Objective:To develop a bacterial endotoxin detection method suitable for mRNA vaccines based on lipid nanoparticle（LNP）delivery system.Methods:The gel clot limit test outlined in the third section of the Chinese Pharmacopoeia（2020 edition）was utilized. Taking the human herpesvirus type 2 mRNA vaccine as the detection object，the optimal conditions for bacterial endotoxin detection were established by evaluating sample dilution factors，diluents，and other interfering elements. The results were validated using the recombinant C factor method. Finally，the applicability of the established method was validated through testing the respiratory syncytial virus mRNA vaccine and the shingles mRNA vaccine.Results:Increasing the sample dilution factor to 480 times with a magnesium-containing buffer and using 0.06 EU/ml tachypleus amebocyte lysate effectively mitigated the interference caused by LNP on the test results of the human herpesvirus type 2 mRNA vaccine，the respiratory syncytial virus mRNA vaccine，and the shingles mRNA vaccine. The results from recombinant C factor method and the gel clot limit method were consistent.Conclusion:A bacterial endotoxin gel clot limit detection method tailored for human herpesvirus type 2 mRNA vaccine，respiratory syncytial virus mRNA vaccine，and shingles mRNA vaccine is successfully developed based on existing protocols in the Chinese Pharmacopoeia（2020 edition）. This study offers insights for the detection of bacterial endotoxin in LNP-based biological products.

This study investigated the role of the nuclear factor of activated T cells c3 (NFATc3) in vascular smooth muscle cells (VSMCs) during aortic aneurysm and dissection (AAD) progression and the underlying molecular mechanisms. Cytoplasmic and nuclear NFATc3 levels were elevated in human and mouse AAD. VSMC-NFATc3 deletion reduced thoracic AAD (TAAD) and abdominal aortic aneurysm (AAA) progression in mice, contrary to VSMC-NFATc3 overexpression. VSMC-NFATc3 deletion reduced extracellular matrix (ECM) degradation and maintained the VSMC contractile phenotype. Nuclear NFATc3 targeted and transcriptionally upregulated matrix metalloproteinase 9 (MMP9) and MMP2, promoting ECM degradation and AAD development. NFATc3 promoted VSMC phenotypic switching by binding to eukaryotic elongation factor 2 (eEF2) and inhibiting its phosphorylation in the VSMC cytoplasm. Restoring eEF2 reversed the beneficial effects in VSMC-specific NFATc3-knockout mice. Cabamiquine-targets eEF2 and inhibits protein synthesis-inhibited AAD development and progression in VSMC-NFATc3-overexpressing mice. VSMC-NFATc3 promoted VSMC switch and ECM degradation while exacerbating AAD development, making it a novel potential therapeutic target for preventing and treating AAD.

The ability to localize sound sources rapidly allows human beings to efficiently understand the surrounding environment. Previous studies have suggested that there is an auditory "where" pathway in the cortex for processing sound locations. The neural activation in regions along this pathway encodes sound locations by opponent hemifield coding, in which each unilateral region is activated by sounds coming from the contralateral hemifield. However, it is still unclear how these regions interact with each other to form a unified representation of the auditory space. In the present study, we investigated whether functional connectivity in the auditory "where" pathway encoded sound locations during passive listening. Participants underwent functional magnetic resonance imaging while passively listening to sounds from five distinct horizontal locations (-90°, -45°, 0°, 45°, 90°). We were able to decode sound locations from the functional connectivity patterns of the "where" pathway. Furthermore, we found that such neural representation of sound locations was primarily based on the coding of sound lateralization angles to the frontal midline. In addition, whole-brain analysis indicated that functional connectivity between occipital regions and the primary auditory cortex also encoded sound locations by lateralization angles. Overall, our results reveal a lateralization-angle-based representation of sound locations encoded by functional connectivity patterns, which could add on the activation-based opponent hemifield coding to provide a more precise representation of the auditory space.
Humans ; Sound Localization/physiology* ; Male ; Female ; Magnetic Resonance Imaging ; Young Adult ; Functional Laterality/physiology* ; Adult ; Brain Mapping ; Auditory Cortex/physiology* ; Acoustic Stimulation ; Auditory Pathways/physiology* ; Brain/physiology*

Complement C3 plays a critical role in periodontitis. However, its source, role and underlying mechanisms remain unclear. In our study, by analyzing single-cell sequencing data from mouse model of periodontitis, we identified that C3 is primarily derived from periodontal fibroblasts. Subsequently, we demonstrated that C3a has a detrimental effect in ligature-induced periodontitis. C3ar^-/- mice exhibited significantly less destruction of periodontal support tissues compared to wild-type mice, characterized by mild gingival tissue damage and reduced alveolar bone loss. This reduction was associated with decreased production of pro-inflammatory mediators and reduced osteoclast infiltration in the periodontal tissues. Mechanistic studies suggested that C3a could promote macrophage polarization and osteoclast differentiation. Finally, by analyzing single-cell sequencing data from the periodontal tissues of patients with periodontitis, we found that the results observed in mice were consistent with human data. Therefore, our findings clearly demonstrate the destructive role of fibroblast-derived C3 in ligature-induced periodontitis, driven by macrophage M1 polarization and osteoclast differentiation. These data strongly support the feasibility of C3a-targeted interventions for the treatment of human periodontitis.
Animals ; Osteoclasts/cytology* ; Periodontitis/metabolism* ; Cell Differentiation ; Mice ; Fibroblasts/metabolism* ; Macrophages ; Disease Models, Animal ; Complement C3/metabolism* ; Humans ; Disease Progression ; Mice, Inbred C57BL ; Male ; Mice, Knockout

Objective:To develop a bacterial endotoxin detection method suitable for mRNA vaccines based on lipid nanoparticle（LNP）delivery system.Methods:The gel clot limit test outlined in the third section of the Chinese Pharmacopoeia（2020 edition）was utilized. Taking the human herpesvirus type 2 mRNA vaccine as the detection object，the optimal conditions for bacterial endotoxin detection were established by evaluating sample dilution factors，diluents，and other interfering elements. The results were validated using the recombinant C factor method. Finally，the applicability of the established method was validated through testing the respiratory syncytial virus mRNA vaccine and the shingles mRNA vaccine.Results:Increasing the sample dilution factor to 480 times with a magnesium-containing buffer and using 0.06 EU/ml tachypleus amebocyte lysate effectively mitigated the interference caused by LNP on the test results of the human herpesvirus type 2 mRNA vaccine，the respiratory syncytial virus mRNA vaccine，and the shingles mRNA vaccine. The results from recombinant C factor method and the gel clot limit method were consistent.Conclusion:A bacterial endotoxin gel clot limit detection method tailored for human herpesvirus type 2 mRNA vaccine，respiratory syncytial virus mRNA vaccine，and shingles mRNA vaccine is successfully developed based on existing protocols in the Chinese Pharmacopoeia（2020 edition）. This study offers insights for the detection of bacterial endotoxin in LNP-based biological products.

ObjectiveTo evaluate the efficacy and safety of Lianhua Qingke tablets in the treatment of acute bronchitis in children with the syndrome of phlegm-heat obstructing lung. MethodA randomized， open， parallel controlled， and multi-center clinical study was conduted. Children with acute bronchitis （syndrome of phlegm-heat obstructing lung） were randomly assigned to an observation group and a control group. The control group received routine basic treatment， and the observation group was treated with Lianhua Qingke Tablets on the basis of routine basic treatment. After 7 days of treatment， the clinical efficacy， TCM efficacy， time to symptom disappearance， time to cough disappearance， and clinical safety were compared between the two groups. ResultA total of 248 children were included （124 in the observation group and 124 in the control group）. After 7 days of treatment， the total response rate in terms of clinical efficacy in the observation group was 96.8% （120/124）， which was higher than that （90.3%， 112/124） in the control group （Z=-5.034， P<0.01）. The total response rate in terms of TCM syndrome in the observation group was 97.6% （121/124）， which was higher than that （93.5%， 116/124） in the control group （χ²=-5.326， P<0.01）. The scores of physical signs and TCM symptoms in the observation group were lower than those in the control group at the time of taking medicine for 3 days and 7 days （P<0.01）. The time to symptom disappearance and the time to cough disappearance in the observation group were shorter than those in the control group （P<0.01）. Drug-related adverse reactions occurred in neither group. ConclusionLianhua Qingke tablets demonstrate a definite effect on acute bronchitis in children with the syndrome of phlegm-heat blocking lung. The tablets can significantly shorten the course of disease and relieve cough and TCM symptoms， with high safety， which is worthy of clinical application and promotion.

Objective To evaluate the efficacy of XELOX regimen as neoadjuvant chemotherapy in the treatment of stage Ⅱ and Ⅲ colon cancer.Methods The clinical data of 50 patients with clinical stage Ⅱ(T4)Ⅲ colon cancer who underwent laparoscopic radical resection at general surgery department of our hospital from January 1,2012 to January 1,2021 were retrospectively analyzed.Patients were divided into neoadjuvant chemotherapy group(NACT)and adjuvant chemotherapy group(ACT)according to whether they received neoadjuvant chemotherapy with XELOX regimen.The general clinical data,adverse reactions of chemotherapy,surgical complications,operation time,intraoperative blood loss,hospitalization time,hospitalization cost,negative conversion rate of tumor markers,tumor remission rate,tumor downstaging rate,tumor response grade after chemotherapy,postoperative disease-free survival curve,and overall survival curve were retrospectively analyzed and compared among the groups.Results There were no significant differences in operative complications,postoperative exhaust time and hospital stay between NACT group and ACT group(P＞0.05).The adverse reactions of chemotherapy,the negative conversion rate of postoperative CEA and CA19-9,the duration of operation,the amount of bleeding,and the hospitalization cost in NACT group were significantly better than those in ACT group(P＜0.05).In terms of DFS and OS survival curves,with the extension of time,the decline of the NACT survival curve was smaller than that of the ACT group,and there was a significant difference in DFS survival curve(P＜0.05),but no significant difference in OS survival curve(P＞0.05).Conclusion XELOX neoadjuvant chemotherapy is safe and effective in the treatment of stage Ⅱ(T4)and stage Ⅲcolon cancer.

Objective:To investigate the effects of antibacterial absorbable suture closure in the repair of small range of bone defect wounds due to deep sternal wound infection after median thoracotomy.Methods:This study was a retrospective non-randomized clinical controlled study. A total of 32 patients (20 males and 12 females, aged (58±11) years) who met the inclusion criteria and underwent closure with antibacterial absorbable sutures (hereinafter referred to as direct closure surgery) admitted to Guangdong Provincial People's Hospital of Southern Medical University (hereinafter referred to as our hospital) from October 2017 to December 2021 were included in direct closure group. A total of 39 patients (27 males and 12 females, aged (59±11) years) who met the inclusion criteria and received bilateral pectoralis major muscle flap packing repair admitted to our hospital from January 2015 to January 2020, were included in muscle flap packing group. In the two groups, sternal infected wounds were thoroughly debrided during stage Ⅰ surgery, followed by wound repair during stage Ⅱ surgery. The width of sternal cross-section defects after debridement was less than 1 cm for patients in the two groups. For patients in direct closure group, stage Ⅱ wound repair involved intermittent sutures to the anterior sternal plate or full-thickness sternum with a total of 6 or 7 double sternal sutures. Relevant data including the duration of the stage Ⅱ wound repair surgery and the volume of blood loss during surgery, length of hospital stay, and bacterial wound infection of patients in the two groups were recorded. The postoperative complications and wound healing of patients in the two groups were recorded. During follow-up, the wound infection or recurrence of patients in the two groups and the sternal healing of patients in direct closure group were observed.Results:Compared with those in muscle flap packing group, the duration of stage Ⅱ wound repair surgery and length of hospital stay of patients in direct closure group were significantly shorter (with t values of 13.61 and 6.25, respectively, P<0.05), and there was no statistically significant difference in intraoperative blood loss volume of the stage Ⅱ wound repair surgery between the two groups ( P>0.05). The main bacterial infection in the two groups was Staphylococcus. In direct closure group, one patient had exudation in the wound two weeks post-operation, however the wound healed well after two weeks of conservative dressing changes; the wounds of the other patients healed well. In muscle flap packing group, 5 patients had postoperative complications, of which one patient died, and the wounds of 4 patients healed after dressing change or reoperation; the wounds of the other patients healed well. There was no statistically significant difference in complication incidence of patients between the two groups ( P>0.05). During the follow-up of 22-45 months, there was no re-infection or recurrence in the wound of patients in direct closure group and surviving patients in muscle flap packing group, the sternum of patients in the direct closure group achieved anatomical union. Conclusions:Direct closure surgery can not only effectively repair sternal cross-sectional defects with width below 1 cm due to deep sternal wound infections after median thoracotomy, but can also significantly shorten the operation time and duration of hospitalization.

Objective:To compare the efficacy of transcatheter arterial embolization (TAE) with laparotomy in the treatment of severe liver injury.Methods:A retrospective cohort study was conducted to analyze the clinical data of 48 patients with severe liver injury admitted to 909th Hospital of Joint Logistics Support Force (Affiliated Dongnan Hospital of Xianmen University Medical College) from December 2013 to June 2020, including 28 males and 20 females; aged 16-75 years [(45.7±6.2)years]. There were 25 patients with grade III, 15 grade IV and 8 grade V according to the American Association for the Surgery of Trauma (AAST) classification. After general treatments such as infusion and hemostasis, TAE was performed in 26 patients (TAE group) and laparotomy in 22 patients (laparotomy group). The operation time and length of hospital stay were compared between the two groups. Erythrocyte, hemoglobin and serum creatinine were compared before operation and at postoperative 1 day. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were analyzed before operation and at postoperative 1, 3, 7 days. Complications were observed.Results:All patients were followed up for 12-60 months [(17.1±9.1)months]. The operation time and length of hospital stay were (65.7±9.2)minutes and (21.6±6.6)days in TAE group, significantly shorter than (162.5±28.1)minutes and (31.5±7.4)days in laparotomy group ( P<0.05 or 0.01). There was no significant difference between the two groups referring to erythrocyte, hemoglobin and serum creatinine before operation and at postoperative 1 day (all P>0.05). There was no significant difference in ALT and AST between the two groups before operation (all P>0.05). TAE group showed ALT level of 1 154(884, 1 698)U/L, (975.3±400.9)U/L and (403.4±232.9)U/L at postoperative 1, 3, 7 days, significantly lower than 2 053(1 965, 2 132)U/L, (1 604.1±188.2)U/L and (915.3±160.5)U/L in laparotomy group (all P<0.05). TAE group showed AST level of (1 313.2±542.0)U/L, 525(302, 971)U/L and 174(84, 324)U/L at postoperative 1, 3, 7 days, significantly lower than (1 962.9±245.4)U/L, 1 478(1 089, 1 677)U/L and 837(674, 1 006)U/L in laparotomy group ( P<0.05 or 0.01). The complication rate was 26.9% (7/26) in TAE group, significantly lower than 59.1% (13/22) in laparotomy group ( P<0.05). Conclusion:For severe liver injury, TAE can significantly shorten operation time and length of hospital stay, accelerate the recovery of liver function and reduce the complication rate in comparison with laparotomy.