ObjectiveTo systematically and objectively characterize the pharmacological effects of Fufang Biejia Ruangan pills （FBRP） in the intervention of alcoholic liver disease （ALD） using acute and chronic ALD mouse models and to elucidate its molecular mechanisms. MethodFifty SPF-grade male BALB/c mice were randomly divided into the normal group， model group， and FBRP low-， medium-， and high-dose groups （9.6， 19.2， 38.4 mg·kg^-1）. Except for the normal group， the remaining groups were given 56° white wine by gavage to establish the acute ALD model， with samples collected after 4 weeks. Thirty SPF-grade male C57BL/6N mice were randomly divided into the normal group， model group， and FBRP medium-dose group （19.2 mg·kg^-1）. The chronic ALD mouse model was established using the Lieber-DeCarli method over a 10-week period. Inflammatory markers in liver tissues were assessed using hematoxylin-eosin （HE）， Sirius Red， oil red O staining， and enzyme-linked immunosorbent assay （ELISA）. Intoxication behaviors of each group were objectively evaluated through sobering-up time， net-catching， and pole-climbing tests. Further bioinformatics analyses based on clinical transcriptomic data were conducted to identify key targets and molecular mechanisms of FBRP in alleviating ALD through liver-brain dialogue， with experimental validation by ELISA， Western blot， and immunohistochemical staining. ResultCompared with the normal group, the levels of aspartate aminotransferase （AST） and alanine aminotransferase （ALT） in liver tissues of mice in the acute and chronic ALD model groups were significantly increased （P<0.05）. Compared with the model group， the levels of AST and ALT in liver tissue of mice in FBRP groups were significantly decreased （P<0.05）. Compared with the normal group， the time of grasping the net and climbing the pole in the acute ALD model group was significantly decreased within 4 weeks （P<0.01）. Compared with the model group， the grasping and climbing time of FBRP high dose groups increased significantly within 4 weeks （P<0.05）. Compared with the normal group， the expression of high mobility group protein B1 （HMGB1） protein in liver tissue and prefrontal lobe tissue of mice in the chronic ALD model group was significantly increased （P<0.01）. Compared with the model group， the expression of HMGB1 protein in FBRP medium dose group was significantly decreased （P<0.05，P<0.01）. Compared with the normal group， the expression of brain-derived neurotrophic factor （BDNF） protein and the release of γ-aminobutyric acid （GABA） in the prefrontal cortex of the model group were significantly decreased （P<0.01）. Compared with the model group, the expression of BDNF protein and the release of GABA in the FBRP medium dose group were significantly increased （P<0.05）. ConclusionThis study revealed that FBRP improved key pathological changes in ALD by modulating liver-brain dialogue mediated by the HMGB1-BDNF axis. These findings provide experimental evidence for the clinical use of FBRP in treating ALD and offer new insights for the development of ALD therapeutic agents.

ObjectiveTo identify the chemical constituents of Ruyi Zhenbaowan in vitro and in vivo. MethodThe chemical constituents of Ruyi Zhenbaowan were identified based on UHPLC-Q Exactive Orbitrap HRMS. A total of 12 male SD rats were randomized into two groups： control （pure water） and Ruyi Zhenbaowan （1.8 g·kg^-1）. The rats were administrated with the suspension of Ruyi Zhenbaowan or pure water by gavage. After 1.5 h， the plasma and cerebrospinal fluid were collected. Chromatographic separation was performed on a Waters ACQUITY UPLC BEH C₁₈ column （2.1 mm × 150 mm， 1.7 μm） with a mixture of 0.1% formic acid aqueous solution （A） and acetonitrile （B） as the mobile phase. Gradient elution was carried out according to the procedure of 0~15 min，97%~80%A；15~30 min ，80%~60%A；30~40 min，60%~30%A；40~45 min，30%~5%A. The ion source was electrospray ionization， and scan range was m/z 100-1 500. The prototype components and the components in the plasma and cerebrospinal fluid were analyzed qualitatively by scanning in positive and negative ion modes and identified by comparison with the data in published literature and the information of standard substances. ResultA total of 126 chemical constituents were identified from the 80% methanol solution of Ruyi Zhenbaowan， and 14 and 7 prototype constituents were detected in the plasma and the cerebrospinal fluid， respectively. In addition， the fragmentation rules of apigenin， apigenin-7-O-glucuronide， galangin， liquiritin， piperine， glycyrrhizic acid， eugenol， gallic acid， and cholic acid were deduced. ConclusionThis study achieved rapid multicomponent characterization and identification of Ruyi Zhenbaowan in vitro and in vivo， providing theoretical support for exploring active substances and performing quality control.l.

Tumor cells use glycolysis to provide material and energy under hypoxic conditions to meet the energy requirements for rapid growth and proliferation， namely the Warburg effect. Even under aerobic conditions， tumor cells mainly rely on glycolysis to provide energy. Therefore， glucose transporter protein 1（GLUT1）， which is involved in the process of glucose metabolism， plays an important role in tumorigenesis， development and drug resistance， and is considered to be one of the important targets in the treatment of malignant tumors. In recent years， research on tumor glucose metabolism has gradually become a hot spot. It has been shown that various factors are involved in the regulation of tumor energy metabolism， among which the role of GLUT1 is the most critical. In this paper， the authors reviewed the latest research progress of GLUT1-targeted traditional Chinese medicine（TCM） active ingredient nano-delivery system in tumor therapy， aiming to reveal the feasibility and effectiveness of this system in the delivery of chemotherapeutic drugs. The GLUT1-targeted TCM active ingredient nano-delivery system can overcome the bottleneck of the traditional targeting strategy as well as the high-permeability long retention（EPR） effect. In summary， the authors believe that the GLUT1-targeted TCM active ingredient nano-delivery system provides a new strategy for targeted treatment of tumors and has a broad application prospect in tumor prevention and treatment.

Objective:To explore the application value of four-dimensional ultrasound combined with maternal serological screening in fetal facial deformities.Methods:A total of 106 pregnant women at mid pregnancy,whose fetuses with suspected fetal facial deformities and who conducted prenatal screening in Sanya Maternal and Child Health Hospital(Sanya Women and Children's Hospital)from January 2020 to December 2022,were selected.All of them underwent four-dimensional ultrasound and maternal serological screening.The results of delivery or induced labor were used as the gold standard of diagnosis to compare the diagnostic values of single four-dimensional ultrasound,single maternal serological screening and the combination of them for fetal facial deformities.Results:The analysis of the receiver operating characteristic(ROC)curve showed that the area under curve(AUC)values of four-dimensional ultrasound,maternal serological screening and the combination of them were respectively 0.932,0.863 and 0.981 in diagnosing fetal facial deformities.Both the sensitivities and accuracies of four-dimensional ultrasound and maternal serological screening were significantly lower than those of the combined diagnosis of them in diagnosing fetal facial deformities,and the differences of them were statistical significance(x2=11.173,0.064,P<0.05),respectively.Conclusion:Four-dimensional ultrasound combines with maternal serological screening can improve the diagnostic accuracy for fetal facial deformities.

Objective:To analyze the genetic sequences of two patients with a rare Ael blood subgroup.Methods:Two female patients undergoing treatment respectively for adenomyoma of the uterus and gastritis at the Second Affiliated Hospital, Yuying Children′s Hospital of Wenzhou Medical University in June 2019 and September 2020 were selected as the study subjects. Their Ael subtypes were identified with a saline tube agglutination assay and absorption-emission assay. Sequence of the ABO gene Ael subtypes was determined by the Sanger method. The impact of genetic variants on the structural stability of N-acetylgalactosaminyl transferase (GTA) was analyzed with PyMOL software by constructing a structure predicted model. Results:Both patients were determined as Ael blood subgroup. Sequencing result of patient 1 was ABO* O.01.02/ ABO* O.01.02, which has resulted in a p. Thr88Profs*31 amino acid substitution. The sequencing result of patient 2 was ABO* Ael.06/ ABO* O.01.02, in which c. 425C>T and c. 467C>T variants in exon 7 have led to p. Met142Thr and p. Pro156Leu substitutions. Prediction of the protein model speculated that the p. Met142Thr not only can change the binding of GTA protein with water molecules, but also the local hydrogen bond network of GTA, which may lead to decreased enzymatic activity. By contrast, the p. Pro156Leu variant has trivial effect on the structural stability of GTA. Conclusion:The molecular structure of Ael subtypes can be diverse. The genotypes of the two patients have been respectively determined as ABO* O.01.02/ ABO* O.01.02 with a G261 deletion and ABO* Ael.06/ ABO* O.01.02.

Objective:To establish a high-performance liquid chromatography (HPLC) characteristic spectrum of Radix Actinidiae Chinensis decoction pieces, standard decoction and formula granules; To simultaneously determine the contents of index components; To study the transfer law of formula granules based on the extraction amount, content of Catechin and characteristic spectrum of standard decoction.Methods:15 batches of Radix Actinidiae Chinensis decoction pieces were collected, the standard decoction was prepared, and the extraction amount was measured. The HPLC fingerprints of decoction pieces, standard decoction and formula granules were established, and the characteristic peaks were calibrated and attributed. The content of Catechins in decoction pieces, standard decoction and formula granules was measured, and the transfer rate law was calculated. The yield rate, the common peak transmission number of fingerprints, the content and the transfer rate were the main evaluation indexes, and the law of the transmission of the magnitude value was analyzed.Results:The paste yield of 15 batches of Radix Actinidiae Chinensis standard decoction ranged from 3.9%-6.3%. A total of 4 characteristic peaks were calibrated in the feature map, and peak 2 was identified as Protocatechuic acid and peak 4 as Catechin; 6 characteristic peaks were detected in Actinidia chinensis Planchon decoction pieces, standard decoction and formula granules, and their relative retention times were all within the specified range. The content of Catechins in Radix Actinidiae Chinensis decoction pieces was 0.4%-1.4%, which was 3.8%-4.9% in formula granules. The transfer rate of decoction pieces-standard decoction was 13.6%-38.3%, and the decoction-formula granules was 15.5%-21.2%.Conclusions:The mass transfer between Radix Actinidiae Chinensis decoction pieces, standard decoction and formula granules has a good migration. The formula granules also have a good correlation and consistency with the standard decoction, which indicating that the preparation process of Radix Actinidiae Chinensis is reasonable and feasible.

BACKGROUND: Significant brain volume deviation is an essential phenotype in children with neurodevelopmental delay (NDD), but its genetic basis has not been fully characterized. This study attempted to analyze the genetic factors associated with significant whole-brain deviation volume (WBDV). METHODS: We established a reference curve based on 4222 subjects ranging in age from the first postnatal day to 18 years. We recruited only NDD patients without acquired etiologies or positive genetic results. Cranial magnetic resonance imaging (MRI) and clinical exome sequencing (2742 genes) data were acquired. A genetic burden test was performed, and the results were compared between patients with and without significant WBDV. Literature review analyses and BrainSpan analysis based on the human brain developmental transcriptome were performed to detect the potential role of genetic risk factors in human brain development. RESULTS: We recruited a total of 253 NDD patients. Among them, 26 had significantly decreased WBDV (<-2 standard deviations [SDs]), and 14 had significantly increased WBDV (>+2 SDs). NDD patients with significant WBDV had higher rates of motor development delay (49.8% [106/213] vs . 75.0% [30/40], P  = 0.003) than patients without significant WBDV. Genetic burden analyses found 30 genes with an increased allele frequency of rare variants in patients with significant WBDV. Analyses of the literature further demonstrated that these genes were not randomly identified: burden genes were more related to the brain development than background genes ( P  = 1.656e -9 ). In seven human brain regions related to motor development, we observed burden genes had higher expression before 37-week gestational age than postnatal stages. Functional analyses found that burden genes were enriched in embryonic brain development, with positive regulation of synaptic growth at the neuromuscular junction, positive regulation of deoxyribonucleic acid templated transcription, and response to hormone, and these genes were shown to be expressed in neural progenitors. Based on single cell sequencing analyses, we found TUBB2B gene had elevated expression levels in neural progenitor cells, interneuron, and excitatory neuron and SOX15 had high expression in interneuron and excitatory neuron. CONCLUSION Idiopathic NDD patients with significant brain volume changes detected by MRI had an increased prevalence of motor development delay, which could be explained by the genetic differences characterized herein.
Child ; Humans ; Neurodevelopmental Disorders/epidemiology* ; Genetic Testing ; Phenotype ; Brain/pathology* ; Genetic Background ; SOX Transcription Factors/genetics*

BACKGROUND: Previous studies have shown that lymph node metastasis only occurs in some mixed ground-glass nodules (mGGNs) which the pathological results were invasive adenocarcinoma (IAC). However, the presence of lymph node metastasis leads to the upgrading of tumor-node-metastasis (TNM) stage and worse prognosis of the patients, so it is important to perform the necessary evaluation before surgery to guide the operation method of lymph node. The aim of this study was to find suitable clinical and radiological indicators to distinguish whether mGGNs with pathology as IAC is accompanied by lymph node metastasis, and to construct a prediction model for lymph node metastasis. METHODS: From January 2014 to October 2019, the patients with resected IAC appearing as mGGNs in computed tomography (CT) scan were reviewed. All the lesions were divided into two groups (with lymph node metastasis or not) according to their lymph node status. Lasso regression model analysis by applying R software was used to evaluate the relationship between clinical and radiological parameters and lymph node metastasis of mGGNs. RESULTS: A total of 883 mGGNs patients were enroled in this study, among which, 12 (1.36%) showed lymph node metastasis. Lasso regression model analysis of clinical imaging information in mGGNs with lymph node metastasis showed that previous history of malignancy, mean density, mean density of solid components, burr sign and percentage of solid components were informative. Prediction model for lymph node metastasis in mGGNs was developed based on the results of Lasso regression model with area under curve=0.899. CONCLUSIONS Clinical information combined with CT imaging information can predict lymph node metastasis in mGGNs.
Humans ; Lymphatic Metastasis ; Lung Neoplasms ; Adenocarcinoma ; Lymph Nodes ; Social Group

Pulmonary hypertension (PH) is an insidious pulmonary vasculopathy with high mortality and morbidity and its underlying pathogenesis is still poorly delineated. The hyperproliferation and apoptosis resistance of pulmonary artery smooth muscle cells (PASMCs) contributes to pulmonary vascular remodeling in pulmonary hypertension, which is closely linked to the downregulation of fork-head box transcriptional factor O1 (FoxO1) and apoptotic protein caspase 3 (Cas-3). Here, PA-targeted co-delivery of a FoxO1 stimulus (paclitaxel, PTX) and Cas-3 was exploited to alleviate monocrotaline-induced pulmonary hypertension. The co-delivery system is prepared by loading the active protein on paclitaxel-crystal nanoparticles, followed by a glucuronic acid coating to target the glucose transporter-1 on the PASMCs. The co-loaded system (170 nm) circulates in the blood over time, accumulates in the lung, effectively targets the PAs, and profoundly regresses the remodeling of pulmonary arteries and improves hemodynamics, leading to a decrease in pulmonary arterial pressure and Fulton's index. Our mechanistic studies suggest that the targeted co-delivery system alleviates experimental pulmonary hypertension primarily via the regression of PASMC proliferation by inhibiting cell cycle progression and promoting apoptosis. Taken together, this targeted co-delivery approach offers a promising avenue to target PAs and cure the intractable vasculopathy in pulmonary hypertension.

Objective:To investigate the effect of early selective reduction of twin pregnancies or spontaneous reduction on the improvement of clinical outcome in IVF-ET treatment.Methods:The clinical data of 6535 cycles(6535 cases)of single or twin pregnancy obtained from fresh and frozen-thawed embryo transfer(FET)of IVF or ICSI-ET in the Department of Reproductive Medicine,Women and Children's Hospital Affiliated to Xiamen Univer-sity from November 2011 to December2021 were analyzed retrospectively,The propensity score matching(PSM)method(1∶4)was used to include the study subjects.The surgical reduction group and the spontaneous single-ton group were included in 128 cycles and 510 cycles,respectively.The spontaneous reduction group and the ini-tial singleton group were included in 511 cycles and 2038 cycles,respectively.The pregnancy outcomes of each groups were compared.Results:After PSM,the number of transferred embryos in the surgical and spontaneous reduction group was higher than that in the initial singleton group.The gestational week of delivery was earlier than that of the initial singleton group,and the birth weight of the newborn was lower than that in the initial single-ton group.The rate of premature delivery and low birth weight was higher than that in the initial singleton group,and the difference was statistically significant(P＜0.05).The incidence of pregnancy complications(postpartum hemorrhage,gestational diabetes,hypertensive disorders of pregnancy)and the rate of cesarean section were not significantly different(P＞0.05).The abortion rate and neonatal birth defect rate were missing after adjusting the PSM model.According to the calculation before adjustment,the early abortion rate of the surgical reduction group and the spontaneous reduction group were significantly lower than that in the initial singleton group(P＜0.05),and the late abortion rate of the spontaneous reduction group was lower than that in the initial singleton group,the difference was statistically significant(P＜0.05).There was no significant difference in the neonatal birth defect rate between the surgical/spontaneous reduction group and the initial singleton group(P＞0.05).Conclusions:Patients with multiple pregnancies caused by assisted reproductive technology(ART)may have a relatively poor prognosis in the perinatal period even if the effect of early pregnancy reduction surgery or spontaneous reduction is limited.Reducing the number of embryos transferred cannot completely reverse the pregnancy outcome.Limiting the number of embryos transferred is the fundamental solution to the adverse outcomes of ART assisted pregnancy.