Objective To investigate the regulatory mechanism of transforming growth factor-β1 (TGF-β1) in different types of mitral valvular disease (MVD) with atrial fibrillation (AF). Methods From August 2011 to August 2012, patients with moderate to severe MVD accompanied by AF who required mitral valve replacement at the Department of Cardiovascular Surgery, West China Hospital, Sichuan University, were included. Based on echocardiographic results, patients were divided into two groups: a mitral regurgitation (MR) with AF (MR-AF) group and a mitral stenosis (MS) with AF (MS-AF) group. Left atrial tissue samples were collected during surgery. Techniques such as enzyme-linked immunosorbent assay, real-time fluorescence quantitative polymerase chain reaction, immunohistochemistry, and Western blotting were used to detect key molecules in the TGF-β1/JNK pathway. Results Sixteen patients were enrolled. There were 8 patients in the MR-AF group, including 5 males and 3 females, with an average age of (41.38±11.19) years; and 8 patients in the MS-AF group, including 6 males and 2 females, with an average age of (43.12±5.30) years. The left atrial volume load was higher in MR-AF patients, while the left atrial pressure load was higher in MS-AF patients. In MS-AF patients, the relative expression levels of MAPK9, JUN, CASP3, BAX, and BCL2 mRNA in left atrial tissues were significantly upregulated. The serum TGF-β1 protein level and the relative expression levels of p-JNK, p-c-Jun, and Caspase-3 proteins in the left atrial tissues of the MR-AF group were higher. Myocardial cell damage was more severe in the MS-AF group, and the protein expression level of Bcl-2 was higher. Conclusion Different MVD have distinct hemodynamic characteristics. The myocardium of the left atrium in MR-AF patients is more prone to apoptosis, possibly through the activation of the TGF-β1/JNK signaling pathway.

BACKGROUND:Pediatric flexible flatfoot is a common foot deformity that often leads to foot pain and reduced quality of life.OBJECTIVE:To explore the relationship between radiographic parameters and clinical efficacy of subtalar arthroereisis in the treatment of pediatric flexible flatfoot.METHODS:A retrospective study was conducted on 56 pediatric patients(mean age of 11.8 years)who underwent subtalar arthroereisis at Guangzhou Orthopedic Hospital between January 2022 and May 2023.All patients underwent detailed radiographic examinations and clinical evaluations before and after surgery,including the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score and Visual Analog Scale score.Paired t-tests and independent t-tests were used to compare changes in radiographic parameters and clinical scores before and after surgery.Correlation analyses were conducted to evaluate the relationship between radiographic parameters and clinical outcomes.RESULTS AND CONCLUSION:(1)All radiographic parameters significantly improved during the 8 to 12-month follow-up after surgery(P＜0.001).(2)Clinical evaluation results indicated that the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score significantly improved from 66.2±6.0 preoperatively to 91.3±5.8 postoperatively,and the Visual Analog Scale score significantly decreased from 3.1±0.8 preoperatively to 1.3±0.8 postoperatively(P＜0.001).(3)Independent t-tests showed a significant difference in postoperative the first metatarsal angle and Visual Analog Scale score grades(P=0.043),with a smaller the first metatarsal angle associated with less postoperative pain;preoperative lateral arch angle showed a significant difference between the"excellent"and"good"groups in postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot scores(P=0.033),suggesting that a smaller preoperative posterior arch angle might predict better postoperative foot function recovery.(4)Correlation analysis showed that preoperative posterior arch angle(r=-0.486,P＜0.01)and heel pitch angle(r=-0.344,P＜0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,while preoperative medial longitudinal arch angle(r=0.293,P＜0.05)was significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.Postoperative medial longitudinal arch angle(r=0.331,P＜0.05)and lateral arch angle(r=0.387,P＜0.01)were significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,whereas postoperative Bohler's angle(r=-0.272,P＜0.05),posterior arch angle(r=-0.461,P＜0.01),and heel pitch angle(r=-0.318,P＜0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.(5)It is concluded that subtalar arthroereisis is significantly effective in correcting pediatric flexible flatfoot,and improvements in radiographic parameters are closely related to clinical efficacy.Preoperative and postoperative radiographic evaluations can serve as important reference indicators for predicting postoperative clinical outcomes,guiding clinicians to optimize treatment plansand rehabilitation programs.

BACKGROUND:Pediatric flexible flatfoot is a common foot deformity that often leads to foot pain and reduced quality of life.OBJECTIVE:To explore the relationship between radiographic parameters and clinical efficacy of subtalar arthroereisis in the treatment of pediatric flexible flatfoot.METHODS:A retrospective study was conducted on 56 pediatric patients(mean age of 11.8 years)who underwent subtalar arthroereisis at Guangzhou Orthopedic Hospital between January 2022 and May 2023.All patients underwent detailed radiographic examinations and clinical evaluations before and after surgery,including the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score and Visual Analog Scale score.Paired t-tests and independent t-tests were used to compare changes in radiographic parameters and clinical scores before and after surgery.Correlation analyses were conducted to evaluate the relationship between radiographic parameters and clinical outcomes.RESULTS AND CONCLUSION:(1)All radiographic parameters significantly improved during the 8 to 12-month follow-up after surgery(P＜0.001).(2)Clinical evaluation results indicated that the American Orthopaedic Foot & Ankle Society Ankle-Hindfoot score significantly improved from 66.2±6.0 preoperatively to 91.3±5.8 postoperatively,and the Visual Analog Scale score significantly decreased from 3.1±0.8 preoperatively to 1.3±0.8 postoperatively(P＜0.001).(3)Independent t-tests showed a significant difference in postoperative the first metatarsal angle and Visual Analog Scale score grades(P=0.043),with a smaller the first metatarsal angle associated with less postoperative pain;preoperative lateral arch angle showed a significant difference between the"excellent"and"good"groups in postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot scores(P=0.033),suggesting that a smaller preoperative posterior arch angle might predict better postoperative foot function recovery.(4)Correlation analysis showed that preoperative posterior arch angle(r=-0.486,P＜0.01)and heel pitch angle(r=-0.344,P＜0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,while preoperative medial longitudinal arch angle(r=0.293,P＜0.05)was significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.Postoperative medial longitudinal arch angle(r=0.331,P＜0.05)and lateral arch angle(r=0.387,P＜0.01)were significantly positively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot,whereas postoperative Bohler's angle(r=-0.272,P＜0.05),posterior arch angle(r=-0.461,P＜0.01),and heel pitch angle(r=-0.318,P＜0.01)were significantly negatively correlated with postoperative American Orthopaedic Foot & Ankle Society Ankle-Hindfoot.(5)It is concluded that subtalar arthroereisis is significantly effective in correcting pediatric flexible flatfoot,and improvements in radiographic parameters are closely related to clinical efficacy.Preoperative and postoperative radiographic evaluations can serve as important reference indicators for predicting postoperative clinical outcomes,guiding clinicians to optimize treatment plansand rehabilitation programs.

With the rapid development of the biopharmaceutical field, the efficient and simultaneous extraction of multiple biological components from biological samples has become a critical process for advancing scientific research. The ability to simultaneously extract various molecular components such as metabolites, DNA, RNA, and proteins is pivotal for multi-omics studies, which aim to comprehensively understand the molecular mechanisms of biological systems. Traditional methods often extract these components separately, leading to challenges such as sample loss, time consumption, contamination, and inconsistencies across different data types. In contrast, simultaneous extraction techniques address these issues by maintaining the consistency of each biological component’s physiological state, improving data reliability and facilitating integration across omic platforms. This review systematically summarizes recent advances in simultaneous extraction technologies, focusing on methods such as methanol/chloroform extraction, TRIzol reagent extraction, and modified Folch extraction, which have shown significant promise in improving the efficiency and integrity of biological sample preparation. These methods offer various advantages, such as reduced sample volume requirements, decreased contamination risk, and enhanced extraction consistency, which are crucial for studies involving small sample sizes or precious clinical specimens. Among these, methanol/chloroform extraction stands out for its simplicity, low cost, and ability to extract a wide range of biological molecules. However, it does face limitations, such as its inefficiency in extracting lipids and potential RNA contamination. On the other hand, the TRIzol reagent method has become a widely adopted technique due to its ability to simultaneously isolate RNA, proteins, and metabolites from the same sample. Despite its effectiveness, the TRIzol method has limitations in RNA quality, especially when handling complex samples or those with high protein content. Modified Folch extraction, which combines liquid-liquid extraction with commercial kits, offers a highly efficient way to extract polar metabolites, lipids, RNA, DNA, and proteins from small tissue samples. This method has proven advantageous in terms of extraction yield, especially for challenging or rare samples, although it requires precise handling to avoid cross-contamination between phases. The integration of automated platforms, microfluidics, and high-throughput systems is another exciting avenue for improving simultaneous extraction. Automation facilitates large-scale, reproducible sample processing with minimal human error, while microfluidics provides high precision in sample handling and enables real-time monitoring of extraction efficiency. These innovations not only enhance the speed and reproducibility of sample preparation but also open new possibilities for single-cell analysis, where sample volumes are often limited, and extraction efficiency is critical. In addition to the technical aspects, the review also highlights the importance of optimizing extraction protocols for specific sample types, such as clinical tissues, plants, and microorganisms. For example, the challenge of extracting multiple components from cancer tissues, where sample degradation and contamination risks are high, can be mitigated by carefully selecting extraction reagents and minimizing sample handling steps. Similarly, in plant studies, where metabolite diversity is vast, the simultaneous extraction methods must be optimized to account for the unique composition of plant tissues, which often include complex secondary metabolites and cell wall components. Looking forward, the development of more efficient and standardized simultaneous extraction methods will be crucial for advancing multi-omics research. There is a growing need for protocols that can be tailored to specific research needs, ensuring both reproducibility and flexibility in diverse applications. Additionally, combining these extraction methods with high-resolution analytical techniques such as mass spectrometry and next-generation sequencing will further enhance the potential of multi-omics studies to provide comprehensive insights into biological systems. As these technologies continue to evolve, their application in personalized medicine, environmental research, and agriculture holds great promise for addressing critical scientific challenges. In conclusion, while simultaneous extraction technologies have made significant strides, several challenges remain in optimizing extraction efficiency, ensuring reproducibility, and reducing costs. Future research should focus on refining extraction protocols, developing innovative extraction reagents, and expanding the scope of these methods to cater to a broader range of biological samples. Ultimately, the continued integration of these advanced techniques will revolutionize the way biological samples are prepared, analyzed, and understood in the context of multi-omics research.

With the rapid development of the biopharmaceutical field, the efficient and simultaneous extraction of multiple biological components from biological samples has become a critical process for advancing scientific research. The ability to simultaneously extract various molecular components such as metabolites, DNA, RNA, and proteins is pivotal for multi-omics studies, which aim to comprehensively understand the molecular mechanisms of biological systems. Traditional methods often extract these components separately, leading to challenges such as sample loss, time consumption, contamination, and inconsistencies across different data types. In contrast, simultaneous extraction techniques address these issues by maintaining the consistency of each biological component’s physiological state, improving data reliability and facilitating integration across omic platforms. This review systematically summarizes recent advances in simultaneous extraction technologies, focusing on methods such as methanol/chloroform extraction, TRIzol reagent extraction, and modified Folch extraction, which have shown significant promise in improving the efficiency and integrity of biological sample preparation. These methods offer various advantages, such as reduced sample volume requirements, decreased contamination risk, and enhanced extraction consistency, which are crucial for studies involving small sample sizes or precious clinical specimens. Among these, methanol/chloroform extraction stands out for its simplicity, low cost, and ability to extract a wide range of biological molecules. However, it does face limitations, such as its inefficiency in extracting lipids and potential RNA contamination. On the other hand, the TRIzol reagent method has become a widely adopted technique due to its ability to simultaneously isolate RNA, proteins, and metabolites from the same sample. Despite its effectiveness, the TRIzol method has limitations in RNA quality, especially when handling complex samples or those with high protein content. Modified Folch extraction, which combines liquid-liquid extraction with commercial kits, offers a highly efficient way to extract polar metabolites, lipids, RNA, DNA, and proteins from small tissue samples. This method has proven advantageous in terms of extraction yield, especially for challenging or rare samples, although it requires precise handling to avoid cross-contamination between phases. The integration of automated platforms, microfluidics, and high-throughput systems is another exciting avenue for improving simultaneous extraction. Automation facilitates large-scale, reproducible sample processing with minimal human error, while microfluidics provides high precision in sample handling and enables real-time monitoring of extraction efficiency. These innovations not only enhance the speed and reproducibility of sample preparation but also open new possibilities for single-cell analysis, where sample volumes are often limited, and extraction efficiency is critical. In addition to the technical aspects, the review also highlights the importance of optimizing extraction protocols for specific sample types, such as clinical tissues, plants, and microorganisms. For example, the challenge of extracting multiple components from cancer tissues, where sample degradation and contamination risks are high, can be mitigated by carefully selecting extraction reagents and minimizing sample handling steps. Similarly, in plant studies, where metabolite diversity is vast, the simultaneous extraction methods must be optimized to account for the unique composition of plant tissues, which often include complex secondary metabolites and cell wall components. Looking forward, the development of more efficient and standardized simultaneous extraction methods will be crucial for advancing multi-omics research. There is a growing need for protocols that can be tailored to specific research needs, ensuring both reproducibility and flexibility in diverse applications. Additionally, combining these extraction methods with high-resolution analytical techniques such as mass spectrometry and next-generation sequencing will further enhance the potential of multi-omics studies to provide comprehensive insights into biological systems. As these technologies continue to evolve, their application in personalized medicine, environmental research, and agriculture holds great promise for addressing critical scientific challenges. In conclusion, while simultaneous extraction technologies have made significant strides, several challenges remain in optimizing extraction efficiency, ensuring reproducibility, and reducing costs. Future research should focus on refining extraction protocols, developing innovative extraction reagents, and expanding the scope of these methods to cater to a broader range of biological samples. Ultimately, the continued integration of these advanced techniques will revolutionize the way biological samples are prepared, analyzed, and understood in the context of multi-omics research.

Methods: Six finite element models of T12 burst fractures were developed to simulate short-segment stabilization under normal or osteoporotic bone conditions, with/without augmentation screws at the fractured vertebra. Spinal stiffness, implant stresses, and axial displacement/micromotion of the bony defect were measured and compared under mechanical loading. Results: Osteoporotic models exhibited a greater range of motion (ROM) than normal bone. All six-screw constructs reduced ROM across all motions compared with traditional four-screw models. Osteoporotic fracture models gained greater benefit from intermediate screw augmentation at the fracture vertebra, which also lowered axial displacement/micromotion. In six-screw models, rod stress increased while pedicle screw stress decreased. Intermediate screws at fractured vertebrae produced similar changes in stress distribution across all fixation models, regardless of bone quality. Conclusions Our findings may facilitate implant selection for osteoporotic burst fractures, supporting the use of more rigid fixation sixscrew constructs to reduce the risk of mechanical failure and postoperative re-collapse.

Background: Aspiration pneumonia (AP), which can be caused by gastric content inhalation into the lower airways, causes acute lung injury (ALI) through complex mechanisms, including inflammation, oxidative stress, and apoptosis. Here, we evaluated the efficacy of exosomes derived from human placental mesenchymal stem cells (hpMSCs) in mitigating ALI in a murine model of AP. We also investigated the role of hsa-let-7i-5p, the most abundant miRNA in hpMSC-derived exosomes, in this respect. Methods: Adult male C57BL/6 mouse AP models were administered hpMSC-derived exosomes (APExo group) or phosphate-buffered saline (AP group) intra-tracheally. After 48 h, the mice were euthanized and evaluated. The effects of hsa-let-7i-5p were assessed by specific inhibition or overexpression. Results: Compared with the APExo group, the AP group exhibited significantly greater ALI, as evidenced by histological damage, increased lung injury scores, impaired lung function, increased leukocyte infiltration, and elevated tissue edema (all P < 0.05). The untreated AP group also showed more inflammation, characterized by nuclear factor-κB upregulation, macrophage M1 polarization, and cytokine level elevation (tumor necrosis factor-α, interleukin-1β, and interleukin-6), as well as increased oxidation and activation of the apoptosis pathway (all P < 0.05). Notably, the therapeutic effects of hpMSC-derived exosomes were compromised by specific inhibition of hsa-let-7i-5p. Furthermore, engineered exosomes derived from genetically modified RAW264.7 overexpressing hsa-let-7i-5p demonstrated therapeutic effects against AP similar to those obtained with hpMSC-derived exosomes. Conclusions In a murine AP model, intra-tracheal administration of hpMSC-derived exosomes has ALI-mitigating effects, involving inflammation, oxidation, and apoptosis modulation, with hsa-let-7i-5p playing a pivotal mediating role.

Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Background and aims Alcohol-associated liver disease (ALD) is a leading cause of liver-related morbidity and mortality worldwide, with no currently effective treatment. ALD is caused by excessive lipid buildup, which eventually triggers inflammation and fibrosis in the liver. Activation of hepatic Kupffer cells (KCs) and macrophages drives liver inflammation, which can worsen alcohol-induced liver injury. The autophagy-lysosome system is crucial for macrophages to support their innate immune functions. Transcription factor EB (TFEB) is a key regulator of autophagy and lysosomal biogenesis, but the role of macrophage TFEB in ALD development is unknown. The aim of this study was to evaluate the effects of Gao-binge alcohol consumption on myeloid cell TFEB and elucidate the role of myeloid TFEB in ALD. Methods Two-to-three-month-old male and female LysM Cre− (WT) and LysM Cre+ Tfeb Flox/Flox (f/f) (myeloid-Tfeb KO) mice were subjected to chronic alcohol feeding plus an acute binge following the Gao-binge model. Serum alanine aminotransferase, aspartate aminotransferase, triglycerides, and cholesterol content were determined using biochemical assays. Total hepatic protein content and messenger RNA (mRNA) levels of autophagy-related proteins and inflammatory markers were determined using immunoblotting, immunohistochemistry, and real-time quantitative polymerase chain reaction (RT-qPCR). Isolated hepatic infiltrating macrophages and KCs from mice given intragastric ethanol infusions were analyzed by Western blot for TFEB and autophagy-related protein content. Raw 264.7 macrophages were treated with ethanol, lipopolysaccharide (LPS), and LPS plus ethanol to examine nuclear TFEB translocation using immunofluorescence. Results We found that TFEB levels were higher in macrophage/KC cells than in hepatocytes and cholangiocytes. While ethanol feeding increased serum alanine aminotransferase and aspartate aminotransferase levels, as well as hepatic triglyceride levels, no significant differences were observed between WT and myeloid-Tfeb KO mice. The number of F4/80-positive KCs/macrophages was similar in all four experimental groups, but hepatic neutrophil infiltration increased in alcohol-fed myeloid-Tfeb KO mice. LPS or ethanol alone induced nuclear TFEB translocation only moderately in Raw 264.7 macrophages. Conclusions Our findings suggest that myeloid TFEB is dispensable for alcohol-induced liver injury in mice.

It is regarded that the disease location of refractory facial paralysis is in the channel sinews of the face， with its primary pathogenesis characterized by a combination of deficiency and stasis of the channel sinews. The integration of repeated shallow needling method and fire acupuncture can first remove stagnation within the channel sinews， and second utilize the warming effect of fire to reinforce yang， stimulate meridian qi， and nourish the channel sinews. This approach balances both supplementation and drainage manipulation， aligning with the underlying pathogenesis of deficiency and stasis combination. In clinical practice， diagnostic methods should be applied flexibly to accurately identify the affected channel sinews. The severity of facial symptoms， the size and mobility of the paralyzed facial muscles， as well as the depth and size of the reactive points identified through palpation， should be considered when determining the extent of the condition. By adjusting the appropriate level of stimulation， the fire acupuncture with repeated shallow needling method could effectively improve facial muscle morphology and function， promoting recovery from the disease.