ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect the reference sample solution， plasma， and tissue samples of Jinshui Liujunjian under positive and negative ion modes， respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times， secondary mass spectral fragments， reference standards， and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian， including 47 flavonoids， 5 amino acids， 13 iridoids， 16 triterpenoid saponins， etc.， of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components； 50 prototype components were identified in different tissues， among which 13， 10， 7， 21， 11， 6， 14， and 40 prototype components were identified in the heart， liver， spleen， lung， kidney， brain， large intestine， and stomach， respectively. Among them， 7 compounds such as ferulic acid， glycyrrhizic acid， and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian， primarily composed of flavonoids and triterpenoid saponins， along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.

ObjectiveTo elucidate the chemical composition of the reference sample of Jinshui Liujunjian and its distribution characteristics in blood and tissues of rats. MethodsUltra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry（UPLC-Q-TOF-MS/MS） was used to detect the reference sample solution， plasma， and tissue samples of Jinshui Liujunjian under positive and negative ion modes， respectively. Qualitative Analysis 10.0 software and a self-constructed database were employed for primary mass spectrum matching.Compound identification was further validated by comparing retention times， secondary mass spectral fragments， reference standards， and literature data to deduce fragmentation pathways. ResultsA total of 122 compounds were identified in the reference sample of Jinshui Liujunjian， including 47 flavonoids， 5 amino acids， 13 iridoids， 16 triterpenoid saponins， etc.， of which 42 compounds were confirmed by comparison with reference substances. A total of 21 prototype components were identified in blood components； 50 prototype components were identified in different tissues， among which 13， 10， 7， 21， 11， 6， 14， and 40 prototype components were identified in the heart， liver， spleen， lung， kidney， brain， large intestine， and stomach， respectively. Among them， 7 compounds such as ferulic acid， glycyrrhizic acid， and nobiletin were exposed in the target organs of lung and kidney. ConclusionThis study elucidates the material basis of the reference samples of Jinshui Liujunjian， primarily composed of flavonoids and triterpenoid saponins， along with their in vivo distribution characteristics. These findings provide a scientific basis for establishing quality evaluation indicators and offer references for subsequent pharmacodynamic and pharmacokinetic investigations.

ObjectiveTo investigate the possible mechanism by which Zhiqiao Gancao Decoction （枳壳甘草汤， ZGD） delays intervertebral disc degeneration （IDD） based on the Hippo-yes-associated protein （YAP）/transcriptional co-activator with PDZ-binding motif （TAZ） signaling pathway. MethodsA total of 50 SD rats were randomly divided into sham surgery group， model group， low-dose ZGD group， high-dose ZGD group， and high-dose ZGD + inhibitor group， with 10 rats in each group. In the sham surgery group， the rats were pierced in the skin and muscle at the Co6/7/8 segments of the tail with a 21G needle （depth approximately 2 mm） without damaging the intervertebral disc. In the other groups， rats were injected with a 21G needle at the Co6/7/8 segments of the tail to establish an IDD model by piercing the tail intervertebral disc 5 mm. One week after modeling， rats in the low-dose and high-dose ZGD groups were given 6.24 and 12.24 g/（kg·d） of the decoction via gastric gavage， respectively. The high-dose ZGD + inhibitor group was given 12.24 g/（kg·d） of the decoction and an intraperitoneal injection of YAP/TAZ inhibitor Verteporfin 10 mg/kg. The sham surgery and model groups were given 5 ml/（kg·d） of normal saline via gavage. The gavage was given once a day， and the intraperitoneal injection was given every other day. After 4 weeks of continuous intervention， the pathological changes of the tail intervertebral discs were observed using HE staining， Oil Red O-Green staining， and Toluidine Blue staining. Immunohistochemistry was used to detect the expression of aggrecan and MMP3 in the nucleus pulposus. TUNEL fluorescence staining was performed to detect apoptosis in the nucleus pulposus， and the apoptosis rate was calculated. Western blot was used to detect the Hippo-YAP/TAZ signaling pathway， including YAP， phosphorylated YAP （p-YAP）， phosphorylated MST1/2 （p-MST1/2）， phosphorylated TAZ （p-TAZ） and apoptosis-related proteins， such as Cleaved Caspase 3， P53， Bcl-2 and Bax. ResultsCompared with sham surgery group， the rats in the model group showed significant degenerative changes in the intervertebral disc. The levels of aggrecan， Bcl-2， and YAP proteins in the nucleus pulposus decreased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate increased （P < 0.01）. Compared with the model group， the drug intervention groups showed partial recovery in intervertebral disc degeneration. The levels of aggrecan， Bcl-2， and YAP proteins increased， while the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and the apoptosis rate decreased （P＜0.05 or P＜0.01）. The high-dose ZGD group showed more significant recovery in intervertebral disc degeneration compared to the low-dose ZGD group， with a decrease in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and an increase in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. Compared with the high-dose ZGD group， the high-dose ZGD + inhibitor group showed a reduced recovery in intervertebral disc degeneration， with an increase in the levels of p-MST1/2， p-YAP， p-TAZ， P53， Bax， Cleaved Caspase 3， MMP3 proteins， and apoptosis rate， and a decrease in the levels of aggrecan， Bcl-2， and YAP proteins （P＜0.05 or P＜0.01）. ConclusionZGD may delay intervertebral disc degeneration by inhibiting the phosphorylation of YAP in the nucleus pulposus， maintaining the function of the Hippo-YAP/TAZ signaling pathway， and reducing apoptosis of nucleus pulposus cells.

BACKGROUND: Diabetic foot is a complex condition with high incidence, recurrence, mortality, and disability rates. Current treatments for diabetic foot ulcers are often insufficient. This study was conducted to identify potential therapeutic targets for diabetic foot. METHODS: Datasets related to diabetic foot and diabetic skin were retrieved from the Gene Expression Omnibus database. Differentially expressed genes (DEGs) were identified using R software. Enrichment analysis was conducted to screen for critical gene functions and pathways. A protein interaction network was constructed to identify node genes corresponding to key proteins. The DEGs and node genes were overlapped to pinpoint target genes. Plasma and chronic ulcer samples from diabetic and non-diabetic individuals were collected. Western blotting, immunohistochemistry, and enzyme-linked immunosorbent assays were performed to verify the S100 calcium binding protein A9 (S100A9), inflammatory cytokine, and related pathway protein levels. Hematoxylin and eosin staining was used to measure epidermal layer thickness. RESULTS: In total, 283 common DEGs and 42 node genes in diabetic foot ulcers were identified. Forty-three genes were differentially expressed in the skin of diabetic and non-diabetic individuals. The overlapping of the most significant DEGs and node genes led to the identification of S100A9 as a target gene. The S100A9 level was significantly higher in diabetic than in non-diabetic plasma (178.40 ± 44.65 ng/mL vs. 40.84 ± 18.86 ng/mL) and in chronic ulcers, and the wound healing time correlated positively with the plasma S100A9 level. The levels of inflammatory cytokines (tumor necrosis factor-α, interleukin [IL]-1, and IL-6) and related pathway proteins (phospho-extracellular signal regulated kinase [ERK], phospho-p38, phospho-p65, and p-protein kinase B [Akt]) were also elevated. The epidermal layer was notably thinner in chronic diabetic ulcers than in non-diabetic skin (24.17 ± 25.60 μm vs. 412.00 ± 181.60 μm). CONCLUSIONS S100A9 was significantly upregulated in diabetic foot and was associated with prolonged wound healing. S100A9 may impair diabetic wound healing by disrupting local inflammatory responses and skin re-epithelialization.
Calgranulin B/therapeutic use* ; Diabetic Foot/metabolism* ; Humans ; Datasets as Topic ; Computational Biology ; Mice, Inbred C57BL ; Animals ; Mice ; Protein Interaction Maps ; Immunohistochemistry

Stathmin 1 (STMN1) is a microtubule-binding cytoplasmic phosphoprotein that promotes microtubule depolymerization or inhibits microtubule assembly, thereby regulating cytoskeletal organization and cell cycle progression. STMN1 is upregulated in a variety of malignant tumors, where it drives proliferation, invasion, metastasis, and angiogenesis through classic pathways such as nuclear factor-κB (NF-κB), mitogen-activated protein kinase (MAPK), and ferroptosis. STMN1 can also modulate the function of immune cells, thereby influencing antitumor immunity. Clinical data show that its high expression correlates positively with tumor drug resistance and poor prognosis, suggesting that STMN1 has potential as a tumor biomarker and therapeutic molecular target with important clinical significance.
Humans ; Stathmin/metabolism* ; Neoplasms/genetics* ; Biomarkers, Tumor/metabolism* ; NF-kappa B/metabolism* ; Cell Proliferation ; Drug Resistance, Neoplasm

Objective:To explore the genetic etiology of a pedigree with intellectual disability and explore its pathogenesis.Methods:A Chinese pedigree which had presented at the Henan Provincial People′s Hospital in March 2023 was selected as the study subject. Clinical data of the pedigree were collected, along with peripheral venous blood samples from its members. Whole exome sequencing (WES) was carried out, and candidate variants were verified by Sanger sequencing. Amniotic fluid was collected for prenatal diagnosis. This study was approved by the Medical Ethics Committee of the Henan Provincial People′s Hospital (No. 2019-134).Results:Both the proband (a 6-year-old male) and his mother (30 years old) had various degrees of intellectual and motor impairment. WES revealed that the proband has harbored a de novo heterozygous c. 2563_2567dup (p.Lys856fs) variant of the UBE3A gene, while his mother, maternal grandmother and fetus had all harbored a novel heterozygous c. 409+ 1G>A variant of the RNF13 gene. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), both variants were predicted to be pathogenic (PVS1+ PS1+ PM2_Supporting; PVS1+ PM2_Supporting+ PP3). Conclusion:Based on the clinical manifestations and the result of genetic testing, the heterozygous c.2563_2567dup (p.Lys856fs) variant of the UBE3A gene probably underlay the intellectual disability and developmental delay in the proband, whilst the heterozygous c. 409+ 1G>A variant of the RNF13 gene may underlie the intellectual disability in the proband′s mother and grandmother. Above results have enabled genetic counseling and prenatal diagnosis for this pedigree.

Objective:To systematically evaluate and integrate the midwifery care needs of parturition, to provide reference for formulating effective measures to improve obstetric care.Methods:A computer search was conducted on Cochrane Library, PubMed, Web of Science, Embase, China National Knowledge Infrastructure, China Biomedical Literature Database and Wanfang Database for qualitative research on midwifery care needs of parturition. The search period was from the establishment of the database to April 2024. The quality of the included literature was evaluated using the qualitative research quality evaluation criteria of the Australian Joanna Briggs Institute Evidence based Health Care Center. A thematic synthesis was used for result integration.Results:A total of 16 articles were included, and 63 major research results were extracted and summarized into 9 new categories, forming 4 integrated results: physiological comfort needs, respect needs, professional nursing support needs and social network harmonious needs.Conclusions:The needs of midwifery care during childbirth are diverse and need help and support from various aspects. It is suggested that nursing staff should pay attention to the experience and needs of childbirth, improve the management plan of nursing during childbirth, improve the cognitive experience of childbirth and promote natural childbirth.

Background::Hypothermia therapy has been suggested to attenuate myocardial necrosis; however, the clinical implementation as a valid therapeutic strategy has failed, and new approaches are needed to translate into clinical applications. This study aimed to assess the feasibility, safety, and efficacy of a novel selective intracoronary hypothermia (SICH) device in mitigating myocardial reperfusion injury.Methods::This study comprised two phases. The first phase of the SICH was performed in a normal porcine model for 30 minutes ( n = 5) to evaluate its feasibility. The second phase was conducted in a porcine myocardial infarction (MI) model of myocardial ischemia/reperfusion which was performed by balloon occlusion of the left anterior descending coronary artery for 60 minutes and maintained for 42 days. Pigs in the hypothermia group ( n = 8) received hypothermia intervention onset reperfusion for 30 minutes and controls ( n = 8) received no intervention. All animals were followed for 42 days. Cardiac magnetic resonance analysis (five and 42 days post-MI) and a series of biomarkers/histological studies were performed. Results::The average time to lower temperatures to a steady state was 4.8 ± 0.8 s. SICH had no impact on blood pressure or heart rate and was safely performed without complications by using a 3.9 F catheter. Interleukin-6 (IL-6), tumor necrosis factor-α, C-reactive protein (CRP), and brain natriuretic peptide (BNP) were lower at 60 min post perfusion in pigs that underwent SICH as compared with the control group. On day 5 post MI/R, edema, intramyocardial hemorrhage, and microvascular obstruction were reduced in the hypothermia group. On day 42 post MI/R, the infarct size, IL-6, CRP, BNP, and matrix metalloproteinase-9 were reduced, and the ejection fraction was improved in pigs that underwent SICH.Conclusions::The SICH device safely and effectively reduced the infarct size and improved heart function in a pig model of MI/R. These beneficial effects indicate the clinical potential of SICH for treatment of myocardial reperfusion injury.

Objective:To investigate the factors influencing the right heart dysfunction in myloproliferative neoplasm (MPN) patients with BCR::ABL fusion gene-negative.Methods:A retrospective case-control study was conducted. A total of 130 MPN patients with BCR::ABL fusion gene-negative admitted to the Second People's Hospital of Lianyungang from January 2020 to December 2022 were selected as the study objects. The general data, laboratory indexes and cardiac function parameters were collected. All patients were divided into the control group (non-right heart function injury, 96 cases) and the observation group (right heart function injury, 34 cases) according to whether there was right heart dysfunction judged by ultrasonic cardiogram. Multivariate logistic regression analysis was used to analyze the independent influencing factors of right heart dysfunction in MPN patients with BCR:: ABL fusion gene-negative. Taking right heart dysfunction judged by ultrasonic cardiogram as the gold standard, the receiver operating characteristic (ROC) curve was drawn to predict right heart dysfunction according to independent influencing factors, and the diagnostic efficacy of the factors was analyzed.Results:The median age was 57 years old (range 19-76 years); among the 130 patients, 62 cases were male and 68 cases were female. There were 69 cases of primary thrombocytosis, 35 cases of polycythemia vera and 26 cases of primary myelofibrosis. The proportions of patients with age > 60 years old, hypertension, embolism history, pulmonary hypertension in the observation group were higher than those in the control group, and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in the composition of patients with gender, body mass index > 28 kg/m2, smoking, drinking, diabetes, hyperlipidemia and different pathological types between the two groups (all P > 0.05). The white blood cell count, neutrophil count, basophil count, monocyte count, red blood cell count, hematocrit (Hct), CD34+ cell count and platelet count in the observation group were higher than those in the control group, and the differences were statistically significant (all P < 0.05). There were no statistically significant differences in lymphocyte count, eosinophilic count, triglyceride, total cholesterol, high density lipoprotein cholesterol, low density lipoprotein cholesterol and high sensitive C-reactive protein levels between the two groups (all P > 0.05). The cardiac ejection time in the observation group was shorter than that in the control group, but the transverse diameter of right atrium, anteroposterior diameter of right ventricle, anterior wall thickness of right ventricle, isovolumic systole time, isovolumic diastole time and right ventricular Tei index in the observation group were all higher than those in the control group, and the differences were statistically significant (all P < 0.001). Multivariate logistic regression analysis showed that age > 60 years (OR = 1.520, 95% CI: 1.250-1.692, P = 0.002), embolism history (OR = 1.765, 95% CI: 1.302-2.020, P = 0.001), pulmonary arterial hypertension (OR = 1.555, 95% CI: 1.303-1.702, P = 0.001), elevated Hct (OR = 1.900, 95% CI: 1.587-2.269, P = 0.002), the increased density of CD34+ cell (OR = 1.400, 95% CI: 1.158-1.630, P = 0.001), and increased Tei index of right ventricle (OR = 2.269, 95% CI: 1.700-3.568, P = 0.001) were independent risk factors of right heart dysfunction in MPN patients with BCR::ABL fusion gene-negative. ROC curve analysis showed that the area under the curve of age > 60 years old, embolism history, pulmonary arterial hypertension, Hct, the density of CD34+ cell, and Tei index of right ventricle, in predicting right heart dysfunction in MPN patients with BCR-ABL fusion gene-negative was 0.780 (95% CI: 0.690-0.925), 0.657 (95% CI: 0.533-0.740), 0.728 (95% CI: 0.660-0.813), 0.619 (95% CI: 0.510-0.708), 0.777 (95% CI: 0.720-0.809), 0.822 (95% CI: 0.749-0.886), respectively. The area under the curve of the 6 combined items was 0.930 (95% CI: 0.850-0.987). The sensitivity and specificity were 89.69% and 96.38% respectively when the optimum critical value was reached.Conclusions:Age > 60 years old, embolism history, pulmonary arterial hypertension, elevated Hct, the increased density of CD34+ cell and increased Tei index of right ventricle are independent risk factors of right heart dysfunction in MPN patients with BCR::ABL fusion gene-negative.

Objective To explore the molecular mechanism of lipopolysaccharide(LPS)on the contraction of pregnant uterine smooth muscle at tissue and cellular levels.Methods C57BL/6J mice at 16 days of gestation were randomly divided into control group and LPS group.The mice in LPS group were intraperitoneally injected with 20 μg in LPS solution to establish the model of preterm birth,and the mice in control group were intraperitone-ally injected with the same amount of normal saline.Isolated uterine muscle strips were used to detect changes in the contractile function of the tissue,as well as changes in the expression and function of the contraction key signa-ling molecule TWIK-related K+channel 1(TREK-1).Primary cultured pregnant mouse uterine smooth muscle cells were used to detect the expression of TREK-1 under the regulation of LPS.Results The contractility of mouse u-terine tissues was significantly enhanced by LPS,and the protein expression of TREK-1,a key signal for contrac-tion,was significantly reduced,and activation of TREK-1 resulted in a significant down-regulation of the enhanced contractility of mouse uterine tissues in the LPS group.However,there was no significant difference in the expres-sion of TREK-1 protein,which was highly expressed in the smooth muscle of pregnant mice,when LPS acted on the primary uterine smooth muscle cells of pregnant mice.Conclusion Uterine contractility is enhanced in pregnant mice uterine tissues by inhibiting TREK-1 expression and function in response to LPS,and it may be one of the mechanisms by which LPS induces preterm labor.However,the effect of LPS on TREK-1 on mouse pregnant uter-ine smooth muscle cells may be realized through intercellular signaling and not directly on uterine smooth muscle cells.This further suggests that the animal and histological experiments cannot be completely replaced by isolated cell experiments in the study of inflammatory preterm labor.