ObjectiveTo construct a stable monoclonal human embryonic kidney 293 （HEK293） cell line expressing recombinant human coagulation factor Ⅶ （rhFⅦ） and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells， and monoclonal stable transfected cell lines were selected using geneticin （G418）. The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction （RT-PCR）. The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay （ELISA）， and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence， indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8， and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was （1.27±0.09） mg/L， and the highest procoagulant activity was （380.29±13.80）%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.

Bile acids (BAs) serve not only as key facilitators of lipid absorption but also as crucial signaling molecules regulating glucose and lipid metabolism, inflammation, and overall energy homeostasis. Aging profoundly alters BA metabolism, characterized by shifts in biosynthetic pathways, compositional changes, disrupted receptor-mediated signaling, and alterations in gut microbiota interactions. These age-related changes contribute to the onset and progression of metabolic conditions, including type 2 diabetes, obesity, metabolic dysfunction-associated fatty liver disease, and neurodegenerative disorders. An increased abundance of hydrophobic and cytotoxic BAs has been associated with systemic inflammation, metabolic rigidity (disruption of metabolic flexibility), and organ dysfunction. Targeting BA signaling—through pharmacological modulation of farnesoid X receptor and Takeda G protein-coupled receptor 5 or microbiota-directed therapies—offers promising strategies to mitigate aging-related metabolic decline. A deeper understanding of how BA metabolism evolves over the lifespan may unveil novel interventions to promote healthy aging and prevent age-related disease.

Objective To investigate the predictive value of multimodal ultrasound parameters combined with systemic inflammation response index(SIRI)and neutrophil-to-lymphocyte ratio(NLR)for efficacy of neoadjuvant chemotherapy(NAC)in patients with breast cancer(BC).Methods A total of 132 BC patients in the Provincial Hospital Affiliated to Shandong First Medical University from March 2022 to March 2024 were selected as the study subjects,and they were divided into effective group(n=100)and ineffective group(n=32)according to the NAC results.Multimo-dal ultrasound parameters,SIRI,and NLR before NAC treatment were compared between the two groups.Multivariate Logistic regression analysis was used to explore the influencing factors of ineffective NAC in BC patients.Results After NAC treatment,among 132 BC patients,23 cases achieved complete remission,77 cases achieved partial remission,29 cases had stable disease,and 3 cases had disease progression.The elasticity score and resistance index(RI)in the effective group were significantly lower than those in the ineffective group,while the peak intensity(PI),area under the curve(AUC),and peak systolic velocity(PSV)were significantly higher than those in the ineffec-tive group(P＜0.05).The neutrophil count and monocyte count in the ineffective group were sig-nificantly higher than those in the effective group,while the lymphocyte count was significantly lower than that in the effectivegroup(P＜0.05).Before NAC treatment,the SIRI and NLR in the inef-fective group were significantly higher than those in the effective group(P＜0.05).The proportions of patients with non-triple-negative breast cancer and lymph node metastasis in the ineffective group were significantly higher than those in the effective group(P＜0.05).Multivariate Logistic regres-sion analysis showed that high RI,high SIRI,high NLR,high elasticity score,low PSV,low PI,low AUC,non-triple-negative breast cancer,and lymph node metastasis were risk factors for ineffec-tive NAC in BC patients(P＜0.05).Receiver operating characteristic(ROC)curve analysis showed that the AUC for predicting the efficacy of NAC in BC by the combination of multimodalultra-sound parameters,SIRI,and NLR was 0.929,which was significantly higher than that of the three independent predictors alone(P＜0.05).Conclusion The combination of multimodal ultrasound parameters,SIRI,and NLR has a high predictive value for the efficacy of NAC in BC and may serve as effective predictors for the efficacy of NAC.

Objective:Postoperative neurosurgical bacterial meningitis (PNBM) has been frequently reported, but fewer studies have focused on the contemporaneous comparison of clinical features of PNBM caused by different pathogenic bacteria. This study aimed to simultaneously investigate the clinical characteristics and outcomes of PNBM by Gram-positive bacterial(GPB) or Gram-negative bacterial (GNB) infection.Methods:Inpatients with PNBM at our institution were recruited between February 2013 and October 2023. These PNBM patients were categorized into two groups: GPB infection and GNB infection. Data from electronic medical records were collected and analyzed.Results:A total of 401 patients with PNBM were finally included, with 78 (19.5%) having GPB infections and 323 (80.5%)having GNB infection. The average age of the patients was 56 years, and 55.1% were male. Compared to the GPB group, PNBM patients with GNB infection had significantly higher SOFA and APACHE Ⅱ scores, higher proportions of hyperthermia (body temperature>39°C) and altered consciousness, increased ratios of postoperative cerebral hemorrhage or intracranial aneurysm, as well as greater needs for ICU treatment and mechanical ventilation (all P <0.05). The proportions of inflammatory indicators such as blood CRP and PCT≥2 ng/mL, alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBIL) were significantly higher in the GNB group (all P<0.05). In contrast, The concentrations of hemoglobin and albumin were substantially lower in this group(both P <0.05). Additionally, the cerebrospinal fluid in the GNB group showed significantly higher nucleated cell counts, protein concentration, and adenosine deaminase concentration, and but lower glucose level (all P <0.001). A total of 426 bacterial strains were isolated, with 343 strains (80.5%) being GNB and 83 strains (19.5%) being GPB. Among these, 25 (6.2%) patients had 2 or more gram-positive or gram-negative bacterial infections. The proportions of multidrug-resistant (MDR) bacteria and intrathecal treatment were higher in the GNB group (80.5% vs. 68.7%, 36.5% vs. 2.6%, respectively), while the ratio of correct empirical antibiotic treatment was significantly lower (30.3% vs. 80.0%) (all P <0.05). In terms of outcomes, the length of stay in the ICU was significantly longer in the GNB group [(median (interquartile range, IQR): 11.5 (5.25,22.75) vs. 17.0 (9.0,30.0), P <0.01)], and the rate of septic shock (9.3% vs. 2.6%), poor prognosis (GCS≤8 at discharge) (65.9% vs. 32.1%), and 28-day hospital mortality rate (34.4% vs. 10.3%) were significantly higher compared to the GPB group (all P <0.05). However, there were no differences in 7-day hospital mortality and total hospitalization time. Conclusions:Gram-negative bacterial infections are more prevalent than Gram-positive bacterial infections in PNBM, and they are also associated with more severe symptoms, abnormal cerebrospinal fluid findings, higher severity, and more treatment difficulty. Despite comparable short-term (7-day) mortality rates between Gram-positive and Gram-negative bacterial infections, Gram-negative bacterial infections result in higher medium- to long-term (14-day and 28-day) case-fatality rates among patients with post-neurosurgical bacterial meningitis and are associated with overall poorer prognosis, warranting greater attention from clinicians.

Objective To investigate the levels of gross α and gross β activities in different water types within a 40-kilometer radius around the Zhangzhou Nuclear Power Plant prior to its operation. Methods In 2018, drinking water samples were collected from the area surrounding the nuclear power plant during both the wet and dry seasons, including source water, treated water, tap water, and well water. The gross α and gross β activity concentrations were measured using a low-background α/β counter, followed by statistical analysis. Results A total of 80 water samples from different sources around the Zhangzhou Nuclear Power Plant were collected. The average gross α and gross β activity concentrations during the wet season were (0.110 ± 0.036) Bq/L and (0.643 ± 0.028) Bq/L, respectively, while those during the dry season were (0.124 ± 0.032) Bq/L and (0.624 ± 0.026) Bq/L, respectively. There were no significant differences in the gross α and gross β activity concentrations between the wet and dry seasons for the overall sample set (P > 0.05). However, there were statistically significant differences in the gross α and gross β activity concentrations between the wet and dry seasons for source water and well water (Z_wet = −2.005, −2.123; Z_dry = −1.943, −3.090; P < 0.05). Conclusion The radioactivity levels in different water types within various ranges around the Zhangzhou Nuclear Power Plant before its operation were determined. The measured activity concentrations were at the same level as those from previous investigations in other regions of Fujian Province.

Bio-mineralization has emerged as a promising strategy to enhance vaccine immunogenicity. This study optimized the calcium phosphate (CaP) mineralization process of foot-and-mouth disease virus-like particles (FMD VLPs) to achieve high mineralization efficiency and scalability. Key parameters, including concentrations of Ca²⁺, HPO₄^2-, NaCl, and VLPs, as well as stirring speed, were systematically optimized. Stability of the scaled-up reaction system and immunogenicity of the mineralized vaccine were evaluated. Optimal conditions [25.50 mmol/L Ca(NO₃)₂, 15 mmol/L Na₂HPO₄, 300 mmol/L NaCl, 0.75 mg/mL VLPs, and 1 500 r/min] yielded CaP-mineralized VLPs (VLPs-CaP) with high mineralization efficiency, uniform morphology, and a favorable particle size. Scaling up the reaction by 25 folds maintained consistent mineralization efficiency and particle characteristics. Immunization in mice demonstrated that VLPs-CaP induced higher titers of specific antibodies and neutralizing antibodies than unmineralized VLPs (P < 0.05). Higher IgG2a/IgG1 ratio and enhanced IFN-γ secretion (P < 0.05) further indicated robust cellular immune responses. We establish a stable and scalable protocol for VLPs-CaP, providing a theoretical and technical foundation for developing high-efficacy VLPs-CaP vaccines.
Vaccines, Virus-Like Particle/immunology* ; Immunogenicity, Vaccine ; Calcium Phosphates/chemistry* ; Foot-and-Mouth Disease Virus ; Biomineralization ; Particle Size ; Animals ; Mice ; Antibodies, Neutralizing/blood* ; Antibodies, Viral/blood* ; Immunity, Cellular

Objective:To observe the efficacy and safety of combined lienal polypeptide injection therapy in the treatment of Mycoplasma pneumoniae pneumonia（MPP）in children aged 3 to 14 years old in multiple clinical centers.Methods:A randomized，controlled，multi-center clinical study design was adopted.A total of 240 hospitalized children aged 3 to 14 years old with MPP from 7 hospitals from September 1，2023 to January 31，2024 were included.According to the severity of pneumonia，they were divided into the mild MPP group with 80 cases and the severe MPP/refractory MPP（SMPP/RMPP）group with 160 cases，and then randomly divided into the control group and the experimental group at a ratio of 1 ∶1，using the random number table method.After screening，subjects entered a treatment period of 5 to 7 days.The control group was treated with azithromycin，while the experimental group was treated with azithromycin plus lienal polypeptide injection .The recovery of lung CT，length of hospital stay，duration of fever，cough score，whether mild cases developed into severe or refractory cases，duration of hormone use，use of intravenous immunoglobulin（IVIG），bronchoscopy treatment，and immune function were observed between the two groups to evaluate the efficacy of lienal polypeptide injection.Adverse events after medication，vital signs，blood routine，urine routine，liver function，myocardial enzymes，renal function，and electrocardiogram were observed to evaluate the safety. Results:A total of 231 subjects have completed the trial in the 7 hospitals，including 118 cases in the experimental group and 113 cases in the control group.Main observation index：the rate of lung CT aggravation in the experimental group was lower than that in the control group（2.6% vs 15.3%， P<0.01），and the difference was statistically significant.Secondary indexes：there were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and immunoglobulin between the two groups（all P>0.05）.However，the rate of cases of plastic bronchitis（PB）found under bronchoscopy in the experimental group was lower than that in the control group（0 vs 18.8%， P=0.03），and the difference was statistically significant.Among the mild MPP（72 cases），there were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and the improvement rate of lung CT between the two groups（all P>0.05）.However，compared with the control group，the rate of cases developing into SMPP/RMPP in the experimental group was less（24.3% vs 48.6%， P=0.03），and the difference in IgG before and after treatment was small［0.53（-0.04，1.18）g/L vs 1.33（0.48，2.25）g/L， P=0.01］.Among the SMPP/RMPP cases（159 cases），the rate of cases of PB found under bronchoscopy in the experimental group was less than that in the control group（0 vs 20%， P=0.04），and the rate of cases with aggravated lung CT in the experimental group was less than that in the control group（1.3% vs 19.5%， P<0.01），and the improvement rate of lung CT in the experimental group was higher than that in the control group（88.8% vs 75.3%， P=0.03），with statistically significant differences.There were no statistically significant differences in the length of hospital stay，duration of fever，cough score，duration of hormone use，whether IVIG treatment was used，the number of bronchoscopy treatment cases，and immunoglobulin between the two groups（all P>0.05）.Two cases in the experimental group developed rashes，which improved after the drug was discontinued.There were no serious adverse reactions such as abnormal vital signs like dyspnea and cyanosis due to the use of lienal polypeptide injection.There were no obvious changes in blood routine，liver function，myocardial enzymes，renal function，electrocardiogram，and urine routine values before and after medication compared with the baseline. Conclusion:The combined use of lienal polypeptide injection in the treatment of MPP in children can reduce the probability of the transformation from mild cases to SMPP/RMPP，reduce the rate of aggravation of the image findings，promote the absorption of lung inflammation，reduce the rate of PB found under bronchoscopy，and has good safety.

Objective:To investigate the effects of different doses of rosuvastatin on lipid metabolism and left ventricular function in patients with type 2 diabetes mellitus (T2DM) accompanied by coronary heart disease (CHD).Methods:A prospective study was conducted involving 63 patients with T2DM accompanied by CHD who were treated at Jinan 2 nd People's Hospital and The Fifth People's Hospital of Jinan from January 2022 to June 2023. In a randomized case-control study, patients were assigned to either a control group ( n = 31, receiving 10 mg/d of rosuvastatin) or an observation group ( n = 32, receiving 20 mg/d of rosuvastatin) using a random number table method. After 8 weeks of treatment, the therapeutic effects were evaluated, and blood lipid levels and left ventricular function were recorded before and after treatment. Plasma levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP), homocysteine (Hcy), C-reactive protein (CRP), and the distance covered in a 6-minute walk test were compared between the two groups. Patients were followed for 1 year to monitor adverse reactions and the occurrence of adverse cardiovascular events. Results:The effective treatment rate in the observation group was significantly higher than that in the control group [90.63% (29/32) vs. 70.97% (22/31), χ2 = 3.95, P < 0.05). Prior to treatment, there were no statistically significant differences between the two groups regarding blood lipid levels, cardiac ultrasound indicators, serum levels of NT-proBNP, Hcy, and CRP, the distance covered in the 6-minute walk test, and the incidences of adverse reactions and adverse cardiovascular events (all P > 0.05). After treatment, both groups showed a reduction in total cholesterol and low-density lipoprotein cholesterol (both P < 0.05), with the observation group demonstrating significantly lower levels ( t = 10.54, 14.01, both P < 0.001). Additionally, both groups showed an increase in high-density lipoprotein cholesterol ( P < 0.05), but the observation group achieved significantly higher levels ( t = -14.07, P < 0.001). Both groups also exhibited an increase in left ventricular ejection fraction ( P < 0.05), with a greater improvement in the observation group ( t = -2.34, P < 0.05). The left ventricular end-diastolic diameter decreased in both groups ( P < 0.05), but the observation group had a smaller diameter ( t = 2.78, P < 0.05). Levels of NT-proBNP, Hcy, and CRP decreased in both groups (all P < 0.05), with the observation group showing significantly lower levels ( t = 6.55, 3.94, 6.38, all P < 0.001). Finally, the distance covered in the 6-minute walk test increased in both groups ( P < 0.05), with the observation group covering a greater distance ( t = -4.32, P < 0.001). Conclusions:High-dose rosuvastatin can effectively improve lipid metabolism and left ventricular function in patients with T2DM complicated by CHD. It significantly reduces levels of NT-proBNP, Hcy, and CRP, enhances exercise capacity, and demonstrates high safety, indicating a high potential for clinical application.

Objective:To explore the effect of non-invasive intervention on preventing maternal breast tenderness, and to provide references for clinicians to select the optimal non-invasive intervention for the prevention of maternal breast tenderness.Methods:Literature focused on the effect of different non-invasive interventions to prevent maternal breast tenderness by randomized controlled trials design was searched in the databases, including China National Knowledge Infrastructure, VIP, Wanfang, SinoMed, PubMed, Web of Science, Embase and Cochrane Library, from inception to April 8, 2024. Two researchers independently screened the literature, extracted the data and evaluated the quality of the literature. Stata 16.0 was used to conduct a network Meta-analysis.Results:A total of 21 studies involving 2 298 women, covering 5 non-invasive interventions (low-frequency pulse electrotherapy, breast massage combined with low-frequency pulse electrotherapy, breast massage and acupoint massage, breast massage and acupoint massage combined with ear point pressure bean, breast massage and acupoint massage combined with external application of Chinese medicine). The network meta-analysis results showed that, compared with routine care, five non-invasive interventions had statistically significant differences in reducing the incidence of maternal breast tenderness ( OR values ranging from 0.20 to 0.39, all P<0.05). Breast massage combined with low-frequency pulse electrotherapy ranked first in terms of effectiveness on preventing of breast tenderness. Conclusions:All five non-invasive interventions are effective on preventing maternal breast tenderness, with breast massage combined with low-frequency pulse electrotherapy being the most effective.

Purpose This study aims to analyze the clinical,pathological,and molecular genetic characteristics of nodal follicular helper T-cell lymphoma,follicular type(nTFHL-F).Methods 7 cases of nTFHL-F were re-viewed.Clinical data were collected,tissue morphology was summarized,and immunohistochemical staining and mo-lecular testing were performed.Results The median age of patients was 62 years with a male-to-female ratio of 6:1.The initial symptoms included neck lymphadenectasis in 6 cases and abdominal discomfort in one.Six cases were in ad-vanced stages,while 1 case was in the localized stage.The tumors exhibited a vague,irregular follicular nodular pat-tern,without significant polymorphic inflammatory background or high endothelial vascular proliferation.Five cases showed a progressive transformation resembling germinal center pattern,and two cases exhibited a follicular lymphoma-like growth pattern.Tumor cells presented three distinct morphologies:centrocyte-like appearance,monocytoid B cell-like appearance,and atypical cells with abundant,transparent cytoplasm.Tumor cells expressed at least three follicu-lar T-cell markers.Testing for ITK::SYK gene fusion was negative in all cases(0/7).Next generation sequencing i-dentified mutations in TET2 gene in two cases(2/2),the RHOA gene in one case(1/2),and VAV1 gene in one case(1/2).The follow-up duration ranged from 2 to 64 months,with three deaths(3/7),and a median survival time of 37 months.Conclusion nTFHL-F predominantly occurs in middle-aged to elderly males,presenting with advanced clinical stages,and has a poor prognosis.nTFHL-F is closely associated with nodal follicular helper T-cell lymphoma,angioimmunoblastic type,and mya coexist within the same individual.