Objective: To propose an improved method for constructing the internal quality control (IQC) framework for ELISA assays and validate its efficacy by statistically analyzing IQC data from nine blood center laboratories. Methods: 1) IQC data was collected from nine blood centers and analyzed using a domestic HBsAg ELISA detection kit as an example. 2) Differences between IQC values across batches within Blood Center 1 were assessed. 3) Statistical analyses were performed on batch usage, number of batches used, days of use, number of QC points, batch-specific means, and coefficients of variation (CV) across all nine centers. 4) Using the improved construction method for IQC framework, provisional and permanent frames were established for batches within Blood Center 1 and Blood Center 9, followed by outlier determination. Results: 1) Statistically significant differences were observed in IQC data between batches within Blood Center 1 (P<0.01). It is recommended that both the control material/reagents and the control chart framework be replaced simultaneously. 2) There were substantial differences among 9 blood centers regarding the control material/reagent lot numbers used, the number of QC runs per batch, and the QC values for identical lots. Therefore, individual laboratories should establish their own IQC chart frameworks. 3) The improved IQC framework construction method for ELISA assays is as follows: provisional frames are established via frame-shifting, using the pre-experimental mean and cumulative coefficient of variation (CV) from the preceding batch. For batches used >20 days with >20 QC points, permanent frames are constructed by aggregating in-control data accumulated over ≥20 days with ≥20 points to calculate cumulative mean and standard deviation. The provisional and permanent frames constructed by this method identified all 26 extreme outliers across Blood Centers 1 and 9 as out-of-control. Among the 218 general outliers, 10 were classified as normal by the provisional frames, while the remainder were designated as warnings or out-of-control. This method effectively monitors assay stability. Conclusion: Based on the statistical analysis of IQC practices across blood centers of varying scales, combined with the inherent characteristics of ELISA assays and the batch-to-batch instability of reagents/QC materials, it is recommended to reconstruct QC charts upon lot changes. The proposed method—utilizing frame-shifting for provisional frames and establishing permanent frames based on cumulative data—is applicable to blood center laboratories of differing sizes and effectively monitors the stability of the ELISA assay process.

ObjectiveTo evaluate the effects and safety of the optimized new Shengmai Powder （优化新生脉散方） on exercise tolerance in patients with chronic heart failure （CHF） of qi deficiency， blood stasis， and fluid retention syndrome. MethodsA randomized， double-blind， placebo-controlled trial was conducted. A total of 78 CHF patients with qi deficiency， blood stasis， and fluid retention syndrome were recruited and randomly assigned to a treatment group （39 cases） and a control group （39 cases）. On the basis of conventional western medical therapy， patients in the treatment group additionally received the optimized new Shengmai Powder granules， while the control group was given an oral placebo of optimized new Shengmai Powder granules. Patients in both groups took 30.6 g each time， twice a day， mixed with water for administration， with a total treatment course of 4 weeks. The primary outcomes were 6-minute walk distance （6MWD） and peak oxygen uptake （Peak VO₂） measured by cardiopulmonary exercise testing. Secondary outcomes included New York Heart Association （NYHA） functional classification， B-type natriuretic peptide （BNP） levels， cardiac function indexes including left ventricular ejection fraction （LVEF）， left ventri-cular end-systolic diameter （LVESD） and left ventricular end-diastolic diameter （LVEDD）， Minnesota Living with Heart Failure Questionnaire （MLHFQ） scores， and scores of four diagnostic information of traditional Chinese medicine （TCM）. All indicators were assessed once before and after treatment respectively. Safety indicators were evaluated， and adverse events during the trial were recorded. ResultsAll patients in both groups were included in the full ana-lysis set （FAS） and safety set （SS）. Compared with baseline， the 6MWD and Peak VO₂ of cardiopulmonary exercise test in the treatment group significantly increased after treatment， while the MLHFQ scores， serum BNP levels and scores of TCM four diagnostic information significantly decreased， and the NYHA cardiac function grade significantly improved （P＜0.01）. After treatment， the 6MWD and Peak VO₂ of cardiopulmonary exercise test， as well as their changes from baseline in the treatment group were higher than those in the control group； the MLHFQ scores， serum BNP levels and scores of TCM four diagnostic information in the treatment group were lower than those in the control group； and the improvement of NYHA cardiac function grade in the treatment group was superior to that in the control group （P＜0.01）. There was no statistically significant differences in all indicators after treatment in the control group （P＞0.05）. The incidence of adverse events was 5.1% （2/39） in the treatment group and 2.6% （1/39） in the control group， with no statistically significant difference between groups （P＞0.05）. ConclusionOn the basis of conventional western medicine treatment， the addition of the optimized new Shengmai Powder can further improve exercise tolerance， cardiac function and quality of life in patients with CHF of qi deficiency， blood stasis and fluid retention syndrome， and show good safety.

OBJECTIVE To evaluate the effect of clinical pharmacists participating in the treatment of chronic heart failure （CHF） based on the clinical pharmacy pathway （CPP）. METHODS Totally 226 CHF patients recruited from August 24th， 2024 to March 14th， 2025， were divided into an observation group and a control group based on the random number table method， with 113 cases in each group. All patients were treated with conventional therapy. The observation group was additionally given CPP management （including pharmaceutical care during hospitalization， the formulation of individualized discharge medication regimens， and pharmaceutical follow-up after discharge）. The cardiac function parameters at admission， at discharge， at 3 and 6 months after discharge， drug use at 6 months after discharge， economic indicators， as well as the readmission rate and mortality rate at 6 months after discharge were compared between the two groups. Morisky Medication Adherence Scale-8 Items （MMAS-8）， Somatic Self-rating Scale （SSS） and Patient Health Questionnaire-9 （PHQ-9） scores were compared at admission， at discharge and at 3 and 6 months after discharge. RESULTS Six months after discharge， 24 patients dropped out. Eventually， 104 patients in the observation group and 98 patients in the control group completed the study. Compared with at admission， New York Heart Association （NYHA） cardiac functional classification， left ventricular ejection fraction （LVEF） and N -terminal pro-B-type natriuretic peptide （NT-proBNP） of both groups of patients at discharge as well as at 3 and 6 months after discharge were significantly improved； moreover， the improvements at 3 and 6 months after discharge were significantly better than those at discharge. Meanwhile， the above indexes （except for NYHA cardiac functional classification at discharge， NT-proBNP and NYHA cardiac functional classification at 3 months after discharge） of the observation group at discharge， at 3 and 6 months after discharge were significantly better than the control group （ P ＜0.05）. The utilization rates of angiotensin converting enzyme inhibitor （ACEI）/angiotensin Ⅱ receptor blocker （ARB）/angiotensin receptor neprilysin inhibitor （ARNI）， the proportion of β-blockers reaching the target dose， the utilization rate of sodium-glucose linked transporter 2 inhibitor （SGLT2i）， and the proportion of SGLT2i reaching the target dose in the observation group were significantly higher than the control group （ P ＜0.05）， and the proportion of drugs and readmission rate were significantly lower than the control group （ P ＜0.05）. Compared with at admission， MMAS-8 scores of the patients in the observation group at discharge， at 3 and 6 months after discharge were significantly increased， while SSS and PHQ-9 scores were significantly lowered （ P ＜0.05）. And all the above scores gradually decreas ed with the extension of discharge time （ P ＜0.05）. CONCLUSIONS Clinical pharmacists can utilize CPP to significantly improve patients’ cardiac function， medication adherence， somatic symptoms and depression. Additionally， they can significantly improve the utilization rates of ACEI/ARB/ARNI and SGLT2i， as well as the proportion of target doses of β-blockers and SGLT2i， while simultaneously reducing readmission rates.

Objective To examine the impact of extreme temperature and drought stress on the survival of Bulinus globosus, so as to provide the theoretical evidence for the genomic research of Bulinus in absence of reference genes. Methods B. globosus snail samples were collected from Kiwani Shehia in Pemba Island, Zanzibar, Tanzania, and offspring snails were obtained through laboratory breeding and reproduction. A total of 120 10-week-old B. globosus snails from the same generation were selected and randomly assigned into four groups, including the high-temperature drought (HD) group, normal temperature drought (D) group, low-temperature drought (LD) group, and the control (C) group, of 30 snails in each group. Snails in HD, D, and LD groups were placed in beakers containing dry soil at the bottom and subsequently housed in climate chambers at 35, 26 ℃, and 10 ℃, respectively, while snails in Group C were maintained in 500 mL petri dishes containing dechlorinated tap water at 26 ℃. Following 3 days of breeding, living snails in each group were collected, and soft tissues were dissected and isolated. Total RNA was extracted from snail soft tissues for library construction, followed by high-throughput sequencing on the Illumina HiSeq 4000 sequencing system. De novo transcriptome assembly was performed using the Trinity software, and the longest transcripts were selected as unigenes. Gene functional annotations of unigenes were conducted using the Diamond software against Gene Ontology (GO) knowledgebase, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database, NCBI non-redundant (NR) protein sequences database, Protein Family (Pfam) database, and UniProtKB/Swiss-Prot (Swiss-Prot) knowledgebase. GO and KEGG enrichment analyses of differentially expressed genes (DEGs) were performed using the topGO and clusterProfiler software, respectively. In addition, four relevant genes were selected for validation using a real-time quantitative PCR (qRT-PCR) assay to verify the reliability of transcriptome sequencing results. Results Following 3 days of breeding, there were 7, 20, 28, and 30 survival B. globosus snails in HD, LD, D, and C groups, with corresponding survival rates of 23.33% (7/30), 66.67% (20/30), 93.33% (28/30), and 100.00% (30/30), respectively (χ² = 52.72, P < 0.001). De novo transcriptome assembly generated 176 942 unigenes, with annotation rates of 0.98%, 13.49%, 26.46%, 12.48%, and 14.39% against GO knowledgebase, KEGG pathway database, NR protein sequences database, Pfam database, and Swiss-Prot knowledgebase, respectively. There were 33 up-regulated and 72 down-regulated genes in Group D, 483 up-regulated and 815 down-regulated genes in Group HD, and 245 up-regulated and 172 down-regulated genes in Group LD relative to in Group C. Following removal of overlapping genes across groups and unmatched genes, 11 candidate genes were identified. GO and KEGG analyses revealed 3 heat shock protein (HSP)-related DEGs in these 11 candidate genes, which were annotated as HSP12.2, HSP70, and HSP20 genes and were all significantly up-regulated in each treatment group. Three immune and nervous system-related DEGs were identified, and were all significantly down-regulated in each treatment group, which were involved in the neural cell adhesion molecule L1-like protein pathway, fibrinogen binding protein pathway, and leukocyte elastase inhibitor-like protein pathway. qRT-PCR assay quantified that the expression trends of four genes related to temperature and drought stress across different treatment groups were highly consistent with transcriptome sequencing data. Conclusion The survival rate of B. globosus significantly reduces under combined stresses of extreme temperature and drought, possibly due to an imbalance in its cellular homeostasis regulatory system.

Objective To establish quality evaluation method of Andrographis paniculata standard decoction by UPLC. Methods 21 batches of Andrographis paniculata standard decoctions were prepared according to the standardization method of TCM decoction pieces. The UPLC characteristic chromatograms analysis method was established. With andrographolide as a reference, quantitative analysis of multi-components by single marker （QAMS） was established for new neoandrographolide, 14 deoxyandrographolide and dehydrated andrographolide. The results were compared with the external standard method （ESM） to determine the accuracy of the method. Results Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition） was used to analyze and compare the characteristic chromatograms, and seven common peaks were determined and five were identified including luteolin-7-O-β-D-glucuronide, andrographolide, neoandrographolide, 14-deoxyandrographolide and dehydroandrographolide. The RSDs of content results of each component by QAMS and ESM were all within 3%. Conclusion The determination method was reliable and accurate, which could be used to reflect the intrinsic quality of Andrographis paniculata standard decoction comprehensively and provide the basis for the quality evaluation of Andrographis paniculata formula granules and other preparations.

Objective To explore the predictive role of pathological risk score of gastric cancer (PRS_GC) in the prognosis of gastric cancer patients, and to further investigate the correlation between PRS_GC and tumor biological behavior. Methods Clinicopathological data were retrospectively collected from 1 120 patients with gastric cancer who underwent surgical resection at Zhongshan Hospital, Fudan University, between April 2006 and November 2019. PRS_GC was calculated using the previously established DeepRisk model, and patients were divided into high and low PRS_GC groups (560 patients per group) based on the median PRS_GC value. The proportions and spatial distances of immune cell subsets in gastric cancer tissues were assessed between the two groups. Data from stomach adenocarcinoma samples in The Cancer Genome Atlas was integrated to analyze PRS_GC-associated genetic mutations and signaling pathways. Results The survival rate of the high PRS_GC group was lower than that of the low PRS_GC group, and the recurrence rate was higher than that of the low PRS_GC group (P<0.000 1). Compared with the low-PRS_GC group, the high-PRS_GC group exhibited increased infiltration of regulatory T cells (Tregs) and neutrophils within the tumor region (P＜0.05), while the infiltration levels of natural killer T cells and cytotoxic T cells were markedly decreased (P＜0.05). Spatial analysis revealed that a shortened spatial distance between CD4⁺ T cells and Tregs was closely associated with poor prognosis in the high-PRS_GC group (P＝0.03). Furthermore, in the high-PRS_GC group, the number of TIM3⁺ cells showed a positive correlation with the infiltration levels of Tregs (r＝0.69, P＝0.01), CD8⁺ T cells (r＝0.61, P＝0.04), and CD4⁺ T cells (r＝0.67, P＝0.02). Gene set enrichment analysis indicated that a high PRS_GC was associated with the activation of pathways related to actin cytoskeleton regulation, the cGMP-PKG pathway, the Hippo pathway, and tumor proteoglycans. Conclusion A high PRS_GC is associated with an immunosuppressive tumor microenvironment, as well as tumor invasion and metastasis. It effectively predicts the prognosis of patients with gastric cancer and, by doing so, provides a potential theoretical basis for developing individualized strategies that combine targeted therapy with immunotherapy.

Prostate cancer is a prevalent malignant tumor of the male genitourinary system in China, with a persistently elevated incidence rate.Given the insidious onset, propensity for metastasis in advanced stages, and poor efficacy of traditional treatments, new hormone therapies have garnered increasing attention in recent years.A number of clinical studies have demonstrated that second-generation androgen receptor inhibitors (SGARIs), including enzalutamide, apalutamide, darolutamide, and rezvilutamide, can be beneficial in different stages of advanced prostate cancer.Furthermore, their early application has been shown to delay disease progression while maintaining a controlled safety profile.At present, SGARIs have increasingly become the primary therapeutic agents for advanced prostate cancer in various stages.This article reviews the clinical research progress of SGARIs in recent years, with the aim of offering valuable clinical reference for clinicians.

Subarachnoid hemorrhage (SAH) is a serious intracranial hemorrhage characterized by acute bleeding into the subarachnoid space. The effects of shikonin, a natural compound from the roots of Lithospermum erythrorhizon, on oxidative stress and blood–brain barrier (BBB) injury in SAH was evaluated in this study. A rat model of SAH was established by endovascular perforation to mimic the rupture of intracranial aneurysms. Rats were then administered 25 mg/kg of shikonin or dimethylsulfoxide after surgery. Brain edema, SAH grade, and neurobehavioral scores were measured after 24 h of SAH to evaluate neurological impairment. Concentrations of the oxidative stress markers superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA) in the brain cortex were determined using the corresponding commercially available assay kits. Evans blue staining was used to determine BBB permeability. Western blotting was used to quantify protein levels of tight junction proteins zonula occludens-1, Occludin, and Claudin-5. After modeling, the brain water content increased significantly whereas the neurobehavioral scores of rats with SAH decreased prominently. MDA levels increased and the levels of the antioxidant enzymes GSH and SOD decreased after SAH. These changes were reversed after shikonin administration. Shikonin treatment also inhibited Evans blue extravasation after SAH. Furthermore, reduction in the levels of tight junction proteins after SAH modeling was rescued after shikonin treatment. In conclusion, shikonin exerts a neuroprotective effect after SAH by mitigating BBB injury and inhibiting oxidative stress in the cerebral cortex.

ObjectiveTo investigate the features and mechanism of action of intestinal flora in patients with nonalcoholic fatty liver disease （NAFLD） and Helicobacter pylori （HP） infection by comparing the changes in intestinal flora between the healthy population， the patients with HP infection， the patients with NAFLD， and the patients with NAFLD and HP infection. MethodsThis study was conducted among the 19 patients with NAFLD （NAFLD group）， 19 patients with HP infection （HP group）， and 19 patients with NAFLD and HP infection （NAFLD+HP group） who were admitted to The Second Affiliated Hospital of Henan University of Science and Technology from March 1， 2023 to April 30， 2024， and 20 individuals undergoing physical examination were enrolled as control group. Fecal samples were collected， total DNA was extracted for PCR amplification， and 16S rDNA sequencing was performed to compare the features of intestinal flora between the four groups. An analysis of variance was used for comparison of continuous data between multiple groups， and the chi-square test was used for comparison of categorical data between multiple groups. The Mann-Whitney U test or the Kruskal-Wallis H test was used for comparison of the species in intestinal flora. ResultsThe NAFLD+HP group showed a tendency of reduction in flora abundance compared with the other three groups. There was a significant difference in flora distribution between the NAFLD+HP group and the NAFLD group and between the NAFLD group and the control group （P<0.05）. At the phylum level， the top three species in the NAFLD+HP group were Firmicutes （59.94%）， Proteobacteria （17.00%）， and Actinobacteria （14.75%）， with an increase in the proportion of Proteobacteria and a reduction in the proportion of Actinobacteria compared with the other three groups. At the genus level， the top five dominant bacteria in the NAFLD+HP group were Bifidobacterium， Streptococcus， Escherichia-Shigella， Agathobacter， and Ruminococcus gnavus_group. Compared with the NAFLD group， the NAFLD+HP group had increases in the abundance of Streptococcus， Veillonella， and Rothia and reductions in the abundance of Dialister and Ruminococcus toraues_group. Compared with the HP group， the NAFLD+HP group had reductions in the abundance of Collinsella， Subdoligranulum， Catenibacterium， and Porphyromonas and increases in the abundance of Citrobacter and Olsenella （all P<0.05）. ConclusionPatients with NAFLD and HP infection have changed in intestinal flora. These flora may be the intestinal microecological factors for HP infection in promoting the development and progression of NAFLD.

Objective This study aims to elucidate the functional mechanism through which seminal plasma exosomal miR-26a-5p and its target genes contribute to idiopathic teratozoospermia,with emphasis on their regulatory pathways in sperm morphogenesis defects,thereby establishing a theoretical foundation for novel diagnostic markers and targeted therapies.Methods A case-control study design was adopted,enrolling 154 male subjects(84 in the teratozoospermia group[TZS]and 70 in the normal control group[NC]).Seminal plasma exosomes were isolated to screen differentially expressed miRNAs,followed by prediction and analysis of target genes and signaling pathways.Dual-luciferase reporter gene assays were employed to validate the direct binding of miR-26a-5p to PTEN.Quantitative real-time PCR and Western blot were used to measure miRNA and protein expression levels.Spearman correlation analysis evaluated relationships between miR-26a-5p,PTEN,and sperm morphological parameters,while ROC curve analysis assessed diagnostic efficacy.Results Seminal plasma exosomal miRNA sequencing identified 11 differentially expressed miRNAs,with miR-26a-5p significantly upregulated in the TZS group(P<0.05)and negatively correlated with the percentage of normal sperm morphology in TZS(r=-0.762,P<0.05).PTEN was confirmed as a direct target of miR-26a-5p,with its expression significantly reduced in the TZS group(P<0.05)and positively correlated with normal sperm morphology(r=0.821,P<0.05).A negative correlation was observed between miR-26a-5p and PTEN(r=-0.878,P<0.05).Dual-luciferase assays demonstrated that miR-26a-5p specifically inhibited PTEN 3'UTR activity(45%reduction in luciferase activity,P<0.05).Functional enrichment analysis revealed that the miR-26a-5p-PTEN axis exacerbates oxidative stress,DNA damage,and abnormal spermatocyte division by regulating the PI3K/AKT/mTOR,p53,Wnt/β-catenin,and NF-κB pathways.ROC analysis showed diagnostic efficacy for teratozoospermia with AUCROC values of 0.785(sensitivity 78.9%,specificity 71.3%)for miR-26a-5p and 0.754(sensitivity 73.3%,specificity 75.1%)for PTEN.Conclusions miR-26a-5p suppresses PTEN through targeted inhibition,activating PI3K/AKT/mTOR signaling pathways while impairing DNA repair functions.This cascade leads to accumulated oxidative stress and sperm morphological abnormalities.The combined pattern of elevated seminal plasma exosomal miR-26a-5p expression and reduced PTEN levels demonstrates diagnostic potential for idiopathic teratozoospermia,with its molecular mechanism providing a theoretical foundation for biomarker development.Targeted silencing of miR-26a-5p,either alone or in combination with AKT/mTOR inhibitors and antioxidant therapy,may represent a novel strategy for improving sperm morphology.