ObjectiveTo explore the clinical efficacy and mechanism of Bupi Qingfei prescription （BPQF） in treating stable bronchiectasis in the patients with syndromes of lung-spleen Qi deficiency and phlegm-heat accumulation in the lungs. MethodsA randomized， double-blind， placebo-controlled trial was conducted. Patients were randomized into BPQF and placebo control （PC） groups. On the basis of conventional Western medicine treatment， the BPQF granules and placebo were respectively administered at 10 g each time， twice a day， for a course of 24 weeks. The TCM symptom scores， Quality of Life Questionnaire for Bronchiectasis （QOL-B） scores， lung function indicators， T lymphocyte subsets， level of inflammatory factors in the sputum， level of neutrophil elastase （NE） in the sputum， and occurrence of adverse reactions were observed before and after treatment in the two groups. ResultsA total of 64 patients completed the study， encompassing 32 in the BPQF group and 32 in the PC group. After treatment， the BPQF group showed decreased TCM symptom scores （P<0.01）， increased QOL-B scores （P<0.01）， and declined levels of tumor necrosis factor （TNF）-α and NE （P<0.05， P<0.01）. The PC group showed decreased TCM symptom （except spleen deficiency） scores （P<0.01）， increased the QOL-B health cognition and respiratory symptom domain scores （P<0.05， P<0.01）， and a declined TNF-α level （P<0.01）. Moreover， the BPQF group had lower TCM symptom （except chest tightness） scores （P<0.05， P<0.01）， higher QOL-B （except treatment burden） scores （P<0.05， P<0.01）， and lower levels of interleukin-6 and TNF-α （P<0.05） than the PC group. Neither group showed serious adverse reactions during the treatment process. ConclusionBPQF can ameliorate the clinical symptoms of stable bronchiectasis patients who have lung-spleen Qi deficiency or phlegm-heat accumulation in the lungs by regulating the immune balance and inhibiting airway inflammatory responses.

OBJECTIVE To explore a model for constructing a platform for medical institution formulation and provide insights for promoting their development. METHODS By systematically reviewing the development status and challenges of medical institution preparations in China， and based on the theory of industry-university-research collaborative innovation， the organizational structure， collaborative processes， and safeguard mechanisms of the platform were designed. RESULTS & CONCLUSIONS Medical institution formulations in China mainly faced challenges such as weak research and development （R&D） capacity， uneven quality standards， and blocked transformation pathways. This study established a full-chain， whole- industry collaborative innovation network covering the government， medical institutions， universities/research institutes， pharmaceutical enterprises， and the market， forming a new “government-industry-university-research-application” five-in-one platform model for medical institution formulations. By establishing mechanisms such as multi-entity collaborative cooperation， full- chain intellectual property management， contribution-based benefit distribution， staged risk-sharing， and third-party evaluation， the model clarified the responsibilities and collaborative pathways of all parties. The new model highlights the whole-process transformation of clinical experience-based prescriptions， enabling precise alignment between clinical needs and technological R&D， as well as between preparation achievements and industrial transformation. While breaking down the barriers of traditional platform construction， it effectively achieves optimal resource allocation and complementary advantages， addresses problems emerging in the development of medical institution preparations， and provides reference value for the formulation of relevant systems.

Background: Influenza A virus (IAV) is a negative-sense RNA virus of the Orthomyxoviridae family and is the etiological agent of a highly contagious acute respiratory disease that can lead to acute lung injury. Objective: To elucidate the molecular mechanisms of IAV infection, an integrative research approach combining gene expression profiling, multinetwork analysis, and in vivo experimental validations was employed. Methods: First, a series of network-based analyses were performed, including protein-protein interaction network construction, weighted gene co-expression network analysis, and subsequent gene set enrichment analysis, to identify the major underlying mechanisms of IAV infection. Following gene expression analysis, core targets, both direct and indirect regulators, were screened. An IAV (H1N1) strain A/PR/8/34-induced acute lung injury mouse model was constructed for in vivo validations. Batch one included two groups to evaluate findings from the multi-network analysis: Mock (n = 10; 5 males and 5 females) and IAV (n = 10; 5 males and 5 females). Batch two included three groups to assess the role of toll-like receptor 3 (TLR3) in IAV infection: Mock (n = 6; 3 males and 3 females), IAV (n = 6; 3 males and 3 females), and TLR3 inhibitor (n = 6; 3 males and 3 females). Body weight was measured on days 0, 3, and 5 after infection. On day 5, lung tissues were collected to assess viral load and histopathological changes. Key targets were examined using enzyme-linked immunosorbent assay, Western blotting, and immunofluorescence staining, both in sera and lung tissues. Results: IAV infection was significantly associated with dysregulation of the immune-inflammation system, such as the LTR, nucle-otide-binding oligomerization domain-(NOD) like receptor, retinoic acid-inducible gene I-like receptor, and nuclear factor kappa-B signaling pathways. Gene set enrichment analysis further indicated that the TLR and necroptosis signaling pathways played crucial roles in the progression of IAV infection (TLR signaling pathway normalized enrichment score = 2.3941, P = 1.00 × 10 ⁻¹⁰; necroptosis normalized enrichment score = 1.9421, P = 6.21 × 10 ⁻⁷). Among the core targets, TLR3 and mixed lineage kinase domain-like protein (MLKL) may regulate gene expression at the transcriptional level (all P < 0.05). In vivo validation using an IAV (PR8) infected acute lung injury mouse model demonstrated increased viral load and lung index, alveolar structural damage, and inflammatory cell infiltration. Immunofluorescence staining exhibited large gaps in Lamin B1 staining and breaches in Emerin signals following IAV-PR8 infection. Expression levels of TLR3, p-receptor-interacting serine/threonine-protein kinase 3 (RIPK3)/RIPK3, and p-mixed lineage kinase domain-like protein (MLKL)/MLKL proteins in lung tissues, as well as proinflammatory factors and mediators in sera, were significantly elevated after IAV infection. Moreover, enhanced neutrophil infiltration (myeloperoxidase) and citrullinated histone H3 (a neutrophil extracellular trap-specific marker), both established indicators of neutrophil extracellular trap formation, were observed. Notably, treatment with a TLR3 inhibitor significantly ameliorated IAV-induced acute lung injury by regulating necroptosis-related targets. Conclusion: Our study provides network-based in vivo evidence that TLR3-receptor-interacting serine/threonine-protein kinase 3-MLKL-mediated necroptosis may underlie IAV-induced acute lung injury and could serve as a potential therapeutic target in severe influenza cases.

ObjectiveTo generate cynomolgus and rhesus monkey models carrying the Booroola fecundity (FecB) variant of bone morphogenetic protein receptor type 1B (BMPR1B) gene (designated BMPR1B^FecB) using adenine base edit technology and to explore genetic improvement strategies for enhancing the reproductive performance of laboratory macaques. MethodsFour single-guide RNAs (sgRNAs) were designed to target the BMPR1B c.746A>G locus in cynomolgus and rhesus monkeys. The sgRNAs were combined with the ABE8e-SpRY base editor, and the resulting editing complexes were microinjected into zygotes of both macaque species. Following in vitro culture of the injected embryos, nested PCR amplification was performed to detect the base-editing efficiency at the target site to screen for highly efficient sgRNAs. Subsequently, normally developing gene-edited embryos were transferred into the oviducts of surrogates. Seven days after birth, ear skin or peripheral blood samples were collected from the newborn monkeys for genotyping and mosaicism analysis, and the growth, development, and health status of the gene-edited monkeys were monitored. ResultsAmong the four candidate sgRNAs, sgRNA4 was identified as achieving 100% editing efficiency at the c.746A>G site. A total of 17 live monkey offspring were obtained via embryo transfer, including 8 cynomolgus monkeys and 9 rhesus monkeys. Genotyping results showed that all 8 newborn cynomolgus monkeys carried the c.746A>G point mutation, with an editing efficiency of 100% (8/8), of which 37.50% (3/8) were mosaics. Among the 9 newborn rhesus monkeys, 8 individuals carried the c.746A>G mutation, with an editing efficiency of 88.89% (8/9), of which 75.00% (6/8) were mosaics. ConclusionThe ABE8e-SpRY base-editing system enables precise editing at the c.746A>G (p.Q249R) locus of the BMPR1B gene in both cynomolgus and rhesus monkey embryos, resulting in the successful establishment of laboratory monkey models carrying the BMPR1B^FecB variant. These models provide a valuable foundation for future studies of the regulatory role of the BMPR1B^FecB variant in reproductive performance at the non‑human primate level, as well as for exploring genetic improvement strategies to enhance reproductive performance in laboratory monkeys.

OBJECTIVE: To explore the effectiveness of the modified Lemaire procedure in anterior cruciate ligament reconstruction (ACLR) in patients with a highly positive pivot shift test. METHODS: The clinical data of 18 patients with anterior cruciate ligament (ACL) rupture and highly positive pivot shift test between April 2020 and September 2022 were retrospectively analyzed. There were 13 males and 5 females with an average age of 28.3 years (range, 17-41 years). Causes of injury included 11 cases of direct violence injury, including 6 cases of traffic accident injury, 4 cases of sports injury, 1 case of falling injury; 7 cases of indirect violence injury, all sports injury. All patients had complete ACL rupture, including 15 acute injuries and 3 old injuries. The preoperative pivot shift test was grade Ⅱ in 9 cases and grade Ⅲ in 9 cases. All patients were treated with ACLR combined with modified Lemaire procedure. The International Knee Documentation Committee (IKDC) score and Lysholm score were used to evaluate the effectiveness before operation and at 3, 6, 12 months after operation. KT-2000 arthrometer was used to measure the anterior stability of the knee joint, and the difference between the healthy and affected sides was recorded. Pivot shift test was used to evaluate the rotational stability of the knee joint. During the follow-up, X-ray films were taken to observe the bone tunnel and internal fixation, and MRI was used to examine the healing of ACL, anterolateral collateral ligament and fibular collateral ligament grafts. RESULTS: All patients completed the operation successfully without complications such as knee joint infection, vascular and nerve injury. All patients were followed up 12-19 months (mean, 13.2 months). After operation, the rotational stability of the knee joint recovered satisfactorily, and there was no adverse symptom such as knee instability and locking at last follow-up. X-ray film and MRI showed that the bone tunnel was anatomically located and healed well, the internal fixation was in good position, and the reconstructed ACL and iliotibial band were continuous and in good tension. The IKDC score, Lysholm score, and the difference of KT-2000 between the healthy and the affected sides significantly improved at 3, 6, and 12 months after operation ( P<0.05). All the indicators further improved with time after operation, except that there was no significant difference in IKDC score between 3 and 6 months after operation and in the difference of KT-2000 between 3 months and 6, 12 months after operation ( P>0.05), and there were significant differences in other indicators between different time points ( P<0.05). Pivot shift test was negative immediately after operation and at last follow-up. CONCLUSION In ACL injuries with a highly positive pivot shift test, ACLR combined with the modified Lemaire procedure can effectively restore anterolateral knee stability, leading to satisfactory knee stability and function in the early postoperative period.
Humans ; Anterior Cruciate Ligament Reconstruction/methods* ; Male ; Female ; Anterior Cruciate Ligament Injuries/physiopathology* ; Adult ; Adolescent ; Retrospective Studies ; Young Adult ; Anterior Cruciate Ligament/surgery* ; Treatment Outcome ; Joint Instability/surgery* ; Knee Joint/physiopathology* ; Range of Motion, Articular

Cells and exosomes derived from them are extensively used as biological carrier systems. Cells demonstrate superior targeting specificity and prolonged circulation facilitated by their rich array of surface proteins, while exosomes, due to their small size, cross barriers and penetrate tumors efficiently. However, challenges remain, cells' large size restricts tissue penetration, and exosomes have limited targeting accuracy and short circulation times. To address these challenges, we developed a novel concept termed exosomal spheres. This approach involved incorporating platelet-derived exosomes shielded with phosphatidylserine (PS) and linked via pH-sensitive bonds for drug delivery applications. The study demonstrated that, compared with exosomes, the exosomal spheres improved blood circulation through the upregulation of CD47 expression and shielding of phosphatidylserine, thereby minimizing immune clearance. Moreover, the increased expression of P-selectin promoted adhesion to circulating tumor cells, thereby enhancing targeting efficiency. Upon reaching the tumor site, the hydrazone bonds of exosome spheres were protonated in the acidic tumor microenvironment, leading to disintegration into uniform-sized exosomes capable of deeper tumor penetration compared to platelets. These findings suggested that exosome spheres addressed the challenges and offered significant potential for efficient and precise drug delivery.

Objective:To compare the efficacy of surgical resection and radiofrequency ablation (RFA) treatment for China liver cancer staging (CNLC) Ia hepatocellular carcinoma (HCC) at difficult-to-reach locations.Methods:A retrospective analysis was conducted on the clinical data of 114 patients with CNLC Ia HCC at Ⅶ、Ⅷ、Ⅳb or Ⅰ segments that were difficult-to-reach locations who were admitted to People's Hospital of Zhengzhou University from December 2018 to December 2023. Among the patients, 85 were males and 29 were females, aged (58.1±1.0) years. The patients were divided into two groups: a RFA group with 31 cases and a surgical resection group with 83 cases. Compare the levels of alanine transaminase (ALT) and aspartate transaminase (AST) before and after surgery, the surgical time, intraoperative blood loss, postoperative hospital stay, postoperative complications, recurrence free survival rate, and cumulative survival rate between the two groups.Results:The comparison of age, gender, ALT, and AST between the two groups showed no statistically significant differences (all P>0.05). The differences in ALT and AST levels before and after surgery in the RFA group were (134.8±38.7) U/L and (195.1±53.9) U/L, respectively, which were significantly lower than those in the surgical resection group [(226.8±17.9) U/L and (229.5±16.2) U/L] ( t=-2.45 and -1.12, P=0.016 and 0.041). The RFA group had shorter operation time [(69.2±11.7) min vs. (210.6±8.9) min], less intraoperative blood loss [(8.7±3.8) ml vs. (238.6±20.8) ml], and shorter postoperative hospital stays [(6.4±1.0) d vs. (13.1±0.4) d] compared to the surgical resection group, with all differences statistically significant (all P<0.05). The overall complication rates were 19.4% (6/31) in the RFA group and 22.9% (19/83) in the surgical resection group, showing no significant difference ( χ2=0.16, P=0.685). No statistically significant diffe-rence was found in recurrence-free survival rates between the two groups ( χ2=0.13, P=0.717). Similarly, there was no statistically significant difference in cumulative survival rates between the groups ( χ2<0.01, P=0.978). Conclusion:For HCC at CNLC Ⅰa in challenging locations, RFA demonstrated shorter operation time and postoperative hospital stay, less intraoperative bleeding, and superior liver function recovery compared with surgical resection, while no significant difference was observed in survival outcomes between the two treatment groups.

Objective:To analyze the efficacy of endovascular treatment for venous outflow tract obstruction after liver transplantation.Methods:A retrospective analysis was conducted on the data of 7 patients with venous outflow tract obstruction after liver transplantation admitted to Beijing Friendship Hospital, Capital Medical University from November 2020 to December 2024. Among them, there were 5 males and 2 females, with the age of (22.3±8.1) years. The primary diseases included 2 cases of Budd-Chiari syndrome, 1 case of hepatic veno-occlusive disease, 1 case of portal veno-hepatic sinus vascular disease, 1 case of ornithine carbamoyltransferase deficiency, 1 case of primary biliary cirrhosis, and 1 case of autoimmune cirrhosis. Analyze the patient's clinical manifestations, obstruction of venous outflow tract, hemoglobin levels within one week before and one week after the operation, endovascular treatment conditions, and intraoperative complications such as abdominal hemorrhage and vascular injury. Ultrasound was used to measure the depth of ascites and pleural effusion. All patients were followed up immediately after the operation through phone calls or follow-up visits. The clinical symptoms, abdominal vascular ultrasound, enhanced CT and survival status of the patients were followed up.Results:All 7 patients were diagnosed with venous outflow tract obstruction by intraoperative angiography, including 1 cases of inferior vena cava obstruction, 2 cases of hepatic vein obstruction, and 4 cases of vena cava combined with hepatic vein obstruction. A total of 12 endovascular treatments were performed on 7 patients. Among them, 4 patients received balloon dilation and angioplasty once, 1 patient received balloon dilation and angioplasty twice, 1 patient underwent hepatic vein stent implantation after 2 hepatic vein balloon dilation and angioplasty, and 1 patient underwent intrahepatic portosystemic shunt via jugular vein after 2 hepatic vein balloon dilation and angioplasty. The abdominal distensionof the patients were all relieved after the operation, the ascites and pleural effusion decreased, and the edema symptoms of the lower extremities disappeared. There were no intraoperative complications. The preoperative hemoglobin of 7 patients was (113.4±34.0) g/L, and the postoperative hemoglobin was (126.6±34.8) g/L, which increased significantly compared with that before the operation, and the difference was statistically significant ( t=-0.71, P=0.038). Seven patients were followed up for 6 to 24 months, with a median of 12 months. None of them had obvious symptoms including abdominal distension. Abdominal ultrasound and CT indicated that the blood flow of the transplanted liver was unobstructed, and no patient died. Conclusion:Venous outflow tract obstruction after liver transplantation can cause severe symptoms. Endovascular treatment is an effective treatment for venous outflow tract obstruction after liver transplantation.

Objective To explore the feasibility of phase-contrast CT for three-dimensional visualization of in vitro intrahepatic vessels and necrotic foci in mouse model of acute liver injury(ALI).Methods Six clean grade male wild-type C57BL/6 mice were randomly divided into model group and control group(each n=3).The mice in model group were fed with overdose of acetaminophen to induce ALI,while those in control group were fed with the same amount of distilled water.After 24 h,the mice were all sacrificed,the livers were harvested and then fixed and dehydrated.Phase-contrast CT images of in vitro liver were acquired,sectional and three-dimensional images were reconstructed.The effect of phase-contrast CT for displaying the outline of liver and internal vessels and necrotic foci were observed,the maximum diameter and volume of necrotic foci were quantitatively analyzed,and the results of phase-contrast CT were compared with pathological findings.Results The original projection and sectional images of phase-contrast CT clearly showed the outline of in vitro liver and internal vasculatures.Necrotic foci within the liver were found in model group,but not in control group.The findings of phase-contrast CT corresponded accurately to those of pathology.Three-dimensional reconstruction images of phase-contrast CT clearly displayed intrahepatic portal vein system and hepatic vein system in both groups,and discontinuous punctate necrotic foci within the liver were found in model group,mainly distributed around hepatic vein,with a median maximum diameter of 18.50 μm and a median volume of 5 870.11 μm3,but was not observed in control group.Conclusion Phase-contrast CT could be used in three-dimensional visualization of intrahepatic vessels and necrotic foci of in vitro liver in mouse model of ALI.

Objective To analyze the influencing factors of tumor-specific mortality in patients with advanced endometrial carcinoma and construct a risk prediction model.Methods A total of 150 patients with advanced endometrial carcinoma admitted to the Fourth Clinical College of Zhejiang Chinese Medical University,Hangzhou First People's Hospital from January 2020 to January 2023 were selected as subjects.Cox regression was employed to analyze factors influencing mortality in advanced endometrial carcinoma.A Logistic regression model was constructed,with receiver operating characteristic(ROC)curve used to evaluate model discrimination,calibration curve for precision assessment,and decision curve for net benefit evaluation.Results Overall 2-year survival rate of 150 patients was 58.67％.The univariate analysis demonstrated that histological grade,lymph node metastasis,tumor maximum diameter,and preoperative carcinoembryonic antigen 125(CA125)levels were all significant risk factors for mortality in advanced endometrial carcinoma(P＜0.05).Multivariate analysis confirmed these parameters as independent prognostic indicators(P＜0.05).A Logistic regression model established using multivariate Cox regression yielded a C index of 0.873(95％CI:0.792-0.951).ROC curve analysis revealed the model's area under the curve for predicting mortality risk factors in advanced endometrial carcinoma was 0.916(95％CI:0.872-0.960).Conclusion The linear chart prediction model constructed based on histological grading,lymph node metastasis,maximum tumor diameter and preoperative CA125 level in this study can effectively predict the death of advanced endometrial carcinoma.