Analysis of Chemical Constituents in Different Parts of Gentiana straminea Based on UPLC-Q-TOF-MS/MS / 中国实验方剂学杂志

ObjectiveUltra-performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry （UPLC-Q-TOF-MS/MS） was used to analyze the chemical constituents in the aerial part and roots of Gentiana straminea from different areas of Qinghai province， and the main chromatographic peaks and differential components of different parts were identified. MethodThe chromatographic separation was performed on a Waters ACQUITY UPLC HSS T3 column （2.1 mm×100 mm， 1.8 μm） with 0.1% formic acid aqueous solution （A）-acetonitrile （B） as the mobile phase for gradient elution （0-1 min， 1%-13%B； 1-5 min， 13%-18%B； 5-7 min， 18%-50%B； 7-9.5 min， 50%-60%B； 9.5-11 min， 60%-99%B； 11-14 min； 99%B； 14-15 min， 99%-1%B； 15-16 min， 1%B）， the column temperature at 40 ℃， and the flow rate of 0.3 mL·min-1. Electrospray ionization （ESI） and negative ion full scan mode were selected for the mass spectrometric conditions to analyze the samples， and the detection range was m/z 50-1 200. Chemical constituents of the aerial part were qualitatively analyzed with the reference substances， literature information and ChemSpider. Principal component analysis （PCA） and orthogonal partial least squares discriminant analysis （OPLS-DA） were used to analyze the classification trend， correlation and differential chemical components between aerial part and roots of G. straminea. ResultA total of 68 components， including 24 iridoids， 13 flavonoids， 8 triterpenoids， 6 xanthones， 5 fatty acids， 4 saccharides， 3 phenolic glycosides， 2 alkaloids， 2 sterols and 1 lignan， were preliminarily identified from the aerial part of G. straminea. Among them， 42 components were firstly reported in 4 Gentiana species included in the 2020 edition of Chinese Pharmacopoeia. Eight differential components were screened out， namely sucrose， maltotriose， loganic acid， shanzhiside methyl ester， 6′-O-β-D-glucosylgentiopicroside， swertiamarin， gentiopicrin and isovitexin. ConclusionThe aerial part of G. straminea is rich in chemical constituents and has good medicinal potential. There were significant differences in the chemical components between the aerial part and roots of G. straminea， and the main differential components were iridoids， which could provide a basis for exploring efficacy differences in different parts of G. straminea.

Effect of different maintenance doses of caffeine citrate on ventilator weaning in very preterm infants with respiratory distress syndrome: a prospective randomized controlled trial / 中国当代儿科杂志

OBJECTIVES@#To study the effect of different maintenance doses of caffeine citrate on the success rate of ventilator weaning in very preterm infants (gestational age of ≤32 weeks) with respiratory distress syndrome (RDS).@*METHODS@#A total of 162 preterm infants with RDS who were admitted to the hospital from January 2016 to December 2018 were enrolled in this prospective trial. These infants had a gestational age of ≤32 weeks and required invasive mechanical ventilation. They were randomly divided into a high-dose caffeine group and a low-dose caffeine group, with 81 infants in each group. Within 6 hours after birth, both groups were given caffeine at a dose of 20 mg/kg. After 24 hours, the high- and low-dose caffeine groups were given caffeine at a maintenance dose of 10 mg/kg and 5 mg/kg, respectively. The two groups were compared in terms of re-intubation rate within 48 hours after ventilator weaning, durations of ventilation and oxygen therapy, enteral feeding, weight gain, and the incidence rates of complications and adverse reactions during hospitalization.@*RESULTS@#The high-dose caffeine group had a significantly lower re-intubation rate within 48 hours after ventilator weaning than the low-dose caffeine group (@*CONCLUSIONS@#A high maintenance dose of caffeine can safely and effectively reduce the incidence rate of apnea after ventilator weaning and the failure rate of ventilator weaning in RDS preterm infants with a gestational age of ≤32 weeks, and therefore, it holds promise for clinical application.

Value of anti-neutrophil cytoplasmic antibody in assessing the severity of bronchiolitis obliterans in children / 中国当代儿科杂志

OBJECTIVE@#To study the value of anti-neutrophil cytoplasmic antibody (ANCA) in assessing the severity of bronchiolitis obliterans (BO) in children.@*METHODS@#A prospective analysis was performed on 59 children who were diagnosed with BO from June 2009 to October 2014. ELISA was used to measure the concentrations of myeloperoxidase (MPO)-ANCA and proteinase 3 (PR3)-ANCA in serum. According to the results of ELISA, the children were divided into three groups: double-negative ANCA (n=22), single-positive ANCA (n=17), and double-positive ANCA (n=20). The three groups were compared in terms of the scores of BO risk factors, clinical symptoms, chest high-resolution computed tomography (HRCT), and lung pathology on admission, as well as the changes in the expression level of ANCA and the scores of clinical symptoms and chest HRCT over time.@*RESULTS@#Compared with the double-negative ANCA group, the double-positive ANCA group had a significantly higher score of BO risk factors (P0.05). The single-positive ANCA and double-positive ANCA groups still had a significantly higher score of clinical symptoms than the double-negative ANCA group (P<0.05).@*CONCLUSIONS@#The expression level of ANCA is correlated with the severity of BO in children and thus has certain clinical significance in disease evaluation.

Expression of Pim1 protein in variant lesions of esophagus and its clinical significance / 新乡医学院学报

Objective To study the expression of Pim1 protein in variant lesions of esophagus and its correlation with the clinicopathological parameters of esophageal cancer patients.Methods The esophageal biopsy specimens of 182 cases,including 37 cases of normal squamous epithelium,43 cases of low-grade intraepithelial neoplasia,31 cases of high-grade intraepithelial neoplasia,71 cases of esophageal squamous cell carcinoma were collected in the Third Affiliated Hospital of Xinxiang Medical University From January 2009 to March 2016.The expression of Pim1 protein in all esophageal tissues was detected by immunohistochemical method,and the correlation between Pim1 protein expression and clinicopathological parameters of esophageal squamous cell carcinoma patients was analyzed.Results The higher expression rate of Pim1 protein in esophageal normal squamous epithelium,low-grade intraepithelial neoplasia,high-grade intraepithelial neoplasia and esophageal squamous cell carcinoma was 24.3％ (9/37),62.8 ％ (27/43),70.9％ (22/31),and 76.1％ (54/71),respectively.The higher expression rate of Pim1 protein in the tissue of low-grade intraepithelial neoplasia,high-grade intraepithelial neoplasia and esophageal squamous cell carcinoma was significanlty higher than that in the esophageal normal squamous epithelium(x2 =11.89,14.79,26.70;P ＜ 0.05);there was no statistic difference of the higher expression rate of Pim1 protein in the tissue of low-grade intraepithelial neoplasia,high-grade intraepithelial neoplasia and esophageal squamous cell carcinoma (P ＜ 0.05).The expression of Pim1 protein was significantly positively correlated with lymph node metastasis and TNM stage(P ＜0.05),but not correlated with age and histological grade of patients (P ＜ 0.05).Conclusion The expression of Pim1 protein is closely related to the occurrence and development of esophageal cancer.

Comparison of differences of Notch1 gene methylation between hyperplastic lesions tissues and breast cancer tissues in Uyghur women / 临床与实验病理学杂志

Purpose To compare the difference of Notch1 methylation in the breast cancer and hyperplastic lesions tissue from Uyghur in Xinjiang. Methods The methylation level of Notch1 gene in Uyghur breast tissues including usual ductal hyperplasia (UDH), atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS), and invasive ductal carcinoma(IDC) were detected by MALDI-TOF-MS technique. The association of the methylation level with clinical pathological characteristics of patients was analyzed. The expression of Notch 1 protein was detected by immunohistochemistry. The relationship between the methylation status and expression was assay. Results The methylation rate of Notch l gene in UDH, ADH, DCIS, and IDC group was gradually decreased (P＜0.05). The 9 CpG sites methylation level of 13 CpG sites are statistically lower in cancer tissues (P＜0.05). The hypomethylation are accompanied with low differentiation, lymph node metastasis and high stage of TNM (P＜0.05). The lower DNA methylation is negatively correlated with the expression of Notch l (P＜0.05). Conclusion There was a differences of the expression and methylation rate of Notch 1 between breast cancer and hyperplastic lesions tissue, and its biological significances needs to be further studied.

Investigation on health status of workers exposed to 7-ACA and 6-APA in an antibiotic enterprise / 中国职业医学

OBJECTIVE: To investigate the health status of workers exposed to 7-aminocephalosporanic acid( 7-ACA) and6-aminopenicillanic acid( 6-APA) in an antibiotics enterprise. METHODS: Using simple random sampling method,207 workers exposed to 7-ACA and 6-APA from an antibiotic production enterprise were selected as the exposed group,and 162 workers with no dust exposure history from the same antibiotic production enterprise were selected as control group. Health examinations were performed. The health status of the workers were analyzed. RESULTS: The detection of symptoms( chest distress,shortness of breath,cough,wheezing,itchy skin) and allergic diseases( bronchial asthma,allergic rhinitis,allergic dermatitis) in exposed group were higher than those in control group( P < 0. 05). The lung function indexes such as forced vital capacity( FVC),forced expiratory volume in first second( FEV1) and FEV1/ FVC in the exposed group were lower than those in control group( P < 0. 05). There was no significant difference in the lung function indexes,respiratory and skin allergy symptoms,and allergic diseases between 7-ACA subgroup and 6-APA subgroup( P < 0. 017).The incidence of bronchial asthma,allergic rhinitis had statistical differences in the length of dust exposure in service workers( P < 0. 05). CONCLUSION: Workers exposed to 7-ACA and 6-APA have a high occurrence rate of respiratory symptoms. These workers also suffered from occupational diseases such as bronchial asthma,allergic rhinitis and others.

Prognostic value of Sox2 expression in digestive tract cancers: A meta-analysis / 华中科技大学学报（医学）（英德文版）

The aim of the present study was to accurately evaluate the association of Sox2 expression with the survival of patients with digestive tract cancers. Relevant literatures were identified by comprehensively searching databases including the Pubmed, Embase, CBMdisc, and Wanfang (up to October 2014). A meta-analysis was performed to clarify the association between Sox2 expression and overall survival or clinicopathological parameters of patients with digestive tract cancers (esophageal, gastric, and colorectal cancers). The results showed a significant association between high Sox2 expression and poor overall survival in patients with digestive tract carcinomas (HR=1.55, 95% CI=1.04-2.31), especially for patients with esophageal cancer (HR=2.04, 95%CI=1.30-3.22), colorectal cancer (HR=1.40, 95% CI=1.04-1.89), and digestive tract adenocarcinoma (HR=1.80, 95% CI=1.12-2.89), for Europeans (HR=1.98, 95% CI=1.44-2.71) or patients who did not receive neoadjuvant treatment (HR=1.73, 95% CI=1.10-2.72). Furthermore, Sox2 over-expression was highly correlated with vascular invasion (OR=1.86, 95% CI=1.25-2.77) and poor differentiation (OR=1.88, 95% CI=1.14-3.08), especially in esophageal and colorectal cancers. In conclusion, Sox2 expression may serve as a novel prognostic factor for patients with digestive tract cancers. Over-expression of Sox2 that is correlated with vascular invasion and poor differentiation suggests poor outcomes of patients with digestive tract cancers.

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>

One-step multiplex RT-PCR for identifying common fusion transcripts in childhood acute lymphoblastic leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To evaluate the efficiency of one-step multiplex RT-PCR for identifying four common fusion transcripts (TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL) in children with acute lymphoblastic leukemia (ALL).</p><p><b>METHODS</b>Total RNA was extracted from bone marrow samples of 76 children who were newly diagnosed with ALL between January 2003 and December 2010. These RNAs were analyzed for TEL/AML1, E2A/PBX1, MLL/AF4 and BCR/ABL by one-step multiplex RT-PCR or common nested-multiplex PCR. The PCR products were confirmed by DNA sequencing.</p><p><b>RESULTS</b>TEL/AML1 was found in 12 cases (the length of products was 298 bp in 9 cases and 259 bp in 3 cases), E2A/PBX1 was found in 3 cases (the length of products was 373 bp), BCR/ABL was found in 1 case (the length of products was 2 124 bp), and MLL/AF4 was found in 7 cases (the length of products was 427 bp in 1 case and 673 bp in 6 cases) using one-step multiplex RT-PCR combined with DNA sequencing. The results were consistent with those using common nested-multiplex PCR.</p><p><b>CONCLUSIONS</b>One-step multiplex RT-PCR may be another alternative for detection of common fusion transcripts in children with ALL.</p>

Seven cases of neonatal Prader-Willi syndrome and a literature review / 中华实用儿科临床杂志

Objective To know more about the features of Prader-Willi syndrome (PWS) in neonates and to promote early interventions for PWS patients.Methods The clinical data from 7 PWS patients being definitely diagnosed by array comparative genomic hybridization(array-CGH) from Jan.2007 to Dec.2012 in Department of Neonatology,Guangzhou Women and Children's Medical Center were collected retrospectively.The data were analyzed with literature being reviewed.Results There were 7 cases described in this study,of which 6 cases were male and 1 case was female.All patients had the features of hypotonia,poor responses,feeding difficulties,less crying or weak crying.Seven cases had 5 or more of the following distinctive facial appearances:prominent forehead,narrow face,almond eyes,small mouth,downturned mouth corners,thin upper lip,and micromandible.Six cases of male children were small scrotum,among them 3 cases were cryptorchidism.Seven cases were diagnosed as PWS by array-CGH.Seven children were fed by using gastric tube,in which 4 cases accepted swallow training after admission,and their gastric tube lasted from 8 to 20 days [(13.0 ±5.1) d],3 cases did not receive continuous tube feeding but swallow training them of from 15 to 35 days [(18.0 ± 4.3) d].Other comprehensive therapy measures included offering the parents a detailed health education,informing and their gastric tube lasted the basic information about the disease,teaching the parents how to facilitate feeding and prevent asphyxia and other aspects of skills.This might increase the baby's passive activities and promote normal development of the baby's nutritional management measures to prevent excessive or inadequate nutritional intake.Multidisciplinary consultation was necessary including the intervention of mass language training,nutrition,neurology,psychiatry,psychology and osteology.Conclusions PWS newborns have the characteristics of hypotonia,poor responses,feeding difficulties,less crying or weak crying,genital hypoplasia and typical facial features.Genetic testing should be necessary for early diagnosis,as early diagnosis is benefitial for early intervention including comprehensive treatment measures such as swallowing training,which may improve the life quality of PWS patients,and improve the prognosis,and prevent growth retardation,obesity and other problems.

Relationship between the methylenetetrahydrofolate reductase gene polymorphism and adverse reactions of high-dose methotrexate in children with acute lymphocytic leukemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the association between methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms and toxicities after high-dose methotrexate (HD-MTX) infusion in children with acute lymphocytic leukemia (ALL).</p><p><b>METHODS</b>MTHFR variants in 52 children with ALL were determined by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis and sequencing. Toxicities of children who received HD-MTX chemotherapy were evaluated according to the National Cancer Institute-Common Toxicity Criteria (NCI-CTC).</p><p><b>RESULTS</b>The children carrying MTHFR 1298AC had a higher risk of developing thrombocytopenia compared with the carriers of the 1298 AA genotype (OR=13.7, 95%CI=1.18-159.36, P=0.036). There was no significant difference in HD-MTX chemotherapy-related adverse effects between the patients with different MTHFR C677T or G1793A genotypes.</p><p><b>CONCLUSIONS</b>MTHFR A1298C polymorohism may associate with the toxicity of HD-MTX chemotherapy in children with ALL.</p>

Effect of methylation inhibitor on EphB4 gene expression, proliferation and apoptosis in CEM cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the regulation of methylation inhibitor 5-aza-2'-deoxycytidine on transcription of EphB4 gene and effects on the proliferation and apoptosis of human acute lymphocyte leukemia cell line CEM.</p><p><b>METHODS</b>Bisulfite sequencing PCR was used to detect CpG island methylation density in EphB4 promoter. The expression of EphB4 mRNA and protein was determined by Q-PCR and Western blot. MTS assay and flow cytometry were used to detect the apoptosis of CEM cells after treatment with different concentrations of 5-aza-2'-deoxycytidine (1.0, 2.5 and 5 μmol/L).</p><p><b>RESULTS</b>Methylation of EphB4 gene promoter was detected in CEM cells (31.4%). The methylation level of EphB4 gene was down-regulated after treatment with various concentrations of 5-aza-2'-deoxycytidine. The EphB4 mRNA and protein expression in CEM cells increased after 5-aza-2'-deoxycytidine treatment. 5-Aza-2'-deoxycytidine significantly inhibited the cell growth in dose and time dependent manners. Early apoptosis rates of CEM cells increased from 4.1% to 24.8% 96 hrs after 5-aza-2'-deoxycytidine treatment. CEM cells in G1 phase decreased from 62.4% to 46.8%, cells in G2 phase increased from 2.1% to 16.2%, and CEM cells were arrested in G2 phase after treatment with 5 μmol/L 5-aza-2'-deoxycytidine for 96 hrs.</p><p><b>CONCLUSIONS</b>5-Aza-2'-deoxycytidine, an inhibitor of specific methylation transferase, can induce expression of the silent EphB4 gene in CEM cells, inhibit the proliferation of leukemia cells and induce cell apoptosis.</p>

Analysis of a 452C/T single nucleotide polymorphism in γ-glutamyl hydrolase gene in children with acute leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the distribution of γ-glutamyl hydrolase gene (GGH) 452C/T genotype and allele frequency in children with acute leukemia (AL) and healthy children.</p><p><b>METHODS</b>Bone marrow samples from 92 children with AL and peripheral blood samples from 124 healthy children were obtained to prepare complementary DNAs (cDNAs). The cDNAs were analyzed for a GGH 452C/T polymorphism by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis (RT-PCR-DGGE) and direct sequencing.</p><p><b>RESULTS</b>The frequencies of the AL patients with TT, CT and CC genotypes were 2.2%, 13.0% and 84.8%, and the frequencies of the control children were 1.6%, 16.9% and 81.5%, respectively. There was no significant difference in GGH genotype or T allele frequency between the two groups (P> 0.05). However, the T allele frequency in Han Chinese children was significantly different from those reported in Japanese, Mexican and African-American populations.</p><p><b>CONCLUSION</b>The frequency of 452C/T polymorphism of GGH gene in Han Chinese children has been determined. The results suggested that an ethnic difference may exist.</p>

Expression and clinical significance of caudal type homeobox transcription factor-2 in adult acute lymphocytic leukemia / 中国实验血液学杂志

This study was aimed to investigate the expression and clinical significance of CDX1, CDX2 and CDX4 genes in acute lymphocytic leukemia (ALL). Expressions of CDX1, CDX2, and CDX4 in 51 adult acute lymphocytic leukemia patients and 14 healthy subjects were detected by reverse transcription polymerase chain reaction (RT-PCR). The results indicated that CDX1, CDX2 and CDX4 were not expressed in 14 healthy persons and 15 CR ALL patients, the positive expression rate of CDX2 gene in de novo ALL patients was 60.8%, while it obviously decreased in patients with complete remission (CR) (p < 0.05); the expression of CDX2 was increased again in relapsed patients (81.8%). When the expression of CDX2 was analyzed in different risk groups of ALL patients, the CDX2 expression rate in high risk (HR) patients was 91.7%, and that in the standard risk (SR) group was 45.7%. Furthermore, analyses of CDX1 and CDX4 expression in series of ALL samples did not show the expression of these genes. In patients with adult ALL at diagnosis and relapse, the CR rate of patients with CDX2 positive expression was lower than that of patients with CDX2 negative expression (p < 0.05). The median survival time in CDX2 positive expression patients was shorter than that in negative expression patient. It is concluded that expression of CDX2 may correlated with pathogenesis and relapse of adult ALL, but the expression of CDX1 and CDX4 don' t associated with pathogenesis and relapse of adult ALL; the CR rate and prognosis of patients with CDX2 positive expression is lower and poor. The expression of CDX2 may be used as a marker for occurrence, relapse and poor prognosis of adult ALL patients.

NB4 cell apoptosis induced by bortezomib combined with As(2)O(3) and its mechanism / 中国实验血液学杂志

This study was aimed to investigate the apoptosis induced by bortezomib combined with As(2)O(3) in APL cell line NB4 and its mechanism. The apoptotic cells were detected by flow cytometry with Annexin V/propidium iodide double staining; the morphology of apoptotic cells was observed by Hoechst staining, Western blot was used to measure activation of caspase-3 and -9 as well as expression of NOXA; the siRNA technique was used to specifically silence NOXA gene; the lipofectamine 2000 was used to transfect pEGFP-Noxa plasmid and pEGFP vacant vector. The results showed that the bortezomib combined with As(2)O(3) could induce significant apoptosis of NB4 cells and activation of caspase 3 and caspase 9, but As(2)O(3) (0.5 µmol/L) alone could not cause marked activation of caspase cascade and apoptosis of NB4 cells. The expression level of NOXA in NB4 cells induced by bortezomib combined with As(2)O(3) was up-regulated; the activation level of caspase-3 and apoptotic rate of NB4 cells treated by bortezomib combined with As(2)O(3) decreased after specifically silencing the NOXA gene. The high expression of NOXA induced by transfection of plasmid could enhance the caspase 3 activity induced by As(2)O(3) alone. It is concluded that bortezomib can enhance sensitivity of NB4 cells to apoptosis induced by As(2)O(3) which may be related with up-regulation of proapoptotic protein NOXA.

Association of single nucleotide polymorphism of reduced folate carrier gene with susceptibility to acute leukemia / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the allele and genotype frequencies of reduced folate carrier gene (RFC) 80G/A polymorphism in Chinese patients with acute leukemia (AL) and healthy control children, and to provide clue for association between the single nucleotide polymorphism (SNP) of RFC and the occurrence of AL.</p><p><b>METHODS</b>Bone marrow samples from 98 childhood patients with AL and peripheral blood samples from 135 healthy children were obtained to prepare complementary DNAs (cDNAs). The cDNAs were analyzed for the polymorphisms in RFC 80G/A by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis (RT-PCR-DGGE) and direct sequencing.</p><p><b>RESULTS</b>The A allele frequencies of the AL patients and control children were 0.515 and 0.415, respectively (P< 0.05). Chi-square test confirmed a statistical significance of the association between RFC80 G/A and AL.</p><p><b>CONCLUSION</b>RFC 80AA or GA genotype may contribute to increasing the susceptibility to AL.</p>

Interleukin-18-induced transdifferentiation in renal proximal tubular cells is mediated by activation of p38MAPK pathway / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To explore the effect of p38MAPK signaling pathway in interleukin-18-induced transdifferentiation in renal proximal tubular cells.</p><p><b>METHODS</b>Human proximal tubular epithelial cell line (HK-2 cells) was cultured in vitro. After preincubated with SB203580 (0, 5, 10, 20 micromol/L) for 30 minutes, cells were exposed to IL-18 (100 ng/ml) for 24, 48 and 72 hours respectively. The expressions of a-smooth actin (alpha-SMA) in cultured HK-2 cells were assessed by RT-PCR and ELISA.</p><p><b>RESULTS</b>IL-18-induced expressions of a-SMA mRNA and protein were inhibited obviously by a dose-dependent manner when HK-2 cells were incubated with SB203580 (0, 5, 10, 20 micromol/L) and IL-18 (100 ng/ml) for different time (P < 0.05).</p><p><b>CONCLUSION</b>IL-18-induced transdifferentiation of renal tubular epithelial cells (RTECs) is suppressed obviously by blocking p38MAPK signaling pathways. IL-18-induced transdifferentiation of RTECs is probably mediated, at least in part, through the activation of p38MAPK signaling pathways.</p>

Analysis of clinical features and gene mutations in two Chinese pedigrees with late-onset methylmalonic acidemia, cblC type / 中华儿科杂志

<p><b>OBJECTIVE</b>CblC is the most common type of methylmalonic acidemia with homocysteinemia. MMACHC is the coding gene. This study aimed at understanding clinical features and gene mutations in 2 Chinese pedigrees who had late-onset methylmalonic acidemia complicated with homocysteinemia.</p><p><b>METHOD</b>The clinical data of 2 cases were analyzed. The MMACHC gene mutation was detected using polymerase chain reaction (PCR) and DNA sequencing.</p><p><b>RESULT</b>The age of onset was 13 years and 12 years, respectively. They both presented with nervous system symptoms. The main clinical features were developmental retardation and degradation, including motion, speech and intelligence. One patient complained of anemia. The other patient was misdiagnosed as having a viral encephalitis. Both patients showed remarkable elevation of methylmalonic acid and homocysteine levels in urine. Both had received therapy with vitamin B(12). The symptoms were rapidly relieved. The follow-up till now showed apparent improvement in the 2 cases. Three mutations in the MMACHC gene were found in the two Chinese pedigrees. Both patients were compound heterozygotes of two mutant alleles: one patient had a G-to-A transition at nucleotide 482 (G482A) that caused an arginine-to-glutamine substitution at position 161 of the protein (R161Q), and a deletion of AAG at nucleotide 658_660 (658_660delAAG) which resulted in lysine deleting at position 220 of the protein (K220del); the other patient had a G482A and a G-to-A transition at nucleotide 609 (G609A) that caused a tryptophan-to-termination codon substitution at position 203 of the protein (W203X). Otherwise, the authors also detected parents of two families. Each had a heterozygote of one mutation.</p><p><b>CONCLUSION</b>Late-onset methylmalonic acidemia patients had a variety of clinical manifestation, the first symptom was mainly abnormality of nervous system. One case was accompanied with hematological abnormalities. Two patients were vitamin B(12) responsive. In this study, the mutations were all detected on the fourth exon, the G482A mutation was probably associated with late-onset cases.</p>

Status of iron metabolism and erythropoietic proliferation in children with various genotypes of thalassemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the status of iron metabolism and erythropoietic proliferation in children with various genotypes of thalassemia.</p><p><b>METHODS</b>Serum concentrations of ferritin (SF), transferrin receptor (sTfR) and erythropoietin (EPO) were measured in 158 children with thalassemia. The differences in the concentrations of the three indices among children with different genotypes of thalassemia were compared. The correlations of the hemoglobin level with sereum SF, sTfR and EPO levels were assessed.</p><p><b>RESULTS</b>Among the 158 children with thalassemia, 52(32.9%) were diagnosed with alpha-thalassemia minor, 27(17.1%) with HbH disease, 59(37.4%) with beta-thalassemia minor, 13(8.2%) with beta-thalassemia major, and 7(4.4%) with combining alpha beta thalassemia. The SF levels in children with HbH disease or beta-thalassemia major were significantly higher than those in the other thalassemia groups (P<0.01). The sTfR levels in children with beta-thalassemia major were the highest when compared with those in the other thalassemia groups (P<0.05). The EPO levels in children with beta-thalassemia major were also the highest when compared with those in the other thalassemia groups (P<0.01). There was a negative correlation between hemoglobin and EPO levels in children with HbH disease (r=-0.656, P<0.01) and beta-thalassemia major (r=-0.641; P<0.05).</p><p><b>CONCLUSIONS</b>The status of iron metabolism and erythropoietic proliferation is different in children with different genotypes of thalassemia. A combined measurement of SF, sTfR and EPO may reflect the status of erythropoietic proliferation.</p>