyarrhythmia on electrocardiogram/Holter monitor, and positive results on anxiety scale). Secondary syndromes include chest tightness, shortness of breath, feeling heavy and weak in the body, sweating, poor appetite, constipation, greasy white tongue coating, wiry pulse, slippery pulse, or knotted and intermittent pulse.
Drugs, Chinese Herbal/therapeutic use* ; Humans ; Delphi Technique ; Anxiety/complications* ; Tachycardia/psychology* ; Female ; Male ; Middle Aged ; Adult ; Aged

BACKGROUND: Biomarkers-based prediction of long-term risk of acute coronary syndrome (ACS) is scarce. We aim to develop a risk score integrating clinical routine information (C) and plasma biomarkers (B) for predicting long-term risk of ACS patients. METHODS: We included 2729 ACS patients from the OCEA (Observation of cardiovascular events in ACS patients). The earlier admitted 1910 patients were enrolled as development cohort; and the subsequently admitted 819 subjects were treated as validation cohort. We investigated 10-year risk of cardiovascular (CV) death, myocardial infarction (MI) and all cause death in these patients. Potential variables contributing to risk of clinical events were assessed using Cox regression models and a score was derived using main part of these variables. RESULTS: During 16,110 person-years of follow-up, there were 238 CV death/MI in the development cohort. The 7 most important predictors including in the final model were NT-proBNP, D-dimer, GDF-15, peripheral artery disease (PAD), Fibrinogen, ST-segment elevated MI (STEMI), left ventricular ejection fraction (LVEF), termed as CB-ACS score. C-index of the score for predication of cardiovascular events was 0.79 (95% CI: 0.76-0.82) in development cohort and 0.77 (95% CI: 0.76-0.78) in the validation cohort (5832 person-years of follow-up), which outperformed GRACE 2.0 and ABC-ACS risk score. The CB-ACS score was also well calibrated in development and validation cohort (Greenwood-Nam-D'Agostino: P = 0.70 and P = 0.07, respectively). CONCLUSIONS CB-ACS risk score provides a useful tool for long-term prediction of CV events in patients with ACS. This model outperforms GRACE 2.0 and ABC-ACS ischemic risk score.

Objective To investigate the effects of Caenorhabditis elegans(C.elegans)endogenous U6 promoters on dpy-10 gene editing efficiency.Methods We screened endogenous U6 small nuclear RNA(snRNA)genes of C.elegans from the WormBase database and constructed 14 editing plasmids targeting dpy-10 by replacing the U6r07e5.16 promoter in the pSX524 plasmid(Peft-3::cas9::tbb-2 terminator::U6 r07e5.16::dpy-10 sgRNA)through molecular cloning.Gene editing was performed in wild-type C.elegans using a standardized microinjection protocol.Gene editing efficiency and the high-efficiency gene editing index were quantified based on the screening of dpy-10 mutant phenotypes in the F1 progeny.Results A total of 15 U6 snRNA genes(r07e5.16,f35c11.9,t20d3.13,k09b11.15,k09b11.16,w05b2.8,c28a5.7,f54c8.9,k09b11.11,k09b11.12,k09b11.14,t20d3.12,f54c8.8,f54c8.10,and k09b11.13)were identified from the WormBase database.Based on the editing efficiency and high-efficiency gene editing index,the activity of these promoters was evaluated,and 4 U6 promoters(w05b2.8,c28a5.7,f54c8.9,and k09b11.11)were found to have significantly enhanced gene editing success rates,outperforming other promoters,including U6r07e5.16 and U6k09b11.12,which are commonly used in the C.elegans research community.Notably,the gRNAF+E scaffold did not show superior editing efficiency over the gRNA scaffold when paired with the optimal U6w05b2.8 promoter.Conclusion In this study,U6 promoters that significantly improve gene editing efficiency in C.elegans are identified and the critical role of promoter optimization in CRISPR-Cas9 systems is highlighted.These findings provide a valuable foundation for improving genome editing strategies and offer new ideas for optimizing the CRISPR technology applied in nematode research.

Objective To optimize the real-time quantitative polymerase chain reaction(RT-qPCR)data analysis process through mathematical principles by replacing the biased 2-??CT method with a more rigorous 2-CT method,thereby improving the accuracy of gene expression quantification analysis.Methods Essentially,the CT value serves as the exponent in a base-2 exponential equation within the logic of comparative CT method.In the traditional 2-??CT method,the arithmetic means of raw CT and ΔCT values are directly calculated and the exponential nature of CT data is overlooked,which may introduce systematic bias to the calculation results.We propose a new method,entitled the 2-CT method,in which all calculations are based on the transformation of CT values into 2-CT.This includes computing the relative initial expression levels of target and reference genes within each sample,the relative abundance of the target gene,and its fold change across groups.Statistical comparisons are then performed based on fold change values.By strictly adhering to the exponential nature of of CT values,the biases introduced by arithmetic averaging at the CT or ΔCT level are avoided.We applied this method to multiple RT-qPCR datasets to evaluate the differences between the traditional 2-??CT and the proposed 2-CT methods in gene expression quantification,as well as the effect of the differences.Results In the original dataset from LIVAK and SCHMITTGEN,the two methods produced similar results.However,in the cadmium exposure experiment,findings from the 2-??CT method indicated that 8-hour cadmium exposure caused an increase of irg-6 gene expression in Caenorhabditis elegans from 1.314-fold to 7.125-fold(P=0.000 2).In contrast,findings from the 2-CT method showed a fold change from 1.0 to 4.124(P=0.001 5),a 70%difference between the two methods.Conclusion The2-CT method provides a mathematically more rigorous approach that more accurately reflects gene expression changes,particularly in experiments with high CT variability.It offers a more reliable computational paradigm for quantitative gene expression analysis.

Objective:To investigate the inhibitory effect of tripartite motif-containing 25 (TRIM25) on the replication of Japanese encephalitis virus (JEV) in cells and its molecular mechanism.Methods:Human glioma cells (U251 cells) and Kunming mice were infected with JEV, and then the cells and brain tissue samples were collected. The transcription levels of six TRIM genes were detected by real-time PCR, and the expression of TRIM25 in cells was detected by Western blot. U251 and A549 cells overexpressed with TRIM25 and U251 cells knocked out with TRIM25 gene were constructed. Cells were infected with JEV, and the replication of JEV was detected by viral plaque assay, real-time PCR and Western blot. The interaction of TRIM25 with viral proteins was investigated by co-immunoprecipitation (Co-IP) and indirect immunofluorescence assay. The expression of IFN-β in overexpressed TRIM25 cells was detected by real-time PCR and ELISA.Results:JEV infection promoted the expression of TRIM25 in cells and mouse brain tissues. TRIM25 overexpression restricted JEV replication in U251 and A549 cells, while TRIM25 knockout enhanced JEV replication. TRIM25 overexpression upregulated the level of IFN-β in cells. TRIM25 interacted with JEV capsid protein and promoted the degradation of capsid protein.Conclusion:TRIM25 can inhibit the replication of JEV in cells by upregulating IFN-β and promoting the degradation of JEV C protein.

Objective:This study aimed to investigate the regulatory role and mechanism of myosin heavy chain 9 (MYH9) in the invasion of Zika virus (ZIKV) into human glioma cells (U251).Methods:Utilizing CRISPR/Cas9 technology, MYH9-knockout U251 cells (U251-MYH9 KD) were constructed. Following ZIKV infection, the protein expression levels, RNA load, and viral titer of ZIKV were detected through western blot (WB), Real-time fluorescence quantitative polymerase chain reaction (qPCR), and plaque formation assays, respectively. The infection efficiency of ZIKV in U251 cells treated with the MYH9 inhibitor blebbistatin was assessed. The binding and internalization efficiency of ZIKV were measured in U251-MYH9 KD cells. The interaction between MYH9 and the ZIKV envelope protein (E) was studied using co-immunoprecipitation (Co-IP). The effects of soluble MYH9 recombinant protein and anti-human MYH9 antibodies on ZIKV infection were evaluated by qPCR and plaque formation assays. Results:It was found that knockout or inhibition of MYH9 significantly suppressed ZIKV infection in U251 cells. MYH9 knockout notably inhibited the binding and internalization of ZIKV in U251 cells. MYH9 interacted with the ZIKV E protein, and both MYH9 recombinant protein and anti-human MYH9 antibodies, by blocking the binding of ZIKV E protein to cell surface MYH9, inhibited ZIKV infection in U251 cells in a dose-dependent manner.Conclusions:MYH9 facilitates ZIKV invasion into U251 cells through interaction with the ZIKV E protein.

Objective To observe the application effect of bedside contrast-enhanced ultrasound combined with gas-water alternating injection method in severe patients with nasointestinal catheterization.Methods A total of 150 severe patients who were admitted to intensive care unit(ICU)and emergency intensive care unit(EICU)and required nasointestinal catheterization were collected.Patients were separated into the blind insertion method group(catheterization without other auxiliary equipment,n=50),the ultrasound method group(ultrasound-guided catheterization,n=53),and the combined method group(bedside contrast-enhanced ultrasound combined with gas-water alternating injection method,n=47)according to the wishes of patients or their families.The catheterization success rate,feeding standard-reaching rate within one week,catheterization time and incidence of adverse events were compared between three groups.Kappa test was used to compare the consistency of three methods with X-ray examination results after catheterization.Receiver operating characteristic(ROC)curve was used to evaluate the catheterization effect.Results The catheterization success rate and the feeding standard-reaching rate within one week were increased successively in the blind insertion method group,the ultrasound method group and the combined method group,and the catheterization time shortened in turn(P＜0.05).The consistency of blind insertion method,ultrasound method,combined method with X-ray examination was strong(Kappa=0.730),very strong(Kappa=0.835)and very strong(Kappa=0.911),respectively.Results of ROC curve showed that the areas under the ROC curve of the blind insertion method group,the ultrasound method group and the combined method group increased in turn,which were 0.838(95%CI:0.661-1.000),0.918(95%CI:0.763-1.000)and 0.988(95%CI:0.959-1.000),and the combined method group had the best catheterization effect.There were no significant differences in the incidence of adverse events such as hiccup,abdominal distension,diarrhea,aspiration and gastrointestinal bleeding between the three groups(P>0.05).Conclusion Bedside contrast-enhanced ultrasound combined with gas-water alternating injection method can improve the success rate of nasointestinal catheterization in severe patients,shorten the catheterization time and without increasing the incidence of adverse events.

To explore the effects of different blood loss on liver and kidney function,blood lactic acid levels,and oxidative stress indexes after hemorrhagic shock resuscitation in dogs,10 healthy Chinese rural dogs were randomly divided into 1.5 h blood loss resuscitation group(HSRA group)and 3.5 h blood loss resuscitation group(HSRB group).The changes in liver and kidney function,blood lactic acid,and oxidative stress-related indexes were detected at 2,6,24,48 and 72 h after re-suscitation.The results showed that the liver function indexes of TBIL,ALT,and AST in the HSRB group were higher than those in the HSRA group at each time point after resuscitation.There was no significant change in renal function indexes between the two groups.The level of Lac in the HSRB group was significantly higher than that in the HSRA group at 2 and 6 h after resuscitation.CAT activity in the HSRB group was significantly lower than that in the HSRA group at 2 h after resuscitation.GSH-px activity in the HSRB group was significantly lower than that in the HSRA group at 2,6 and 24 h after resuscitation.SOD activity in the HSRB group was significantly lower than that in the HSRA group at 24 h after resuscitation.MDA content in the HSRB group was sig-nificantly higher than that in the HSRA group at 2,6,24 and 48 h after resuscitation.The results showed that HS could cause liver injury and oxidative stress after resuscitation,and the degree of liver injury and oxidative stress injury in dogs increased with the prolongation of blood loss.

AIM:To investigate the involvement of HIF-1α/VEGF regulation in its anti-angiogenesis effects using adjuvant-induced arthritis(AIA)rats.METHODS:AIA rats were orally treated by QLY ex-tract for 24 days.After sacrifice,the joints were subjected to histological examination,while the blood was used in ELISA or biochemical tests.In ad-dition,HUVEC cells were treated by QLY in vitro.MTT,wound-healing and tube-formation experi-ments were then performed.Expression of some relevant proteins in cells were investigated.RE-SULTS:Compared to the healthy controls,obvious synovial invasion and angiogenesis occurred in AIA rats.Blood levels of HIF-1α,VEGF,PDGF,and TGF-β1 were increased,while the ratio of MDA/SOD was decreased a lot.After QLY treatment,all these ab-normalities were attenuated.In vitro experiments,QLY showed notable potentials in inhibiting prolifer-ation,migration and tube-constructing abilities of HUVEC cells.Furthermore,it suppressed the ex-pression of p-MEK/MEK and p-ERK/ERK.CONCLU-SION:QLY can reduce the pathological functions of vascular endothelial cells by inhibiting HIF-1α/VEGF,and it consequently eased AIA-related abnor-mal angiogenesis in AIA rats'joints.