Research advances in limb-girdle muscular dystrophy type 2Q / 中国当代儿科杂志

Limb-girdle muscular dystrophy (LGMD) is a group of muscular dystrophies with predominantly proximal muscular weakness, and some genes associated with this disease have been identified at present. LGMD type 2Q (LGMD2Q) is a subtype of LGMD and is associated with PLEC gene mutation. Major phenotypes of PLEC gene mutation include epidermolysis bullosa with late-onset muscular dystrophy and epidermolysis bullosa with other lesions. LGMD2Q without skin lesions is rarely reported. This article reviews the pathogenic gene PLEC and clinical manifestations of LGMD2Q, so as to deepen the understanding of the pathogenic gene and phenotype of LGMD2Q.

A review on muscle-specific microRNAs as the biomarker for Duchenne muscular dystrophy / 中国当代儿科杂志

MicroRNA (miRNA) is a non-coding single-stranded RNA with a length of approximately 22 nucleotides and is mainly responsible for the regulation of gene expression at the post-transcriptional level. At present, miRNA have become potential biomarkers for various diseases such as tumor, leukemia, and nervous system disease. Muscle-specific microRNAs are enriched in the skeletal muscle of patients with Duchenne muscular dystrophy (DMD) and also play an important role in the pathogenesis of DMD. Creatine kinase has limited specificity in the diagnosis of DMD since its level is not significantly associated with disease severity, and therefore, it is of great clinical significance to explore whether muscle-specific microRNAs can be used as ideal biomarkers for DMD. This article reviews the research advances in this field.

Effects of light intensities on growth,physiological characteristic and chemical composition of Viola yedoensis / 中国中药杂志

The study is aimed to investigate the effects of light intensities on growth,photosynthetic physiology,antioxidant systems and chemical composition of Viola yedoensis and provide cultivation references for V.yedoensis.Five groups of V.yedoensis were planted under five light intensities conditions,namely 100%,80%,50%,35%,5%of full sunlight,and then morphological index,growth,chlorophyll fluorescence parameters,photosynthetic parameters and antioxidant enzyme system indexes were measured during harvest.The results showed that there was no significant difference in the biomass of V.yedoensis among 35% -100%full sunlight,but the biomass of those were significantly higher than that in the 5%full sunlight treatment(P<0.05).The net photosynthetic rate,transpiration rate,stomatal conductance,intercellular CO_2 concentration and water use efficiency increased firstly and then decreased with the decrease of light intensity;F_m,F_v/F_mand Yield in 5% full sunlight treatment were significantly lower than those in the other four groups(P<0.05).The structure of chloroplast was normal under light intensity ranged from 50%to 100% full sunlight.The lamellar concentration of chloroplast matrix decreased and the starch granules decreased in 35% full sunlight treatment,and the margin of lamellar layer of chloroplast and substrate were blurred,and the starch granules were small and the number of starch granules decreased significantly under 5% full sunlight.MDA content in 5%full sunlight treatment was significantly higher than those in the other four groups(P<0.05).The total coumarin content and total flavonoid content decreased with the decrease of light intensity.In summary,the light in-tensity range suitable for the growth of V.yedoensis is wide(ranging from 35% to 100% full sunlight).The content of flavonoids and coumarins is positively correlated with light intensity.

A prospective cohort study on the risk of HBV infection in preschool children without immune response to hepatitis B vaccine / 中华疾病控制杂志

@# Objective To evaluate the risk of hepatitis B virus(HBV) infection among preschool children who were the non-responders to hepatitis B vaccine in future. Methods A prospective cohort study was conducted. Children aged 2 to 5 years were selected from 64 kindergartens.These children were inoculated three doses of hepatitis b vaccine at 0, 1 and 6 months after birth. Hepatitis B surface antigen (HBsAg)and Hepatitis B surface antibody (anti-HBs)were detected during the period from March to May 2015. The children who were HBsAg negative were enrolled in the study. The subjects were divided into exposure group (anti-HBs negative) and control group (anti-HBs positive) . The follow-up began on June 1, 2015 and ended on June 1, 2016. Serum HBsAg of children in the cohort was then collected and detected from June 1 to 30, 2016. At the end of the study, the HBsAg positive rates between two groups were compared. Results 83 children who received hepatitis B vaccine again during the follow-up period were excluded from 1 907 non-responders. The actual number in non-responders group was 1 824. 151 children were lost at the end of the study. The actual number of follow-up was 1 673 and 5 children were found to be positive for HBsAg and the infection rate was 0.30% (5/1673). In the respondent goup, 2 054 were enrolled and followed. Finally, 140 children were lost and none of the remaining 1 914 people were HBsAg positive at the end of the study. HBsAg positive rate was higher in the non-responder group than in the responder group (P=0.023). Conclusion There is a risk of HBV infection in the children who are non-responders to hepatitis B vaccine in future.

Clinical and genetic features of limb-girdle muscular dystrophy type 1B: a case report / 中国当代儿科杂志

This article reports a case of limb-girdle muscular dystrophy type 1B (LGMD1B) caused by a novel splicing heterozygous mutation in the LMNA gene. The proband presented with progressive aggravation of weakness in walking. There was no atrophy of the scapular muscles and the lower-extremity proximal muscles, with normal muscle tension of the extremities, grade 4 muscle strength in the upper and lower extremities, and positive Gower sign. The level of creatine kinase was 779 U/L. Muscle hematoxylin-eosin staining showed muscular dystrophy, and there was no significant reduction in the expression of Lamin A protein. Second-generation sequencing revealed a novel splicing heterozygous mutation, c.810+2T>C, in the LMNA gene, while this locus was normal in his parents. GERP++RS software predicted that the mutation site was highly conservative. Human Splice Finder and Spliceman software predicted that the mutation might be a pathogenic mutation. ExPASy software predicted that the new amino acid sequence became shorter. There were two sequences of mRNA in the patient's muscle: one was the normal sequence, which accounted for 92.2%; the other was partial intron 4 retention, which was the abnormal splice variant accounting for 7.8%. LGMD1B is a type of autosomal dominant inherited myopathy caused by a mutation in the LMNA gene located on the autosomal 1q22. This study extends the mutation spectrum of the LMNA gene and provides help to the diagnosis of LGMD1B.

Infantile hypophosphatasia caused by a novel compound heterozygous mutation: a case report and pedigree analysis / 中国当代儿科杂志

This article reported the clinical features of one child with infantile hypophosphatasia (HPP) and his pedigree information. The proband was a 5-month-old boy with multiple skeletal dysplasia (koilosternia, bending deformity of both radii, and knock-knee deformity of both knees), feeding difficulty, reduction in body weight, developmental delay, recurrent pneumonia and respiratory failure, and a significant reduction in blood alkaline phosphatase. Among his parents, sister, uncle, and aunt (other family members did not cooperate with us in the examination), his parents and aunt had a slight reduction in alkaline phosphatase and his aunt had scoliosis; there were no other clinical phenotypes or abnormal laboratory testing results. His ALPL gene mutation came from c.228delG mutation in his mother and c.407G>A compound heterozygous mutation in his father. His aunt carried c.228delG mutation. The c.407G>A mutation had been reported as the pathogenic mutation of HPP, and c.228delG mutation was a novel pathogenic mutation. Hypophosphatasia is caused by ALPL gene mutation, and ALPL gene detection is an effective diagnostic method. This study expands the mutation spectrum of ALPL gene and provides a theoretical basis for genetic diagnosis of this disease.

BMP9 effectively induces osteogenic differentiation of immortalized calvarial mesen-chymal progenitor cells / 军事医学

Objective To investigate the effect of adenovirus-bone morphogenic protein 9 ( Ad-BMP9 ) on osteogenic differentiation of immortalized calvarial mesenchymal progenitor cells ( iCALs ) .Methods iCALs were infected with adenoviral vectors encoding BMP-9 or green fluorescent protein ( GFP) and the early osteogenic differentiation was assessed by detecting alkaline phosphatase (ALP) activity after being cultured for 3, 5 and 7 days.14 days after infection, alizarin red S staining was performed to study the formation of osteogenic calcium nodules .The expression of osteogenic marker genes Runx2 and OCN was assessed by quantitative real-time ( RT )-PCR and Western blotting .Results Significant increases in ALP activity and in the expressions of Runx 2 and OCN were detected in BMP-9 treated iCALs compared with GFP-treated cells(P<0.05).Meanwhile, alizarin red S staining showed that more mineralized nodules were found in the BMP-9 induced group .Conclusion BMP-9 can promote the osteogenic differentiation of iCALs .

Advance in therapy for Duchenne/Becker muscular dystrophy / 中国当代儿科杂志

Duchenne/Becker muscular dystrophy (DMD/BMD) is the most common X-linked recessive inherited neuromuscular disease, characterized by progressive muscle weakness. Mutations in the dystrophin gene are responsible for this disease. Treatment for this disease has always been a topic of interest. With the development of diagnosis and treatment technology of molecular biology, promising therapies have been developed. This review article summarizes the advance in traditional therapy, cell transplantation and gene therapy for this disease.

Role of platelet-derived growth factor after hemorrhagic shock in rats / 局解手术学杂志

Objective To investigate platelet-derived growth factor ( PDGF ) protection on blood flow and mitochondrial function of hemorrhagic shock rats. Methods Ninety-six SD rats were randomly divided into six groups including shock group, lactated ringer's solution (LR) resuscitation group,PDGF treatment groups(1,3. 5,7,15μg/kg). Laster-Doppler and oxygen concentration determination method were applied to observe the protective effect of PDGF treatment on animal survival,blood flow and mitochondrial function in liver and kidney. Re-sults As compared with LR resuscitation group,PDGF treatment increased animal survival rate and also improved blood fiow of liver and kindy,mitochondrial respiration control ration(RCR),of which the group with 3. 5μg/kg had the best result. Conclusion This finding sug-gests that PDGF may be a potential agent to treat acute critical such as hemorrhagic shock.

Effect and mechanism of high-dose Vitamin B3 on granulopoiesis in normal rat / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the effect and mechanism of high dose Vitamin B3 on granulopoiesis in normal rat.</p><p><b>METHODS</b>Twenty one healthy SD rats were randomly divided into three groups: the Vitamin B3 group (Vit B3 500 mg·kg⁻¹·d⁻¹, × 7 d), the rhG-CSF group (rhG-CSF 25 μg·kg⁻¹·d⁻¹, × 7 d) and the normal saline group (2 ml/d, × 7 d). The peripheral blood cell counts were analyzed by automatic blood cell counter before (day 0) treatment, the third day (day 3) and the seventh day (day 7) after administration of drugs, respectively. The concentration of serum nicotinamide adenine dinucleotide (NAD⁺) level was measured by enzymatic cycling assay before and after drugs treatment. The expressions of G-CSF, G-CSFR, SIRT1, C/EBPα, C/EBPβ, C/EBPε and NAMPT mRNA were detected by reverse transcription real-time fluorescent quantitative PCR.</p><p><b>RESULTS</b>The neutrophil counts increased significantly after 7 days of Vitamin B3 and rhG-CSF treatment compared with that of control group [(1.64 ± 0.19) × 10⁹/L, (1.88 ± 0.37)× 10⁹/L vs (0.86 ± 0.18) × 10⁹/L, P<0.01]; the level of serum NAD⁺ increased significantly [(0.96 ± 0.08) nmol/L, (0.65 ± 0.12) nmol/L vs (0.36 ± 0.15) nmol/L, P<0.01]; the expression of G-CSF, G-CSFR, SIRT1, C/EBPα, C/EBPε and NAMPT mRNA in bone marrow mononuclear cells were increased significantly compared with that of control group (P<0.01).</p><p><b>CONCLUSION</b>High dose of Vitamin B3 may play an important role in increasing absolute neutrophil count in healthy rat under steady state, and the mechanism may be dependent on NAMPT-NAD⁺-SIRT1 signaling pathways.</p>

Bone marrow-derived mesenchymal stem cells regulate the function of Th17/Treg in peripheral blood of severe asthmatic children / 中国病理生理杂志

AIM: To investigate the regulatory function of bone marrow-derived mesenchymal stem cells (MSCs) on T helper 17 cells (Th17) and regulatory T cells (Treg) in peripheral blood of severe asthmatic children . METHODS:MSCs were isolated , cultured and identified in vitro.MSCs digested with mitomycin were cocultured with T lymphocytes (TLC) at different ratios (1∶1, 1∶2, 1∶10 and 1∶20) from severe asthmatic children for 72 h.The prolifera-tion of TLC was measured by CCK-8 method.In the coculture system of the 1∶2 ratio and the single TLC system , the super-natant levels of interleukin-17 (IL-17) and transforming growth factor-β(TGF-β) were measured by ELISA.The mRNA expression of retinoic acid-related orphan nuclear receptor C (RORC) and forkhead box protein 3 (Foxp3) in TLC was de-tected by qRT-PCR.RESULTS:After cocultured with MSCs , the proliferation of TLC decreased significantly in a dose-dependent manner (P0.05).CONCLUSION: MSCs suppresses Th17 polarization of naive peripheral blood CD 4 +T cells and matures Th17 cells secreting IL-17, which may ef-fectively revise Th17/Treg imbalance of asthma .

FGFR3 gene mutation analysis of achondroplasia and hypochondroplasia families / 临床儿科杂志

Objective To screen the sequence of fibroblast growth factor receptor 3 (FGFR3) genes in children with dys-chondroplasia and their family members for searching the mutations. Methods The sequence of exon 10 and exon 13 in muta-tion hot spot region of FGFR3 gene in seven families was analyzed using polymerase chain reaction (PCR) and DNA sequenc-ing technology. Results The c.1138G>A missense mutation in exon 10 was found in 4 probands who were diagnosed as achon-droplasia (ACH), while this mutation was absent in their parents. The c.1620C>A missense mutation in exon 13 was found in one girl and her mother who both were diagnosed as hypochondroplasia (HCH) with mild symptoms. Neither mutation men-tioned above was found in the other two probands. Conclusions Through detecting the mutation in exon 10, exon 13 of FGFR3 gene, most patients of ACH or HCH can be finally diagnosed. However, it is necessary to perform the mutation screening on the other zones of FGFR3 gene and on other related genes for a few cases.

Report of a case of severe congenital neutropenia with literature review / 临床儿科杂志

Objective To investigate the clinical features and pathogenesis of severe congenital neutropenia (SCN) by detecting the gene mutation of a SCN patient suspected by clinical diagnosis. Methods The intravenous anticoagulant and clin-ical data and laboratory results of this child were collected;the phagocyte and oxidation function of neutrophils were evaluated by flow cytometry;ELANE, HAX1, WAS, GFI1, CSF3R and CXCR4 genes were screened by PCR amplification and sequencing. Results The neutrophil function of this patient was normal; sequencing results revealed no mutation occurred in ELANE, HAX1, WAS, GFI1, CSF3R and CXCR4;and granulocyte colony-stimulating factor (G-CSF) can obviously enhance the level of neutrophils. Conclusion SCN is a kind of genetic heterogeneity syndrome associated with multiple gene mutations, gene diag-nosis will contribute to understanding of the pathogenesis of the disease and provide theoretical basis for treatment. Though more and more pathogenic genes were found to be connected with SCN, the cases of unknown mutation still account for a large proportion of this disease.

New progress in genetic mechanisms of congenital neutropenia / 中华实用儿科临床杂志

Congenital neutropenia (CN) is a heterogeneous group of disorders in the inherited bone marrow failure syndromes (IBMFS),characterized by a severe decrease in the number of blood neutrophils,and a maturation arrest of bone marrow progenitor cells mainly at the promyelocyte/myeloid stage,and most of congenital neutropenia patients are susceptibility to bacterial infections in clinically.In recent years,the mutations in ELANE,GFI1,HAX1,G6PC3,WAS,CSF3R and so on are associated with the occurrence and development of CN,and with the further research,more new virulence gene will be found gradually.

Nicotinamide phosphoribosyltransferase in inlfammation and cancer / 临床儿科杂志

Nicotinamide phosphoribosyltransferase (NAMPT) is the rate-limiting enzyme in the salvage pathway for the biosynthesis of nicotinamide adenine dinucleotide (NAD+) from nicotinamide. NAMPT is also a cytokine that inhibits the apoptosis of neutrophils under various inlfammatory stimuli, regulates various diseases and closely associates with the progression and prognosis of cancers. However, it is still not clear whether the cytokine-like function of NAMPT is interrelated with the biosynthesis enzyme activity of NAD+. This article aims to provide novel insights for inflammation and cancers treatment by reviewing the function of NAMPT in inflammation, carcinogenesis, cancer progression and its inhibitors, APO866/FK866.

Effect of Intranasal T-bet Plasmid Transfer on Th1/Th2 Balance in Spleen MNC in a Established Murine Allergic Asthmatic Model / 中山大学学报(医学科学版)

[Objective] This study was designed to determine Th1, Th2 cell numbers and investigate T-bet mRNA, GATA-3 mRNA expression of spleen MNC in a mufine asthmatic model which intended to understand effect of airway T-bet plasmid gene transfer on Th differentiation. [ Methods] A mouse asthmatic model was established by sensitization with ovalbumin (OVA). Thirty-two C57BL/6 mice were divided into four groups (8 mice in each group): the normal control group (group A ), the asthmatic model group (group B), the pcDNA3 plasmid group (group C), the pcDNA3-T-bet group (group D). All animals were sensitized and challenged with OVA, except group A normal saline was applied. The group C was intranasally administered 50 μg pcDNA3 plasmid at 24 h before intranasal challenges, and the 50 μg pcDNA3-T-bet plasmid for the mice of group D. We investigated Th1 and Th2 cell numbers by FACS and T-bet, GATA-3mRNA expression of spleen mononuclear cells (MNC) by semi-quantitative PCR in the four groups. [Result] Th1 percent in spleen MNC of pcDNA3-T-bet treated mice was significantly increased ([2.29±1.551% vs. [1.93±1.141%, P<0.05), while Th2 percent was significantly decreased ([0.93±0.64]% vs. [1.63±0.59]%), compared with that of the asthmatic control group mice by FACS. Spleen MNC was detected a high level of T-bet mRNA expression (0.53±0.027 vs. 0.28±0.035, P<0.05) and a low level of GATA-3 mRNA expression (0.24±0.022 vs. 0.58±0.038, P<0.05) after pcDNA3-T-bet treatment by RT-PCR. There was no significant change between the pcDNA3 plasmid group and the asthmatic model group. [Conclusion] The intranasal transfer of pcDNA3-T-bet plasmid was effective in modulating the imbalance of Th1/Th2 in mice asthma model, which provides a novel therapeutic strategy for transferring transcriptional factor in allergic asthma.

Rapid diagnosis of 21 trisomy syndrome by fluorescence quantitative polymerase chain reaction / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To establish a method of fluorescence quantitative PCR to detect 21 trisomy syndrome.</p><p><b>METHODS</b>At first, using one pair of primer to simultaneously amplify different fragments of two highly homologous genes of the human liver-type phosphofructokinase located on chromosome 21 (PFKL-CH21) and the human muscle-type phosphofructokinase located on chromosome 1 (PFKM-CH1). Then, staining the PCR products of these homologous genes with SYBR Green I, comparing the fluorescence intensities of the bands after electrophoresis, and analyzing the data.</p><p><b>RESULTS</b>The relative fluorescence intensity ratios of PFKL-CH21/PFKM-CH1 in 21 trisomy syndrome and normal individuals were 1.58+/-0.17 (mean+/-SD) and 1.00+/-0.05 (mean+/-SD), respectively; the difference between the two groups was highly significant.</p><p><b>CONCLUSION</b>SYBR Green I fluorescence quantitative polymerase chain reaction is an acurate, rapid, safe and practical approach for the detection of 21 trisomy syndrome.</p>