ObjectiveTo investigate the anti-Alzheimer's disease （AD） effect of Dipsacus asper（DA） in the Caenorhabditis elegans model， and decipher the underlying mechanism via the peroxisome proliferator-activated receptor α （PPARα）/transcription factor EB （TFEB） pathway. MethodsFirst， transgenic AD C. elegans individuals were assigned into the blank control， model， positive control （WY14643， 20 µmol·L^-1）， and low-， medium-， and high-dose （100， 200， and 400 mg·L^-1， respectively） DA groups. The amyloid β-42 （Aβ₄₂） formation in the muscle cells， the paralysis time， and the deposition of amyloid β-protein （Aβ） in the head were detected. The lysosomal autophagy in the BV2 cell model was examined by Rluc-LC3wt/G120A. The expression levels of lysosomal autophagy-related proteins LC3Ⅱ， LC3I， LAMP2， and TFEB were detected by Western blot. Real-time quantitative polymerase chain reaction （Real-time PCR） was employed to determine the mRNA levels of autophagy-related genes beclin1 and Atg5 and lysosome-related genes LAMP2 and CLN2 downstream of PPARα/TFEB. A reporter gene assay was used to detect the transcriptional activities of PPARα and TFEB. Immunofluorescence was used to detect the fluorescence intensity of PPARα， and the active components of the ethanol extract of DA were identified by UPLC-MS. RCSB PDB， Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）， and Autodock were used to analyze the binding between the active components and PPARα-ligand-binding domain （LBD）. ResultsCompared with the model group， the positive control group and 200 and 400 mg·L^-1 DA groups showed prolonged paralysis time （P<0.05）， and all the treatment groups showed decreased Aβ deposition in the head （P<0.01）. DA within the concentration range of 50-500 mg·L^-1 did not affect the viability of BV2 cells. In addition， DA enhanced the autophagy flux （P<0.05）， up-regulated the mRNA levels of beclin1， Atg5， LAMP2， and CLN2 （P<0.05， P<0.01）， promoted the nuclear translocation of TFEB （P<0.05）， increased LAMP2 expression and autophagy flux （P<0.05， P<0.01）， and enhanced the transcriptional activities of PPARα and TFEB （P<0.01）. The positive control group and 200 and 400 mg·L^-1 DA groups showed enhanced fluorescence intensity of PPARα in the BV2 nucleus （P<0.01）. UPLC-MS detected nine known compounds of DA， from which 8 active components of DA were screened out. The docking results suggested that a variety of components in DA could bind to PPARα-LBD and form stable hydrogen bonds. ConclusionDA may reduce the pathological changes in AD by regulating the PPARα-TFEB pathway.

[Objective] To study the molecular biological mechanism for a case of ABO blood group B subtype, and perform three-dimensional modeling of the mutant enzyme. [Methods] The ABO phenotype was identified by the tube method and microcolumn gel method; the ABO gene of the proband was detected by sequence-specific primer polymerase chain reaction (PCR-SSP), and the exon 6 and 7 of the ABO gene were sequenced and analyzed. Homologous modeling of Bw.03 glycosyltransferase (GT) was carried out by Modeller and analyzed by PyMOL2.5.0 software. [Results] The weakening B antigen was detected in the proband sample by forward typing, and anti-B antibody was detected by reverse typing. PCR-SSP detection showed B, O gene, and the sequencing results showed c.721 C>T mutation in exon 7 of the B gene, resulting in p. Arg 241 Trp. Compared with the wild type, the structure of Bw.03GT was partially changed, and the intermolecular force analysis showed that the original three hydrogen bonds at 241 position disappeared. [Conclusion] Blood group molecular biology examination is helpful for the accurate identification of ambiguous blood group. Homologous modeling more intuitively shows the key site for the weakening of Bw.03 GT activity. The intermolecular force analysis can explain the root cause of enzyme activity weakening.

[Objective] To retrospectively analyze the detection results of alanine aminotransferase (ALT) unqualified repeat blood donors in Guangzhou, so as to provide evidence for further expanding the repeat blood donor pool, reducing the rate of blood discarding and improving the qualified rate of blood test. [Methods] Blood donors with unqualified ALT in Guangzhou Blood Center from January 2018 to April 2024 were selected as the research objects. The past blood donation and population characteristics were analyzed according to the number of blood donations and ALT unqualified times. [Results] Among repeat blood donors with previous ALT disqualification, 99.5% to 99.7% did not have reactive markers for transfusion-transmitted diseases (TTD), which was higher than the rate among first-time blood donors with unqualified ALT (95.8%) (P<0.05). The rate of single-item ALT disqualification in repeat blood donors was higher in males than in females (P<0.05); it also varied by age (18-25 years > 26-35 years > 36-45 years > over 45 years) (P<0.05); and by quarter (third and fourth quarters > first and second quarters) (P<0.05). The ALT unqualified rate was significantly higher whole blood donors than that of platelet donors and returning blood donors (P<0.05). The overall ALT level (51.0 U/L), individual ALT level (56.0 U/L) and individual ALT unqualified rate (66.7%) of repeat blood donors with multiple ALT disqualifications were higher than those of repeat blood donors with single-item ALT disqualifications (26.0 U/L, 38.5 U/L, and 33.3%, respectively) (P<0.05). Moreover, as the number of ALT disqualifications increased, the overall level of ALT in repeat blood donors also increased (P<0.05), and the average level of individual ALT and individual ALT unqualified ratio tended to increase. Repeat blood donors with frequent ALT disqualifications had higher ALT levels (69.0 U/L). [Conclusion] The ALT unqualified rates of repeat blood donors were mostly non-specific elevation without TTD. Repeat blood donors with multiple ALT disqualifications tend to have continuous high ALT. Moreover, and with the increase of ALT disqualifications times, the overall ALT levels the average individual ALT levels and individual ALT unqualified rates showed an increasing trend.

ObjectiveTo investigate the current status of emergency management for severe mental disorders in Shanghai, and to provide countermeasures and suggestions for the establishment of a sound emergency management system for severe mental disorders and the enhancement of emergency management capability. MethodsA questionnaire survey and qualitative interviews were used to conduct an investigation into the emergency management in 17 district-level mental illness prevention and control institutions in Shanghai, which includes the basic situation of emergency management for severe mental disorders, the construction of emergency response teams and personnel, emergency preparedness drills and training, emergency management plans and rules and regulations, and problems encountered in emergency management. ResultsIn terms of emergency management mechanism and basic situation, resources such as personnel allocation, security funds and green channel were well equipped in each district-level mental illness prevention and control institution in Shanghai. However, the equipment of some hardware facilities was still insufficient to some extent. Therefore, further improvement on the emergency management mechanism for severe mental disorders was needed. With regard to the construction of emergency team and personnel allocation, the majority were those aged between 35‒<45 years old, with a bachelor’s degree, and more than 10 years of working experience. For example, 90.27% staff in district-level mental illness prevention and control institution had a bachelor’s degree or above, which was higher than that among the staff in community-level (73.60%); staff majored in clinical medicine in district-level institution accounted for the proportion at 52.71%, higher than that among the staff in community-level (28.86%); 57.24% staff in district-level institution had an intermediate professional title, higher than that among the staff in community-level (42.28%); and 69.90% staff in district-level institution had more than 10 years of working experience, higher than that among the staff in community-level (43.62%). In the aspect of emergency drills and training, all district-level mental illness prevention and control institutions in Shanghai had a high demand for emergency training, and the weak aspects mainly focused on lack of emergency service protocols, skills of addressing technical challenges, and construction of effectiveness evaluation system. Moreover, the teaching methods were primarily centered on case analysis, simulation drills, interactive discussions, and so forth. Concerning emergency management plans and rules and regulations, all districts in Shanghai had relatively established well-developed systems for emergency response plans, emergency response leadership groups, and emergency response operational task forces for severe mental disorders. About half of the institutions had established other rules and regulations related to emergency management of severe mental disorders in addition to emergency plans. ConclusionShanghai has initially established an emergency management system for severe mental disorders, but it is still fragile in specialized training for emergency management of severe mental disorders, construction of emergency management mechanisms, and the building-up of grassroots emergency teams. Further priorities should include strengthening emergency management training, enhancing the construction of emergency management personnel teams, and gradually establishing a more comprehensive and integrated emergency management mechanism for severe mental disorders.

OBJECTIVE: To explore the molecular mechanism of a case with RhD-- phenotype. METHODS: A proband with RhD-- phenotype who attended the clinic of the First Affiliated Hospital of Zhengzhou University on January 29, 2024 was selected as the study subject. Peripheral blood samples were collected from the proband (8 mL) and her close relatives (father, mother and brother; 3 mL each) for Rh phenotyping and irregular antibodies testing with gel card and test tube methods. Direct agglutination reaction and absorption-elution test were used to detect the c antigen on the red blood cells of the proband. PCR-sequence specific primers (PCR-SSP) typing and gene sequencing were used to determine the RHCE gene of the proband and her relatives. The origin of the proband's variant was traced by pedigree analysis. Three-dimensional structural models of the wild-type RhCE*cE protein and the RhD-- phenotype protein were constructed to predict the alterations of the RhD-- phenotype protein caused by the variant. The procedures of this study were approved by the Medical Ethics Committee of the First Affiliated Hospital of Zhengzhou University (Ethics No.: 2023-KY-0870-003). RESULTS: The red blood cells of the proband did not agglutinate with anti-C, anti-c, anti-E, and anti-e. The result of the serum irregular antibody test was negative. The results of direct agglutination reaction and absorption-elution test of the proband were both negative. Her Rh blood group was identified as RhD--. The results of the Rh blood grouping of her close relatives were normal. PCR-SSP detection showed that the RHCE genotypes of the proband and her close relatives were cE/cE and Ce/cE, respectively. Gene sequencing analysis showed that the RHCE genotypes of the proband and her close relatives were RHCE*cE (c.365C>A)/RHCE*cE (c.365C>A) and RHCE*Ce/RHCE*cE (c.365C>A), respectively. Pedigree analysis revealed that the variants in the proband were inherited from her father and mother, respectively. Homology modeling of RhCE*cE protein showed that the RhD-- type peptide chain with a significantly shortened C-terminal was encoded by only 121 amino acid resides, which was 296 amino acid resides shorter compared to the wild-type RhCE*cE peptide chain encoded by 417 amino acid residues. CONCLUSION Above results revealed the molecular biological mechanism of a RhD-- phenotype. The c.365C>A variant in the RHCE gene has rendered the RHCE*cE alleles invalid, which ultimately led to the RhD-- phenotype.
Humans ; Rh-Hr Blood-Group System/chemistry* ; Female ; Phenotype ; Male ; Alleles ; Pedigree ; Base Sequence ; Molecular Sequence Data ; Adult

OBJECTIVE: To investigate the clinical phenotypes and genetic variant characteristics in children with epilepsy. METHODS: A total of 91 children with epilepsy admitted to the Women's and Children's Hospital Affiliated to Ningbo University from July 2021 to October 2022 were selected as the study subjects. Peripheral blood samples were collected from the children for whole exome sequencing. Candidate genetic variants were validated by Sanger sequencing and copy number variation sequencing (CNV-seq). The clinical phenotypes and treatment outcomes of the children with epilepsy were followed up, and an analysis of the relationship between genotype and phenotype was conducted. This study was approved by the Women's and Children's Hospital Affiliated to Ningbo University (Ethics No.: EC2020-048). RESULTS: Among the 91 children with epilepsy, 21 cases (23.08%, 21/91) were found to carry pathogenic or likely pathogenic variants. Of these, 18 cases had involved single base variant or insertional deletion, while 3 cases involved copy number variations. The gene with the highest detection rate was PRRT2 (38.10%, 8/21). Among the children with genetic variants, 47.62% (10/21) had onset during infancy, with 8 diagnosed with Benign familial infantile epilepsy (BFIE), 8 with Developmental epileptic encephalopathy (DEE), and 3 with Epileptic encephalopathy (EE). One case of Dravet syndrome (DS) and one case of Infantile spasms (IS) were also noted. The clinical manifestations of children were diverse and primarily included generalized tonic-clonic seizures and focal seizures. Among them, 52.38% (11/21) had exhibited cluster seizures, 23.81% (5/21) showed fever sensitivity, and 14.29% (3/21) experienced status epilepticus. After pharmacological treatment, 42.86% (9/21) of children had achieved complete seizure control, while 61.90% (13/21) had intellectual disability and 19.05% (4/21) had co-morbid autism spectrum disorder. CONCLUSION Pathogenic or likely pathogenic variants were identified in 23.08% of the pediatric epilepsy cases, with the PRRT2 gene being the most frequently involved. Among children carrying genetic variants, 47.62% had seizure onset during infancy. Genetic factors are an important cause of epilepsy, and early genetic testing may facilitate precise diagnosis, treatment, and prognostic evaluation.
Humans ; Female ; Male ; Epilepsy/genetics* ; Child, Preschool ; Child ; Phenotype ; Genotype ; DNA Copy Number Variations/genetics* ; Infant ; Membrane Proteins/genetics* ; Nerve Tissue Proteins/genetics* ; Adolescent ; Exome Sequencing

Objective: To investigate the characteristics of schizophrenic patients using long-acting antipsychotic medications, so as to provide the basis for applicable population of long-acting antipsychotic medications. Methods: Data of schizophrenic patients using long-acting antipsychotic medications in Shanghai City from June 2020 to June 2022 were collected through Shanghai Mental Health Information Management System, and demographic characteristics, illness and medication use of patients were descriptively analyzed. Results: A total of 2 684 schizophrenic patients using long-acting antipsychotic medications were included in the study, had a mean age of (46.92±12.39) years, with 1 246 males (46.42%) and 1 438 females (53.58%). There were 1 397 unemployed cases, accounting for 52.05%; 1 429 cases with an educational level in junior high school or below, accounting for 53.24%; 1 301 unmarried cases, accounting for 48.47%; 832 cases in poverty, accounting for 31.00%. The caregivers of patients were mainly their parents, with 1 507 cases accounting for 56.15%. The courses of illness were mainly ≤10 years and >10-20 years, with 860 cases each, both accounting for 32.04%; 1 963 cases with incomplete self-awareness, accounting for 73.14%; 1 570 cases hospitalized at least once, accounting for 58.49%. There were 2 486 cases with continuous medication, accounting for 92.62%. The main method of taking medication was given by others, with 1 947 cases accounting for 72.54%. The medication adherence was mainly taking medication on time and in the right amount, with 2 437 cases accounting for 90.80%. Conclusion The main characteristics of schizophrenic patients using long-acting antipsychotic medications are young, unmarried, and unemployed adults, with incomplete self-awareness, continuous medication and medication given by others.

Objective To generate and identify the Itgbl1(integrin beta-like)promoter-driven Cre knock-in mouse line.Methods Itgbll-Cre knock-in mice were generated using clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(Cas9)gene editing.The Itgbl1-Cre mice were crossed with the Cre reporter ROSALSL-tdTomato)mice to detect the expression profile of Cre activity.The tdTomato expression pattern across tissues and cell-specific markers were used to identify the cell types of Itgbl1-expressing cells and their progeny.Results and Conclusion tdTomato was specifically expressed in mucous acinar cells of the sublingual gland,pancreatic islet cells,and gastric endocrine cells.In addition,tdTomato expression was also found in some of the neurons of the retina and brain,as well as in a few cells in the serosal layer of the intestine,articular cartilage,periosteum,and bone marrow.The first Itgbl1-Cre recombinase transgenic mouse line was established,which can specifically label the mucous acinar cells of the sublingual gland.

Objective:To evaluate the dosimetric characteristics, safety and effectiveness of magnetic resonance-guided fractionated stereotactic radiotherapy (FSRT) for brain tumors.Methods:Clinical data of 8 brain tumor patients treated with magnetic resonance-guided FSRT in the Radiotherapy Department of the First Medical Center of the PLA General Hospital from July 2023 to February 2024 were retrospectively analyzed. Online adaptive radiotherapy was adopted for all patients. Adapt-to-position (ATP) or adapt-to-shape (ATS) radiotherapy was chosen by radiologists. Each adaptation was initiated after the radiotherapy plan was re-examined. The radiotherapy fractionation plan was 21-30 Gy/3-5 F. Clinical characteristics, radiotherapy plans and plan parameters were analyzed by statistical description. Median ( Q1， Q3) was used to describe continuous data and percentage was used to describe categorical data. Results:In this study, 9 lesions were treated a total of 41 times, including 20 times (49%) of ATP plan and 21 times (51%) of ATS plan. The median target area coverage rate was 95.1% (95%, 99.8%), the median target area maximum dose rate was 1.15 (1.07, 1.31), the median conformity index (CI) was 0.75 (0.69, 0.86), the median homogeneity index (HI) was 1.09 (1.06, 1.21), and the median gradient index (GI) was 4.73 (3.36, 8.45), respectively. After ATS plan, the median reduction in gross target volume (GTV) was 8.22 cm3 (1.2, 10.1 cm3), and the median reduction in brain tissue V12 Gy was 30.46 cm3 (8.34, 31.13 cm3).The median follow-up was 3.2 months (1.4, 6.1 months). No radiation necrosis was found in any patient. There were 2 cases of acute brain edema during radiotherapy (both were mild). Except for 1 case who died due to systemic disease progression, the remaining patients had no local recurrence, and achieved good quality of life. Conclusions:The parameters of the treatment plan of magnetic resonance-guided FSRT are generally acceptable. The adaptive plan can effectively reduce the dose of normal brain tissues. It is safe and feasible to use the magnetic resonance-guided FSRT for brain tumors.

Aim To investigate the risk factors of death after acute Stanford type A aortic dissection(ATAAD)complicated with malperfusion syndrome(MPS).Methods 244 patients with ATAAD complicated with MPS who ad-mitted to Nanchong Central Hospital from June 2020 to June 2023 were selected as the study objects.The postoperative survival of the patients was followed up and they were classified into survival group(156 cases)and death group(88 ca-ses).After propensity score matching(PSM)was applied in 1 ∶1 matching,there were 54 cases in both groups.Uni-variate and Logistic regression analysis was performed to analyze the risk factors of postoperative death in patients with ATA-AD complicated with MPS.Area under curve(AUC)of receiver operating characteristics(ROC)was used to analyze the prognosis of ATAAD complicated with MPS.The prediction model was established by using the regression equation y=1-1/(1+e-z)and the stability of the model was verified by cross-checking method.Results After matching,compared with the survival group(n=54),in the death group(n=54),the proportion of sex(male),the proportion of alcohol con-sumption,acute physiology and chronic health status Ⅱ(APACHE Ⅱ)score,sequential organ failure(SOFA)score,al-anine aminotransferase(ALT),aspartate aminotransferase(AST),total serum bilirubin(TSB),cholinesterase,serum creatinine(SCr),blood urea nitrogen(BUN),N-terminal pro-brain natriuretic peptide(NT-proBNP),D-dimer(D-D),white blood cell(WBC),neutrophile granulocyte(NEU),fibrinogen degradation product(FDP),platelet(PLT),fi-brinogen(FIB),C-reactive protein(CRP),hypersensitive troponin,operation time,ICU stay time,ventilator stay time,hospital stay,distal extremity hypoperfusion,renal hypoperfusion were significantly increased(P＜0.05).Logistic analy-sis displayed that gender(male),history of drinking,NT-proBNP ≥271.86 ng/L,D-D≥0.74 mg/L and NEU≥13.06× 109 L-1 were independent risk factors in ATAAD patients complicated with MPS for postoperative death(P＜0.05).The combination of NT-proBNP,D-D,gender(male),alcohol drinking history and NEU(referred to as"five factors")had the highest value in predicting ATAAD patients with MPS.The AUC of its ROC curve was 0.979(95％CI:0.937～0.984),the sensitivity was 94.3％,and the specificity was 91.8％,which was higher than the independent predictor.The best critical value predicted by the five factors was 5.02.The survival rate of the group＞5.02 was significantly high-er than that of the group ≤5.02.Log Rank test P＜0.01.A prediction model was established based on the important factors of postoperative death in ATAAD patients with MPS.The results showed that the model had good prediction accu-racy.Conclusion NT-proBNP≥271.86 ng/L,D-D≥0.74 mg/L,gender(male),history of alcohol consumption,and NEU≥×109 L-1 were independent risk factors for long-term prognosis in patients with ATAAD combined with MPS,and their combined application could effectively increase the accuracy of prognosis assessment.