ObjectiveTo investigate the role of paraventricular nucleus (PVN) corticotropin releasing hormone (CRH) neurons in chronic restraint stress (CRS)-induced anxiety-like behavior. And whether exercise relieves chronic restraint stress-induced anxiety through PVN CRH neurons. MethodsTwenty 8-week-old male C57BL/6J mice were randomly divided into control (Ctrl) group and chronic restraint stress (CRS) group. The open field test (OFT) and elevated plus maze (EPM) were used to evaluate anxiety-like behavior of the mice. Food intake was recorded after CRS. Immunofluorescence staining was used to label the expression of c-Fos expression in PVN and calculate the co-expression of c-Fos and CRH neurons. We used chemogenetic activation of PVN CRH neurons to observed the anxiety-like behavior. 8-week treadmill training (10-16 m/min, 60 min/d, 6 d/week) were used to explore the role of exercise in ameliorating CRS-induced anxiety behavior and how PVN CRH neurons involved in it. ResultsCompared with Ctrl group, CRS group exhibited significant anxiety-like behavior. In OFT, the mice in CRS groups spent less time in center area (P<0.001). In EPM, the time in open arm in CRS group were significantly decreased (P<0.001). Besides, food intake was also suppressed in CRS group compared with Ctrl group (P<0.05). Compared with Ctrl group, CRS significantly increase c-Fos expression in PVN and most of CRH neurons co-express c-Fos (P<0.001). Chemogenetic activation of PVN CRH neurons induced anxiety-like behavior (P<0.05) and inhibited feeding behavior (P<0.01). Exercise relieves chronic restraint stress-induced anxiety (P<0.001) and relieved the anorexia caused by chronic restraint stress (P<0.05). Aerobic exercise inhibited the CRS labeled c-Fos in PVN CRH neurons (P<0.001). Furthermore, ablation of PVN CRH neurons attenuated CRS induced anxiety-like behavior. ConclusionCRS activated PVN CRH neurons, induced anxiety-like behavior and reduced food intake. 8-week exercise attenuated CRS-induced anxiety-like behavior through inhibiting PVN CRH neuron. Ablation of CRH PVN neurons ameliorated CRS-induced anxiety-like behavior. These finding reveals a potential neural mechanism of exercise-relieving CRS-induced anxiety-like behavior. This provides a new idea and theoretical basis for the treatment of anxiety and related mental disorders.

ObjectiveTo investigate the role and mechanism of DNA repair regulation in the process of hepatocellular carcinoma （HCC） recurrence. MethodsHCC tissue samples were collected from the patients with recurrence within two years or the patients with a good prognosis after 5 years， and the Tandem Mass Tag-labeled quantification proteomic study was used to analyze the differentially expressed proteins enriched in the four pathways of DNA replication， mismatch repair， base excision repair， and nucleotide excision repair， and the regulatory pathways and targets that play a key role in the process of HCC recurrence were analyzed to predict the possible regulatory mechanisms. The independent samples t-test was used for comparison of continuous data between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. ResultsFor the eukaryotic replication complex pathway， there were significant reductions in the protein expression levels of MCM2 （P=0.018）， MCM3 （P=0.047）， MCM4 （P=0.014）， MCM5 （P=0.008）， MCM6 （P=0.006）， MCM7 （P=0.007）， PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the nucleotide excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019）， RFC4 （P=0.002）， RFC5 （P<0.001）， and LIG1 （P=0.042）； for the base excision repair pathway， there were significant reductions in the protein expression levels of PCNA （P=0.019） and LIG1 （P=0.042） in the HCC recurrence group； for the mismatch repair pathway， there were significant reductions in the protein expression levels of MSH2 （P=0.026）， MSH6 （P=0.006）， RFC4 （P=0.002）， RFC5 （P<0.001）， PCNA （P=0.019）， and LIG1 （P=0.042） in recurrent HCC tissue. The differentially expressed proteins were involved in the important components of MCM complex， DNA polymerase complex， ligase LIG1， long patch base shear repair complex （long patch BER）， and DNA mismatch repair protein complex. The clinical sample validation analysis of important differentially expressed proteins regulated by DNA repair showed that except for MCM6 with a trend of reduction， the recurrence group also had significant reductions in the relative protein expression levels of MCM5 （P=0.008）， MCM7 （P=0.007）， RCF4 （P=0.002）， RCF5 （P<0.001）， and MSH6 （P=0.006）. ConclusionThere are significant reductions or deletions of multiple complex protein components in the process of DNA repair during HCC recurrence.

Objective To study the effects of Ginkgo biloba extract(GBE)on the proliferation and apoptosis of human cerebral adventitia fibroblasts(HBVAFs)by regulating the phosphatidylinositol 3 kinase(PI3K)/protein kinase B(AKT)signaling pathway.Methods HBVAFs cells were divided into blank group(normal culture,no treatment),experimental-L,-M,-H groups(25,50 and 100 mg·L-1 GBE treatment for 24 h)and combined group(100 mg·L-1 GBE treatment and 2 nmol·L-1 insulin-like growth factor-1 treatment for 24 h).Cell proliferation was detected by thiazole blue assay,cell apoptosis was detected by flow cytometry,and the protein expressions of phosphorylated-PI3K(p-PI3K),PI3K、phosphorylated-AKT(p-AKT)and AKT were detected by Western blot.Results The cell survival rates of experimental-L,experimental-M,experimental-H,combined and blank groups were(66.78±8.21)％,(53.51±7.40)％,(42.33±6.01)％,(75.69±5.87)％and(96.77±12.80)％;the apoptosis rates were(24.83±2.47)％,(30.07±3.06)％,(37.02±4.66)％,(16.71±2.11)％and(8.99±0.19)％;p-PI3K/PI3K ratios were 0.75±0.07,0.62±0.06,0.43±0.04,0.72±0.05 and 0.90±0.09;p-AKT/AKT ratios were 0.70±0.07,0.60±0.06,0.38±0.03,0.69±0.06 and 0.90±0.09,respectively.The above indexes of the experimental-L,-M,-H groups were compared with the blank group,and the above indexes of the combined group were compared with the experimental-H group,the differences were statistically significant(all P＜0.05).Conclusion GBE can reduce the proliferation of HBVAFs,promote apoptosis,and its mechanism is related to the inhibition of PI3K/AKT signaling pathway.

AIM To prepare the soluble microneedles of Aconitum brachypodum Diels alkaloids.METHODS Centrifugal molding method was adopted in the preparation of soluble microneedles.With chondroitin sulfate consumption,PVP K120 consumption and 40%ethanol consumption as influencing factors,piercing rate as an evaluation index,the formulation was optimized by Box-Behnken response surface method,after which the morphology,piercing performance,drug content and in vitro transdermal performance were investigated.RESULTS The optimal formulation was determined to be 123 mg for chondroitin sulfate consumption,298 mg for PVP K120 consumption,and 2.4 mL for 40%ethanol consumption,the piercing rate was 98.3%.The soluble microneedles were yellow and square patch with conoid needle,which could pierce aluminum foil and rat skin,along with the drug content of(0.94±0.025)mg.The soluble microneedle group demonstrated the accumulative permeability rate of 91.4%within 24 h,which was higher than that in the gel ointment group,and the permeability accorded with Higuchi equation.CONCLUSION The soluble microneedles of A.brachypodum alkaloids exhibit good mechanical strength,which can achieve effective transdermal delivery of drugs.

Objective:To evaluate the feasibility of non-invasive prenatal testing (NIPT) for the screening of fetal chromosome aneuploidies in twin pregnancies.Methods:A total of 2 745 women with twin-pregnancies were subjected for NIPT screening. Chromosomal karyotyping and chromosomal microarray analysis (CMA) were carried out on amniotic fluid samples from those with a high risk for fetal chromosome aneuploidies, and the diagnosis and pregnancy outcome were followed up. The sensitivity, specificity, positive predictive value and false positive rate of the NIPT were calculated.Results:Compared with other chromosomal abnormalities, NIPT had a higher efficacy for trisomy 21 and sex chromosomal aneuploidy (SCA) in twin pregnancies (with sensitivity being 100%, 100%, and specificity being 99.93%, 99.9%, respectively). It is difficult to evaluate the efficacy for trisomies 18 and 13 due to the limited data. For chromosome microdeletions and microduplications spanning 15 ~ 21 Mb, NIPT also had a certain detection rate. Compared with women with natural conception, NIPT had a higher detection rate for those with twin pregnancies by assisted reproduction ( P < 0.05). Conclusion:It is feasible to use NIPT for the detection of chromosome aneuploidies in women with twin pregnancies.

ObjectiveTo systematically analyze international disability data policies and standards, as well as the application of disability data in policymaking, service optimization and inclusive social development, and to clarify the importance of international disability data policies, standard systems and disability data application for the development of disability-related services. MethodsThrough the analysis of policy content and research on the data standard system, this study explored the disability data policy framework, standard system and technical path of data interoperability and integration of international organizations including the United Nations (United Nations Statistics Division and United Nations Children's Fund), World Health Orgnization, United Nations Educational Scientific and Cultural Organization, and International Labour Organization. ResultsInternational organizations established disability data policy frameworks based on their respective mandates, involving data and service development, data standards, data governance, and data application. The international community established a disability data standard system for disability data collection, coding, exchange, interoperability, statistical analysis, data fusion and application. Building a standardized disability data standard system based on the framework of international health classification standards such as International Classification of Functioning, Disability and Health, and International Classification of Diseases, Eleventh Revision would ensure the consistency of cross-national disability data policies, and the interoperability and comparability of disability data, promoting the development of data-driven disability-related services, accurately identifying the service needs of people with disabilities, and optimizing service provision, thereby improving the quality of life and social participation of people with disabilities. ConclusionThe construction and implementation of international disability data policies and data standards have promoted the standardization and interoperability of disability data. With the application of big data, artificial intelligence and blockchain technologies in disability data, international cooperation and cross-industry data fusion in the field of disability data have been promoted, further promoting the development of data-driven disability services, ensuring equal opportunities for people with disabilities to enjoy service resources, and improving the coverage and quality of disability services.

Objective To optimize the ethanol extraction technology parameters of Jinlei Compound through orthogonal experiment combined with beetle antennae search(BAS)-back propagation(BP)neural network.Methods On the basis of the optimal extraction concentration obtained by single factor investigation,the ratio of solid to liquid,extraction time and extraction times were set as the orthogonal test factors.The entropy weight method was used to calculate the comprehensive scores of the yield of luteolin,kaempferol,swertianin and dry paste.Then,the BAS-BP neural network model was established,and the optimum extraction process was predicted by the BAS.Results BAS-BP neural network optimized Jinlei Compound alcohol extraction process was as follows:solid-liquid ratio 1:10,extraction time of 0.5 h,extraction times of 3,the comprehensive score was 96.352 6.The optimal process parameters obtained by orthogonal design were:solid-liquid ratio 1:10,extraction for 0.5 h,extraction for 3 times,the comprehensive score 90.988 0.The comprehensive score of BAS-BP neural network model was slightly better than that of orthogonal experiment,but the difference between the two was small.The optimal extraction process of Jinlei Compound was determined by comprehensive production practice as the ratio of solid to liquid 1:10,extraction for 0.5 h,extraction for 3 times.Conclusion The optimized process based on BAS-BP neural network has higher extraction efficiency and good stability,which can provide reference for subsequent development and quality control.

Objective:To compare the predictive value of different cardiac ultrasound indexes in mechanical ventilation withdrawal / weaning outcome of intensive care unit (ICU) respiratory failure.Methods:Patients who received mechanical ventilation for >48 hours and were to be withdrawn due to respiratory failure in the Department of Critical Care Medicine, 921st Hospital of the People′s Liberation Army from October 2020 to March 2023 were continuously selected, and their general information, mechanical ventilation conditions, and relevant weaning inspection indicators were recorded. Left ventricular ejection fraction (LVEF), left ventricular outflow tract velocity time integral (LVOT-VTI), left heart Tei index (TI), ratio of early mitral valve diastolic peak blood flow velocity to early mitral ring diastolic peak motion velocity (E/e′) were measured by ultrasound. According to the weaning outcome, the receiver operating characteristic (ROC) curves of different cardiac ultrasound indexes were plotted to evaluate the predictive value of the withdrawal outcome.Results:A total of 83 patients were included, of which 62 were successfully withdrawn and 21 were unsuccessful. There were significant differences in mechanical ventilation time, rapid shallow breathing index (RSBI), LVEF, VTI, left cardiac TI and E/e′ between the successful group and the failed group (all P<0.05). Multivariate regression analysis suggested that mechanical ventilation time and E/e′ were independent factors affecting the outcome of patients with respiratory failure ( OR=0.603, 0.282, all P<0.05). The area under ROC curve of E/e′ predicting mechanical ventilation withdrawal/ weaning failure was 0.844(95% CI: 0.729-0.959), which was superior to the RSBI and other cardiac ultrasound indexes. Conclusions:Echocardiography has predictive value for the outcome of mechanical ventilation of respiratory failure, and E/e′, which reflects the diastolic function of the heart, is the most effective predictor, but it is not suitable for single index prediction.

Objective To investigate the impacts of long non-coding RNA-lung adenocarcinoma metastasis-associated transcripton-1(lncRNA MALAT1)on apoptosis,autophagy and phosphatidylinositol 3 kinase(PI3K)/protein kinase B(Akt)/mammalian rapasin target protein(mTOR)pathway in granulosa cells of polycystic ovary syndrome(PCOS).Methods The expression levels of lncRNA MALAT1 in PCOS ovarian tissue,normal ovarian tissue,human ovarian granulosa cells(KGN)and normal ovarian epithelial cells IOSE80 were detected by qRT-PCR.KGN cells were cultured in vitro and grouped into the control group,the si-NC group,the si-MALAT1 group,the pc-NC group,the pc-MALAT1 group and the pc-MALAT1+740Y-P(PI3K activator)group.qRT-PCR was applied to detect the expression level of MALAT1 mRNA in cells of each transfection group.CCK-8 was applied to detect cell viability.Flow cytometry was applied to detect cell apoptosis rate.Transmission electron microscopy was applied to observe autophagosomes in KGN cells.Western blot assay was applied to detect expression levels of B-cell lymphoma 2(Bcl-2),Bcl-2 associated X protein(Bax),microtubule-associated protein Ⅱ light chain 3(LC3 Ⅱ),microtubule-associated protein Ⅰ light chain 3(LC3Ⅰ),Beclin1,p-PI3K,PI3K,p-Akt,Akt,p-mTOR and mTOR proteins in cells.Results The expression level of MALAT1 mRNA in PCOS ovarian tissue was lower than that in normal ovarian tissue,and that in KGN cells was lower than that in normal ovarian epithelial IOSE80 cells.A small number of autophagosomes were observed in KGN cells in the control group.After interfering MALAT1 expression,OD450 values(24,48 and 72 hours)and the Bcl-2 protein expression of KGN cells were reduced,and the apoptosis rate,the expression of Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin1,p-PI3K/PI3K,p-Akt/Akt and p-mTOR/mTOR proteins were increased(P＜0.05).The number of autophagosomes increased.Changes of the above indexes were reversed after overexpression of MALAT1(P＜0.05).Compared with the pc-MALAT1 group,the OD450 values(24,48,72 h),and the expression of Bcl-2 protein were obviously reduced in the pc-MALAT1+740Y-P group,while the apoptosis rate,number of autophagosomes,the expression of Bax,LC3 Ⅱ/LC3 Ⅰ,Beclin1,p-PI3K/PI3K,p-Akt/Akt,and p-mTOR/mTOR proteins were obviously increased(P＜0.05).The number of autophagosomes increased.Conclusion Overexpression of MALAT1 may inhibit apoptosis and autophagy in PCOS granulosa cells by inhibiting the PI3K/Akt/mTOR signaling pathway.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.