Vaccination is a crucial strategy for the prevention and control of infectious diseases. Virus-like particles (VLPs), composed of structural proteins, have garnered significant attention as a novel type of vaccine due to their excellent safety and immunogenicity. However, similar to most vaccine antigens, VLPs exhibit insufficient thermal stability, which not only restricts the widespread application of vaccines but also increases the risk of vaccine inactivation. This study aims to enhance the stability and shelf life of VLPs derived from type A foot-and-mouth disease virus (FMDV) by employing vacuum freeze-drying technology. The optimal lyoprotectant formulation was determined through single-factor and combinatorial screening. Subsequently, the correlation between the immunogenicity of the freeze-dried vaccine and the content of FMDV VLPs was evaluated via a mouse model. The stability of FMDV VLPs before and after freeze-drying was further assessed by storing them at 4, 25, and 37 ℃ for varying time periods. Results indicated that the lyoprotectant formulation No.1, composed of 7.5% trehalose, 0.1% Tween 80, 50 mmol/L glycine, 1% sodium glutamate, and 3% polyvinylpyrrolidone (PVP), effectively preserved the content of FMDV VLPs during the vacuum freeze-drying process. The immunization trial in mice revealed that the levels of specific antibodies, immunoglobulin G1 (IgG1), interleukin-4 (IL-4), and neutralizing antibodies induced by freeze-dried FMDV VLPs were comparable to those induced by non-freeze-dried FMDV VLPs. The heat treatment results showed that the storage periods of freeze-dried FMDV VLPs at 4, 25, and 37 ℃ were significantly longer than those of non-freeze-dried FMDV VLPs. In conclusion, the selected lyoprotectant formulation effectively improved the stability of FMDV VLPs vaccines. This study provides valuable insights for enhancing the stability of novel subunit vaccines.
Freeze Drying/methods* ; Animals ; Foot-and-Mouth Disease Virus/immunology* ; Mice ; Vaccines, Virus-Like Particle/chemistry* ; Foot-and-Mouth Disease/immunology* ; Vacuum ; Drug Stability ; Mice, Inbred BALB C ; Viral Vaccines/immunology*

OBJECTIVE: To investigate the biomechanical characteristics of Schatzker type Ⅱ tibial plateau fracture fixed by different bone grafting methods and internal fixations. METHODS: Twenty-four embalmed specimens of adult knee joint were selected to make Schatzker type Ⅱ tibial plateau fracture models, which were randomly divided into 8 groups (groups A1-D1 and groups A2-D2, n=3). After all the fracture models were restored, non-structural iliac crest bone grafts were implanted in group A1-D1, and structural iliac crest bone grafts in groups A2-D2. Following bone grafting, group A was fixed with a lateral golf locking plate, group B was fixed with lateral golf locking plate combined compression bolt, group C was fixed with lateral tibial "L"-shaped locking plate, and group D was fixed with lateral tibial "L"-shaped locking plate combined compression bolt. Compression and cyclic loading tests were performed on a biomechanical testing machine. A distal femur specimen or a 4-cm-diameter homemade bone cement ball were used as a pressure application mould for each group of models. The specimens were loaded with local compression at a rate of 10 N/s and the mechanical loads were recorded when the vertical displacement of the split bone block reached 2 mm. Then, compressive and cyclic loading tests were conducted on the fixed models of each group. The specimens were compression loaded to 100, 400, 700, and 1 000 N at a speed of 10 N/s to record the vertical displacement of the split bone block. The specimens were also subjected to cyclic loading at 5 Hz and 10 N/s within the ranges 100-300, 100-500, 100-700, and 100-1 000 N to record the vertical displacement of the split bone block at the end of the entire cyclic loading test. The specimens were subjected to cyclic loading tests and the vertical displacement of the split bone block was recorded at the end of the test. RESULTS: When the vertical displacement of the collapsed bone block reached 2 mm, the mechanical load of groups A2-D2 was significantly greater than that of groups A1-D1 ( P<0.05). The mechanical load of groups B and D was significantly greater than that of group A under the two bone grafting methods ( P<0.05); the local mechanical load of group D was significantly greater than that of groups B and C under the structural iliac crest bone grafts ( P<0.05). There was no significant difference ( P>0.05) in the vertical displacement of the split bone blocks between the two bone graft methods when the compressive load was 100, 400, 700 N and the cyclic load was 100-300, 100-500, 100-700 N in groups A-D. However, the vertical displacement of bone block in groups A1-D1 was significantly greater than that in groups A2-D2 ( P<0.05) when the compressive loading was 1 000 N and the cyclic load was 100-1 000 N. The vertical displacement of bone block in group B was significantly smaller than that in group A, and that in group D was significantly smaller than that in group C under the same way of bone graft ( P<0.05). CONCLUSION Compared with non-structural iliac crest bone grafts implantation, structural iliac crest bone grafts is more effective in preventing secondary collapse of Schatzker type Ⅱ tibial plateau fracture, and locking plate combined with compression bolt fixation can provide better articular surface support and resistance to axial compression, and the lateral tibial "L"-shaped locking plate can better highlight its advantages of "raft" fixation and show better mechanical stability.
Humans ; Bone Transplantation/methods* ; Tibial Fractures/physiopathology* ; Fracture Fixation, Internal/instrumentation* ; Biomechanical Phenomena ; Bone Plates ; Ilium/transplantation* ; Adult ; Tibia/surgery* ; Bone Cements ; Knee Joint/surgery* ; Male ; Tibial Plateau Fractures

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.

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.

Objective To investigate the protective effect of Qianggukangwei Recipe(QG)on myotube cell atrophy induced by simulated microgravity effect(SMG).Methods The induced differentiation mouse myotube cells(C2C12 cells)were divided into six groups.Then the 48h simulated microgravity effect(SMG)model of mouse myotube cells was established by three-dimensional gyrotron(48h-SMG group).The cells in 3 groups were treated with 2.50,1.25,0.625 mg/mL QG.The cells in the left group were treated with 2.70 μg/mL alendronate sodium as positive control group.The levels of oxidative and inflammatory factors in myotube cells were detected by kits.The protein expression levels of NF-κB/IκB protein decomposition pathway and IGF-1/PI3K/Akt protein synthesis pathway were detected by Western blotting.The IGF-1/PI3K/Akt pathway was verified by 7 μmol/L of IGF-1 inhibitor.Results As compared with the control group,oxidation and inflammation levels in myotube cells increased significantly(P<0.05).The protein expression levels of IKK,NF-κB,and MURF-1 were significantly increased by 19.9%±6.1%,189.3%±15.9%,445.0%±46.1%,and IκB was significantly decreased by 26.5%±0.1%(P<0.05).The levels of IGF-1,PI3K,p-Akt/Akt and mTOR in the IGF-1/PI3K/Akt pathway were decreased by 23.6%±0.5%,30.7%±2.7%,13.3%±1.1%,21.0%±1.6%(P<0.05).The three doses of QG could and reduce the level of cellular oxidative stress and inflammation(P<0.05).QG could also activate the IGF-1/PI3K/Akt pathway and inhibit the NF-κB/IκB pathway.Among them,the high dose of QG(2.50 mg/mL)significantly decreased the protein expression levels of IKK,NF-κB and MURF-1 by 39.9%±2.4%,48.6%±0.1%,70.0%±2.1%(P<0.05),and significantly increased the expression levels of IGF-1,PI3K,mTOR and Akt phosphorylation by 43.1%±1.1%,100.0%±7.7%,71.8%±2.5%,95.2%±12.8%(P<0.05).Conclusion QG can significantly alleviate myotube cell atrophy induced by SMG effect by regulating relevant protein synthesis and degradation pathways.

Objective To investigate the role of sphingosine kinase-1(SPHK1)in regulating migration and invasion of gastric cancer(GC)cells.Methods TIMER2.0,GEPIA and HPA databases were used to investigate SPHK1 expression in GC,and its association with prognosis of the patients was analyzed using Kaplan-Meier Plotter database.In 40 clinical GC and adjacent tissue samples,SPHK1 and MKI67 expressions were detected with immunohistochemistry,Western blotting,and RT-qPCR.Gene enrichment pathway analysis was conducted to explore the biological functions of SPHK1.In HGC-27 and MGC-803 cells,the effects of lentivirus-mediated SPHK1 knockdown or overexpression on cell migration and invasion and expressions of key proteins in the nuclear factor-κB(NF-κB)signaling were evaluated using cell scratch test,Transwell assays and Western blotting.The changes in tumorigenic capacity of the transfected GC cells were evaluated in nude mice.Results SPHK1 was highly expressed in GC tissues in negative correlation with overall survival,overall survival after progression,and relapse-free survival of the patients(all P<0.001).In clinical GC samples,SPHK1 and MKI67 expressions showed a positive correlation(P=0.00049)and were both significantly up-regulated(P<0.001).Gene enrichment pathway analysis suggested the involvement of SPHK1 in cell adhesion,migration,angiogenesis and the NF-κB pathway(P<0.05).In the cell experiment,SPHK1 knockdown significantly decreased while SPHK1 overexpression enhanced migration and invasion abilities of the GC cells.SPHK1 positively regulated the expressions of phosphorylated P65(P-P65),VEGFA and IL-17,and blocking the NF-κB pathway by PDTC significantly lowered migration and invasion ability of the cells.In nude mice,the GC cells with SPHK1 knockdown resulted in significantly reduced tumor size and mass,while the SPHK1-overexpressing cells showed enhanced tumorigenicity.Conclusion SPHK1 regulates migration and invasion of GC cells via the NF-κB signaling pathway and may serve as a potential diagnostic marker for GC progression.

Objective To investigate the role of sphingosine kinase-1(SPHK1)in regulating migration and invasion of gastric cancer(GC)cells.Methods TIMER2.0,GEPIA and HPA databases were used to investigate SPHK1 expression in GC,and its association with prognosis of the patients was analyzed using Kaplan-Meier Plotter database.In 40 clinical GC and adjacent tissue samples,SPHK1 and MKI67 expressions were detected with immunohistochemistry,Western blotting,and RT-qPCR.Gene enrichment pathway analysis was conducted to explore the biological functions of SPHK1.In HGC-27 and MGC-803 cells,the effects of lentivirus-mediated SPHK1 knockdown or overexpression on cell migration and invasion and expressions of key proteins in the nuclear factor-κB(NF-κB)signaling were evaluated using cell scratch test,Transwell assays and Western blotting.The changes in tumorigenic capacity of the transfected GC cells were evaluated in nude mice.Results SPHK1 was highly expressed in GC tissues in negative correlation with overall survival,overall survival after progression,and relapse-free survival of the patients(all P<0.001).In clinical GC samples,SPHK1 and MKI67 expressions showed a positive correlation(P=0.00049)and were both significantly up-regulated(P<0.001).Gene enrichment pathway analysis suggested the involvement of SPHK1 in cell adhesion,migration,angiogenesis and the NF-κB pathway(P<0.05).In the cell experiment,SPHK1 knockdown significantly decreased while SPHK1 overexpression enhanced migration and invasion abilities of the GC cells.SPHK1 positively regulated the expressions of phosphorylated P65(P-P65),VEGFA and IL-17,and blocking the NF-κB pathway by PDTC significantly lowered migration and invasion ability of the cells.In nude mice,the GC cells with SPHK1 knockdown resulted in significantly reduced tumor size and mass,while the SPHK1-overexpressing cells showed enhanced tumorigenicity.Conclusion SPHK1 regulates migration and invasion of GC cells via the NF-κB signaling pathway and may serve as a potential diagnostic marker for GC progression.

Objective To comprehensively review the clinical research on the treatment of hyperlipidemia with traditional Chinese Medicine(TCM)through the evidence mapping,and to understand the distribution of evidence in this field. Methods Databases including CNKI,Wangfang,VIP,SinoMed,PubMed,Cochrane Library,and Embase were searched from January 2004 to December 2023 to collect clinical studies,systematic reviews/meta-analyses,guidelines and clinical pathways related to the treatment of hyperlipidemia with TCM. The results were analyzed and displayed in charts and graphs according to the screening criteria,and the Assessment of Multiple Systematic Reviews 2 (AMSTAR 2) tool and the Preferred Reporting Item for Systematic Review and Meta-analysis of Chinese herbal medicine (PRISMA-CHM) were used to evaluate the quality of the systematic review/meta-analysis. Results A total of 1223 studies were included in the analysis according to Population,Intervention,Comparison,Outcome and Study design(PICOS) principles,involving 920 RCTs,249 non-RCTs,49 systematic reviews/meta-analyses,and 5 guidelines/expert consensus. In recent years,the overall number of clinical research publications has shown a downward trend. Hyperlipidemia frequently occurs in middle-aged and elderly people,and age of onset tends to be younger. The sample size of randomized controlled studies is mostly concentrated in 60-300 cases. There are many types of clinical treatment regimens for the treatment of hyperlipidemia with TCM,among which TCM decoction (50.13％) and Chinese patent medicine (38.41％) account for a relatively high proportion,and TCM exercise therapy (0.51％) is the lowest treatment. Jiangzhi Decoction has attracted more attention in trial group of TCM decoction,while Xuezhikang Capsule has attracted more attention in trial group of Chinese patent medicine. In terms of methodological design,199 papers(21.63％) explicitly mentioned the method of generating random sequence,17 papers(1.85％) mentioned allocation concealment,37 papers (4.02％) mentioned blinding. The control group was dominated by the statins,including simvastatin and atorvastatin. The outcome indicators mainly include the total effective rate,TCM syndrome score,blood lipid level,coagulation index,and adverse reactions,while the attention of TCM characteristic efficacy,inflammation,oxidative stress,and vascular endothelial index were low. The methodological and reporting quality of the systematic review/Meta-analysis were generally not high. AMSTAR-2 evaluation was extremely low,and the average PRISMA-CHM score was 15. Conclusion TCM has certain advantages in the treatment of hyperlipidemia,but there is a lack of high-quality evidence-based proof,and more high-quality clinical studies are still needed to further provide evidence supports in the future. It has been suggested that more large-sample and multi-center clinical studies should be carried out in the future. We should formulate systematic reviews/Meta analysis and guidelines/expert consensus according to the guidelines of clinical practice issues,also consult international standards and regulations,enhance normativity and reliability to improve the quality of their evidence.

Objective:To analyze the epidemiological characteristics of hospitalized patients diagnosed with sepsis in a large class Ⅲ general hospital in Southwest China in a period of 2 years, and to explore the risk factors related to death in patients with sepsis.Methods:A retrospective study was conducted to select patients with sepsis admitted to Sichuan Provincial People's Hospital from September 1, 2021 to August 31, 2023, and general characteristics such as gender, age, discharge diagnosis, discharge department, hospitalization cost, length of stay, and prognosis during hospitalization were collected. The baseline of two groups of patients were compared, and the risk factors of in-hospital cause of death in patients with sepsis were analyzed by multivariate Logistic regression.Results:A total of 3 568 patients with sepsis were included with median age of 58 (35, 74) years old. Of all patients, there were 2 147 males (60.17%). The median length of hospitalization was 13 (8, 24) days, and the median hospitalization cost was 3.98 (1.87, 8.83) ten thousand yuan. The departments with more than 100 cases of sepsis in 2 years were central intensive care unit (ICU), pediatrics department, nephrology department, emergency medicine department, emergency intensive care unit (EICU), infectious department, respiratory medicine department, hematology department, neonatal care unit and emergency surgical department. A total of 1 210 patients (33.91%) admitted to ICU (including central ICU and EICU). The hospitalization cost of ICU patients were higher [6.7 (3.1, 15.5) ten thousand yuan], the hospitalization duration was longer [9 (3, 17) days], and the mortality was higher [35.29% (427/1 210)]. Among 3 568 patients with sepsis, 448 died and 3 120 survived during hospitalization. The age, male proportion and hospitalization cost of patients with sepsis in the death group were significantly higher than those in the survival group [age (years old): 75 (60, 86) vs. 57 (30, 71), male proportion: 67.86% (304/448) vs. 59.07% (1 843/3 120), hospitalization cost (ten thousand yuan): 6.7 (3.0, 16.9) vs. 3.7 (1.8, 8.1)], the ratio of diabetes mellitus was significantly lower than that of survival group [4.91% (22/448) vs. 10.45% (326/3 120)], the length of hospitalization was shorter than that of survival group [days: 10.0 (3.0, 19.0) vs. 13.0 (8.0, 24.0)], the differences were statistically significant (all P < 0.01). Multivariate Logistic regression analysis showed that male [odds ratio ( OR) = 0.75, 95% confidence interval (95% CI) was 0.59-0.96], elder ( OR = 1.04, 95% CI was 1.03-1.05) and diabetes ( OR = 0.32, 95% CI was 0.19-0.54) were independent risk factors for in-hospital death in patients with sepsis (all P < 0.05). Conclusions:Sepsis is a heavy burden in Southwest China, especially for ICU, with high mortality, high hospitalization costs, and heavy economic burden on patients and society. Male, elder and diabetes were independent risk factors for in-hospital death of sepsis patients.

ObjectiveTo establish the specific chromatogram and thin layer chromatography（TLC） of Qingxin Lianziyin（QXLZY） benchmark samples， in order to clarify the key quality attributes and provide a reference for the quality evaluation of QXLZY. MethodHigh performance liquid chromatography（HPLC） specific chromatogram of QXLZY benchmark samples was developed by using a YMC Hydrosphere C₁₈ column（4.6 mm×250 mm， 5 μm） with the mobile phase of acetonitrile（A）-0.2% formic acid aqueous solution（B） for gradient elution（0-10 min， 5%-20%A； 10-20 min， 20%A； 20-25 min， 20%-24%A； 25-40 min， 24%-30%A； 40-55 min， 30%-50%A； 55-65 min， 50%-100%A； 65-75 min， 100%A； 75-75.1 min， 100%-5%A； 75.1-90 min， 5%A）， and the detection wavelength was 360 nm. Ultra-high performance liquid chromatography-linear ion trap/orbitrap mass spectrometry（UHPLC-LTQ-Orbitrap MS） with electrospray ionization（ESI） was used to identify the components of QXLZY benchmark samples by accurate relative molecular weight and multilevel MS fragment ion information， the detection conditions were positive and negative ion modes and data dependency scanning mode. TLC identification methods for Ophiopogonis Radix， Lycii Cortex， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma in QXLZY were established. ResultA total of 15 characteristic peaks were identified from Glycyrrhizae Radix et Rhizoma， Plantaginis Semen and Scutellariae Radix， and the relative standard deviations of the retention times of 15 characteristic peaks in 15 batches of QXLZY benchmark samples were≤3% with peak 8（baicalin） as the reference peak. A total of 100 compounds， including flavonoids， organic acids， saponins， amino acids and others， were identified in the benchmark samples by UHPLC-LTQ-Orbitrap MS. The established TLC had good separation and was suitable for the identification of Ophiopogonis Radix， Lycii Cortex， Nelumbinis Semen， Poria， Astragali Radix and Ginseng Radix et Rhizoma in QXLZY. ConclusionThe material basis of QXLZY benchmark samples is basically determined by MS designation and source attribution. The established specific chromatogram and TLC of QXLZY are simple， stable and reproducible， which can provide a reference for the development and quality control of QXLZY.