ObjectiveTo investigate the mechanism by which Shengjiang Pingchuan Zhike pills （SPZP） alleviate airway injury in the rat model of cough variant asthma （CVA） via regulating autophagy through activating the adenosine monophosphate-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR）/Unc-51-like autophagy activating kinase 1 （ULK1） signaling pathway based on transcriptomics， thus providing experimental evidence for the clinical application of SPZP. MethodsForty SPF-grade SD rats were enrolled in the present study and randomly via the random number table method assigned into four groups： a blank group， a model group， a SPZP （2 160 mg·kg^-1·d^-1） group， and a positive drug （montelukast sodium， MTL， 0.9 mg·kg^-1·d^-1）group， with 10 rats in each group. After one week of adaptive feeding， the rat model of cough variant asthma （CVA） was established by two-week sensitization with ovalbumin （OVA） and aluminium hydroxide， followed by capsaicin atomization challenge. Rats in the blank group received an equal volume of normal saline during modeling. Upon successful modeling， rats in the SPZP group and positive drug group were administrated with SPZP suspension and MTL solution， respectively， while those in the blank and model groups received the same dose of normal saline by gavage. The whole intervention lasted for 21 consecutive days. After the intervention， general behaviors of rats in each group were observed. Pulmonary function indicators were subsequently detected. Hematoxylin-eosin （HE） staining was performed to visualize the pathological morphology of the lung tissue， and periodic acid-Schiff （PAS） staining was adopted to evaluate airway mucus secretion. Transcriptomic sequencing was conducted to screen differentially expressed genes in the lung tissue across groups. The levels of inflammatory cytokines including interleukin （IL）-4， IL-5， immunoglobulin E （IgE）， and interferon-γ （IFN-γ） in the bronchoalveolar lavage fluid （BALF） were quantified by enzyme-linked immunosorbent assay （ELISA）. Immunofluorescence staining was used for detecting the co-localization of autophagy-related protein Beclin-1 and phosphorylated mammalian target of rapamycin （p-mTOR）. Transmission electron microscopy was employed to count autophagosomes and observe mitochondrial morphological changes in the lung tissue. Western blot assay was utilized to measure the expression levels of AMPK/mTOR/ULK1 axis-related proteins， including AMPK， phosphorylated adenosine monophosphate-activated protein kinase （p-AMPK）， mTOR， phosphorylated mammalian target of rapamycin （p-mTOR）， ULK1， and phosphorylated Unc-51-like autophagy activating kinase 1 （p-ULK1） as well as the autophagy biomarkers Beclin-1 and selective autophagy adaptor protein p62 in lung specimens. ResultsCompared with the blank group， the model group exhibited markedly elevated cough frequency accompanied by wheezing and stridor， dull fur， poor mental activity， diminished motor activity， accelerated respiratory rate， and weakened sensory response and mobility to external stimuli， accompanied by a declining tendency of body mass. Meanwhile， the model group exhibited decreased dynamic lung compliance， increased airway resistance （P<0.01）， severe pathological damage and excessive mucus secretion in the lung tissue （P<0.01）， and elevated levels of IL-4， IL-5， and IgE and declined level of IFN-γ in the BALF （P<0.01）. Ultrastructural observation revealed damaged mitochondrial morphology and a sharp reduction in the number of autophagosomes in the lung tissue. In addition， the expression of p-AMPK， p-ULK1， and Beclin-1 were downregulated， whereas p-mTOR and p62 were highly expressed （P<0.05， P<0.01） in the model group. Gene Ontology （GO） and Kyoto Encyclopedia of Genes and Genomes （KEGG） enrichment analyses demonstrated significant enrichment of AMPK， mTOR， and autophagy-related pathways. Compared with the model group， both the positive drug group and SPZP group obtained visible improvements in general behavioral manifestations and obvious recovery of pulmonary function indicators （P<0.05）， alleviated pulmonary pathological lesions and suppressed excessive airway mucus secretion （P<0.05）， recovered levels of inflammatory cytokines in the BALF （P<0.05）， restored mitochondrial morphology， increased autophagosomes， upregulated expression levels of p-AMPK， p-ULK1， and Beclin-1， and downregulated expression levels of p-mTOR and p62 （P<0.05）. ConclusionSPZP can effectively ameliorate airway inflammation and improve the pulmonary function in the rat model of CVA by activating the AMPK/mTOR/ULK1 signaling pathway， enhancing autophagy in the lung tissue， and inhibiting excessive airway mucus secretion and immune inflammatory imbalance.

OBJECTIVE: To evaluate the effectiveness of functional perforator flaps utilizing the superficial circumflex iliac artery as a vascular pedicle, as well as chimeric iliac bone flaps, in the reconstruction of composite tissue defects in the hand and foot. METHODS: A retrospective review of the clinical data from 13 patients suffering from severe hand or foot injuries, treated between May 2019 and January 2025, was conducted. The cohort comprised 8 males and 5 females, with ages ranging from 31 to 67 years (mean, 48.5 years). The injuries caused by mechanical crush incidents (n=9) and traffic accidents (n=4). The distribution of injury sites included 8 cases involving the hand and 5 cases involving the foot. Preoperatively, all patients exhibited bone defects ranging from 2.0 to 6.5 cm and soft tissue defects ranging from 10 to 210 cm². Reconstruction was performed using functional perforator flaps based on the superficial circumflex iliac artery and chimeric iliac bone flaps. The size of iliac bone flaps ranged from 2.5 cm×1.0 cm×1.0 cm to 7.0 cm×2.0 cm×1.5 cm, while the size of the soft tissue flaps ranged from 4 cm×3 cm to 15 cm×8 cm. In 1 case with a significant hand defect, a posterior interosseous artery perforator flap measuring 10.0 cm×4.5 cm was utilized as an adjunct. Likewise, an anterolateral thigh perforator flap measuring 25 cm×7 cm was combined in 1 case involving a foot defect. All donor sites were primarily closed. Postoperative flap survival was monitored, and bone healing was evaluated through imaging examination. Functional outcomes were assessed based on the location of the defects: for hand injuries, grip strength, pinch strength, and flap two-point discrimination were measured; for foot injuries, the American Orthopaedic Foot & Ankle Society (AOFAS) score, visual analogue scale (VAS) score, Maryland Foot Score, plantar pressure distribution and gait symmetry index (GSI) were evaluated. RESULTS: All flaps survived completely, with primary healing observed at both donor and recipient sites. All patients were followed up 6-18 months (mean, 12.2 months). No significant flap swelling or deformity was observed. Imaging examination showed a bone callus crossing rate of 92.3% (12/13) at 3 months after operation, and bone density recovered to more than 80% of the healthy side at 6 months. The time required for bone flap integration ranged from 2 to 6 months (mean, 3.2 months). One patient with a foot injury exhibited hypertrophic scarring at the donor site; however, no major complication, such as infection or bone nonunion, was noted. At 6 months after operation, grip strength in 8 patients involving the hand recovered to 75%-90% of the healthy side (mean, 83.2%), while pinch strength recovered to 70%-85% (mean, 80%). Flap two-point discrimination ranged from 8 to 12 mm, approaching the sensory capacity of the healthy side (5-8 mm). Among the 5 patients involving the foot, the AOFAS score at 8 months was 80.5±7.3, VAS score was 5.2±1.6. According to the Maryland Foot Score, 2 cases were rated as excellent and 3 as good. Gait analysis at 6 months after operation showed GSI above 90%, with plantar pressure distribution closely resembling that of the contralateral foot. CONCLUSION The use of functional perforator flaps based on the superficial circumflex iliac artery, combined with chimeric iliac bone flaps, provides a reliable vascular supply and effective functional restoration for the simultaneous repair of composite bone and soft tissue defects in the hand or foot. This technique represents a viable and effective reconstructive option for composite tissue defects in these anatomical regions.
Humans ; Male ; Middle Aged ; Female ; Perforator Flap/transplantation* ; Adult ; Plastic Surgery Procedures/methods* ; Hand Injuries/surgery* ; Aged ; Retrospective Studies ; Foot Injuries/surgery* ; Ilium/transplantation* ; Iliac Artery/surgery* ; Soft Tissue Injuries/surgery* ; Bone Transplantation/methods* ; Treatment Outcome

Neurological disorders are an important cause of global human disease burden， but the mechanisms of the development and progression of neurological disorders remain unclear， and most studies have shown that they are associated with the expression of mutations in human genes. MEF2D， as an important transcription factor in the MEF2 family， plays an important role in both physiological and pathological processes. Related studies have found that MEF2D can be involved in neuronal survival， regulate neuronal responses to stimuli， and participate in the pathogenic process of neurological disorders. This article reviews the research advances in MEF2D in common neurological disorders.

Listeria monocytogenes (Lm) is zoonotic pathogen that can cause listeriosis, and vaccine is one of the effective methods to prevent this pathogen infection. In this study, we developed a novel vaccine that is a mixture of inactivated bacteria and Montanide™ ISA 61 VG, a mineral oil adjuvant, and evaluated the safety and immune response characteristics of this vaccine. The mice immunized with the ISA 61 VG adjuvant had high safety, and it could induce significantly higher titer of anti-listeriolysin O (LLO) antibody and higher value of IgG2a/IgG1 ratio compared with the group without the adjuvant. In particular, it could provide 100% immune protection against lethal doses of Lm challenge in mice. In summary, ISA 61VG adjuvant significantly enhanced the ability of inactivated listeria vaccine to induce humoral and cellular immune responses, thereby enhanced the protective immune response in the host, and it is a potential vaccine candidate for the prevention of Lm infection in humans and animals.
Adjuvants, Immunologic ; pharmacology ; Animals ; Hemolysin Proteins ; immunology ; pharmacology ; Immunity, Cellular ; drug effects ; Listeria monocytogenes ; immunology ; Listeriosis ; prevention & control ; Mice ; Mice, Inbred BALB C ; Vaccines, Inactivated ; immunology

The ongoing pandemic of Coronavirus disease 19 (COVID-19) is caused by a newly discovered β Coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). How long the adaptive immunity triggered by SARS-CoV-2 can last is of critical clinical relevance in assessing the probability of second infection and efficacy of vaccination. Here we examined, using ELISA, the IgG antibodies in serum specimens collected from 17 COVID-19 patients at 6-7 months after diagnosis and the results were compared to those from cases investigated 2 weeks to 2 months post-infection. All samples were positive for IgGs against the S- and N-proteins of SARS-CoV-2. Notably, 14 samples available at 6-7 months post-infection all showed significant neutralizing activities in a pseudovirus assay, with no difference in blocking the cell-entry of the 614D and 614G variants of SARS-CoV-2. Furthermore, in 10 blood samples from cases at 6-7 months post-infection used for memory T-cell tests, we found that interferon γ-producing CD4
Adaptive Immunity/physiology* ; Adult ; Aged ; Antibodies, Neutralizing/blood* ; COVID-19/immunology* ; Cohort Studies ; Female ; Humans ; Immunoglobulin G/blood* ; Male ; Middle Aged ; SARS-CoV-2/immunology* ; T-Lymphocytes/physiology* ; Time Factors ; Viral Proteins/immunology*

Objective: To explore the clinicopathological characteristics of primary hepatic neuroendocrine neoplasm (HNEN) and metastatic HNEN from digestive tract, to screen the risk factors of hepatic metastasis of gastroenteropancreatic neuroendocrine neoplasm (GEP-NEN) and to analyze the differences between primary and metastatic HNEN in clinical features, diagnosis, treatment and prognosis. Methods: From January 2010 to June 2017, the clinical data of 182 patients with HNEN admitted at The First Affiliated Hospital of Zhengzhou University were retrospectively analyzed, including 39 cases of primary HNEN, 129 cases of metastatic HNEN and 14 cases of HNEN with unknown primary lesions. Chi-square test and t test were performed to analyze the pathologic characteristics among groups. Logistic regression method was used to analyze the risk factors of hepatic metastasis. Kaplan-Meier method and log-rank test were used for survival analysis. Cox model was used for the prognostic multivariate survival analysis. Results: Metastatic HNEN from digestive tract was more common in male (70.5%, 91/129). The case number of serological tumor biomarkers neuronspecific enolase and alpha-fetoprotein positive in primary HNEN were two cases and one case, respectively, and the positive rates in metastatic HNEN were 37.2% (32/86) and 6.4% (7/110). Most primary HNEN was single lesion (61.5%, 24/39), while multiple lesions were more common in metastatic HNEN (78.3%, 90/115). Primary HNEN mainly occurred in the right lobe of the liver (44.7%, 17/38), while metastatic HNEN located simultaneously in the left and right lobes of the liver (68.4%, 78/114). There were significant differences between primary HNEN and metastatic HNEN in tumor number, pathological grading, location of tumors and maximum diameter of tumors (χ²=21.264, 11.696, 19.461 and 4.547, all P<0.05). The median survival time of patients with primary HNEN and metastatic HNEN were 17.0 months and 10.0 months, and there was a significant difference in survival curves between the two groups (χ²=7.235, P=0.007). The type of hepatic tumors (primary or metastatic)(P=0.002), pathological grading of hepatic tumors (P=0.044), lymph node metastasis (P=0.024), the growth pattern of tumors (P<0.01) and treatment methods (P=0.018) were the independent factors for the prognosis of patients. Conclusions There are significant differences between primary HNEN and metastatic HNEN in tumor number, size and location. The type of hepatic tumors, pathological grading, lymph node metastasis, growth pattern of tumors and treatment methods are the independent factors for the prognosis of patients. Early topical treatment and combination treatment can help to prolong survival time of HNEN patients.

Objective: To investigate the expression and clinical significances of ubiquitin conjugating enzyme E2 T (UBE2T) in hepatocellular carcinoma (HCC), and explore the role of UBE2T gene expression in migration and invasion of HCC cells. Methods: The expression of UBE2T mRNA in 54 pairs of HCC tissues and the matched noncancerous liver tissues was analyzed by real-time fluorescent quantitative PCR. The expression level of UBE2T protein in 233 cases of HCC tissues was detected by immunohistochemistry, then the relationship between UBE2T expression and clinicopathological features of HCC patients was analyzed. The recombinant lentiviral vector pWPXL-UBE2T contained UBE2T gene sequences was constructed and further transfected into Hep3B and SMMC-7721 cells. In the other hand, the recombinant lentiviral vector GV248-shUBE2T-1 and GV248-shUBE2T-2 targeted UBE2T gene were transfected into MHCC-LM3 and SK-Hep1 cells. The forced expression and knockdown of UBE2T gene were validated by real-time fluorescent quantitative PCR and Western blotting. Afterwards, the effects of UBE2T gene overexpression and silencing on migratory and invasive abilities of HCC cells were detected by Transwell chamber assays, respectively. Results: The UBE2T mRNA level in HCC tissues was significantly higher than that in the matched noncancerous liver tissues (P < 0.000 1). The expression of UBE2T protein was correlated with intrahepatic metastasis of HCC (P = 0.004) in 233 patients with HCC. Compared with the control group, the forced expression of UBE2T gene markedly promoted both migration and invasion of Hep3B and SMMC-7721 cells (both P < 0.01); while the knockdown of UBE2T gene obviously inhibited the migration and invasion of MHCC-LM3 and SK-Hep1 cells as compared with the negative control group (both P < 0.05). Conclusion: High expression level of UBE2T in HCC tissues is correlated with the presence of intrahepatic metastasis, and the overexpression of UBE2T gene can promote the migration and invasion of HCC cells.

The active copper-containing carbon nanodots were prepared by hydrothermal method, and then characterized by fluorescence spectroscopy and UV-visible absorption spectroscopy.Subsequently, a highly sensitive and selective electrochemical biosensor was fabricated on the basis of this synthesized carbon nanodots with electro-deposition technique.The electrode behavior was investigated by cyclic voltammetry, electrochemical impedance spectroscopy, and differential pulse voltammetry.Furthermore, the catalysis mechanism was studied.The experimental results indicated that the biosensor exhibited a strong electrocatalytic activity toward the oxidation of uric acid (UA).What′s more, the interference from ascorbic acid and dopamine was eliminated effectively.Under the optimum conditions, there were linear relationships between the anodic peak current and the concentration of UA (1.00-300.0 μmol/L), and the limit detection was 0.30 μmol/L (S/N=3).The prepared biosensor had advantages such as easy fabrication, strong anti-interference ability, high sensitivity, and wide detection range, and could be used for real sample detection.

The stroke rehabilitation evaluation system has been designed and developed in view of the present practice status of clinical rehabilitation medicine. The system not only implements patient information collection and rehabilitation evaluation, but also outputs individual rehabilitation program automatically according to evaluation outcome.
Humans ; Outcome Assessment (Health Care) ; methods ; Software Design ; Stroke Rehabilitation