Objective:To summarize the clinical features of epilepsy and (or) developmental delay associated with KCNB1 gene variants in children.Methods:A case series study was conducted on 24 children with KCNB1 gene variants associated with epilepsy and (or) developmental delay who were treated at the Children′s Medical Center of Peking University First Hospital and the Department of Neurology of Shenzhen Children′s Hospital from July 2015 to June 2024. The manifestations of seizures, electroencephalogram (EEG) and genetic test results of those children were analyzed.Results:All the KCNB1 gene variants were de novo, involving 20 different variation, including 15 missense variations, 3 frameshift variations and 2 nonsense variations. There were 7 novel variations. Among the 24 developmental and epileptic encephalopathy children, there were 14 boys and 10 girls. The last follow-up age ranged from 9 months to 13 years and 9 months. Seizures were present in 21 children (88%), with onset ranging from 1 month to 7 years, and 76% (16/21) began before 2 years of age. The seizure types included focal seizures in 15 children (71%), epileptic spasms, myoclonic seizures, and generalized tonic-clonic seizures in 6 children respectively, atypical absence seizures in 4 children, and myoclonic atonic seizures in 1 child. Seventeen children (81%) had a cluster of seizures and 5 had a history of focal status epilepticus with impaired consciousness. All 24 children had varying degrees of developmental delay, with 3 presenting solely developmental delay. EEG abnormalities were present in all the 21 children with seizures, including focal or multifocal discharges in 20 children, generalized discharges in 10 children, hypsarrhythmia in 2 children, and electrical status epilepticus during sleep in 3 children. Magnetic resonance imaging abnormalities were found in 5 of the 24 children. Among the 21 children with seizures, 57% (12/21) achieved seizure control.Conclusions:KCNB1 gene variants are predominantly de novo missense variation. Most affected children present with epilepsy, though some may exhibit only developmental delay. Epilepsy often begins before 2 years of age, with focal seizures being the most common type. About 80% of patients experience clustered seizures. Although most patients achieve seizure control, they still exhibit varying degrees of developmental delay, consistent with developmental epileptic encephalopathy.

Objective To investigate the serum levels of soluble programmed death-1 (sPD-1) and soluble programmed death-ligand 1 (sPD-L1) in chronic hepatitis B (CHB) patients with clinical cure, the correlation between programmed death-1 (PD-1) and lymphocytes by flow cytometry, and the recovery of hepatitis B virus (HBV)-specific immunity. Methods A total of 26 CHB patients with clinical cure, 26 treatment-naïve CHB patients, and 26 healthy controls who were diagnosed at the outpatient service of Peking University First Hospital from January to May of 2022 were enrolled, and related clinical data and peripheral blood samples were collected. ELISA was used to measure the serum levels of sPD-1 and sPD-L1, and flow cytometry was used to measure the expression of PD-1 in peripheral blood lymphocytes. CHB patients with clinical cure were compared with the treatment-naïve CHB patients and the healthy controls. The Kruskal-Wallis H test was used for comparison of non-normally distributed continuous data between three groups, and the chi-square test was used for comparison of categorical data between groups. The Pearson correlation analysis or the Spearman correlation analysis was used to investigate the correlation between two continuous variables. Results For the 26 CHB patients with clinical cure, the mean time of antiviral therapy was 8.33 years, with entecavir as the antiviral drug. The CHB patients with clinical cure had significantly higher levels of sPD-1 and sPD-L1 than the healthy controls ( P < 0.05) and significantly lower percentages of PD-1 + cells/lymphocytes and PD-1 + CD8 + T cells/lymphocytes than the treatment-naïve CHB patients ( P < 0.05). In the treatment-naïve CHB patients, the serum levels of sPD-1 and sPD-L1 were moderately negatively correlated with HBsAg level ( r =-0.524 and -0.583, both P < 0.05). The serum levels of sPD-1 and sPD-L1 were moderately positively correlated with PD-1 + CD8 + T cells/lymphocytes ( r =0.535 and 0.419, both P < 0.05). In the CHB patients with clinical cure, the serum levels of sPD-1 and sPD-L1 were not correlated with age, sex, alanine aminotransferase, T cells/lymphocytes, CD8 + T cells/lymphocytes, PD-1 + T cells/lymphocytes or PD-1 + CD8 + T cells/lymphocytes (all P > 0.05). Conclusion The serum levels of sPD-1 and sPD-L1 in treatment-naïve CHB patients are mainly associated with exhausted CD8 + T cells in peripheral blood, while there is no significant correlation between serum sPD-1/sPD-L1 and exhausted CD8 + T cells in peripheral blood in CHB patients with clinical cure.

Objective:To analyze and reveal the genetic evolution and variation of influenza viruses in cases of co-infection in Guangdong Province.Methods:Throat swab samples were collected from four cases of H1N1pdm and H3N2 co-infection for viral isolation. The isolated strains were subjected to antigen analysis and neuraminidase inhibitor susceptibility test. High-throughput sequencing was used to detect the sequences of strains in three throat swab samples and one virus strain, and then genetic variations were analyzed.Results:Four influenza viruses were isolated with one strain of H1N1pdm and three of H3N2 subtype, and all of them were genetically similar to the vaccine strain in 2022-2023. The HA genes of H1N1pdm and H3N2 strains belonged to clade 6B.1A.5a.2a and 2a.3a.1, respectively. The isolated strains belonged to the same clade as the strains prevalent in Guangdong during the same period. No drug-resistant variations were detected in N1 or N2 gene, and the isolated strains were sensitive to oseltamivir and zanamivir.Conclusions:H1pdm subtype had stronger replication ability than H3 subtype in the influenza viruses isolated from co-infected cases. H1N1pdm and H3N2 subtype influenza viruses were genetically similar to the strains circulating in Guangdong at the same time. The isolated H1N1pdm and H3N2 strains were sensitive to both oseltamivir and zanamivir, indicating that they could continue to be used in the treatment of influenza virus infections caused by one or two genotypes.

Objective: To evaluate the impact of KIT D816 mutation on the salvage therapy in relapsed acute myeloid leukemia (AML) with t(8;21) translocation. Method: The characteristics of the first relapsed AML with t(8;21) translocation from 10 hospitals were retrospectively collected, complete remission (CR₂) rate after one course salvage chemotherapy and the relationship between KIT mutation and CR₂ rate was analyzed. Results: 68 cases were enrolled in this study, and 30 cases (44.1%) achieved CR₂. All patients received KIT mutation detection, and KIT D816 mutation was identified in 26 cases. The KIT D816 positive group had significantly lower CR₂ compared with non-KIT D816 group (23.1% vs 57.1%, χ²=7.559, P=0.006), and patients with longer CR₁ duration achieved significantly higher CR₂ than those with CR₁ duration less than 12 months (74.1% vs 31.9%, χ²=9.192, P=0.002). KIT D816 mutation was tightly related to shorter CR₁ duration. No significant difference of 2 years post relapse survival was observed between KIT D816 mutation and non-KIT D816 mutation group. Conclusion KIT D816 mutation at diagnosis was an adverse factor on the salvage therapy in relapsed AML with t(8;21) translocation, significantly related to shorter CR1 duration, and can be used for prediction of salvage therapy response. KIT D816 mutation could guide the decision-making of salvage therapy in relapsed AML with t(8;21) translocation.

Objective To explore the effect of long-term low dose ionizing radiation on telomere length in adults. Methods Forty female residents aged more than 55 years old from high level natural background radiation area in Yangjiang city and forty age-matched female residents from control area in Enping city were selected by quota sampling as high background group and control group, respectively. Genomic DNA was isolated from their peripheral blood. Telomere length was determined using real time q-PCR. The t-test was used to compare the square roots of the means of two groups. The individuals were divided into four groups based on ages ( 55 -, 60 -, 65 - and ≥70 ) and four groups based on BMI ( <18. 5, 18. 5-23. 99, 24. 0 -27. 99 and ≥28. 0). After adjusting age and BMI, multivariate linear regression analysis was performed to study the relationship between telomere length and cumulative exposure dose. The individuals were divided into longer telomere length group (≥2 ) and shorter telomere length group ( <2). Logistic regression analysis was performed to study the relationship between telomere length and cumulative exposure dose. Results The average cumulative dose was(169. 52 ± 27. 43)mSv for high background group and(47. 52 ± 6. 50)mSv for control group. The telomere length of high background group was( 1. 98 ± 1. 25 ) , shorter than that of control group ( 2. 69 ± 1. 44 ) with statis-tically significant difference(t=2. 24, P <0. 05). The multivariate linear regression analysis revealed that the effect of cumulative dose on telomere length was not significant ( P>0. 05 ) . Association between telomere length and cumulative dose was explored through Logistic regression, and odds ratio was taken as 0. 992(95% CI, 0. 985-0. 999 ) . There was a weak inverse association between telomere length and cumulative dose, because the odds ratio ( OR) was very close to 1. Conclusions No obvious dose-effect relationship between telomere length of residents and cumulative radiation doses was found. But the long-term low dose ionizing radiation may lead to the shortening of the telomere length in adults.

Objective To investigate the distribution situation of human papillomavirus (HPV) genotypes profile in cervical cells among women in Xuzhou area and its clinical significance .Methods 23 kinds of HPV DNA were extracted in cervical cell samples from 8 010 women in Xiuzhou area .The gene‐chips technique of PCR combined with reverse dot blot was adopted to detect the HPV genotypes .Results Among 8010 cervical cell samples ,there were 1 852 HPV infected cases ,the total HPV infection rate was 23 .12% ,the HPV infection rates of single type accounted for 17 .17% and its predominant types were 16 type (4 .35% ) ,followed by 58 type (2 .12% ) and 52 type (1 .82% ) ,The detection rate of multiple HPV infection was 5 .96% ,in which the predominant types were HPV16+58(4 .40% ) ,16+52(2 .94% ) ,11+16(2 .52% ) .Conclusion The single HPV infection of HPV16 ,58 ,52 and the multiple HPV infection of HPV16+58 ,16+52 ,11+16 are the main genotypes of cervical cells among women in Xuzhou area , this gene chip technique is suitable for the cervical cell sample ,its once detection can detect 23 kinds of HPV genotypes with high specificity and high sensitivity ,which has an important significance for the molecular epidemiologic survey study of HPV genotypes distribution among women in our country .

Objective To compare the genotype distribution of HPV in cervical cells of natural crowd and tissues of cervical in‐traepithelial neoplasia(CINⅢ grade) and cervical carcinomas patients .Methods PCR and gene‐chip technology were utilized for the genotype detection of 23 kinds of HPV in cell specimens from 1 047 women of natural crowd (normal group) and tissue specimens from 173 cases of cervical intraepithelial neoplasia(precancerosis group) and 133 cases of patients with cervical carcinoma (cervical carcinoma group) .Results There were 109 ,159 and 121 cases of HPV positive specimens respectively in normal group ,precancer‐osis group and cervical carcinoma group ,and the HPV infection rates were 10 .41% (109/1 047) ,91 .91% (159/173) and 90 .98%(121/133) ,respectively .Conclusion PCR and gene‐chip technology can be used to detect HPV genotypes in cervical cells and cer‐vical tissues specimens .

Objective To compare the genotypes distribution of human papillomavirus (HPV ) infection in tissues of cervical cancers and cervical intraepithelial neoplasias (CIN ) and its clinical significance .Methods The polymerase chain reaction (PCR) and the gene-chips technique were utilized for the detection of 23 kinds of HPV genotypes in the tissue specimens from 192 cases of cervical intraepithelial neoplasia (CIN) and 85 cases of cervical cancers .And the related data of all subjects were analyzed .Results In 192 cases of CIN ,the total positive rate of HPV was 82 .29% (158/192) ,the positive rate of single genotype infection was 46 .88% (90/192) and the positive rate of multiple genotypes infection was 35 .42% (68/192);In 85 cases of cervical cancers ,the to-tal infection rate of HPV was 88 .24% (75/85) ,the positive rate of single genotype infection was 65 .88% (56/85) and the positive rate of multiple genotypes infection was 22 .35% (19/85) .Conclusion PCR combined with the gene-chips technique can be used in the detection of the tissue samples of cervical lesions ,once detection can detect 23 kinds of HPV genotypes with high sensitivity and strong specificity ,which has very important significance to the prevention and treatment of cervical cancer and precancerous lesions and the their vaccine research .

Objective To confirm whether or not let-7b and miR-199a were significantly associated with malignant melanoma growth and proliferation. Methods An over -expression plasmid and an inhibitor, which targeted on let-7b and miR-199a, was constructed. B16F10 cells were divided into seven groups: control group, let-7b plasmid group, miR-199a plasmid group, empty plasmid group, let-7b inhibitor group, miR-199a inhibitor group, inhibitor control group. Foreign gene was transfected into B16F10 cells, let-7b and miR-199a expression were validated from RNA level, protein level and cell level. Results The relative let-7b or miR-199a gene expression of the let-7b plasmid group (3.8776±0.1372)and miR-199a plasmid group (2.8660±0.2821)were significantly higher than control group (P＜0.05), the relative let-7b or miR-199a gene expression of the let-7b inhibitor group (0.2057±0.0263) and miR-199a inhibitor group(0.2656±0.0253) were significantly lower than control group(P＜0.05). The cyclinD1 expression of the let-7b plasmid group(2.023±0.315) and let-7b inhibitor group (1.857±0.377) were significantly higher than control group (0.997±0.041) (P＜0.05), whereas, the Met expression of themiR-199a plasmid group (5.19±0.309) and miR-199a inhibitor group (4.87±0.044) were significantly higher than control group (2.2±0.198) (P＜0.05). The let-7b plasmid group and miR-199a plasmid group B16F10 cell growth rate were slower than control group, especially on the third day after transfection, the growth rate gradually dropped to the lowest value (P＜0.05). In addition, the apoptosis rates of the let-7b plasmid group and miR-199a plasmid group reach to (11.8±1.19)% and (11.3±1.59)%,which were significantly higher than control group (P＜0.05). Conclusions let-7b and miR-199a may be a negative regulator on the B16F10 cell growth and proliferation.

BACKGROUND:Bone marrow is the main source of mesenchymal stem cells(MSCs)at present,but its application has been limited,because of some reasons such as inconvenience of isolation,and the quantity of cells decreases with human increased age.Umbilical cord as a new source of MSCs has been widespread concerned recently.OBJECTIVE:To explore the approach of isolating and culturing MSCs from Wharton's jelly of human umbilical cord,and the methods of identifying the surface antigens and the differentiation potential.METHODS:MSCs were isolated and amplified via tissue-cultivation,and cultured by FasGrow medium.Morphology of MSCs from Wharton's jelly of human umbilical cord was observed under the optical microscope.Its immunophenotypes were detected using immunohistochemistry.The differentiation of MSCs into the osteoblasts was determined utilizing Gomori calcium-cobalt alkaline phosphatase staining,von Kossa calcium node staining,and tetracyclinefluorescence labeling.The differentiation of MSCs into the adipocytes was detected using oil red O staining.RESULTS AND CONCLUSION:MSCs were easily obtained from Wharton's jelly of human umbilical cord via the proposed approach.The primary cells grew up to 70%-80% confluence after 12-16 days of culture,and meanwhile the undifferentiated state was maintained and proliferation was stabilized after passage.The cell cycle of double increase was about 2 days,and proliferation in vitro reached twenty generation above.Surface antigen analysis showed that CD44,CD105,CD133,MHC-I were positively expressed,while CD34,CD45 were negatively expressed.Experiments of differentiation in vitro indicated that the obtained cells were capable of differentiating into fat,osteoblast and nerve-like cells.