Objective: To systematically analyze the confirmatory positivity of different combinations of HBsAg screening results in blood testing, providing data to support the optimization of blood donor eligibility management. Methods: A retrospective analysis was conducted on blood screening data from 174 266 voluntary blood donor samples at the Chongqing Blood Center between October 2021 and September 2022. Samples with inconsistent results between the two HBsAg enzymelinked immunosorbent assays (ELISA) and individual donor nucleic acid testing (NAT) were confirmed using an electrochemiluminescence immunoassay (ECLIA) and a neutralization test. The detection efficacy of four different HBsAg ELISA reagents was compared using the HBsAg-confirmed positive samples. Results: A total of 767(0.44%) HBV-reactive (HB-sAg and/or HBV DNA reactive) samples were detected. Among them, 344 samples with discordant serological and NAT results were collected, of which 64(18.6%) were confirmed positive by neutralization test. Additionally, 5 samples that were neutralization-negative but double-reactive for HBsAg and HBV DNA were confirmed as positive according to FDA guidance, resulting in a total of 69(20.1%) confirmed HBsAg-positive samples. There were significant differences in the neutralization test confirmation rates among different screening result categories (P<0.05): The group with dual HBsAg reagent reactivity (double reactive) & NAT-negative had the highest confirmation rate (96.9%, 31/32); the group reactive to only reagent 2 (single reactive) had a rate of 25.7% (29/113); while the confirmation rates for samples reactive to only reagent 1 and samples with isolated HBV DNA positivity were extremely low [0(0/34) and 2.4%(4/165), respectively]. The four commercial reagents showed significant differences in their ability to detect confirmed positive samples that were initially single reactive (P<0.05). Conclusion: Given the performance variations among HBsAg screening reagents, thorough performance verification is essential before implementation. When NAT is negative, dual HBsAg reactivity in screening can serve as a basis for confirming infection and directly deferring blood donors. However, confirming infection in donors with single HBsAg reactivity is more challenging, necessitating supplementary tests to rule out infection risk.

Objective:To investigate the expression characteristics of Zeste enhancer of Zeste homolog 2 (EZH2) in breast cancer tissue and its influence on tumor progression and prognosis.Methods:Transcriptome data of breast cancer tissue and normal breast tissue adjacent to cancer as well as clinical data of patients were obtained from the cancer genome atlas (TCGA) database, gene expression comprehensive database and European genome phenotype archives database, and the difference of EZH2 expression was analyzed using TIMER 2.0 platform. The survival information of breast cancer patients was obtained from the Kaplan Meier Plotter database, and the overall survival time, relapse free survival time and distant metastasis free survival time of breast cancer patients with low EZH2 expression and high EZH2 expression were compared. Select 14 nude mice were selected and randomly divided into si-EZH2 group and control group, with 7 mice in each group.MCF7 culture suspensions transfected with EZH2 knockdown plasmid and control plasmid were inoculated for corresponding group. The body mass and tumor volume of two groups of nude mice inoculated with MCF7 cells were compared at different times. On the 28th day, the nude mice were euthanized and the tumors were dissected to compare the tumor mass of the two groups of nude mice. The normally distributed quantitative data was represented by xˉ ± s. Two independent sample t-tests were used for comparison between two groups, repeated measures ANOVA was used for comparison of body mass and tumor volume between two groups of nude mice at different times, and Bonferroni test was used for pairwise comparison. The comparison of survival rates was conducted using log rank test. Results:A total of 1085 breast cancer tissues and 291 normal adjacent breast tissues were included in the TCGA database. EZH2 expression in breast cancer tissues was higher than that in normal adjacent breast tissues ( P<0.05). In the Kaplan Meier Plotter database, the total survival time, relapse free survival time, and distant metastasis free survival time of breast cancer patients in the EZH2 overexpression group were shorter than those in the EZH2 low expression group ( P=0.013, <0.001, <0.001). After 7 days of inoculation with MCF7 culture suspension, significant subcutaneous tumors were observed on the left back of both groups of nude mice. On the first day, there were no statistically significant difference in body mass between the two groups of nude mice ( P>0.05); On day 7, 13, 19, 25, and 28, the body mass and tumor volume of both groups of nude mice gradually increased (nude mouse body mass: within group F=29.31, P<0.001, between groups F=234.32, P<0.001, Finteraction=16.83, P<0.001; Tumor volume: within group F=34.00, P<0.001, between groups F=193.17, P<0.001, Finteraction=35.61, P<0.001). And the body mass of the siEZH2 group nude mice was higher than that of control group (all P<0.05). On days 19, 25, and 28, tumor the volume of the siEZH2 group nude mice was smaller than that of control group (all P<0.05). On the 28th day, the mass of tumors dissected in the siEZH2 group of nude mice was lower than that in the control group [(0.30±0.07) g vs. (0.61±0.14) g, t=5.16, P<0.001]。 Conclusions:EZH2 is highly expressed in breast cancer tissues and is significantly associated with poor prognosis. Knockdown of EZH2 can significantly inhibit the proliferation and tumor formation of breast cancer cells.

Objective To investigate the relationship between high DNA stainability(HDS)and routine semen parameters.Methods Semen samples were collected from 396 men of childbearing age who were admitted to our department.Correlations of HDS with routine semen parameters and age were analyzed.Multiple linear regression analysis was performed to identify the routine semen parameters that had the greatest influence on HDS.The correlation of HDS and DNA fragmentation index(DFI)with routine semen parameters and age in 244 patients with teratozoospermia was analyzed.The 244 patients were divided into extremely severe,severe,moderate,and mild teratozoospermia groups,and differences in HDS,DFI,and routine semen parameters were compared among the four groups.Results HDS was negatively correlated with total sperm count,sperm concentration,sperm progressive motility,and normal sperm morphology rate(NSMR)(P<0.01).After adjusting for potential confounders,including total sperm count,sperm concentration,sperm progressive motility,NSMR,and DFI,NSMR had the most significant negative effect on HDS(P<0.05).In the 244 patients with teratozoospermia,HDS was negatively correlated with NSMR(P<0.01)and positively correlated with the percentage of sperm head and tail abnormalities(P<0.05),while DFI was positively correlated with the percentage of sperm tail abnormalities(P<0.01).There was a significant difference in the percentage of sperm head abnormalities among the four teratozoospermia groups.The more severe the malfor-mation,the higher the percentage of sperm head abnormalities was(P<0.001).HDS in the extremely severe teratozoospermia group was significantly higher than that in the mild and moderate teratozoospermia groups(P<0.05).No significant differences in DFI were found among the four groups(P>0.05).Conclusion HDS was closely correlated with routine semen parameters and was a crucial biomarker for assessing sperm quality,particularly the extent of sperm head abnormalities.

ObjectiveTo investigate the protective effect and mechanism of action of flavonoid combination of Alpiniae Officinarum Rhizoma （A. officinarum） against Helicobacter pylori （H. pylori） gastritis in mice. MethodsAfter acclimatization for one week， 56 SPF-grade healthy C57BL/6J mice were gavaged with mixed antibiotics for three consecutive days. They were randomly divided into a normal group， model group， positive drug group （triple therapy group）， and low- and high-dose groups （100， 200 mg·kg^-1） of flavonoid combination of A. officinarum. The H. pylori gastritis mice model was established by gavage with H. pylori bacterial suspension in each group except for the normal group. After successful modeling， mice were administrated with corresponding drugs once a day for two weeks. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in gastric tissue. Rapid urease test paper was used to detect the positive rate of H. pylori. Silver staining was used to observe the H. pylori adherence on the surface of gastric tissue. Immunohistochemistry was used to detect the protein expression of interleukin-8 （IL）-8 and myeloid differentiation factor （MyD88） in gastric tissue. The serum levels of IL-6， tumor necrosis factor-α （TNF-α）， IL-8， and IL-1β were detected by enzyme-linked immunosorbent assay （ELISA）. The expressions of phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） protein were detected by Western blot. ResultsCompared with those in the normal group， mice in the model group had lower gastric weight coefficients， higher pH of gastric juice， 100% H. pylori infection rate， and significantly changed gastric histopathology. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the model group were significantly elevated， and the serum levels of inflammatory factors IL-6， TNF-α， IL-8， and IL-1β were significantly up-regulated in mice. Compared with that in the model group， the gastric weight coefficient of mice in each treatment group of the flavonoid combinations of A. officinarum was elevated （P<0.01）， and the pH of gastric juice was reduced （P<0.01）. The infection rate of H. pylori was reduced. The expressions of IL-8 and MyD88 proteins in the gastric tissue of mice in the treatment groups were significantly reduced （P<0.01）， and the serum levels of inflammatory factors IL-6， TNF-α， IL-8， and IL-1β were significantly reduced in a dose-dependent manner （P<0.01）. The flavonoid combinations of A. officinarum down-regulated the expression of PI3K and Akt proteins in H. pylori gastritis-infected cells （P<0.01）. ConclusionThe protective effect of flavonoid combinations of A. officinarum against H. pylori gastritis is associated with the inhibition of H. pylori infection rate and regulation of PI3K/Akt signaling pathway， resulting in inhibiting the release of inflammatory factors.

Objective:To explore the differences in active components between Hedysari Radix and Astragali Radix; To explore the material basis and therapeutic functions of Hedysari Radix in promoting blood circulation and nourishing blood.Methods:The research materials used in this study were Hedysari Radix and Astragali Radix from Wudu District, Longnan City, Gansu Province. The contents of seven resource-based components-total sugars, total polyphenols, total flavonoids, crude proteins, total saponins, total alkaloids, and total dietary fiber were measured according to standard methods. Additionally, an HPLC method was established to quantify 21 active compounds. In vitro antioxidant experiments were conducted to assess the scavenging ability of DPPH and ABTS free radicals.Results:The contents of resource-based components in Hedysari Radix were higher than in Astragali Radix. Among the 21 active components, the characteristic active components of Astragali Radix, such as calycosin, formononetin, calycosin glucoside and so on, were high in contents. The contents of paeoniflorin and Medicinol in Astragali Radix were low or not detected, and the contents in Hedysari Radix were 98.828 μg/g and 103.986 μg/g, and the antioxidant capacity was also better than Astragali Radix.Conclusions:The basic components of Hedysari Radix for promoting blood circulation and nourishing blood are paeoniflorin, Medilac, vanillic acid, ferulic acid and formononetin, which can provide a scientific basis for the determination of the characteristic components and the improvement of the main functions of Hedysari Radix.

OBJECTIVE To investigate the effect and mechanism of Jingangteng capsules in the treatment of non-alcoholic fatty liver disease （NAFLD）. METHODS Thirty-two SD rats were randomly divided into normal group and modeling group. The modeling group was fed a high-fat diet to establish a NAFLD model. The successfully modeled rats were then randomly divided into model group， atorvastatin group[positive control， 2 mg/（kg·d）]， and Jingangteng capsules low- and high-dose groups [0.63 and 2.52 mg/（kg·d）]， with 6 rats in each group. The pathological changes of the liver were observed by hematoxylin-eosin staining and oil red O staining. Enzyme-linked immunosorbent assay was performed to determine the serum levels of triglycerides （TG）， total cholesterol （TC）， high-density lipoprotein cholesterol （HDL-C）， low-density lipoprotein cholesterol （LDL-C）， alanine transaminase （ALT）， aspartate transaminase （AST）， tumour necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， IL-6， IL-18. 16S rDNA amplicon sequencing and metabolomics techniques were applied to explore the effects of Jingangteng capsules on gut microbiota and metabolisms in NAFLD rats. Based on the E-mail：591146765@qq.com metabolomics results， Western blot analysis was performed to detect proteins related to the nuclear factor kappa-B （NF-κB）/NOD-like receptor family protein 3 （NLRP3） signaling pathway in the livers of NAFLD rats. RESULTS The experimental results showed that Jingangteng capsules could significantly reduce the serum levels of TG， TC， LDL-C， AST， ALT， TNF-α， IL-1β， IL-6， IL-18， while increased the level of HDL-C， and alleviated the hepatic cellular steatosis and inflammatory infiltration in NAFLD rats. They could regulate the gut microbiota disorders in NAFLD rats， significantly increased the relative abundance of Romboutsia and Oscillospira， and significantly decreased the relative abundance of Blautia （P＜0.05）. They also regulated metabolic disorders primarily by affecting secondary bile acid biosynthesis， fatty acid degradation， O-antigen nucleotide sugar biosynthesis， etc. Results of Western blot assay showed that they significantly reduced the phosphorylation levels of NF-κB p65 and NF-κB inhibitor α， and the protein expression levels of NLRP3， caspase-1 and ASC （P＜0.05 or P＜0.01）. CONCLUSIONS Jingangteng capsules could improve inflammation， lipid accumulation and liver injury in NAFLD rats， regulate the disorders of gut microbiota and metabolisms， and inhibit NF-κB/NLRP3 signaling pathway. Their therapeutic effects against NAFLD are mediated through the inhibition of the NF-κB/NLRP3 signaling pathway.

Objective To investigate the relationship between high DNA stainability(HDS)and routine semen parameters.Methods Semen samples were collected from 396 men of childbearing age who were admitted to our department.Correlations of HDS with routine semen parameters and age were analyzed.Multiple linear regression analysis was performed to identify the routine semen parameters that had the greatest influence on HDS.The correlation of HDS and DNA fragmentation index(DFI)with routine semen parameters and age in 244 patients with teratozoospermia was analyzed.The 244 patients were divided into extremely severe,severe,moderate,and mild teratozoospermia groups,and differences in HDS,DFI,and routine semen parameters were compared among the four groups.Results HDS was negatively correlated with total sperm count,sperm concentration,sperm progressive motility,and normal sperm morphology rate(NSMR)(P<0.01).After adjusting for potential confounders,including total sperm count,sperm concentration,sperm progressive motility,NSMR,and DFI,NSMR had the most significant negative effect on HDS(P<0.05).In the 244 patients with teratozoospermia,HDS was negatively correlated with NSMR(P<0.01)and positively correlated with the percentage of sperm head and tail abnormalities(P<0.05),while DFI was positively correlated with the percentage of sperm tail abnormalities(P<0.01).There was a significant difference in the percentage of sperm head abnormalities among the four teratozoospermia groups.The more severe the malfor-mation,the higher the percentage of sperm head abnormalities was(P<0.001).HDS in the extremely severe teratozoospermia group was significantly higher than that in the mild and moderate teratozoospermia groups(P<0.05).No significant differences in DFI were found among the four groups(P>0.05).Conclusion HDS was closely correlated with routine semen parameters and was a crucial biomarker for assessing sperm quality,particularly the extent of sperm head abnormalities.

Objective:To develop a training program for general practitioners (GPs) on carotid plaque screening using portable intelligent point-of-care ultrasonography (POCUS).Methods:A draft training program for GPs on carotid plaque screening using POCUS was initially formulated through focus group discussions involving experts in ultrasonography and general practice. Two rounds of Delphi consultation were conducted with 15 ultrasound experts from 6 secondary and tertiary public hospitals in Shanghai in February and April 2023. The analytic hierarchy process (AHP) was applied to assign weights to the indicators in the final program.Results:The final program consisted of 4 first-level indicators, 10 second-level indicators, and 47 third-level indicators. Among the first-level indicators,"training objects" had the lowest weight (7.69%), while the other three indicators were equally weighted (30.77% each). The top 3 third-level indicators by combined weight were "mastering key diagnostic points of carotid plaque"(10.26%),"mastering carotid plaque screening using portable POCUS"(6.84%), and "primary care hands-on training sessions"(6.39%).Conclusion:A structured training program for GPs has been successfully developed for carotid plaque screening using portable intelligent POCUS, comprising 4 first-level, 10 second-level, and 47 third-level indicators.

Objective:To evaluate the efficacy and safety of Neuroform Atlas stent-assisted coil embolization in patients with middle cerebral artery bifurcation aneurysms.Methods:A retrospective analysis was performed; the clinical data of 46 patients with middle cerebral artery bifurcation aneurysms accepted Neuroform Atlas stent-assisted coil embolization in First Affiliated Hospital of Zhengzhou University, Beijing Tiantan Hospital Affiliated to Capital Medical University, First Affiliated Hospital of Harbin Medical University, Zhujiang Hospital of Southern Medical University and First Affiliated Hospital of Chongqing Medical University from January 2022 to March 2024 were collected. There were 28 ruptured aneurysms (60.87%) and 18 unruptured aneurysms (39.13%). Follow-up was performed for more than 3 months; Raymond-Roy grading was used to evaluate the aneurysm embolization immediately after embolization and during follow-up; perioperative hemorrhagic or ischemic complications were recorded; modified Rankin Scale (mRS) was used to evaluate the prognosis of the patients at discharge and during follow-up (mRS score≤2: good prognosis, and mRS score>2: poor prognosis).Results:Coil embolization was successful in all 46 patients. DSA immediately after embolization showed that 41 patients (89.13%) had completely occluded aneurysms (Raymond-Roy grading I), 2 patients (4.35%) had residual aneurysm neck (Raymond-Roy grading Ⅱ) and 3 patients (6.52%) had partially occluded aneurysms (Raymond-Roy grading Ⅲ). Perioperative complications occurred in 5 patients, including 2 with postoperative cerebral infarction, 1 with hydrocephalus, 1 with postoperative pneumonia leading to respiratory failure, and 1 with stent thrombosis during embolization. Both at discharge and 3 months after embolization, 43 patients (93.48%) had good prognosis and 3 patients (6.52%) had poor prognosis. No obvious ischemic complications (such as stent restenosis) or hemorrhagic complications (such as re-rupture of the aneurysms) were found in all patients. Thirty patients (65.22%) had imaging follow-up for 6-12 months: 26 (86.67%) had Raymond-Roy grading I, 3 (10.00%) had Raymond-Roy grading II, and 1 (3.33%) had Raymond-Roy grading III.Conclusion:Neuroform Atlas stent-assisted coil embolization has good short-term efficacy and high safety in middle cerebral artery bifurcation aneurysms, but long-term follow-up observation is still needed to verify its efficacy.