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.

In this study, an established ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS) method was combined with multivariate statistical analysis to investigate the commonality and difference of main chemical components in the medicinal parts of Paeonia lactiflora from different cultivars; in addition, a high performance liquid chromatography(HPLC) method was established to simultaneously determine the content of eight active components in Paeoniae Radix Alba. Non-targeted analysis was carried out by UPLC-Q-TOF-MS on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 μm) column with a gradient elution of 0.1% aqueous formic acid(A)-acetonitrile(B) as the mobile phase at a flow rate of 0.2 mL·min~(-1). The column temperature was 30 ℃, and an electrospray ionization source was used to acquire mass spectrometry data in positive and negative ion modes. According to the accurate molecular weight and fragment ion information provided by multi-stage mass spectrometry and by comparison with reference substances and literature reports, thirty-six identical components were identified in Paeoniae Radix Alba from different cultivars with positive and negative ion modes. In the negative ion mode, two groups of samples were well separated; specifically, seventeen components with significant differences in content were screened and identified, and one component unique in "Bobaishao" was obtained. Quantitative analysis was conducted by high-performance liquid chromatography(HPLC) on an Agilent HC-C_(18)(4.6 mm×250 mm, 5 μm) column with a gradient elution of 0.1% aqueous phosphoric acid(A)-acetonitrile(B) as the mobile phase at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃ and the detection wavelength was at 230 nm. An HPLC method was developed for the simultaneous determination of eight active components(gallic acid, oxypaeoniflorin, catechin, albiflorin, paeoniflorin, galloylpaeoniflorin, 1,2,3,4,6-O-pentagalloylglucose, benzoyl-paeoniflorin) in Paeoniae Radix Albaa from different cultivars. Satisfactory linearity was achieved within the investigated linear ranges and with fine coefficients(r>0.999 0), and the methodological investigation showed that the method had good precision, repeatability and stability. The mean recoveries were 90.61% to 101.7% with RSD of 0.12% to 3.6%(n=6). UPLC-Q-OF-MS provided a rapid and efficient qualitative analytical method for the identification of the chemical components in Paeoniae Radix Alba, and the developed HPLC method was simple, rapid and accurate, which could provide a scientific basis for the evaluation of the germplasm resources and herbal quality of Paeoniae Radix Alba from different cultivars.
Chromatography, High Pressure Liquid ; Paeonia ; Acetonitriles

Interleukin-1 receptor associated kinase 4 (IRAK-4), acting as a serine threonine kinase, is considered as a key signal node for the transduction of IL-1R family and TLRs signal pathway. Studies have found that IRAK-4 has a hand in many signal pathways, involving the inflammatory response of human joints, intestines, liver and nervous system, as well as other autoimmune diseases. It is also one of the causes of drug resistance of some cancer cells. Therefore, IRAK-4 tends to be an effective therapeutic target for inflammatory diseases and cancer. The prospects for the development of drugs in this pathway is to develop novel IRAK-4 small molecule inhibitors and investigate their safety and effectiveness, enrich the clinical treatment of inflammatory and cancer diseases finally. This paper classified and summarized the latest research progress on small molecule inhibitors of IRAK-4 signaling pathway according to structures of the compounds, in order to provide assistances and references for the research and development of related drugs.

METTL3 and METTL14 are two components that form the core heterodimer of the main RNA m6A methyltransferase complex (MTC) that installs m6A. Surprisingly, depletion of METTL3 or METTL14 displayed distinct effects on stemness maintenance of mouse embryonic stem cell (mESC). While comparable global hypo-methylation in RNA m6A was observed in Mettl3 or Mettl14 knockout mESCs, respectively. Mettl14 knockout led to a globally decreased nascent RNA synthesis, whereas Mettl3 depletion resulted in transcription upregulation, suggesting that METTL14 might possess an m6A-independent role in gene regulation. We found that METTL14 colocalizes with the repressive H3K27me3 modification. Mechanistically, METTL14, but not METTL3, binds H3K27me3 and recruits KDM6B to induce H3K27me3 demethylation independent of METTL3. Depletion of METTL14 thus led to a global increase in H3K27me3 level along with a global gene suppression. The effects of METTL14 on regulation of H3K27me3 is essential for the transition from self-renewal to differentiation of mESCs. This work reveals a regulatory mechanism on heterochromatin by METTL14 in a manner distinct from METTL3 and independently of m6A, and critically impacts transcriptional regulation, stemness maintenance, and differentiation of mESCs.
Animals ; Mice ; Methylation ; Chromatin ; Histones/metabolism* ; RNA, Messenger/genetics* ; Methyltransferases/metabolism* ; RNA/metabolism*

Morphine is a frequently used analgesic that activates the mu-opioid receptor(MOR),which has prominent side effects of tolerance.Although the inefficiency of morphine in inducing the endocytosis of MOR underlies the development of morphine tolerance,currently,there is no effective therapy to treat morphine tolerance.In the current study,we aimed to develop a monoclonal antibody(mAb)precisely targeting MOR and to determine its therapeutic efficacy on morphine tolerance and the underlying molecular mechanisms.We successfully prepared a mAb targeting MOR,named 3A5C7,by hybridoma technique using a strategy of deoxyribonucleic acid immunization combined with cell immunization,and identified it as an immunoglobulin G mAb with high specificity and affinity for MOR and binding ability to antigens with spatial conformation.Treatment of two cell lines,HEK293T and SH-SY5Y,with 3A5C7 enhanced morphine-induced MOR endocytosis via a G protein-coupled receptor kinase 2(GRK2)/β-arrestin2-dependent mechanism,as demonstrated by immunofluorescence staining,flow cytometry,Western blotting,coimmunoprecipitation,and small interfering ribonucleic acid(siRNA)-based knock-down.This mAb also allowed MOR recycling from cytoplasm to plasma membrane and attenuated morphine-induced phosphorylation of MOR.We established an in vitro morphine tolerance model using differentiated SH-SY5Y cells induced by retinoic acid.Western blot,enzyme-linked immunosorbent assays,and siRNA-based knockdown revealed that 3A5C7 mAb diminished hyperactivation of adenylate cyclase,the in vitro biomarker of morphine tolerance,via the GRK2/β-arrestin2 pathway.Furthermore,in vivo hotplate test demonstrated that chronic intrathecal administration of 3A5C7 significantly alle-viated morphine tolerance in mice,and withdrawal jumping test revealed that both chronic and acute 3A5C7 intrathecal administration attenuated morphine dependence.Finally,intrathecal electroporation of silencing short hairpin RNA illustrated that the in vivo anti-tolerance and anti-dependence efficacy of 3A5C7 was mediated by enhanced morphine-induced MOR endocytosis via GRK2/β-arrestin2 pathway.Collectively,our study provided a therapeutic mAb,3A5C7,targeting MOR to treat morphine tolerance,mediated by enhancing morphine-induced MOR endocytosis.The mAb 3A5C7 demonstrates promising translational value to treat clinical morphine tolerance.

Objective: To measure and analyze parameters of the three-dimensional (3D) model of the condylar movement envelope surface (ES) and provide a reference for the design of the temporomandibular prosthesis. Methods: Thirty-four healthy adults aged (25.4±2.8) years were recruited from the Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology as subjects. There were 9 males and 25 females, most of them were university students and others outside the university have received undergraduate education or above. Condylar movement ES were obtained using the previous method on the 3D trajectory of condyle. The simulations of condylar movement were projected according to opening movements, protrusion movements, and lateral movements respectively. The total area of the ES and the area proportions of models formed by above different mandibular movements were measured and calculated. The adults' head and maxillofacial 3D models reconstructed by cone beam CT were registered with ES in Geomagic Studio. The inner and outer poles of the condyle, the corrective sagittal axis of the ES, and the median sagittal plane (MSP) were calibrated in registration models using Geomagic Studio, and the parameters were measured as follows: the anteroposterior and medialateral diameters of the condyle, the anteroposterior diameters and the transverse diameters (anterior, middle and posterior parts) of the ES, the angles between the corrective sagittal axis of the ES and MSP (ES-MSP). Pearson correlation analysis was performed by SPSS 24.0. Results: The total area of the ES was (760±133) mm², the opening movement part accounted for (63.3±15.2)%, the protrusion movement part accounted for (14.9±9.6)%, the lateral movement part accounted for (21.8±13.3)%. Parameter measurements were as follows: mediolateral diameters of condyle was (19.8±2.3) mm; anteroposterior diameter of the ES was (21.2±3.1) mm, the transverse diameters (anterior, middle and posterior parts) of the ES were (20.6±2.4), (20.4±2.4), (22.0±2.6) mm, respectively; the transverse diameters of the ES were about 2 mm larger than that of the condyle. The angle between the corrective sagittal axis of ES and the MSP was 6.8°±6.2°. The coefficient of variation (CV) in these parameters showed: CV of the transverse diameters (anterior, middle and posterior parts) of the ES and mediolateral diameter of the condyle were 0.98, 0.99, 0.93, respectively (P<0.001). CV of aera of ES and mediolateral diameter of the condyle was 0.64 (P=0.002). CV of aera of ES and anteroposterior diameter of ES was 0.62 (P=0.004). Conclusions: The 3D envelope surface model formed by mandibular opening movements accounted for the largest proportion of the ES. The corrective sagittal axis of the ES was at an angle to the MSP, the transverse diameters (anterior, middle and posterior parts) of the ES were approximately 2 mm larger than the mediolateral diameter of the condyle, the transverse diameters (anterior, middle and posterior parts) of the ES were highly positively correlated with the mediolateral diameter of the condyle.
Humans ; Adult ; Male ; Female ; Mandibular Condyle ; Cone-Beam Computed Tomography ; Mandible ; Movement ; Temporomandibular Joint

Vaccination is an effective way to reduce the morbidity and mortality of infectious diseases. As a needle-free transcutaneous immunization (NF-TCI) vaccination technology, microneedles (MNs), composed of multiple micro-needles orderly attached to a substrate, can overcome the problems of low immune efficiency, poor compliance and waste of resources that exists in the conventional vaccination by injection, thus becoming a research hotspot in biomedicine. The microneedle vaccine can directly break through the stratum corneum barrier of the skin without touching nerves and blood vessels in the dermis, and effectively delivers the vaccine to the immune cells in the skin tissue to initiate the immune response of the body, thus triggering strong humoral and cellular immune processes. Vaccine delivery via the MNs system possesses the advantages of high safety, satisfying immune effect and practical economy, and shows great prospect in the prevention and treatment of infectious diseases and antineoplastic therapy. This article reviews the development background of MNs in transcutaneous immunization, the types of vaccine delivery, the factors affecting the immune effect, the problems to be solved and development direction in the future.

ObjectiveTo explore the effect of health locus of control on the early rehabilitation after anterior cruciate ligament（ACL） reconstruction. MethodsFrom July 2019 to October 2019， a prospective cohort study of 155 ACL patients receiving reconstruction （male=124 and female=31） in our hospital was conducted. The general data questionnaire， MHLC-C， Tegner activity score， IKDC Score， Lysholm Score and Y-Balance Test were used for further analysis. The correlation between HLC and early rehabilitation after ACL reconstruction was explored by Wilcoxon signed-rank tests， correlation analysis and Logistics regression analysis. ResultsPositive correlations were found between the internality health locus of control （IHLC） and the IKDC score （r_3m=0.77， r_6m=0.70， P<0.001）， as well as the Lysholm scores （r_3m=0.68， r_6m=0.64， P<0.001） and the Tegner activity score （r_3m=0.24， r_6m=0.46， P<0.05） in 3 and 6 months after surgery， and higher IHLC score indicated a better y-balance test outcome［OR 95%CI=0.86（0.76， 0.97）， P=0.016］. Chance health locus of control （CHLC） was negatively correlated with the IKDC score （r_3m=-0.71， r_6m=-0.67， P<0.001）， the Lysholm score （r_3m=-0.49， r_6m=-0.43， P<0.001） and the Tegner activity score （r_3m=-0.22， r_6m=-0.35， P<0.05） in 3 and 6 months after surgery， and higher CHLC score indicated worse outcome of y-balance test ［OR 95%CI=1.26（1.12， 1.41）， P<0.001］. There was a negative correlation between the Powerful others health locus of control （PHLC） and the IKDC score （r_3m=-0.51， r_6m=-0.50， P<0.001）， the Lysholm scores （r_3m=-0.36， r_6m=-0.40， P<0.001）， but there was no correlation with the Tegner activity score in 3 and 6 months after surgery（P>0.05）. The risk of poor y-balance test increased by higher score of PHLC ［OR 95%CI=1.74（1.29， 2.34）， P<0.001］. ConclusionA significant correlation was found between the health locus of control and the early rehabilitation effect after ACL reconstruction. Higher internality health locus of control scores indicated a better rehabilitation outcome， while higher scores of external loci of control indicated higher risk of worse rehabilitation outcome.

Objective:To evaluate the changes in proteome in hippocampus and bioinformatics analysis in mice with perioperative neurocognitive disorders (PND).Methods:Clean-grade healthy male C57BL/6 mice, aged 15 months, weighing 30-35 g, were divided into 2 groups ( n=9 each) using a random number table method: control group (group C) and group PND.The model of PND was established by performing open tibial fracture with intramedullary fixation under isoflurane anesthesia in anesthetized mice.The Morris water maze test, open field test and fear conditioning test were performed at 1 day before operation and at 1, 3 and 7 days after operation.At 1, 3 and 7 days after operation, 3 mice with worst cognitive performance in each cognitive function assessments were sacrificed in group P, and three mice were randomly sacrificed in group C. The hippocampal tissues were then obtained, the expression of differentially expressed proteins was identified by high-performance liquid chromatography-mass spectrometry, and Gene Ontology (GO) functional analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed to analyze the differentially expressed proteins. Results:Compared with group C, the escape latency at different time points was significantly prolonged, and the percentage of time spend on target quadrant and the percentage of freezing time in fear conditioning test were decreased in group P ( P<0.05). There were 21 differentially expressed proteins, of which 12 proteins showed up-regulated expression and 9 proteins showed down-regulated expression.The GO functional analysis showed that the differentially expressed proteins were involved in the process such as the metabolism, signal transmission, regulation of biological processes, formed cell components such as synapses and organelles, and were related to molecular function such as binding and transportation.KEGG signaling pathway analysis showed that there were also differences in MAPK signaling pathway, ErbB signaling pathway, AMPK signaling pathway and the transport of SNARE protein in vesicle and etc. Conclusion:There are 21 differentially expressed proteins in the hippocampus of PND mice, and these proteins are involved in the pathophysiological process probably related to PND such as neuroinflammatory responses, abnormal synaptic structure, mitochondrial dysfunction and decreased autophagy.

Objective: To compare the clinical effects of autologous semitendinosus tendon and allogenic tendon arthroscopic anatomical reconstruction of anterior talofibular ligament (ATFL) combined with calcaneofibular ligament (CFL) in the treatment of chronic lateral ankle instability. Methods: A retrospective analysis was made of 55 patients with chronic lateral ankle instability who underwent arthroscopic reconstruction of ATFL combined with CFL from January 2012 to June 2017. A total of 28 cases were treated with autologous semitendinosus tendon (autologous group), including 19 males and 9 females, with an average age of 28.5±8.03 years (range, 16-46 years). A total of 27 cases were treated with allogenic tendon (allogenic group), including 17 males and 10 females, with an average age of 27.48±7.89 years (range, 16-46 years). ATFL/CFL was reconstructed by the same method in both groups. The reconstruction methods were the same between the groups. The talus and calcaneus were fixed with absorbable compression nails. Results: The operation duration in the autologous group was 94.07±7.83 min, which was longer than that in the allogeneic group 63.56±7.96 min (t=14.51, P<0.001). Fever days 5.26±0.90 days in allogeneic group were longer than 2.46±0.74 days in autologous group (t=-12.55, P<0.001). Wound healing duration in allogeneic group was 13.44±3.33 days longer than that in autologous group 10.32±2.34 days (t=-4.01, P<0.001). In the autologous group, 28 cases were followed up for 34.54±16.04 months, and 27 cases in the allograft group were followed up for 42.74±17.79 months. The mean AOFAS score improved from 63.64±11.20 before operation to 90.21±4.48 after operation in the autologous group, and that improved from 63.93±10.59 before operation to 89.56±5.15 after operation in the allogeneic group with no significant difference between the two groups after operation (t=0.506, P=0.615). The mean VAS score decreased from 5.79±1.79 before operation to 1.54±1.35 after operation in the autologous group, and from 5.89±1.78 before operation to 2.04±1.32 after operation in the allogeneic group. There was no significant difference between the two groups after operation (t=-1.396, P=0.168). Tegner score increased from 4.07±1.39 to 6.43±1.14 in the autologous group and from 3.85±1.06 to 6.52±0.85 in the allogeneic group with no significant difference between the two groups after operation (t=-0.333, P=0.740). Stress radiographic showed that the talar tilt angle decreased from 15.60°±3.86° to 6.01°±2.64° in the autologous group, 16.99°±3.78° to 7.14°±3.34° in the allogeneic group, and there was no significant difference between the two groups after operation (t=-1.382, P=0.171). Anterior talar displacement reduced from 10.82±3.12 mm to 4.03±1.69 mm in the autologous group, from 10.10±2.02 mm to 4.17±1.52 mm in the allogeneic group, and there was no significant difference between the two groups after operation (t=-0.326, P=0.746). No donor tendon dysfunction was found in the autologous group. At the end of follow-up, there was no difference in ankle dorsiflexion, plantar flexion and hind foot mobility between autologous group and allogeneic group. Conclusion Arthroscopic autologous tendon and allogeneic tendon reconstruction of AFTL combined with CFL can obtain satisfactory short-term results. The autologous tendon group was superior to the allogeneic group in terms of fever, wound healing time. However, there was no significant difference in clinical effects between the two groups.