Recent clinical research on the application of liquid biopsy in neuroblastoma / 中国当代儿科杂志

Neuroblastoma (NB) is the most common extracranial solid tumor in children and has the features of high recurrence rate and low survival rate, and therefore, early diagnosis, treatment response evaluation, and recurrence monitoring are of great significance for NB patients. Liquid biopsy refers to the detection of cells and nucleic acids in fluid specimens, mainly blood. It is noninvasive and can overcome tumor heterogeneity, thus making it possible to achieve the early diagnosis and dynamic detection of NB. This review introduces the latest advances in clinical research on the application of liquid biopsy in NB.

Design and application of female infant diapers / 中国实用护理杂志

Objective:To design and manufacture the female infant diaper, and to explore the application effect of the diaper in routine urine collection of female infants.Methods:Female infants hospitalized in department of Orthopedic Rehabilitation of Children′s Hospital of Zhejiang University School of Medicine from July 2020 to June 2021 were selected as the research objects. According to the chronological order, 148 female infants admitted from July to December 2020 were selected as the control group, and 154 female infants admitted from January 2021 to June 2021 were selected as the experimental group. In the experimental group, urine routine was collected by female infants diapers, while in the control group, urine routine was collected by traditional disposable urine collection bag. The time required for routine urine collection, the number of urine collection bags, the incidence of perineal skin injury and the satisfaction of parents were observed.Results:The time required for routine urine collection was (16.86 ± 12.58) h in the experimental group and (28.76 ± 22.20) h in the control group. The difference between the two groups was statistically significant ( t=5.70, P<0.05). The number of female infant diapers in experimental group was (1.69 ± 0.67); The number of disposable urine collection bags used in the control group was (2.76 ± 1.32), and the difference between the two groups was statistically significant ( t=8.81, P<0.05). In the experimental group, the incidence of perineal skin injury was 0. In the control group, the incidence of perineal skin injury was 24.8%(42/148). The difference between the two groups was statistically significant ( Z=-9.78, P<0.01). The satisfaction of parents in the experimental group was 98.7% (152/154), witch higher than that in the control group was 79.7% (118/148), and the difference was statistically significant ( Z=-9.68, P<0.01). Conclusions:Compared with the traditional disposable urine collection bag, the diaper collection for female infants can shorten the time required for routine urine collection, reduce the number of urine collection tools, reduce the incidence of perineal lesions in children, improve the satisfaction of parents of children, reflect the people-oriented nursing concept, and provide greater clinical convenience.

Changes of intestinal flora in children with acute lymphoblastic leukemia before and after chemotherapy / 中国当代儿科杂志

OBJECTIVES@#To examine the changes of intestinal flora in children newly diagnosed with acute lymphoblastic leukemia (ALL) and the influence of chemotherapy on intestinal flora.@*METHODS@#Fecal samples were collected from 40 children newly diagnosed with ALL before chemotherapy and at 2 weeks, 1 month, and 2 months after chemotherapy. Ten healthy children served as the control group. 16S rDNA sequencing and analysis were performed to compare the differences in intestinal flora between the ALL and control groups and children with ALL before and after chemotherapy.@*RESULTS@#The ALL group had a significant reduction in the abundance of intestinal flora at 1 and 2 months after chemotherapy, with a significant reduction compared with the control group (P<0.05). Compared with the control group, the ALL group had a significant reduction in the diversity of intestinal flora before and after chemotherapy (P<0.05). At the phylum level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Actinobacteria at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05) and a significant increase in the relative abundance of Proteobacteria at 1 and 2 months after chemotherapy (P<0.05). At the genus level, compared with the control group, the ALL group had a significant reduction in the relative abundance of Bifidobacterium at 2 weeks, 1 month, and 2 months after chemotherapy (P<0.05); the relative abundance of Klebsiella in the ALL group was significantly higher than that in the control group at 1 and 2 months after chemotherapy and showed a significant increase at 1 month after chemotherapy (P<0.05); the relative abundance of Faecalibacterium in the ALL group was significantly lower than that in the control group before and after chemotherapy and showed a significant reduction at 2 weeks and 1 month after chemotherapy (P<0.05). The relative abundance of Enterococcus increased significantly at 1 and 2 months after chemotherapy in the ALL group (P<0.05), and was significantly higher than that in the control group (P<0.05).@*CONCLUSIONS@#The diversity of intestinal flora in children with ALL is significantly lower than that in healthy children. Chemotherapy significantly reduces the abundance of intestinal flora and can reduce the abundance of some probiotic bacteria (Bifidobacterium and Faecalibacterium) and increase the abundance of pathogenic bacteria (Klebsiella and Enterococcus) in children with ALL.

Study on mechanism of curcumol against liver fibrosis based on autophagy and apoptosis of hepatic stellate cells / 中国中药杂志

The present study clarified the molecular mechanism of curcumol against liver fibrosis based on its effects on the autopha-gy and apoptosis of hepatic stellate cells. The hepatic stellate cells were divided into a blank control group, a transforming growth factor-β1(TGF-β1)(10 ng·mL~(-1)) group, and low-(12.5 mg·L~(-1)), medium-(25 mg·L~(-1)), and high-dose(50 mg·L~(-1)) curcumol groups. The effect of curcumol on the viability of hepatic stellate cells induced by TGF-β1 was detected by the MTT assay kit. The apo-ptosis in each group was determined by flow cytometry. Real-time fluorescence-based quantitative PCR(RT-PCR) was employed for the detection of mRNA expression of α-smooth muscle actin(α-SMA), type Ⅰ collagen(collagen Ⅰ), and type Ⅲ collagen(collagen Ⅲ). Western blot was used to detect the protein expression of p62, microtubule-associated protein 1 light chain 3(LC3), beclin1, B cell lymphoma 2(Bcl-2), and Bcl-2-associated X protein(Bax). Transmission electron microscopy(TEM) was used to observe cell morphology and autophagosome formation in each group. The autophagic flux was observed after cell infection with adenovirus under double fluorescence labeling. The cell viability assay revealed that compared with the TGF-β1 group, the curcumol groups showed significantly decreased cell viability. The apoptosis assay showed that the apoptosis rates of the curcumol groups were significantly higher than that of the TGF-β1 group. RT-PCR indicated that the mRNA expression of α-SMA, collagenⅠ, and collagen Ⅲ in the curcumol groups was significantly lower than that of the TGF-β1 group. Western blot showed that the expression of p62, LC3, beclin1, Bcl-2, and Bax in the curcumol groups was significantly different from that in the TGF-β1 group. As demonstrated by TEM, compared with the TGF-β1 group, the curcumol groups showed significantly increased autophagosomes. The detection of autophagic flow by the adenovirus under double fluorescence labeling showed that autolysosomes in the curcumol groups were significantly increased compared with those in the TGF-β1 group. Curcumol can induce the autophagy and apoptosis of hepatic stellate cells, which may be one of its anti-liver fibrosis mechanisms.

Effect of electroacupuncture on motor function and gait in patients with post-stroke spasticity in lower limbs / 中国针灸

OBJECTIVE@#To observe the effect on motor function, spasticity degree, muscle strength and the relevant parameters of three-dimensional gait analysis in the patients with post-stroke spasticity in the lower limbs treated with the combined therapy of electroacupuncture (EA) and muscle electricity biofeedback or the simple muscle electricity biofeedback therapy on the base of rehabilitation medicine.@*METHODS@#A total of 60 patients with post-stroke spasticity in the lower limbs were randomized into an EA + biofeedback group, a biofeedback group and a rehabilitation group, 20 cases in each one. In the rehabilitation group, the basic rehabilitation training was provided, 45 min each time. In the biofeedback group, on the base of the treatment as the rehabilitation group, the biofeedback therapy was added, 30 min each time. In the EA + biofeedback group, besides the treatment as the biofeedback group, acupuncture was supplemented at Futu (ST 32), Liangqiu (ST 34), Zusanli (ST 36) and Fenglong (ST 40), etc, and EA was applid at Zusanli (ST 36) and Taichong (LR 3) with continuous wave and 5 Hz in frequency. In each group, the treatment was given once daily, 5 times a week, for 6 weeks totally. Separately, before and after treatment, the score of Fugle-Meyer assessment (FMA), the score of clinical spasticity index (CSI) in the lower limbs and the strength of the anterior tibial muscle on the affected side were assessed, and the spatial-temporal parameters (step frequency and steep speed) in the three-dimensional gait analysis and the kinematic parameters (maximum dorsal flexion and maximum plantar flexion of ankle joint on the affected side) were measured in the patients of three groups.@*RESULTS@#After treatment, FMA score was increased as compared with that before treatment in all of three groups (P<0.05). FMA score in the EA + biofeedback group and the biofeedback group was higher than the rehabilitation group respectively (P<0.05). CSI score in the EA + biofeedback group and the biofeedback group was lower than that before treatment respectively (P<0.05), and lower than the rehabilitation group (P<0.05). After treatment, the step frequency and speed were all improved and the angles of maximum dorsal flexion and maximum plantar flexion of ankle joint on the affected side were all increased as compared with those before treatment in the patients of three groups separately (P<0.05). The step frequency and speed, as well as the angles of maximum dorsal flexion and maximum plantar flexion of ankle joint on the affected side in either the EA + biofeedback group or the biofeedback group were all higher than the rehabilitation group (P<0.05), and the step speed in the EA + biofeedback group was higher than the biofeedback group (P<0.05). After treatment, the strength of the anterior tibial muscle on the affected side was increased as compared with that before treatment in the patients of each group (P<0.05); and the strength of the anterior tibial muscle in the EA + biofeedback group and the biofeedback group was larger than the rehabilitation group (P<0.05).@*CONCLUSION@#On the base of rehabilitation treatment, the combined regimen of EA and muscle electricity biofeedback therapy and the simple muscle electricity biofeedback therapy all effectively strengthen the motor functions and reduce spasticity as well as improve step frequency, step speed and the range of motion of ankle joint in the patients with post-stroke spasticity in the lower limbs. Regarding the gait improvement, the combined regimen of EA and muscle electricity biofeedback is better than the simple muscle electricity biofeedback.

Germacranolide sesquiterpenes from Carpesium cernuum and their anti-leukemia activity / 中国天然药物

In this study, three new germacranolide sesquiterpenes (1-3), together with six related known analogues (4-9) were isolated from the whole plant of Carpesium cernuum. Their structures were established by a combination of extensive NMR spectroscopic analysis, HR-ESIMS data, and ECD calculations. The anti-leukemia activities of all compounds towards three cell lines (HEL, KG-1a, and K562) were evaluated in vitro. Compounds 1-3 exhibited moderate cytotoxicity with IC

Prognostic Factors Analysis of Children with Hemophagocytic Lymphohistiocytosis / 中国实验血液学杂志

OBJECTIVE@#To analyze the risk factors affecting prognosis of children with hemophagocytic lymphohistiocytosis (HLH).@*METHODS@#The clinical manifestations and laboratory data of 143 HLH children who met the HLH-2004 diagnostic criteria in Shenzhen Children's Hospital from January 2009 to May 2017 were retrospectively analyzed, and the independent factors affecting prognosis were also analyzed.@*RESULTS@#The median age of 143 HLH children was 1.9 (0.1-14.3) years old, and the median follow-up time was 6.7 years (1 day - 11.9 years). The overall survival rate of 1 month, 1 year, and 10 years was (87.4±5.5)%, (81.1±6.5)%, and (81.1±6.5)%, respectively. The deaths occurred within 1 year after onset. Multivariate analysis showed that central nervous system (CNS) involvement (P=0.047), low hemoglobin (P=0.002), prolonged activated partial thromboplastin time (APTT) (P<0.001), high triglyceride (P=0.005) were all the independent risk factors affecting survival of the children. Receiver operating characteristic curve indicated that APTT (AUC=0.753, P<0.001) was more valuable than other risk factors in predicting death of the children. The cut-off value of APTT was 56.6 s, and the sensitivity and specificity of which was 55.6% and 89.7%, respectively.@*CONCLUSION@#Hypohemoglobinemia, prolonged APTT, hypertriglyceridemia, and CNS involvement the risk factors affecting prognosis of HLH, and prolonged APTT shows a strong predictive value for death.

Clinical features of sleep-disordered breathing in children with neuromuscular disease / 中国当代儿科杂志

OBJECTIVE@#To study the clinical features of sleep-disordered breathing (SDB) in children with neuromuscular disease (NMD).@*METHODS@#A retrospective analysis was performed on the medical data of 18 children who were diagnosed with NMD and underwent polysomnography (PSG) (NMD group). Eleven children without NMD who had abnormal sleeping habit and normal sleep structure on PSG were enrolled as the control group. The two groups were compared in terms of the daily and nocturnal symptoms of SDB, incidence rate of obstructive sleep apnea (OSA), pulmonary function, end-tidal partial pressure of carbon dioxide (PetCO@*RESULTS@#In the NMD group, 16 children (89%) had related daily and nocturnal symptoms of SDB, and the youngest age was 1 year at the onset of such symptoms. Compared with the control group, the NMD group had significant reductions in total sleep time and sleep efficiency (@*CONCLUSIONS@#There is a high proportion of children with SDB among the children with NMD, and SDB can be observed in the early stage of NMD, which results in the damage of sleep structure and the reduction in sleep efficiency. Respiratory events are mainly obstructive events, and oxygen reduction events are mainly observed during REM sleep.

Inhibitory Effect and Mechanism of Jingulian Capsule on Human Breast Cancer MDA-MB-231 Cells / 中国实验方剂学杂志

Objective:To observe effect of Jingulian capsule on the proliferation of human breast cancer MDA-MB-231 cells and investigate its action mechanism against triple negative breast cancer （TNBC）. Method:The ingredients of Jingulian capsule were identified by ultra-performance liquid chromatography-tandem mass spectrometry（UPLC-MS/MS）. The inhibitory effect of Jingulian capsule at different doses （0.125，0.25，0.5，1，and 2 g·L^-1） against the proliferation of MDA-MB-231 cells were detected by methyl thiazolyl tetrazolium （MTT） assay. After treatment for 24 h， the morphological changes in nuclear apoptosis of MDA-MB-231 cells were detected by Hoechst 33258 staining. The effect of different concentrations of Jingulian capsule on the apoptosis and cycle of MDA-MB-231 cells after different treatment time were determined by flow cytometry. The protein expression levels of Poly-ADP-ribose polymeras （PARP）， proto-oncogene c-Myc， cyclin B₁， and phosphorylated extracellular signal-regulated kinase （p-ERK） in each group were assayed by Western blot. Result:A total of 113 compounds in Jingulian capsule were identified by UPLC-MS/MS. As revealed by MTT assay，compared with blank group，Jingulian capsule （0.125，0.25，0.5，1，2 g·L^-1） significantly inhibited viability of MDA-MB-231 cells （<italic>P</italic><0.01）， with the half maximal inhibitory concentration （ IC₅₀） of（0.13±0.02）g·L^-1. According to flow cytometry，compared with the blank group，Jingulian capsule at 1 g·L^-1 significantly induced the apoptosis of MDA-MB-231 cells （<italic>P</italic><0.05）and Jingulian capsule at 0.5， 1 g·L^-1 obviously increased the number of MDA-MB-231 cells in S phase （<italic>P</italic><0.05，<italic>P</italic><0.01）. The results of Western blotting demonstrated that the protein expression levels of PARP，c-Myc，and cyclin B₁ in 0.5， 1 g·L^-1Jingulian capsule groups were remarkably down-regulated as compared with those in the blank group（<italic>P</italic><0.01）， and the protein expression level of p-ERK in 1 g·L^-1Jingulian capsule group was also down-regulated （<italic>P</italic><0.01）. Conclusion:Jingulian capsule is able to inhibit the proliferation of MDA-MB-231 cells，induce S phase cell cycle arrest， and promote their apoptosis， which may be related to the inactivation of the MAPK signaling pathway.

Pediatric Airway Foreign Body Retrieval During the Prevention and Control of 2019 Novel Coronavirus / 中华急诊医学杂志

The prevention and control of 2019 novel coronavirus (2019-nCoV) is currently the primary task of all industries in China. People can be infected through respiratory droplets, airborne and close contact. Pediatric airway foreign body is a common otorhinolaryngology emergency, especially occurred in 1 to 3-year-old children. It usually causes complications like airway obstruction, suffocation and pneumonia, which may become an acute threat to life. The common practice in otorhinolaryngology emergency room is to perform direct laryngoscope, bronchoscope and foreign body removal. Medical staff need to be closely contacted with these children during surgical operation, who may produce massive droplets and aerosols during examination. With a combination of characteristics of surgical operation, this article intends to provide advices on diagnosis and treatment of airway foreign body removal for pediatric otorhinolaryngology colleagues during the prevention and control of 2019-nCoV. Adjustments could be made subsequently due to changes of the epidemic situation and the recognition of 2019-nCoV.

Pediatric airway foreign body retrieval during the prevention and control of 2019 novel coronavirus / 中华急诊医学杂志

The prevention and control of 2019 novel coronavirus (2019-nCoV) is currently the primary task of all industries in China. The virus infection is mainly transmitted by respiratory droplets, airborne and close contact. Pediatric foreign body in the respiratory tract is a common otorhinolaryngology emergency, especially occurred in 1-3 years old children, and usually causes airway obstruction, suffocation and pneumonia, which may become an acute threat to life. The principle treatment in otorhinolaryngology emergency room is direct laryngoscope, bronchoscope and foreign body removal. Due to the close contact between the relevant medical staff and child during the operation, a large number of droplets and aerosols can be produced during the reactive cough of the child. Combined with the characteristics of the operation, this article intends to provide advices on diagnosis and treatment of airway foreign body removal for pediatric otorhinolaryngology colleagues during the prevention and control of 2019-nCoV. Adjustments could be made subsequently due to changes of the epidemic situation and the recognition of 2019-nCoV.

Latest advances in minimal residual disease evaluation in B-cell lymphoproliferative disease / 中国当代儿科杂志

The clearance of cancer cells is closely associated with the prognosis of various hematologic malignancies. Clinical studies have shown that minimal residual disease (MRD) can directly reflect the clearance of cancer cells, but the tools for MRD detection need to be improved. This article reviews the latest advances in the MRD detection by digital polymerase chain reaction and next-generation sequencing in B-cell lymphoproliferative disease.

Daxx overexpression inhibits AngⅡ-induced proliferation and migration in vascular smooth muscle cells / 南方医科大学学报

OBJECTIVE@#To construct a recombinant lentiviral expression vector pCDH-Daxx-EGFP to investigate the effect of Daxx on the proliferation of vascular smooth muscle cells (VSMCs).@*METHODS@#The recombinant lentiviral expression vector pCDHDaxx-EGFP was constructed using PCR-based accurate synthesis method. After identification by sequencing and enzyme digestion, the recombinant lentiviral vector was contransfected into 293T cells with lentivirus packaging vector. The recombinant lentivirus particles were collected and purified to infect VSMCs, whose expression of Daxx was detected with Western boltting. The cells infected with the empty vector pCDH-EGFP or pCDH-Daxx-EGFP were incubated in serum-free medium or in the presence of angiotensin Ⅱ (AngⅡ). The cell viability was determined with MTT assay, and the cell cycle changes were analyzed with flow cytometry. The cell migration ability was assessed using a scratch wound healing assay. The expression of p-Akt protein in the cells was detected using Western blotting.@*RESULTS@#Double enzyme digestion and sequencing confirmed successful construction of the recombinant plasmid. Compared with the cells infected with the empty vector, the cells infected with pCDH-Daxx-EGFP exhibited significantly increased expressions of Daxx protein ( < 0.05). AngⅡ treatment of the cells infected with the pCDH-Daxx-EGFP, as compared with the cells infected with the empty vector, significantly lowered the cell viability, S phase cell ratio and cell migration ability ( < 0.05), and significantly decreased the expression level of p-Akt protein ( < 0.05).@*CONCLUSIONS@#We successfully constructed the recombinant lentiviral vector pCDH-Daxx-EGFP and overexpressed Daxx in primary cultured VSMCs using this vector. Daxx overexpression can inhibit AngⅡ-induced proliferation and migration in VSMCs probably by regulating p-Akt protein.

Protective Effect of Syringaresinol on Excitatory Damage Induced by Sodium Glutamate in SH-SY5Y Cells / 中国实验方剂学杂志

Objective: To establish a model for the injury of human neuroblastoma cell (SH-SY5Y) induced by sodium glutamate, and to observe the protective effect of syringaresinol on cell damage from Viscum liquidambaricolum hayataon, and to explore its mechanism. Method: Construction of SH-SY5Y cell injury model using sodium glutamate.The experiment was divided into normal cell group, injury model group (sodium glutamate 50 mmol·L-1, sodium glutamate 50 mmol·L-1 + DMSO),syringaresinol experimental group (6.25, 12.5, 25 μmol·L-1), by cell counting, cell morphology observation, Annexin V-FITC/PI apoptosis detection, ROS reactive oxygen species detection, mitochondrial membrane potential, and Western blot, evaluation of syringaresinol on glutamate-induced neuronal excitability injury neuroprotective activity. Result: Compared with normal group, the cell survival rate of the model group was significantly decreased (PPPPP-1) showed a concentration-dependent increase in cells. Survival rate (PPPPPConclusion: Syringaresinol has significant protective activity against excitatory damage induced by sodium glutamate in SH-SY5Y neurons, the mechanism may be through anti-oxidative stress, repairing mitochondrial function and DNA damage to significantly reduce sodium glutamate-induced neuronal apoptosis.

The Lymphatic Drainage Pattern of Internal Mammary Sentinel Lymph Node Identified by Small Particle Radiotracer ((99m)Tc-Dextran 40) in Breast / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The purpose of this study was to detect the lymphatic drainage pattern of internal mammary area and verify the concept of internal mammary sentinel lymph node (IM-SLN) in breast. MATERIALS AND METHODS: A small particle radiotracer ((99m)Tc-Dextran 40) was prepared and tested. (99m)Tc-Dextran 40 was injected into intraparenchyma at the sound breast by a modified radiotracer injection technique. Subsequently, dynamic single-photon emission computed tomography (SPECT), computed tomography (CT), and SPECT/CT combination images were performed to identify the radioactive lymph vessels and internal mammary lymph nodes (IMLNs). The direction of lymph drainage and the location of the IMLNs were identified in the SPECT/CT imaging. RESULTS: The radiochemical purity of (99m)Tc-Dextran 40 was > 95%. (99m)Tc-Dextran 40 could drainage into first, second, and third lymph node and the radioactive lymph node could be detected by the γ detector in the animal experiment. After (99m)Tc-Dextran 40 injecting into intraparenchyma, 50.0% cases (15/30) were identified the drainage lymphatic vessels and radioactive IMLNs by SPECT. The drainage lymphatic vessel was found from injection point to the first IMLN (IM-SLN) after 10.5±0.35 minutes radiotracer injection, and then (99m)Tc-Dextran 40 was accumulated into the IM-SLN. The combination imaging of SPECT/CT showed the second IMLN received the lymph drainage from the IM-SLN. The lymphatic drainage was step by step in the internal mammary area. CONCLUSION: The lymph was identified to drain from different regions of the breast to IM-SLN, and then outward from IM-SLN to other IMLN consecutively. It demonstrated the concept of the IM-SLN and provided more evidences for the application of internal mammary sentinel lymph node biopsy.

Pulmonary rehabilitation of chronic obstructive pulmonary disease / 中国医师杂志

There is a high incidence of adult chronic obstructive pulmonary disease (COPD),which induces high rate of disability,and heavy financial burden in China.As the core of comprehensive treatments of COPD,lung rehabilitation can improve the quality of life and reduce the cost of treatment.This article introduces not only the definition of COPD lung rehabilitation and its recent states and progress,but also the related indications of different lung rehabilitation techniques with the formulations and resolutions of rehabilitation programs.

Cloning and characterization of an aromatic amino acid aminotransferase (ArAT) gene involved in tropane alkaloid biosynthesis from Hyoscyamus niger / 药学学报

Tropane alkaloids are anticholinergic drugs widely used clinically. Biosynthesis of tropane alkaloids in planta involves a step of transamination of phenylalanine. Based on the sequenced transcriptomes of lateral roots and leaves of Hyoscyamus niger, we found three annotated aromatic amino acid aminotransferases, which were respectively named HnArAT1, HnArAT2 and HnArAT3. Sequence analysis showed that HnArAT3 had highest similarity with the reported Atropa belladonna AbArAT4, which was involved in tropane alkaloid (TA) to provide the precursor of the phenyllactic acid moiety. Tissue expression pattern analysis indicated that HnArAT3 was specifically expressed in lateral roots, where is the organ synthesizing tropane alkaloids. Then, method of virus induced gene silencing (VIGS) was used to characterize the function of HnArAT3 in H. niger. Gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had lower expression levels of HnArAT3 than the non-transgenic control, and HPLC analysis of alkaloids demonstrated significant decrease in the contents of hyoscyamine, anisodamine and scopolamine in planta. These results suggested that HnArAT3 was involved in the phenyllactic acid branch of TA biosynthetic pathway. Molecular cloning and functional identification of HnArAT3 laid the foundation for further understanding of TA biosynthesis and metabolic regulation, and also provided a new candidate gene for engineering biosynthetic pathway of tropane alkaloids.

Brain injury after induction chemotherapy in children with acute lymphoblastic leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the changes in brain injury after the induction chemotherapy in children with acute lymphoblastic leukemia (ALL) by cranial MRI.</p><p><b>METHODS</b>The clinical data and cranial MRI results of 62 children with ALL who were hospitalized from March 2014 to June 2015 were analyzed retrospectively.</p><p><b>RESULTS</b>Before chemotherapy, MRI showed bone marrow infiltration of the skull in 33 patients (53%); the children with WBC<20×10(9)/Lhad a significantly lower incidence rate of bone marrow infiltration of the skull than those with WBC≥20×10(9)/L (16 patients/42% vs 17 patients/71%; P<0.05), and the high-risk group had a significantly higher incidence rate of bone marrow infiltration of the skull than the non-high-risk group (71% vs 44%; P<0.05). Before chemotherapy, there were 4 cases (7%) of brain atrophy, and 2 cases (3%) of abnormal signals in the sensory conduction bundle. MRI reexamination in 28 patients after 3 months of chemotherapy showed 3 new cases (11%) of brain atrophy and 1 aggravated case of brain atrophy.</p><p><b>CONCLUSIONS</b>The children with ALL have bone marrow infiltration of the skull, brain atrophy, and abnormal signals in the sensory conduction bundle before chemotherapy, especially bone marrow infiltration of the skull, and some changes in brain injury disappear after treatment.</p>

Gene promoter methylation in glucose-6-phosphate dehydrogenase deficiency / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the features of methylation in the promoter region of glucose-6-phosphate dehydrogenase (G6PD) gene and the association between gene promoter methylation and G6PD deficiency.</p><p><b>METHODS</b>Fluorescent quantitative PCR was used to measure the mRNA expression of G6PD in 130 children with G6PD deficiency. Sixty-five children without G6PD deficiency served as the control group. The methylation-sensitive high-resolution melting curve analysis and bisulfite PCR sequencing were used to analyze gene promoter methylation in 22 children with G6PD deficiency and low G6PD mRNA expression. The G6PD gene promoter methylation was analyzed in 44 girls with normal G6PD mRNA expression (7 from G6PD deficiency group and 37 from control group).</p><p><b>RESULTS</b>Twenty-two (16.9%) children with G6PD deficiency had relatively low mRNA expression of G6PD; among whom, 16 boys showed no methylation, and 6 girls showed partial methylation. Among the 44 girls with normal G6PD mRNA expression, 40 showed partial methylation, and 4 showed no methylation (1 case in the G6PD group and 3 cases in the control group).</p><p><b>CONCLUSIONS</b>Gene promoter methylation is not associated with G6PD deficiency in boys. Girls have partial methylation or no methylation in the G6PD gene, suggesting that the methylation may be related to G6PD deficiency in girls.</p>

Detection of gene mutation in glucose-6-phosphate dehydrogenase deficiency by RT-PCR sequencing / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Since glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common hereditary hemolytic erythrocyte enzyme deficiency, most cases have single nucleotide mutations in the coding region, and current test methods for gene mutation have some missed detections, this study aimed to investigate the feasibility of RT-PCR sequencing in the detection of gene mutation in G6PD deficiency.</p><p><b>METHODS</b>According to the G6PD/6GPD ratio, 195 children with anemia of unknown cause or who underwent physical examination between August 2013 and July 2014 were classified into G6PD-deficiency group with 130 children (G6PD/6GPD ratio <1.00) and control group with 65 children (G6PD/6GPD ratio≥1.00). The primer design and PCR amplification conditions were optimized, and RT-PCR sequencing was used to analyze the complete coding sequence and verify the genomic DNA sequence in the two groups.</p><p><b>RESULTS</b>In the G6PD-deficiency group, the detection rate of gene mutation was 100% and 13 missense mutations were detected, including one new mutation. In the control group, no missense mutation was detected in 28 boys; 13 heterozygous missense mutations, 1 homozygous same-sense mutation (C1191T) which had not been reported in China and abroad, and 14 single nucleotide polymorphisms of C1311T were detected in 37 girls. The control group showed a high rate of missed detection of G6PD deficiency (carriers) in the specimens from girls (35%, 13/37).</p><p><b>CONCLUSIONS</b>RT-PCR sequencing has a high detection rate of G6PD gene mutation and a certain value in clinical diagnosis of G6PD deficiency.</p>