African swine fever(ASF)is an acute,febrile,and highly fatal disease caused by African swine fever virus(ASFV)in pigs.Given the current lack of commercial vaccines and the continu-ous evolution of ASFV in recent years,the emergence of moderately virulent genotype Ⅱ strains and the introduction of genotype Ⅰ attenuated strains have led to persistent and chronic infections in pigs.Therefore,the detection of specific antibodies against ASFV has become imperative.In this study,we established a competitive chemiluminescence immunoassay(p30-cCLIA)for detecting ASFV p30 antibodies using p30 monoclonal antibodies.By detecting sera with clear negative and positive backgrounds,we determined that the Cut-off value of this method was 50％,with both di-agnostic sensitivity(Dsn)and diagnostic specificity(Dsp)reaching 100％.Under optimal reaction conditions,we screened out an enzyme-labeled stabilizer suitable for p30 monoclonal antibody 16-5E7E8-HRP.Furthermore,the sensitivity of the established p30-cCLIA method was higher than that of the commercial blocking ELISA kit(1∶2 048 vs 1∶512)and exhibited good repeatability.Detection of sera positive for other porcine virus infections showed no cross-reactivity.The estab-lishment of this method provides a powerful tool for early diagnosis of ASF.

African swine fever(ASF)is an acute,febrile,and highly fatal disease caused by African swine fever virus(ASFV)in pigs.Given the current lack of commercial vaccines and the continu-ous evolution of ASFV in recent years,the emergence of moderately virulent genotype Ⅱ strains and the introduction of genotype Ⅰ attenuated strains have led to persistent and chronic infections in pigs.Therefore,the detection of specific antibodies against ASFV has become imperative.In this study,we established a competitive chemiluminescence immunoassay(p30-cCLIA)for detecting ASFV p30 antibodies using p30 monoclonal antibodies.By detecting sera with clear negative and positive backgrounds,we determined that the Cut-off value of this method was 50％,with both di-agnostic sensitivity(Dsn)and diagnostic specificity(Dsp)reaching 100％.Under optimal reaction conditions,we screened out an enzyme-labeled stabilizer suitable for p30 monoclonal antibody 16-5E7E8-HRP.Furthermore,the sensitivity of the established p30-cCLIA method was higher than that of the commercial blocking ELISA kit(1∶2 048 vs 1∶512)and exhibited good repeatability.Detection of sera positive for other porcine virus infections showed no cross-reactivity.The estab-lishment of this method provides a powerful tool for early diagnosis of ASF.

Objective:To understand the gene carrying rate of neonatal genetic deafness in Dalian area, and to analyze the pedigree of 6 newborns with positive deafness gene test, to provide a reference basis for preventing genetic deafness.Methods:A total of 1 536 newborns born in Dalian Women′s and Children′s Medical Center (Group) from January to October in 2022 were retrospectively enrolled to detect the 4 genes of hereditary deafness, including GJB2, GJB3, SLC26A4 (PDS) and MT-RNRI (12SrRNA). Among them, 6 newborns with hereditary deafness were tested for NGS Panel gene.Results:A total of 85 deafness gene mutations were detected in 1 536 newborns, with the total carrying rate of 5.53% (85/1 536). Thirty-two cases of GJB2 mutations with carrying rate of 2.08% (32/1 536); 4 cases of GJB3 mutation of 0.26% (4/1 536); 32 cases of SLC26A4 (PDS) gene mutations of 2.08% (32/1 536); 14 cases of MT-RNRI (12SrRNA) mutations with carrying rate of 0.91% (14/1 536); 2 cases had compound heterozygous mutations of GJB2/GJB3, with a carrier rate of 0.13% (2/1 536); 1 cases had compound heterozygous mutations of GJB2/SLC26A4 (PDS), with a carrier rate of 0.07% (1/1 536); 1 case of compound heterozygous mutation in three-gene and a heterozygous mutation in KCNQ4 were detected in NGS Panel testing for hereditary deafness.Conclusions:Homozygous mutation and compound heterozygous mutation are the main factors of autosomal recessive gene deafness, and the NGS Panel gene detection is of great significance for gene traceability and the detection of rare deafness gene.

Objective:To explore the related factors of chromosome abnormality in abortion tissue of patients after in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) and to establish a predictive model of chromosome abnormality, so as to provide more clinical treatment basis. Methods:This was a retrospective case-control study. The abortion tissues of 113 cases of IVF/ICSI assisted pregnancy (106 cases of IVF and 7 cases of ICSI) were collected from the Reproduction and Genetics Laboratory of Dalian Women and Children Medical Center (Group) from June 2022 to August 2023. The samples were detected by copy number variation sequencing (CNV-seq). According to the maternal age, the samples were divided into young pregnant women (<35 years old, n=45) group and elderly pregnant women (≥35 years old, n=68) group. The correlations between different maternal age, male age, duration of infertility, times of previous abortion, number of embryos transferred, gestational age and chromosome abnormalities were analyzed. Results:1) A total of 72 cases [63.72% (72/113)] of abnormal chromosomes were detected in the abortion tissues of patients with IVF/ICSI-assisted pregnancy, of which 61 cases [84.72% (61/72)] were numerically abnormal, most of them were trisomy (53 cases), structural abnormalities were found in 8 cases [11.11% (8/72)], and other complex abnormalities were found in 3 cases [4.17% (3/72)]. 2) The chromosome abnormality rate of abortion tissue in the young pregnant women [51.11% (23/45)] was lower than that in the elderly pregnant women [72.06% (49/68)], and the difference was statistically significant ( P=0.023). 3) In the abortion tissue of elderly pregnant women, univariate analysis showed that the age of the man and times of previous abortion were related to the occurrence of chromosome abnormalities ( P=0.004, P=0.008). 4) Multivariate logistic regression analysis showed that the age of the man and times of previous abortion were the independent risk factors for chromosomal abnormalities in elderly pregnant women ( OR=1.248, 95% CI: 1.064-1.464, P=0.006; OR=0.493, 95% CI: 0.287-0.848, P=0.011), the regression equation was logit (P)=0.222×male age-0.707×times of previous abortion-6.042, area under curve was 0.789, 95% CI: 0.673-0.905, and the maximum value of Jordan index was 0.489. Conclusion:The age of pregnant women was correlated with the occurrence of chromosomal abnormalities in abortion tissues. At the same time, in pregnant women aged 35 years or above, under the condition of fixed abortion rates caused by female factors, an increase in male age and a decrease in previous abortions will lead to an increase in the incidence of chromosomal abnormalities. The model is of good value in predicting chromosome abnormalities.

Objective:To explore the related factors of chromosome abnormality in abortion tissue of patients after in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET) and to establish a predictive model of chromosome abnormality, so as to provide more clinical treatment basis. Methods:This was a retrospective case-control study. The abortion tissues of 113 cases of IVF/ICSI assisted pregnancy (106 cases of IVF and 7 cases of ICSI) were collected from the Reproduction and Genetics Laboratory of Dalian Women and Children Medical Center (Group) from June 2022 to August 2023. The samples were detected by copy number variation sequencing (CNV-seq). According to the maternal age, the samples were divided into young pregnant women (<35 years old, n=45) group and elderly pregnant women (≥35 years old, n=68) group. The correlations between different maternal age, male age, duration of infertility, times of previous abortion, number of embryos transferred, gestational age and chromosome abnormalities were analyzed. Results:1) A total of 72 cases [63.72% (72/113)] of abnormal chromosomes were detected in the abortion tissues of patients with IVF/ICSI-assisted pregnancy, of which 61 cases [84.72% (61/72)] were numerically abnormal, most of them were trisomy (53 cases), structural abnormalities were found in 8 cases [11.11% (8/72)], and other complex abnormalities were found in 3 cases [4.17% (3/72)]. 2) The chromosome abnormality rate of abortion tissue in the young pregnant women [51.11% (23/45)] was lower than that in the elderly pregnant women [72.06% (49/68)], and the difference was statistically significant ( P=0.023). 3) In the abortion tissue of elderly pregnant women, univariate analysis showed that the age of the man and times of previous abortion were related to the occurrence of chromosome abnormalities ( P=0.004, P=0.008). 4) Multivariate logistic regression analysis showed that the age of the man and times of previous abortion were the independent risk factors for chromosomal abnormalities in elderly pregnant women ( OR=1.248, 95% CI: 1.064-1.464, P=0.006; OR=0.493, 95% CI: 0.287-0.848, P=0.011), the regression equation was logit (P)=0.222×male age-0.707×times of previous abortion-6.042, area under curve was 0.789, 95% CI: 0.673-0.905, and the maximum value of Jordan index was 0.489. Conclusion:The age of pregnant women was correlated with the occurrence of chromosomal abnormalities in abortion tissues. At the same time, in pregnant women aged 35 years or above, under the condition of fixed abortion rates caused by female factors, an increase in male age and a decrease in previous abortions will lead to an increase in the incidence of chromosomal abnormalities. The model is of good value in predicting chromosome abnormalities.

Objective:To analyze the results of the joint screening of newborn hearing and deafness genes in Dalian to provide a reference for the prevention and control of hereditary deafness.Methods:Eight hundred and forty-two neonates born in Dalian Women and Children′s Medical Group from January 1, 2022 to May 30, 2022 were screened retrospectively, using AABR (automatic brainstem evoked potential). And 20 mutation sites of common genetic deafness 4 genes , including GJB2, GJB3, SLC26A4 (PDS) and mitochondrial genes associated with drug-induced deafness (MT-RNRI)(12SrRNA), were detected by high-throughput sequencing.Results:Among the 842 newborns, 840 passed hearing screening (99.8%); 36 cases (4.3%) passed the hearing screening but not the hearing loss gene screening; 804 cases passed through the both screening (95.5%); 2 cases (0.24%) failed in the both screening. 38 cases of deafness gene mutations were detected, with a total carrying rate of 4.51% (38/842). Among them, the carrying rates of heterozygous mutations in GJB2, GJB3, SLC26A4 (PDS), MT-RNRI (12SrRNA) were 1.90%, 0.24%, 1.30%, and 0.95%, respectively. The carrying rates of GJB2/GJB3 composite heterozygous mutations were 0.12%.Conclusions:The combined screening of neonatal hearing and deafness genes can reduce the missed rate of hearing screening. The carrier rate of neonatal deafness gene in Dalian is 4.51%, with the highest GJB 2 carrier rate, followed by SLC26A4 (PDS) carrier rate.

OBJE CTIVE To prepare and characterize evodiamine phospholipid complex self-microemulsifying drug delivery system（EVO-PC-SMEDDS），and to investigate its gastric mucosal permeability. METHODS EVO-PC-SMEDDS was prepared ， and particle size ，polydispersity（PDI）and Zeta potential were tested ，and microscopic observation was carried out. The stability of EVO-PC-SMEDDS in simulated gastric liquid with different pH （1.2，2.0，4.0，7.0）was investigated. The entrapment efficiency and drug-loading amount of the preparation were determined ，and the in vitro release was investigated. The gastric mucosal permeability of EVO-PC-SMEDDS was studied by combining rat gastric mucosal tissue and Ussing Chamber technology. RESULTS The particle size of EVO-PC-SMEDDS was （53.63±1.51）nm，PDI and Zeta potential were 0.217±0.017 and （－12.20±0.15）mV，entrapment efficiency was （95.25±0.97）％ and drug-loading amount was （19.30±1.21）mg/g. EVO-PC- SMEDDS exhibited a uniformly dispersed round spherical shape under transmission electron microscope. Stability experiments showed that EVO-PC-SMEDDS exhibited no significant change in particle size ，PDI and Zeta potential under the simulated gastric fluid with different pH and showed excellent stability. Results of in vitro release test showed that compared with evodiamine （EVO），in vitro accumulative release of EVO-PC-SMEDDS were enhanced 6.83-fold，which was in line with the first-order kinetic release model. Results of gastric mucosal permeability showed that gastric mucosal permeation transport ，permeation rate ， permeation flux and area under curve of cumulative permeability of EVO-PC-SMEDDS were higher than those of EVO ， respectively. CONCLUSIONS EVO-PC-SMEDDS is prepared N successfully and shows good stability. It could significantly improve the release behavior and gastric mucosal permeability of EVO.

BACKGROUND: Insufficient cerebral perfusion is suggested to play a role in the development of Alzheimer disease (AD). However, there is a lack of direct evidence indicating whether hypoperfusion causes or aggravates AD pathology. We investigated the effect of chronic cerebral hypoperfusion on AD-related pathology in humans. METHODS: We enrolled a group of cognitively normal patients (median age: 64 years) with unilateral chronic cerebral hypoperfusion. Regions of interest with the most pronounced hypoperfusion changes were chosen in the hypoperfused region and were then mirrored in the contralateral hemisphere to create a control region with normal perfusion. 11C-Pittsburgh compound-positron emission tomography standard uptake ratios and brain atrophy indices were calculated from the computed tomography images of each patient. RESULTS: The median age of the 10 participants, consisting of 4 males and 6 females, was 64 years (47-76 years). We found that there were no differences in standard uptake ratios of the cortex (volume of interest [VOI]: P = 0.721, region of interest [ROI]: P = 0.241) and grey/white ratio (VOI: P = 0.333, ROI: P = 0.445) and brain atrophy indices (Bicaudate, Bifrontal, Evans, Cella, Cella media, and Ventricular index, P > 0.05) between the hypoperfused regions and contralateral normally perfused regions in patients with unilateral chronic cerebral hypoperfusion. CONCLUSION Our findings suggest that chronic hypoperfusion due to large vessel stenosis may not directly induce cerebral β-amyloid deposition and neurodegeneration in humans.
Aged ; Alzheimer Disease/pathology* ; Amyloid beta-Peptides/metabolism* ; Arteries ; Atrophy ; Brain/metabolism* ; Cerebral Cortex/metabolism* ; Cerebrovascular Circulation ; Constriction, Pathologic/pathology* ; Female ; Humans ; Magnetic Resonance Imaging/methods* ; Male ; Middle Aged ; Positron-Emission Tomography/methods*

Background::The pharmacokinetic and clinical behaviors of many proton pump inhibitors (PPIs) in peptic ulcer treatment are altered by CYP2C19 genetic polymorphisms. This non-inferiority study evaluated the efficacy and safety of the novel PPI anaprazole compared with rabeprazole. We also explored the influence of Helicobacter pylori ( H. pylori) infection status and CYP2C19 polymorphism on anaprazole. Methods::In this multicenter, randomized, double-blind, double-dummy, positive-drug parallel-controlled, phase III study, Chinese patients with duodenal ulcers were randomized 1:1 to receive rabeprazole 10 mg + anaprazole placebo or rabeprazole placebo + anaprazole 20 mg once daily for 4 weeks. The primary efficacy endpoint was the 4-week ulcer healing rate assessed by blinded independent review. Secondary endpoints were the proportion of patients with improved overall and individual duodenal ulcer symptoms at 4 weeks. Furthermore, exploratory subgroup analysis of the primary endpoint by H. pylori status and CYP2C19 polymorphism was conducted. Adverse events were monitored for safety. Non-inferiority analysis was conducted for the primary endpoint. Results::The study enrolled 448 patients (anaprazole, n = 225; rabeprazole, n = 223). The 4-week healing rates were 90.9% and 93.7% for anaprazole and rabeprazole, respectively (difference, -2.8% [95% confidence interval, -7.7%, 2.2%]), demonstrating non-inferiority of anaprazole to rabeprazole. Overall duodenal ulcer symptoms improved in 90.9% and 92.5% of patients, respectively. Improvement rates of individual symptoms were similar between the groups. Healing rates did not significantly differ by H. pylori status or CYP2C19 genotype for either treatment group. The incidence of treatment-emergent adverse events was similar for anaprazole (72/220, 32.7%) and rabeprazole (84/219, 38.4%). Conclusions::The efficacy of anaprazole is non-inferior to that of rabeprazole in Chinese patients with duodenal ulcers.Registration::ClinicalTrials.gov, NCT04215653.

OBJECTIVE：To optimize the form ulation of Zuojin pectin c apsules，and to prepare modern Zuojin pectin capsules with protective effects against gastric ulcers. METHODS ：The formulation of Zuojin pectin capsules was optimized with orthogonal test with the contents of pectin ，soluble starch and dextrin as factors ，using formability ，moisture absorption and flow ability as indicators. Zuojin pectin capsule was prepared by wet granulation filling method with Zuojin extract powder as raw material. The contents of palmatine hydrochloride ，berberine hydrochloride ，evodiamine and rutaecarpin were evaluated by HPLC. Basket method was used to investigate the release behavior of the capsule in 0.1 mol/L HCl solution. The gastric ulcer model of rats was established by intragastric administration of 75％ ethanol. Gastric ulcer index ，the inhibition rate of gastric ulcer and the pathological sections were used as indexes to investigate the protective effect of Zuojin pectin capsules （the doses were 54，108， 216 mg/kg）on gastric ulcer. RESULTS ：The optimal formulation of Zuojin pectin capsules included 45％ pectin，12％ soluble starch，27％ dextrin and 1％ xylitol. Results of in vitro drug ， release showed that palmatine hydrochloride and berberine， hydrochloride in Zuojin pectin capsules released 53.76％ and No.54.82％ respectively within 1 h，completely released at about 8 h， and conformed to the zero-order release behavior. 2492109374@qq.com Different doses of Zuojin pectin capsule could improve the ulcer injury of gastric tissue in gastric ulcer model rats to different extent ，and significantly reduced the gastric ulcer index（P＜0.01），significantly increased the inhibition rate of gastric ulcer and the percentage of positive expression area of Schiff ’s iodate staining （P＜0.01）. CONCLUSIONS ：Zuojin pectin capsule with protective effect on gastric ulcer and certain sustained- release effect is successfully prepared.