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.

Objective To investigate the effects of Fuzheng Touxie Prescription on pulmonary injury of Pseudomonas aeruginosa pneumonia(PAP)in rats;To discuss its possible mechanism.Methods A total of 36 SD rats were divided into blank group,model group,piperacillin group and Fuzheng Touxie Prescription low-,medium-and high-dosage groups according to random number table method,with 6 rats in each group.Except for the blank group,0.1 mL of Pseudomonas aeruginosa suspension with a concentration of 3×109 CFU/mL was injected through oral catheterization.In the blank group,0.1 mL sterile saline was injected by oral catheterization.Fuzheng Touxie Prescription low-,medium-and high-dosage groups were gavaged 1.6 mL of Fuzheng Touxie Prescription solution with the concentration of 0.43,0.86 and 1.73 g/mL respectively,the blank group and model group were gavaged equal volume of distilled water,piperacillin group was intraperitoneal injected 0.53 mL piperacillin tazobactam sodium solution with a concentration of 0.11 g/mL,once a day,for consecutive 3 days.HE staining was used to observe the pathological changes of lung tissue,lung injury score was performed and bronchial wall thickness was measured,flow cytometry was used to detect Th17/Treg ratio in peripheral blood,the serum contents of RORγt,FoxP3,TGF-β and inflammatory factors TNF-α,IL-6 and IL-10 were detected by ELISA,the mRNA expression of RORγt,FoxP3 and inflammatory factors in lung tissue were detected by RT-qPCR,the protein expressions of RORγt,FoxP3 and inflammatory factors in lung tissue were detected by Western blot.Results Compared with the blank group,a large amount of inflammatory cell infiltration was observed in lung tissue of the model group rats,accompanied by bronchial wall hyperplasia,alveolar dilation,interstitial edema,and a significant increase in lung injury score and bronchial wall thickness(P<0.01),the proportion of Th17 in peripheral blood increased(P<0.01),the proportion of Treg decreased(P<0.01),and the Th17/Treg ratio increased(P<0.05),the contents of serum RORγt,TNF-α,TGF-β and IL-6 significantly increased(P<0.01),the contents of FoxP3 and IL-10 significantly decreased(P<0.01),the mRNA and protein expressions of RORγt,TNF-α,TGF-β and IL-6 in lung tissue significantly increased(P<0.01),while the mRNA and protein expressions of FoxP3 and IL-10 significantly decreased(P<0.01).Compared with the model group,the alveoli of rats in each treatment group were relatively intact and the structure was clearer,inflammatory cell infiltration,pulmonary interstitial edema and congestion were reduced,and lung injury scores and bronchial wall thickness were reduced(P<0.05,P<0.01),the proportion of Th17 in peripheral blood decreased(P<0.05),the proportion of Treg increased(P<0.05),and the Th17/Treg ratio decreased(P<0.05)in Fuzheng Touxie Prescription high-dosage group and piperacillin group,the contents of serum RORγt,TNF-α,TGF-β and IL-6 significantly decreased in Fuzheng Touxie Prescription medium-and high-dosage groups and piperacillin group(P<0.05,P<0.01),the contents of FoxP3 and IL-10 significantly increased(P<0.05,P<0.01),the mRNA and protein expressions of RORγt,TNF-α,TGF-β and IL-6 in lung tissue significantly decreased(P<0.05,P<0.01),the mRNA and protein expressions of FoxP3 and IL-10 significantly increased(P<0.05,P<0.01).Conclusion Fuzheng Touxie Prescription can improve pulmonary inflammatory symptoms,reduce the level of inflammatory factors and increase the level of anti-inflammatory factors in PAP rats.The mechanism may be through the TGF-β/RORγt/FoxP3 pathway to regulate immune balance.

As a cheap and effective antibiotic,sulfonamides are often used in animal husbandry.However,their residues in animal-derived foodstuffs will threaten human health.Consequently,a high-performance liquid chromatography(HPLC)method integrated with supramolecular solvent microextraction was successfully established for simultaneous quantification of sulfonamide residues sulfachlorpyridazine,sulfamethoxazole,sulfamethoxypyridazine and sulfadimethoxine in milk matrices.This approach exhibited prominent characteristics of operational simplicity,environmental sustainability,and high extraction efficiency.The supramolecular solvents prepared by tributyl octylphosphine tetrafluoroborate and tetrahydrofuran were employed as extraction solvents.The analytes underwent isolation and concentration via dispersive liquid-liquid microextraction(DLLME)prior to quantitative determination using high-performance liquid chromatography(HPLC).The composition and microscopic morphology of the supramolecular solvent were characterized through a series of analytical techniques,including phase diagram,Fourier transform infrared spectroscopy,scanning electron microscopy,and inverted fluorescence microscopy and so on.The density and pH value of supramolecular solvents were determined.The extraction conditions were optimized through the one-factor experiments.The experimental results demonstrated that under the optimal extraction conditions,the four kinds of sulfonamide antibiotics exhibited excellent linearity within respective detection range(R2 ≥ 0.9998)and the limits of detection(LOD)were 0.67-1.45 μg/L.Compared with literature methods,this approach offered some advantages such as simplicity of operation and less reagent consumption,and could be used for analysis and detection of sulfonamide antibiotic residues in milk samples.The present method provided technical support for food safety regulation and paved a new way for the application of supramolecular solvents in the field of extraction and separation.

Objective To verify the efficacy of a domestic human DNA quantification kit based on real-time fluorescence quantitative PCR in detecting the total human DNA concentration,male DNA concen-tration in mixed male/female DNA samples,the degree of DNA degradation and inhibitor tolerance.Methods Samples with different concentrations,different male/female ratios,different concentrations of inhibitors,and different degradation degrees were tested using the domestic human DNA quantification kit based on real-time fluorescence quantitative PCR.This kit was compared with a similar product on the market and was applied to the detection of DNA from real cases.Results This human DNA quan-tification kit can effectively detect human DNA as low as 0.001 65 ng/μL,and 6.25 pg/μL of male DNA in mixed samples with a male-to-female ratio of 1∶15 000.Even when the sample contains as high as 400 ng/μL of humic acid or 1 000 μmol/L of hemin alone,the DNA concentration can still be accurately detected.The degradation index can effectively characterize the degradation degree of the sample.This kit has been successfully applied in forensic practice.Conclusion This human DNA quan-tification kit is accurate and reliable in detection.It can accurately reflect the degradation of DNA and inhibitor tolerance.It has good performance in quantitative accuracy,determination of the male/female ratio in mixed samples,and inhibitor tolerance.It has application potential in forensic case examination.

Objective To investigate the correlation of serum brain natriuretic peptide(BNP)and D-dimer levels with cardiac function in elderly patients with chronic heart failure(CHF).Methods A retrospective analysis was conducted on 91 elderly CHF patients(observation group)admitted in our hospital from January 2021 to November 2024.They were then classified into grade Ⅰ-Ⅱsubgroup(66 cases)and grade Ⅲ-Ⅳ subgroup 25 cases according to NYHA cardiac function grading.Another 84 healthy individuals who underwent physical examinations in our hospital dur-ing the same period were selected as the control group.The levels of serum BNP and D-dimer were detected.Multivariate logistic regression model was used to analyze the risk factors affecting the severity of cardiac function in elderly CHF patients,and Spearman correlation analysis was adopted to analyze the correlation between the levels and cardiac function in elderly patients with CHF.Results Serum D-dimer and BNP levels were significantly higher in the observation group than the control group(P＜0.01),and in the grade Ⅲ-Ⅳ subgroup than the grade Ⅰ-Ⅱ sub-group(P＜0.01).Multivariate logistic regression analysis revealed that age(OR=2.815,95％CI:1.501-5.281,P=0.001),BNP(OR=2.901,95％CI:1.458-5.772,P=0.003)and D-dimer(OR=2.872,95％CI:1.501-5.495,P=0.002)were independent influencing factors for cardiac function in elderly CHF patients(P＜0.01).Spearman correlation analysis showed that serum BNP and D-dimer were negatively correlated with cardiac function(r=-0.324,P=0.023;r=-0.285,P=0.035).Conclusion The serum levels of D-dimer and BNP are significantly increased in elderly CHF patients,and the two levels are related to cardiac function.

Objective:To investigate the selection of treatment methods for renal tumors in pilots as well as the clinical significance of robot-assisted surgery by summarizing the process of robot-assisted surgery in the treatment of renal cell carcinoma in a pilot.Methods:The diagnosis, robot-assisted surgery and aeromedical assessment of a pilot with renal cell carcinoma were reported, and the related literature was reviewed.Results:The patient was a 44-year-old male transporter pilot, who was diagnosed with a left renal mass in the middle-lower pole of the kidney during a routine abdominal CT scan. After detailed preoperative evaluation that ruled out the possibility of distant metastasis and other surgical contraindications, the patient underwent robot-assisted laparoscopic partial nephrectomy in August 2022. The postoperative recovery went well, and renal function remained within normal limits at follow-ups. In March 2023, the pilot was concluded as qualified for flight after aeromedical assessment.Conclusions:Robot-assisted partial nephrectomy can significantly reduce surgical trauma, lower the risk of complications, and maximally preserve renal function. It is a good approach to renal tumors in pilots who can recover quickly.

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.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy with a poor prognosis, despite advancements in treatment. Many patients struggle with relapse or refractory disease. Investigating the role of the super-enhancer (SE) regulated gene ubiquitin-specific protease 20 (USP20) in T-ALL could enhance targeted therapies and improve clinical outcomes. Analysis of histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing (ChIP-seq) data from six T-ALL cell lines and seven pediatric samples identified USP20 as an SE-regulated driver gene. Utilizing the Cancer Cell Line Encyclopedia (CCLE) and BloodSpot databases, it was found that USP20 is specifically highly expressed in T-ALL. Knocking down USP20 with short hairpin RNA (shRNA) increased apoptosis and inhibited proliferation in T-ALL cells. In vivo studies showed that USP20 knockdown reduced tumor growth and improved survival. The USP20 inhibitor GSK2643943A demonstrated similar anti-tumor effects. Mass spectrometry, RNA-Seq, and immunoprecipitation revealed that USP20 interacted with hypoxia-inducible factor 1 subunit alpha (HIF1A) and stabilized it by deubiquitination. Cleavage under targets and tagmentation (CUT&Tag) results indicated that USP20 co-localized with HIF1A, jointly modulating target genes in T-ALL. This study identifies USP20 as a therapeutic target in T-ALL and suggests GSK2643943A as a potential treatment strategy.

OBJECTIVE: Blood culture remains the gold standard for diagnosing bloodstream infections. Clinical laboratories must ensure the quality of blood culture processes from receipt to obtaining definitive results. We examined laboratory analytical indicators associated with positive blood culture results. METHODS: Blood cultures collected from Peking Union Medical College Hospital between January 1, 2020, and December 31, 2022, were retrospectively analyzed. The mode of transportation (piping logistics delivery vs. staff), source of blood cultures (outpatient/emergency department vs. inpatient department), rotation of personnel, and time of reception (8:00-19:59 vs. 20:00-07:59) were compared between blood culture-positive and -negative results. RESULTS: Between 2020 and 2022, the total positive rate of blood culture was 8.07%. The positive rate of blood cultures in the outpatient/emergency department was significantly higher than that in the inpatient department (12.46% vs. 5.83%; P < 0.0001). The time-to-detection of blood cultures was significantly affected by the delivery mode and personnel rotation. The blood culture positive rate of the total pre-analytical time within 1 h was significantly higher than that within 1-2 h or > 2 h ( P < 0.0170). CONCLUSION Laboratory analytical indicators such as patient source, transportation mode, and personnel rotation significantly impacted the positive detection rate or time of blood culture.
Blood Culture/statistics & numerical data* ; Humans ; Retrospective Studies ; Emergency Service, Hospital/statistics & numerical data*