Vitamin B6 related epilepsy is a group of epileptic diseases,seizures in which could not be con-trolled by antiepileptic drugs,but could be controlled or obviously improved by vitamin B6 .It comprises of pyridoxine dependent epilepsy and pyridoxine responsive epilepsy predominantly,and the latter includes vitamin B6 responsive in-fantile spasms,pyridox(am)ine -5′-phosphate oxidase (PNPO)deficiency,hyperphosphatasia mental retardation syndrome (Mabry syndrome)and so on.The clinical presentations of the diseases above are nonspecific,manifesting as refractory seizures onset in neonatal or infantile period,which need to be distinguished from other diseases such as new-born hypoxic ischemic encephalopathy,Ohtahara syndrome and non -ketotic hyperglycinemia.Pyridoxine dependent epilepsy,PNPO deficiency and Mabry syndrome have relative specific biomarkers and known disease -causing genes, which are helpful for diagnosis.It is suggested that pyridoxine or pyridoxal phosphate should be tried first for all patients started seizures early (including all infantile spasms patients),avoiding missing these diseases.And once diagnosed,vi-tamin B6 should be maintained long -term or all the life according to the detailed disease.

ABSTRACT:Objective For preparation of vascularized islets , to isolate and culture human adipose derived stem cells , investigate the role of adipose derived stem cells (ADSCs ) in promoting the proliferation and vascularization of human umbilical vein endothelial cells (HUVECs ) co‐cultured in vitro , and explore its mechanism .Methods ADSCs and HUVECs were isolated by collagenase digestion method ,then cultured ,and identified by morphology ,immunofluorescence or multi‐directional differentiation .The co‐culture system of ADSCs and HUVECs was established , HUVECs cultured alone were set up for control group . The proliferation , vascularization and concentration of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b‐FGF)in the supernatant were compared between the two groups .Results The third generational ADSCs had uniform long spindle fiberous morphology and multi‐directional differentiational function . Immunofluorescence test of surface antigens on ADSCs revealed CD44/CD49d (+ ) ,CD31/CD34 (-) ,on HUVECs CD31/vWF (+ ) . High vascular density was found when co‐cultured in Matrigel of ADSCs and HUVECs than alone of HUVECs .Growth curve shown at days 3 , 4 and 5 of the logarithmic phase , HUVECs count in co‐culture group of ADSCs and HUVECs was (4 .52 ± 0 .31) × 104 ,(7 .18 ± 0 .45) × 104 ,and (8 .23 ± 0 .36) × 104 under indirect co‐culture condition , while that in individual HUVECs group was (2 .71 ± 0 .25) × 104 ,(4 .87 ± 0 .26) × 104 ,and (6 .86 ± 0 .33) × 104 ( P<0 .01) .Population doubling time of HUVECs was shorter in co‐culture group than in individual group .Also ,the OD value of HUVECs was higher in co‐culture group than in individual group when cultured at days 1 ,3 ,5 and 7 ( P<0 .01) .When cultured at days 3 ,7 and 13 ,the concentration of VEGF and b‐FGF in the supernatant was higher in co‐culture group than in individual group ( P< 0 .01 ) . Conclusion ADSCs can promote the proliferation and vascularization of HUVECs in vitro co‐culture conditions by secreting or increasing the HUVECs secretion of VEGF and b‐FGF .

Objective To study the relationship between EZH2 and Ki-67 expression and both in ameloblastoma and normal oral mucosa.Methods EZH2 and Ki-67 were detected by immunohistochemical SP method in 50 cases of ameloblastoma (30 cases primary,20 cases recurrence) and expression in 20 cases of normal oral mucosa.Results The positive expression of EZH2 protein in recurrent ameloblastoma was (36.25±7.24) ％,significantly higher than that of primary ameloblastomas (25.26±4.28) ％ (P ＜ 0.001).Ki-67 in recurrent positive expression in ameloblastoma was (34.96±5.28) ％,significantly higher than that of primary ameloblastomas (29.68±3.27) ％ (P ＜ 0.05).There was positively corTelated on EZH2 and Ki-67 (P ＜ 0.05),but not on the size of tumor and gender (P ＞ 0.05).Conclusions There are high expression of EZH2 and Ki-67 in ameloblastoma,and their play key role in the occurrence and development of ameloblastoma.They can be used as a reference index for the prognosis.The combined detection of EZH2 and Ki-67 protein expression with clinical evidence and its prognosis may contribute to the process of ameloblastoma and recurrence.

Objective To study the insulin‐like growth factor binding protein (IGFBP)‐2 and IGFBP‐6 expression and clini‐cal significance in colorectal adenomas .Methods A collection of Chengde Medical College Hospital from July 2012 to March 2013 after surgical treatment of colorectal cancer confirmed by pathology (colorectal cancer ,CRC) tissue samples of 50 patients ,colorec‐tal adenomas (colorectal adenoma ,CRA) 50 cases ,20 cases of colorectal normal mucosa .Immunohistochemistry and RT‐PCR were used to detect the expression of IGFBP‐2 and IGFBP‐6 protein and mRNA ,combined with clinical and pathological data were statis‐tically analyzed .Results IGFBP‐2 protein positive expression and the amount of mRNA expression in the CRA group compared with normal colorectal mucosa had a rising trend;While compared with CRC group had a tendency to reduce ,and the differences are obvious statistical significance (P<0 .05) ,and IGFBP‐6 protein positive expression in the CRA group compared with normal color‐ectal mucosa had a lower trend;While compared with CRC group amount IGFBP‐6 mRNA expression in the CRA group compared with normal colorectal mucosa had a rising trend;While compared with CRC group had a tendency to reduce ,and the differences were obvious statistical significance (P<0 .05) .In the CRA group ,IGFBP‐2 ,IGFBP‐6 positive expression and the patient′s age , sex ,tumor and the number of parts were no significant statistical difference (P>0 .05) ,but with the degree of hyperplasia had sig‐nificant statistical difference(P<0 .05);In the CRA group ,IGFBP‐2 and IGFBP‐6 expression were negatively related to each other , and the difference was statistically significant (P<0 .01) .Conclusion Colorectal adenomas in normal colorectal mucosa of colorec‐tal cancer to the middle part of the transformation process ,and in its occurrence and development process ,insulin‐like growth factor family (IGFs) and IGFS‐R axis plays an important and irreplaceable role ,so IGFBP‐2 ,IGFBP‐6 may be used as diagnostic colorectal adenomas and early predictors of prognosis ,clinical studies on colorectal adenomas is important .

Objective To isolate and culture human adipose-derived stem cells (hADSCs),investigate the influence of hADSCs on the cellular morphology,survival rate,and function of human islet cells under the in vitro non-contact co-culture conditions,and explore its mechanism.Method hADSCs were isolated by collagenase digestion method,then cultured,and identified by morphology,immunofluorescence and multi-directional differentiation.Adult islet cells were separated and purified by Liberase enzyme and Ficoll 400,then divided into co-culture group and individual group.The cellular growth morphology of islet cells was observed by inverted phase contrast microscope.The survival rate of islet cells,insulin secretory volume,insulin stimulation index and concentration of growth factor in the supernatant were compared between the two groups.Result hADSCs of the third generation showed uniform long spindle fibrocyte-like morphology,and had multi-directional differentiational potentials of osteogenesis and adipogenesis.Immunofluorescence test of surface antigens on hADSCs revealed CD44 + and CD49d +,CD31-,CD34-and CD106-.After 14-day culture,the islet cellular morphology in co-culture group was more intact than that in individual group.The survival rate of islet cells in co-culture group was (82.83 + 2.32) ％,and that in individual group was (53.00 + 2.82) ％ (P＜0.01).Insulin secretory volumes were (23.66 + 2.11) and (7.82 +1.09) mU/L respectively in co-culture group and individual group under high glucose concentration,and 13.22 + 0.77 and 6.40 + 0.44 mU/L respectively under low glucose concentration (P＜0.01 for all).Insulin stimulation index was decreased from 1.67 + 0.10 (at 3rd day) to 1.77 + 0.13 (at 14th day) in co-culture group,and from (1.67 + 0.10) (at 3rd day) to (1.77 + 0.13) (at 14th day) in individual group (P＜0.01).After 14-day culture,the concentrations of HGF,TGF-β,VEGF and bFGF in the supernatant were higher in co-culture group than in individual group (P＜0.01).Conclusion hADSCs were isolated and cultured successfully from adult adipose tissue.They could increase the survival rate and improve the function of islet cells when co-culture with the adult islet cells in vitro through secreting HGF,TGF-β,VEGF and b-FGF.

Purpose To investigate the expression of IGF-Ⅱand IGFBP-6 in co1orecta1 cancer,and to exp1ore the c1inica1 significance in co1orecta1 cancer. Methods IGF-Ⅱand IGFBP-6 were detected by immunohistochemistry and RT-PCR in 50 cases of co1orecta1 cancer(experimenta1 group)and 50 cases of the adjacent mucosa(contro1 group). Results (1)In the experimenta1 group,the ex-pression 1eve1 of IGF-Ⅱprotein and mRNA was significant1y higher than the contro1 group. The expression 1eve1s of IGF-Ⅱprotein and mRNA in co1orecta1 cancer were significant1y 1ower than the contro1 group.(2)The expression 1eve1s of IGF-Ⅱand IGFBP-6 were sig-nificant1y different between different tumor infi1tration depth,1ymph node metastasis,invasion depth and Duke’s stages( P<0. 05), but no difference between genders,age and the degree of tumor differentiation( P>0. 05). Conclusions There is a obvious corre1a-tion between of IGF-Ⅱ and IGFBP-6 in c1inica1 patho1ogica1 parameters in co1orecta1 cancer. Combined detection of the two markers may be the bio1ogica1 indicators of occurrence and prognosis of co1orecta1 cancer,and provide a new scheme for diagnosis and treatment of co1orecta1 cancer.

Objective To explore the genetic etiologies in 2 siblings with different epileptic encephalopathies (EEs) diagnosed as Ohtahara syndrome(OS) and atypical benign partial epilepsy(ABPE) from one family.Methods The 2 brothers were diagnosed at the Pediatric Neurological Clinic of Peking University First Hospital in September 2013,whose clinical data were collected.The coding region of the syntaxin-binding protein 1 gene (STXBP1) and glutamate receptor subunit gene (GRIN2A) were detected by using Sanger sequencing in the 2 siblings.For the elder brother,targeted next-generation sequencing was further performed to detect the genes associated with epilepsy.Results The younger brother manifested focal motor seizures and tonic spasms in cluster at the age of 1 month.Interictal electroencephalogram (EEG) showed suppression-burst pattern.He was diagnosed as OS.The elder brother had seizure onset at age of 6 years old.Focal motor seizure during sleep was his seizure type.His EEG showed interictal discharges in Rolandic area primarily.Electrical status epilepticus during sleep,epileptic negative myoclonus and intellectual disabilities occurred during the course.He was diagnosed as ABPE.Brain magnetic resonance imagines for both of them were normal.Screening of STXBP1 mutations for the younger brother found a de novo heterozygous mutation:c.1672C ＞ T (p.Q558X).Gene detection for the elder brother and the parents showed negative results.Conclusions Coexistence of distinct EEs was reported in 2 brother siblings:the younger brother had OS associated with a novel nonsense mutation in STXBP1,and the elder brother had ABPE without genetic evidence.This study indicated that different pathological mechanisms might exist underlying the two different EEs in a family.

Objective To investigate the relation between the cellular immune function and unexplained recurrent spontaneous abortion(URSA) and the mechanism of active immunotherapy on URSA patients.Methods The flow cytometry(FCM) was used to detect CD3+ ,CD4+ ,CD8+ T lymphocyte and CD16+CD56+ natural killer(NK) cell subsets and the ratio of CD4+/CD8+ of peripheral blood(PBL).112 cases with URSA(76 cases treated with active immunotherapy) and 30 cases of normal fertiled(NF) women were studied.The percentages of T lymphocyte and NK cell subsets before and after therapy were compared among 76 cases wtih URSA treated by active immunotherapy.The rate of next successful pregnancy of URSA patients treated with and without active immunotherapy was compared.Results The percentages of CD3+ and CD16+CD56+ cell subsets as well as the ratio of CD4+/CD8+ of the URSA patients were significantly higher than those of NF cases(P <0.05).After active immunotherapy,the percentages of CD3+ and CD56+CD16+ cell subsets as well as the ratio of CD4+/CD8+ of URSA cases were significantly decreased(P <0.05 ).The rate of next successful pregnancy of URSA cases with and without active immunotherapy were 88.2% and 31.2% respectively,the difference was significant(P< 0.05).Conclusion The changes in the percentages of T lympyocyte and NK cell subsets have something to do with URSA.Active immunotherapy can effectively regulate the cellular immune function and increase the rate of next successful pregnancy of URSA patients.

Objective: To investigate the genotype and phenotype of children with KCNA2 gene related developmental and epileptic encephalopathy (DEE). Methods: Clinical data including the manifestations and electroencephalogram of 8 children with KCNA2 variants treated in the Department of Pediatrics, Peking University First Hospital from March 2017 to June 2019 were collected and analyzed retrospectively. Results: Among the 8 epileptic patients with KCNA2 variants, 5 were males and 3 were females. The age of onset was from 1 day to 11 months. The age at last follow-up ranged from 4 months to 86 months. Two variants including c.1214C>T (loss-of-function) and c.1120A>G (gain-and loss-of-function) were identified. The variant of c.1214C>T was found in six patients (case 1-6). For these patients, the age of onset was from 5 to 11 months and they were characterized by multiple seizure types. All had focal seizures and had normal development before seizure onset with developmental regression after seizure onset. The first electroencephalogram showed epileptic discharges in Rolandic region in two, epileptic discharges in Rolandic region combined with generalized discharge in one, generalized discharge with posterior predominance in two (combined with or transferred to Rolandic region during the course) and epileptic discharges in posterior region combined with generalized discharge in one. And in 5 of them the Rolandic discharges developed into epileptic electrical status (ESES) during sleep. All the six patients were still treated with a combination of multiple antiepileptic drugs. Two of them had seizure controlled at 80 months and 68 months, respectively. The variant of c.1120A>G were identified in two of eight patients (case 7 and 8) and they had seizure onset on the 1st day after birth. Their epileptic seizures were frequent and difficult to control. They had remarkably developmental delay and microcephaly since birth. One case (case 8) had a wide forehead. They had frequent seizures up to the last follow-up. In case 7, the early electroencephalogram showed epileptic discharges in temporal region, and interictal electroencephalogram at 3 months of age showed multifocal discharge with posterior and temporal region predominance. In case 8, the early electroencephalogram was normal and electroencephalogram showed burst suppression at 2 months of age, and it developed epileptiform discharge in posterior region at 1 year of age. Conclusions KCNA2 gene variants can lead to DEE with multiple seizures types. Among them, loss-of-function c.1214C>T is the most common, and these patients have seizure onset at infancy with Rolandic discharges tended to develop into to ESES pattern. The variant of c.1120A>G is a gain-of- and loss-of-function variant, patients with c.1120A>G have seizure onset in neonatal period, the phenotype overlaps with the former but is more severe.

AIM:To study the visual acuity and refractive status of students pupils and middle school students in Shenzhen, and to provide a scientific basis for the prevention and control of myopia.
METHODS:A cluster sampling method was used to select five primary school students(6 737) and three junior middle school students(1 925) from Shenzhen. The visual acuity, anterior segment, fundus, eye position, and refractive status were measured. Information on associated factors for poor vision were also obtained using a questionaire. The risk factors of poor vision and the rate of myopia between grade or gender were analysed by Chi-square test.
RESULTS:The rate of poor vision was 67. 0%. Female, family history of high myopia, long time of continuous look near, short time of outdoor activities were the main risk factors. The rate of emmetropia, hyperopia, astigmatism and myopia were 15. 1%, 11. 3%, 11. 0% and 62. 6% respectively. Emmetropia, hyperopia and astigmatism incidenece rate decreased with age growing, but myopia incidence rate was increased. There were significant differences between adjacent two grades in myopia(χ2=7. 338-45. 018, P<0. 05 ) except the primary grade six and the junior grade one. There were significant differences between boys ( 61. 0%) and girls ( 65. 5%) in myopia(χ2=17. 180, P<0. 05).
CONCLUSION:The rate of poor vison is pretty high in students of Shenzhen aged between 5 to 16 years old, and myopia is the main reason. The development rate of myopia is increased with age. Early management of myopia may play an important role in controlling poor vision in students.