Objective:To explore the mechanism of immunomodulatory effects of methionine-enkephalin(MENK)on dendritic cells(DC).Methods:We used scanning electronic microscope for DC morpholopy,assay for acid phosphatas activity,flow cytometry(FCM) and ELISA to study the effects of DC by MENK.Results:MENK(10-12mol/L) could increase the expression of MHC classⅡ,CD86 and CD40 molecules on DC surface(P

ObjectiveTo explore the influence of clinical symptoms and applicability of discontiguous naikan cognitive therapy(DNCT) among convalescent schizophrenic patients.MethodsApplying DNCT,100 convalescent paranoid schizophrenic patients with convalescent clinical state were consecutively recruited.All the patients were randomly divided into DNCT group and control group and were pretreated with antipsychotic agent therapy,40 patients in DNCT group and 49 patients in control group entered the statistic analysis,11 lost.In DNCT group,the patients received DNCT for successive 28 days.In control group,the patients only received antipsychotic agent therapy.Positive and Negative Syndrome Scale (PANSS),Nurses'Observation Scale for Inpatient Evaluation (NOSIE) were administered to all subjects pre- and post-treatment.ResultsAfter treatment,in the study group,total PANSS scales ( (54.00 ± 10.19 ) vs (45.05 ± 5.28 ),t =5.430,P ＜ 0.01 ),the positive symptom item ((11.00±3.33) vs (9.53 ±1.85),t=3.670,P=0.01),negative symptoms item((12.15 ±4.38) vs (9.40± 2.15 ),t =4.371,P ＜ 0.01 ),general psychopathology item ( (26.90 ± 5.66) vs (22.65 ± 3.07 ) 分,t =4.494,P＜0.01 ) scored lower than before,The difference was statistically significant.PANSS study group after treatment,total scores( (45.05 ±5.28 ) vs (52.04 ± 10.36),t=-3.876 P＜0.01 ),negative symptom item score( t =- 3.789,P ＜ 0.01 ),composite item ( t =2.251,P =0.027 ),the general psychopathology item ( t =- 3.336,P =0.01 ),score significantly lower than the control group.After twelve weeks follow-up study,in the study group,PANSS total scores ( t =4.764,P ＜ 0.01 ),item score of positive symptoms ( t =2.335,P =0.025 ),negative symptoms item score( t =3.083,P =0.004) ),genial psychopathology item score ( t =4.325,P ＜ 0.01 ) was still significantly lower than before treatment,the difference was statistically significant.In study group,after treatment,NOSIE Scale total negative factors scores( t =3.083,P =0.004) were significantly lower than before,total positive factors( t =-2.446,P=0.019),the total estimated factor in the disease scores ( t =-4.730,P ＜ 0.001 )were significantly higher than before treatment.After treatment,in the study group,negative factors ( t =-3.953,P=0.000) were significantly lower than the control group,twelve weeks follow-up,study group total negative factors of NOSIE scale score( t =2.126,P =0.040) was still lower than before treatment,the difference was statistically significant,total positive factor( t =- 2.054,P =0.047 ) still higher than before treatment,the difference was statistically significant.ConclusionDNCT can possibly improve part clinical symptoms of patients with convalescent schizophrenia to a certain extent,especially negative symptom,and the impact remained to the twelve weeks,but need to further prove the effect of naikan cognitive therapy.

AIM: To investigate whether grafting neural stem cells (NSCs) improves the impaired cognitive deficits and spatial recognition after ischemic-hypoxic brain damage (HIBD) in neonatal rats. METHODS: Non-immunosuppressed 7-day-old SD rats were used as research subject and randomly divided into 3 groups: (1) sham group (n=10); (2) HIBD group (n=11); (3) transplant group (n=13). (2) and (3) were anesthetized and subjected to a hypoxic/ischemic injury obtained by combination of left carotid ligation and exposure to 8% oxygen for 2 h. At 3 days post injury, hypoxic-ischemic brain damaged animals were re-anesthetized and randomized to receive stereotactic injection of NSCs prelabeling with BrdU or control media into the hippocampus in the ipsilateral hemisphere. Cognitive (i.e., learning) deficits were assessed at 2 to 4 weeks after transplantation. At the end of the behavioral tests, the animals were killed and evaluated for NSC survival and histopathological analysis. RESULTS: Transplant group showed significantly improved cognitive function in selected tests as compared with HIBD group during the 4-week observation period. They took less time than HIBD group in finding the 3 arms baited with water and had a decreased number of working and reference memory errors in radial maze acquisition tests. Histological analysis showed that transplanted NSCs attenuated CA1 cell loss after HIBD, and NSCs survived for as long as 4 weeks after transplantation and were detected in the hippocampus. CONCLUSION: These data suggest that transplanted NSCs attenuate brain damage and cognitive dysfunction after hypoxic-ischemic brain damage. This approach warrants continued investigation in light of potential therapeutic uses.

Objective: To detect potential variation in an ethnic Han Chinese family affected with late-onset lipid storage myopathy. Methods: Next generation sequencing (NGS) was used to screen disease-related genes in the proband. Suspected mutation was validated with PCR and Sanger sequencing in two patients, their father, and 100 healthy controls. Results: Heterozygous c. 770A>G (p.Tyr257Cys) and c. 1395dupT (p.Gly466Tryfs) mutation were detected in the two patients. Their father was found to be heterozygous for the c. 770A>G (p.Tyr257Cys) mutation, while the c. 1395dupT (p.Gly466Tryfs) variation was not reported previously and not found among the healthy controls. Conclusion Mutations of the ETFDH gene probably underlie the pathogenesis in this family. The novel c. 1395dupT (p.Gly466Tryfs) has enriched the mutation spectrum of EDFDH gene.

BACKGROUND:MicroRNA plays an important role in the process of growth and aging of living body. To know the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons can promote the stem celltransplantation. OBJECTIVE:To investigate the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons. METHODS:(1) The lentiviral vector of let-7d was constructed and transfected into rat bone marrow mesenchymal stem cells. The cells were divided into non-transfected group, negative control group (transfected with FU-RNAi-NC-LV), transfected enhancement group (transfected with let-7d-LV), transfected inhibition group ( transfected with let-7d-inhibition-LV). (2) Rat bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The viability of bone marrow mesenchymal stem cells was determined by MTT method. RESULTS AND CONCLUSION:Under inverted fluorescence microscope, the cells were successful y transfected with let-7d. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in transfected enhancement group were higher than those in the negative control group (P<0.05);while in the inhibition group, they were lower than those in the negative control group (P<0.05). These findings indicate that let-7d can promote the differentiation of bone marrow mesenchymal stem cells into neurons induced by fasudil, and by control ing the expression of let-7d we can influence the differentiation efficiency from bone marrow mesenchymal stem cells to neurons.

OBJECTIVE: To detect potential variation in an ethnic Han Chinese family affected with late-onset lipid storage myopathy. METHODS: Next generation sequencing (NGS) was used to screen disease-related genes in the proband. Suspected mutation was validated with PCR and Sanger sequencing in two patients, their father, and 100 healthy controls. RESULTS: Heterozygous c.770A>G (p.Tyr257Cys) and c.1395dupT (p.Gly466Tryfs) mutation were detected in the two patients. Their father was found to be heterozygous for the c.770A>G (p.Tyr257Cys) mutation, while the c.1395dupT (p.Gly466Tryfs) variation was not reported previously and not found among the healthy controls. CONCLUSION Mutations of the ETFDH gene probably underlie the pathogenesis in this family. The novel c.1395dupT (p.Gly466Tryfs) has enriched the mutation spectrum of EDFDH gene.
Asian Continental Ancestry Group ; Electron-Transferring Flavoproteins ; genetics ; Heterozygote ; High-Throughput Nucleotide Sequencing ; Humans ; Iron-Sulfur Proteins ; genetics ; Lipid Metabolism, Inborn Errors ; genetics ; Muscular Dystrophies ; genetics ; Mutation ; Oxidoreductases Acting on CH-NH Group Donors ; genetics

Objective:To analyze the variations of SGCA gene in two Chinese pedigrees of Han nationality with limb-girdle muscular dystrophy type 2D (LGMD2D) and provide prenatal diagnosis and genetic counseling for subsequent pregnancies within the pedigrees. Methods:This study involved two unrelated patients who were the probands of their pedigrees diagnosed with LGMD2D in the First Affiliated Hospital of Zhengzhou University from June 2017 to January 2018. Genomic DNA was extracted from peripheral blood samples of the probands and their parents. Coding sequences and flanking sequences of 21 LGMD-related genes from the probands were captured and subjected to high-throughput sequencing. Suspected mutations in their parents were detected and validated by Sanger sequencing and/or fluorescence quantitative polymerase chain reaction (PCR). Prenatal genetic diagnosis for high-risk fetuses in the two pedigrees was performed after the causative factors being identified.Results:(1) The proband of pedigree 1 carried compound heterozygous point mutations in SGCA gene with c.218C>G(p.P73R) and c.101G>A(p.R34H) inherited from his father and mother, respectively. Prenatal diagnosis indicated that the second fetus of the family carried the same mutations as the proband, and the family chose to terminate the gestation. (2) The proband of pedigree 2 inherited the compound heterozygous mutations of c.218C>T (p.P73L) and heterozygous deletion of exons 7 and 8 in SGCA gene from his parents. Their second fetus did not carry any of the above mutations and was delivered at full term. Serum creatinase level and physical, motor and mental development of the child were all within the normal range during a two-year follow-up after birth. Conclusions:The heterozygous mutations in SGCA gene are the cause of LGMD2D in the two pedigrees, and c.218C>G(p.P73R) and c.218C>T(p.P73L) are novel mutations. Genetic and prenatal diagnosis based on high-throughput targeted next-generation sequencing can rapidly and accurately detect the mutations responsible for LGMD2D.

OBJECTIVE: To detect potential variant of AR gene in an infant with complete androgen insensitivity syndrome. METHODS: The coding regions and splicing sites of the AR gene were subjected to PCR amplification and direct DNA sequencing. Fluorescence quantitative PCR was also used to detect copy number alterations of exons 2 to 8 of the AR gene. RESULTS: Deletion of exons 2 to 8 was detected in the proband, and the results were verified among the family members. CONCLUSION Hemizygotic deletion of exons 2 to 8 of the AR gene probably underlies the complete androgen insensitivity syndrome in this infant.
Androgen-Insensitivity Syndrome ; genetics ; Base Sequence ; Exons ; Humans ; Infant ; Male ; Polymerase Chain Reaction ; Receptors, Androgen ; genetics

OBJECTIVE: Genetic screening and prenatal diagnosis was performed in eighteen families with high risk of 21-hydroxylase deficiency (21-OHD) to provide valuable information for genetic counseling in these affected families. METHODS: First, multiplex ligation-dependent probe amplification (MLPA) combined with nested-PCR based Sanger sequencing was used to detect CYP21A2 gene mutations in probands and their parents of eighteen families, with seven probands had been dead. Second, paternity test was applied to exclude the possibility of maternal genomic DNA contamination, and fetal prenatal diagnosis is based on the mutations found in proband or parents of the family. RESULTS: Ten mutations were identified in these eighteen families, including large fragment deletion, I2G, E3del8bp, I172N, V281L, E6 cluster, L307Ffs, Q318X, R356W and R484Pfs. All probands were caused by homozygous or compound heterozygous mutations of CYP21A2 gene and their parents were carriers. By comparing short tandem repeat sites contamination of maternal genomic DNA was not found in fetal DNA. Prenatal diagnosis showed that five fetus were 21-OHD patients, four fetus were carriers and the other nine fetus were normal. CONCLUSION CYP21A2 gene mutation is the etiology of 21-OHD. Genetic testing of CYP21A2 could assist physicians in 21-OHD diagnosis and provided genetic counseling and prenatal diagnosis for parents who are at risk for having a child with congenital adrenal hyperplasia.
Adrenal Hyperplasia, Congenital ; diagnosis ; genetics ; Female ; Genetic Testing ; Humans ; Mutation ; Pregnancy ; Prenatal Diagnosis ; Steroid 21-Hydroxylase