Ischemic stroke (IS) is a globally life-threatening disease. Presently, few therapeutic medicines are available for treating IS, and rt-PA is the only drug approved by the US Food and Drug Administration (FDA) in the US. In fact, many agents showing excellent neuroprotection but no blood flow-improving activity in animals have not achieved ideal clinical efficacy, while thrombolytic drugs only improving blood flow without neuroprotection have limited their wider application. To address these challenges and meet the huge unmet clinical need, we have designed and identified a novel compound AAPB with dual effects of neuroprotection and cerebral blood flow improvement. AAPB significantly reduced cerebral infarction and neural function deficit in tMCAO rats, pMCAO rats, and IS rhesus monkeys, as well as displayed exceptional safety profiles and excellent pharmacokinetic properties in rats and dogs. AAPB has now entered phase I of clinical trials fighting IS in China.

AIM:To identify transcriptional differences between the ocular surface ectoderm(OSE)and surface ectoderm(SE)using RNA-seq, and elucidate the OSE transcriptome landscape and the regulatory networks involved in its development.METHODS:OSE and SE cells were differentiated from human embryonic stem(hES)cells. Differentially expressed genes(DEGs)between OSE and SE were analyzed using RNA-seq. Based on the DEGs, we performed gene ontology(GO)analysis, Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis, and protein-protein interaction(PPI)network analysis. Transcription factors(TFs)and hub genes were screened. Subsequently, TF-gene and TF-miRNA regulatory networks were constructed using the NetworkAnalyst platform.RESULTS:A total of 4 182 DEGs were detected between OSE and SE cells, with 2 771 up-regulated and 1 411 down-regulated genes in OSE cells. GO-BP analysis revealed that up-regulated genes in OSE were enriched in the regulation of ion transmembrane transport, axon development, and modulation of chemical synaptic transmission. Down-regulated genes were primarily involved in nuclear division, chromosome segregation, and regulation of cell cycle phase transition. KEGG analysis indicated that up-regulated genes in OSE cells were enriched in signaling pathways such as cocaine addiction, axon guidance, and amphetamine addiction, while down-regulated genes were enriched in proteoglycans in cancer, ECM-receptor interaction, protein digestion and absorption, and cytokine-cytokine receptor interaction. Additionally, compared with SE, 204 TFs(including FOS, EGR1, POU5F1, SOX2, and PAX6)were up-regulated, and 80 TFs(including HAND2, HOXB6, HOXB5, HOXA5, and HOXB8)were down-regulated in OSE cells. Furthermore, we identified 6 up-regulated and 9 down-regulated hub genes in OSE cells, and constructed TF-gene and TF-miRNA regulatory networks based on these hub genes.CONCLUSIONS:The transcriptome characteristics of OSE and SE cells were elucidated through RNA-seq analysis. These findings may provide a novel insight for studies on the development and in vitro directed induction of OSE and corneal epithelial cells.

Abstract: Objective To evaluate the impact of meningococcal vaccine immunization on the incidence of meningococcal meningitis, aiming to refine the local immunization strategies and programs. Methods Data on the reported incidence of meningococcal meningitis in Hebei province for 1970-2023 were collected, and interrupted time-series( ITS) analysis was used to quantitatively analyze the levels and slope change of the incidence of meningococcal meningitis before and after vaccine immunization and before and after inclusion in the Expanded Program on Immunization (EPI). Results The annual average reported incidences of meningococcal meningitis in Hebei province before vaccination (1970-1979), before (1980-2007) and after (2008-2023) the inclusion of the vaccine in the EPI were respectively 20.79 per 100 000, 1.66 per 100 000, and 0.018 per 100 000. The interrupted time-series analysis from 1970 to 2023 showed an initial meningitis incidence rate of 24.12 per 100 000 (t=9.86, P<0.05), with an average annual decrease of 1.07 per 100 000 (t=-2.42, P<0.05). After the introduction of the meningococcal vaccine in 1980, the incidence of meningococcal meningitis decreased quickly with an annual average decline of 18.39 per 100 000, showing a significant short-term intervention effect (t=-2.70, P<0.05); however, the rate of decrease slowed over the long term to 0.13 per 100 000, with the long-term intervention effect not being significant (P>0.05). And the incidence was significantly increased due to the outbreak (t=7.80, P<0.05). From 1980 to 2023, the initial level of incidence was 5.13 per 100 000 (t=8.70, P<0.05), and decreased by 0.23/100 000 per year on average (t=-6.42, P<0.05). After the inclusion of the vaccine in the EPI in 2008, the rate of decrease further slowed down to an average of 0.008 per 100,000 per year, with the long-term intervention effect being significant (t=2.50, P<0.05); the impact of epidemic outbreaks on incidence during this period was not statistically significant (P>0.05). Conclusions Meningococcal meningitis vaccination has led to a general downward trend in the incidence of meningitis in Hebei Province, and has flattened the trend of increased incidence caused by outbreaks. Therefore, long-term maintenance in immunization programs on the meningococcal meningitis is necessary. In addition, it is important to strengthen the monitoring of the distribution of epidemic serogroups in patients and healthy carriers, and to adjust immunization strategies timely based on changes in bacterial populations, selecting and promoting vaccines accordingly for the prevalent strains.

Objective:To investigate the correlation between thyroid hormone resistance and hyperuricemia in euthyroid population.Methods:It was a cross-sectional study. A total of 548 euthyroid subjects who were hospitalized or underwent physical examination in the Heart Center and Health Management Center of the First Affiliated Hospital of Tsinghua University from January 2021 to December 2022 were selected. Thyroid function, uric acid, blood lipid and other indicators were collected in the subjects, and thyroid function parameters were calculated: thyroid-stimulating hormone index (TSHI), thyrotropin T4 resistance index (TT4RI), parametric thyroid feedback quantile-based index (PTFQI), free triiodothyronine/free thyroxine (FT3/FT4), the sum activity of peripheral deiodinases (SPINA-GD) and the secretory capacity of the thyroid gland (SPINA-GT). According to uric acid level, the subjects were divided into high uric acid group and normal group, the clinical characteristics and thyroid function parameters of the two groups were compared, the correlation between thyroid hormone resistance and hyperuricemia was further explored.Results:Compared with the normal group, male proportion (94.4% and 52.5%), smoking rate (5.2% and 21.3%), body mass index [(27.10±3.96) and (24.26±3.42) kg/m 2], waist-to-hip ratio [0.90(0.86, 0.94) and 0.86(0.80, 0.91)], serum creatinine [(85.50±12.27) and (73.77±28.79) μmol/L], total cholesterol [(5.08±0.99) and (4.72±0.86) mmol/L], triglyceride [2.10(4.40, 5.59) and 1.14(0.79, 1.67)mmol/L], low-density lipoprotein [(3.19±0.97) and (2.85±0.84) mmol/L] and homocystine [(15.07±9.13) and (12.50±10.85) μmol/L] were all higher in the high uric acid group, and the level of high-density lipoprotein [1.15(1.43, 2.88) and 1.39(1.16, 1.64) mmol/L] was lower (all P<0.05). In the aspect of thyroid hormone and thyroid function parameters, the FT4[16.90(5.40,17.95) and 16.00(14.30,17.80) pmol/L], FT3[5.56(5.25, 5.94) and 5.22(4.81, 5.63) pmol/L], FT3/FT4 [0.34(0.31, 0.37) and 0.32(0.29, 0.36)], TSHI (2.70±0.50 and 2.58±0.60), PTFQI (0.406±0.332 and 0.335±0.353) and SPINA-GD [3.72(3.41, 4.05) and 3.52(3.18, 4.00) ]were all higher in high uric acid group than those in normal group(all P<0.05). Spearman correlation analysis showed that blood uric acid level was positively correlated with FT4 ( r=0.185), FT3 ( r=0.422), FT3/FT4 ( r=0.16), TSHI ( r=0.134), TT4RI ( r=0.09), PTFQI ( r=0.121) and SPINA-GD ( r=0.157) (all P<0.05). Conclusion:In people with euthyroid function, central resistance to thyroid hormone is correlated with hyperuricemia.

Objective:To investigate the carriage status of Neisseria meningitidis ( Nm) among the healthy population in Hebei Province for the prevention and control of meningitis. Methods:From 2015 to 2022, throat swabs were collected from health people, which were selected by cluster random sampling in 11 cities of Hebei.The positive rate of Nm was detected by bacterial culture. The serogroups of isolated strains were identified.The laboratory detection results of Nm strains, combined with epidemiological survey data, were synthetically analyzed. Results:A total of 20 245 people were investigated; 249 strains of Nm were isolated; the overall Nm carriage rate was 1.23%. The carriage rate was significantly higher in men than in women(χ 2=28.831, P<0.05). The positive rates of Nm in different age groups were significantly different(χ 2=428.018, P<0.05), with the highest rates in the 15-19 year-old group(4.90%, 149/3 042). The positive rates of Nm were significantly different in different regions(χ 2=177.512, P<0.05), with the highest positive rate of Nm in Xingtai, Shijiazhuang, Chengde and Baoding city in sequence. Among the isolated Nm strains, ungroupable serogroups, serogroup B, serogroup C, and serogroup W accounted for 71.49%(178/249), 13.65%(34/249), 6.83%(17/249) and 4.42%(11/249), respectively. Conclusions:The carriage rate of Nm among healthy population is generally low in Hebei Province. It is recommended to continue to strengthen monitoring, pay attention to the changes and distribution characteristics of Nm, and formulate scientific and targeted prevention and control measures of meningococcal disease.

Objective To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly(VM)in fetuses.Methods Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA,and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups.Three miRNAs with significant differential expression between the two groups,whose high expression was associated with VM,were selected for verification with RT-qPCR.Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes AKT3 and CCDC88C.Gene ontology(GO)and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.Results We identified a total of 272 differentially expressed miRNAs in VM cases,including 43 up-regulated and 229 down-regulated miRNAs.The target genes of these differential miRNAs were associated with DNA and transcription factor binding,transmembrane transporter and nucleic acid binding transcription factor activity,and cell developmental process.These miRNAs were mostly enriched in the MAPK,cGMP-PKG and Wnt signaling pathways.Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group(P<0.05),which was consistent with miRNA sequencing results;let-7b-5p expression level was significantly lower in VM group,which was contrary to miRNA sequencing result.Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating AKT3 or CCDC88C expressions.Conclusions The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK,PI3K-Akt,Wnt and cGMP-PKG signaling pathways.

Objective To evaluate the bioequivalence of two formulations of minocycline hydrochloride capsules administered orally after fasting administration and fed administration.Methods An open-label,randomized,two-period,self-crossover design was employed to assess the bioequivalence study.Twenty-eight healthy subjects were enrolled in both fasting and fed groups,with each period involving a single administration of either the reference formulation or the test formulation of 50 mg,separated by a washout period of 7 days.The concentration of minocycline in human plasma was determined by HPLC-MS/MS and was used for calculating pharmacokinetic parameters and evaluating the bioequivalence of the test formulation and reference formulation.Results After oral administration of test and reference formulations of minocycline under fasting condition,the Cmax Values of minocycline were(541±137)ng·mL-1 for the test formulation and(558±140)ng·mL-1for the reference formulation.The AUC0-t values were(8 347±1 986)h·ng·mL-1 for the test and(8 205±1 790)h·ng·mL-1 for the reference.The t1/2 values were(18.2±2.84)h for the test and(18.0±3.05)h for the reference.After oral administration of the test and reference formulations of minocycline under fed condition,the Cmax values of minocycline were(349±72.1)ng·mL-1 for the test and(352±73.2)ng·mL-1for the reference.The AUC0-twere(6 428±1 077)h·ng·mL-1 for the test and(6 588±1 118)h·ng·mL-1 for the reference.The t1/2values were(18.5±3.10)h for the test and(18.4±3.21)h for the reference.Under fasting condition,the 90% confidence intervals for the geometric mean ratios of Cmax,AUC0-t,and AUC0-∞ between the test and reference formulations were(90.84%,101.46% ),(95.2%,102.8% ),and(95.31%,102.71% ),respectively.Under fed conditions,the 90% confidence intervals for the geometric mean ratios of Cmax,AUC0-t,and AUC0-∞ between the test formulation and the reference formulation were(94.71%,103.42% ),(95.40%,99.83% ),and(95.79%,100.02% ),respectively.Conclusions Bioequivalence of the two minocycline formulations was demonstrated after fasting administration and fed administration in a healthy Chinese population.

Objective To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly(VM)in fetuses.Methods Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA,and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups.Three miRNAs with significant differential expression between the two groups,whose high expression was associated with VM,were selected for verification with RT-qPCR.Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes AKT3 and CCDC88C.Gene ontology(GO)and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.Results We identified a total of 272 differentially expressed miRNAs in VM cases,including 43 up-regulated and 229 down-regulated miRNAs.The target genes of these differential miRNAs were associated with DNA and transcription factor binding,transmembrane transporter and nucleic acid binding transcription factor activity,and cell developmental process.These miRNAs were mostly enriched in the MAPK,cGMP-PKG and Wnt signaling pathways.Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group(P<0.05),which was consistent with miRNA sequencing results;let-7b-5p expression level was significantly lower in VM group,which was contrary to miRNA sequencing result.Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating AKT3 or CCDC88C expressions.Conclusions The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK,PI3K-Akt,Wnt and cGMP-PKG signaling pathways.

Objective To evaluate the predictive value of CT-enhanced imaging-based radiomics nomogram for the status of microsatel-lite instability(MSI)in colon cancer.Methods A retrospective analysis was conducted on 129 postoperative colon cancer patients with confirmed MSI status.They were randomly divided into a training group(n=90)and a validation group(n=39)at a ratio of 7:3.Radiomics features were extracted from preoperative CT-enhanced images of the patients.The predictive performance of various machine learning algorithms was evaluated using the area under the curve(AUC).A nomogram model was developed by incorporating clinical independent risk factors,and the model's overall performance was assessed using decision curve analysis(DCA).Results Age and lesion site were identified as prominent independent risk factors and utilized in the construction of a clinical model.The light gradient boosting machine(LightGBM)algorithm was chosen for building a radiomics model.As a joint model,the AUC of the nomogram model of 0.917 in the training group and 0.822 in the validation group.The DCA confirmed the substantial clinical applicability of the nomogram model.Conclusion The CT-enhanced imaging-based radiomics nomogram offers a pioneering and individualized predic-tive approach for determining the MSI status in colon cancer.

Background::Genetic variants of dopaminergic transcription factor-encoding genes are suggested to be Parkinson’s disease (PD) risk factors; however, no comprehensive analyses of these genes in patients with PD have been undertaken. Therefore, we aimed to genetically analyze 16 dopaminergic transcription factor genes in Chinese patients with PD.Methods::Whole-exome sequencing (WES) was performed using a Chinese cohort comprising 1917 unrelated patients with familial or sporadic early-onset PD and 1652 controls. Additionally, whole-genome sequencing (WGS) was performed using another Chinese cohort comprising 1962 unrelated patients with sporadic late-onset PD and 1279 controls.Results::We detected 308 rare and 208 rare protein-altering variants in the WES and WGS cohorts, respectively. Gene-based association analyses of rare variants suggested that MSX1 is enriched in sporadic late-onset PD. However, the significance did not pass the Bonferroni correction. Meanwhile, 72 and 1730 common variants were found in the WES and WGS cohorts, respectively. Unfortunately, single-variant logistic association analyses did not identify significant associations between common variants and PD. Conclusions::Variants of 16 typical dopaminergic transcription factors might not be major genetic risk factors for PD in Chinese patients. However, we highlight the complexity of PD and the need for extensive research elucidating its etiology.