Association between BIM gene and glucocorticoid resistance in children with acute lymphoblastic leukemia / 中国当代儿科杂志

Acute lymphoblastic leukemia (ALL) is the most common malignant hematological disease in childhood. Glucocorticoids are frequently used in the chemoradiotherapy regimen for ALL and can induce the apoptosis of ALL cells through several signaling pathways, but about 10% of ALL children have poor response to glucocorticoids. Studies have revealed that glucocorticoids induce the apoptosis of ALL cells by upregulating the expression of BIM gene, and BIM gene is associated with glucocorticoid resistance in childhood ALL. This article reviews the recent studies on glucocorticoid resistance in childhood ALL, especially the role of BIM and its expression products in this process.

Association of CCR2 gene rs1799864 polymorphism with hemophagocytic lymphohistiocytosis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the association between rs1799864 single nucleotide polymorphism (SNP) of the C-C chemokine receptor 2 (CCR2) gene and susceptibility of hemophagocytic lymphohistiocytosis (HLH) in children.</p><p><b>METHODS</b>The clinical and laboratory data of 86 children diagnosed with HLH between January 2007 and December 2013 were retrospectively reviewed. The CCR2 gene rs1799864 was genotyped by SNaPshot technique in 86 HLH children and 128 healthy controls. The genotypic and allelic frequencies in the two groups were comparatively analyzed.</p><p><b>RESULTS</b>No significant difference either in genotypic or allelic frequencies of rs1799864 polymorphism of the CCR2 gene was observed between HLH patients and controls (P>0.05), but there were significant differences in the age of onset and the periods of temperature and platelet returning to normal after treatment (P<0.05).</p><p><b>CONCLUSIONS</b>There is no association between CCR2 gene rs1799864 polymorphism and the risk for HLH in children. However, the genotypic differences of this polymorphism might be associated with clinical characteristics and prognosis of HLH.</p>

Association between gene polymorphisms of Perforin 1 and hemophagocytic lymphohistiocytosis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate frequency distribution of gene polymorphisms of PRF1 gene in children with hemophagocytic lymphohistiocytosis (HLH), and to explore whether the possible gene polymorphisms of PRF1 gene confer an increased risk of susceptibility to HLH.</p><p><b>METHODS</b>Forty-eight children who were diagnosed with HLH between January 2009 and December 2013 (HLH group) and 100 healthy children (control group) were enrolled in this study. The gene polymorphisms in the coding region of PRF1 gene, which consists of three exons and two introns, were genotyped by PCR, followed by direct sequencing.</p><p><b>RESULTS</b>Three single nucleotide polymorphisms (SNPs) were revealed in the coding sequence of PRF1 in the 48 children with HLH. Seven SNPs were detected in the noncoding sequence. Other two SNPs in the noncoding sequence including rs10999426 and rs10999427 were detected only in 5 healthy children (5%). There was no significant difference in allelic frequencies of all the SNPs above between the HLH and control groups (P>0.05). Haplotype analysis showed there was a pair-wise linkage disequilibrium between rs10999426 and rs10999427 (D=1, r2=1), but there was no significant difference in the distribution of A-T haplotype between the HLH and control groups (P>0.05).</p><p><b>CONCLUSIONS</b>There is no association between gene polymorphisms of PRF1 gene and the susceptibility to HLH. There is a pair-wise linkage disequilibrium between rs10999426 and rs10999427, but a low detection rate of A-T haplotype in healthy children indicates that it might not play a protective role in the development of HLH.</p>

Clinical features of sinusoidal obstruction syndrome: report of 12 cases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To summarize the clinical features of sinusoidal obstruction syndrome (SOS) and to improve the understanding of this disease.</p><p><b>METHODS</b>A retrospective study was performed on the clinical data of 12 children with SOS including clinical manifestations, laboratory results, imaging findings, treatment, and prognosis.</p><p><b>RESULTS</b>Of the 12 cases, 8 were secondary to acute lymphoblastic leukemia, and 4 had undergone hematopoietic stem cell transplantation. Manifestations mainly included abdominal distention (4 cases), hepatomegaly with tenderness (9 cases), splenomegaly (6 cases), and weight increase (10 cases). Biochemical tests revealed varying degrees of liver damage in all cases. In the coagulation function test, the activated partial thromboplastin time (APTT) was prolonged in 7 cases. Out of the 7 patients who underwent serum D-dimer test, 4 showed elevated serum level of D-dimer. In routine blood tests, 4 cases showed decreases in both white blood cells and neutrophils. In addition, varying degrees of thrombocytopenia were observed in 9 cases. Eight patients were subjected to color Doppler ultrasound examination, and diffuse hepatomegaly, inhomogeneous liver parenchyma, unclear or thinner hepatic veins, hydrothorax/ascites, or splenomegaly was observed. Sinusoidal dilatation or hepatic cell infiltration was observed in 2 patients who underwent liver biopsy. Treatments were basically symptomatic and supportive therapies, and the prognosis was good in all patients.</p><p><b>CONCLUSIONS</b>SOS should be considered in children who present with hepatomegaly, sudden weight gain, liver dysfunction and coagulation dysfunction after they have received chemotherapy for leukemia and hematopoietic stem cell transplantation.</p>

XIAP gene mutation screening in children with hemophagocytic lymphohistiocytosis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the prevalence of mutations and sequence variations in X-linked inhibitor of apoptosis (XIAP) gene among Chinese pediatric patients with hemophagocytic lymphohistiocytosis (HLH).</p><p><b>METHODS</b>Sixty-five children who were diagnosed with HLH between January 2009 and December 2012 (case group), as well as 70 healthy children (control group), were enrolled in the study. The exons of XIAP gene (1-1, 1-2, 2-6) were amplified by PCR and directly sequenced. The genotypic and allelic frequencies of single nucleotide polymorphism (SNP) were analyzed.</p><p><b>RESULTS</b>None of the HLH patients showed mutations in these exons of XIAP gene. Only one nonsynonymous SNP, rs5956583 located in exon 5, was observed, but there were no significant differences in the genotypic and allelic frequencies of this SNP between the case and control groups (P>0.05).</p><p><b>CONCLUSIONS</b>HLH caused by XIAP mutations may be rare in children. SNP rs5956583 of XIAP gene may have little contribution to the development of childhood HLH.</p>

Colorimetric detection of norovirus genotype GII by reverse transcription loop-mediated isothermal amplification / 病毒学报

A simple, rapid and sensitive colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) method was established to detect norovirus genotype GII. The method employed a set of six specially designed primers that recognized eight distinct sequences of RNA-dependant RNA polymerase and capsid protein gene for amplification of nucleic acid under isothermal conditions at 65 degrees C for 60 minutes. The amplification process of RT-LAMP was monitored by the addition of HNB (Hydroxy naphthol blue) dye prior to amplification. A positive reaction was indicated by a color change from violet to sky blue and confirmed by agarose electrophoresis. The specificity of the RT-LAMP was validated by detecting several different diarrhea viruses including norovirus genotype GII. The sensitivity was determined by serial dilutions of RNA molecules from in vitro transcription of norovirus genotype GII in parallel with conventional RT-PCR detection. The assay was further evaluated with 93 clinical specimens of diarrhea patients. The results showed that the sensitivity of RT-LAMP was 1 000 copies/microL with a high specificity and the relative sensitivity was at the same level as that of conventional RT-PCR. Positive rate of RT-LAMP in analysis of clinical specimens was approximately the same as that of conventional RT-PCR as well. This colorimetric RT-LAMP assay was potential for rapid detection of norovirus genotype GII on spot due to the observation of visual result with high specificity and sensitivity, time-saving and cost benefit.

Prognostic factors for hemophagocytic lymphohistiocytosis in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the main factors influencing prognosis of hemophagocytic lymphohistiocytosis (HLH) in children by summarizing the clinical features of HLH and investigating the relationship between relevant factors and prognosis.</p><p><b>METHODS</b>The medical data of 63 children with HLH were retrospectively reviewed. Kaplan-Meier method was employed to draw survival curves. Factors influencing prognosis were assessed with Cox univariate analysis, and Cox multivariate analysis was done on statistically significant factors.</p><p><b>RESULTS</b>The 3-year and 5-year survival rates were both 62.9%. The survival rate decreased from 98.4% at 1 day after definite diagnosis to 73.2% at 4 months. Univariate analysis demonstrated only one factor, which was that the condition of platelet recovery after treatment of 2 to 3 weeks was significantly related to prognosis (P=0.002). In children receiving etoposide therapy, temperature recovery after one day of treatment was significantly related to prognosis (P=0.016).</p><p><b>CONCLUSIONS</b>Children with HLH have a satisfactory prognosis, but the survival rate reduces rapidly in the first 4 months after definite diagnosis. Platelet recovery after treatment of 2 to 3 weeks and temperature recovery after one day of treatment are factors influencing prognosis of HLH in children.</p>

A 3-year clinical trial of deferasirox in heavily iron-overloaded patients with Beta-thalassemia major / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of deferasirox in heavily iron-overloaded patients with beta-thalassemia major.</p><p><b>METHODS</b>A single arm, open-label clinical trial was conducted to evaluate the efficacy and safety of deferasirox in the treatment for 23 patients with beta-thalassemia major and heavily iron-overloaded in 3 years follow-up.</p><p><b>RESULTS</b>The 23 patients never received regular chelation before enrolling this trial [the mean baseline of serum ferritin was (5433.96 ± 2873.90) µg/L]. In this trial, a deferasirox dose of 20 mg×kg(-1)×d(-1) could stabilize serum ferritin levels, while of ≥ 30 mg×kg(-1)×d(-1) reduced the levels and achieved negative iron balance. There were no serious adverse events related to the drug. Most common adverse events were mild increases of liver enzyme and serum creatinine levels. Overall, 23 patients could tolerate the drug on schedule and all completed the trial.</p><p><b>CONCLUSION</b>As a new oral iron chelator, deferasirox has a significant efficacy for the treatment of iron overload. The effectiveness is dependent on the courses of treatment and the dose of deferasirox. The single-dose used is safe and tolerated, so deferasirox can remarkably improve life quality of patients.</p>

Efficacy and prognosis analysis in 188 children with acute lymphoblastic leukemia of China / 中华儿科杂志

<p><b>OBJECTIVE</b>To analyze the therapeutic effect and the influencing factors of event-free survival (EFS) of childhood acute lymphoblastic leukemia (ALL) in Xiangya Hospital of Central South University and the First Affiliated Hospital of Guangxi Medical University.</p><p><b>METHODS</b>All the patients adopted chemotherapy according to therapeutic guideline revised by the Subspecialty Group of Hematology, The Society of Pediatrics, Chinese Medical Association for the second-time in 1998 (the Rongcheng ALL-98 Protocol). Kaplan-Meier method was used to estimate the survival rates of 188 patients who received therapy with good compliance. The differences of EFS between groups were assessed by Log-rank test. The independent influencing factors on EFS were analyzed by the Cox proportional hazards regression model.</p><p><b>RESULTS</b>After receiving inductive treatment, 354 of 374 (93.6%) patients demonstrated a complete remission; 188 patients who received complete courses of treatment with good compliance showed (68.1 +/- 5.6)% five-year EFS. Meanwhile, the five-year EFS in standard-risk (SR) group and high-risk (HR) group were (75.2 +/- 6.0)% and (47.6 +/- 11.6)%, respectively. The total relapse rate was 10.6% and the median time to relapse was 13 months. Twenty-nine of 188 patients (15.4%) were dead, and 13 patients (7.0%) died from treatment-related complications. Independent adverse prognostic factors included risk grouping, t (9; 22)/bcr-abl gene and leukocyte count.</p><p><b>CONCLUSIONS</b>The total EFS of childhood ALL patients treated with Rongcheng ALL-98 Protocol in two hospitals was close to 70%. Therefore, it is necessary to evaluate risk factors and consider the grouping in more detail to reduce the treatment-related mortality and to increase the compliance of treatment which can ultimately improve the EFS.</p>

Suicidal cancer vaccine enhances anti-tumor immunotherapeutic effect and its safety in the treatment of ovarian cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the anti-tumor immunotherapeutic effect induced by the suicidalcancer vaccine FC/TK, and to evaluate the safety of this vaccine.</p><p><b>METHODS</b>The suicidal cancer vaccine, named FC/TK, was prepared by fusion of suicide gene (HSVI,-TK gene) -modified ovarian carcinoma NuTu-19 cells with rat bone marrow-derived dendritic cells (DCs). The morphology of FC/TK was evaluated by scanning electron microscopy. The stimulatory effect of FC/TK on T cells was determined by T cell proliferation assay. In immunotherapeutic studies in vivo, Fischer344 rats were injected subcutaneously with NuTu-19 cells, followed by treatment of FC/TK on days 7 and 14, compared to controls treated with irradiated FC/TK, FC or PBS, respectively. Tumor incidence and volume were measured in 90 days after challenge. To determine the killing effect of FC/TK in vivo, TUNEL assays were applied to detect apoptotic cell death in spleen of vaccinated rats with prodrug ganciclovir administration.</p><p><b>RESULTS</b>FC/TK cells were of irregular shape with surface membrane processes. Compared to the control groups, FC/TK significantly promoted T cell proliferation (P <0.01). The rats vaccinated with FC/TK and FC significantly inhibited the tumor growth compared to rats vaccinated with irradiated FC/TK (P <0.05) or with PBS ( P <0.01). The immunotherapeutic effect induced by FC/TK was similar to that using FC. Fluorescence microscopy showed that fluorescein-stained FC/TK cells migrated into spleen also showed to be TUNEL-positive, suggesting that the FC/TK cells were killed by ganciclovir in vivo.</p><p><b>CONCLUSION</b>Our data indicate that suicidal cancer vaccine is an effective and safe therapy for ovarian carcinoma and may serve as a broadly applicable approach for other cancer vaccines in the future.</p>

Detection of Clonal T Cell Receptor ? Gene Rearrangement in Children with Acute Lymphoblastic Leukemia and Its Clinical Significance / 实用儿科临床杂志

Objective To investigate the clonal rearrangement of T cell receptor (TCR) ? gene which was of monoclonality,oligoclona-lity or clonal evolution /subclonality during the course of disease in patients with acute lymphoblastic leukemia (ALL) and its significance.Methods Between Sep. 2004 and Sep. 2007,70 patients with ALL were diagnosed in Department of Pediatrics of the First Affiliated Hospital of Guangxi Medical University,among which 51 cases were boys and 19 cases were girls.Their ages ranged from 2 years to 14 years (average age was 8.5 years).DNA samples were extracted from bone marrow cells or venous blood cells by phenol/phenol-isoamyl alcohol-chloroform/isoamyl alcohol law.DNA was amplified by polymerase chain reaction (PCR).Single strand conformation polymorphism analysis (SSCP) of silver stained technique was employed to detect PCR products.Results The amplification products of 23 cases of the 70 ALL patients were positive,in which 10 cases still had positive results in period of complete remission and had poor sensitivity to chemotherapeutic drugs,4 cases were of oligoclonality/ subclonality,and 2 cases were of clonal evolution with poor prognosis.Conclusions Detecting TCR ? gene rearrangement reflects clonal evolution of leukemia cells.The oligoclonality clonal evolution continues to exist,whose multiplication is the main reason of recurrent ALL.Detecting TCR? gene rearrangement,evaluating ALL of the patient's prognosis,the judgment of recurrence and the development of individualized treatment programs have great guiding significance,which can maximize the possibility of the sick children to make long-term disease-free survival and reduce the side effects of chemotherapy on the long-term basis.

Relationship between Glucose-6-Phosphate Dehydrogenase Deficiency and Viral Hepatitis in Guangxi Area / 实用儿科临床杂志

Objective To study the glucose-6-phosphate dehydrogenase(G6PD)genotypes in the children of viral hepatitis with G6PD deficiency and investigate the relationship between G6PD deficiency and viral hepatitis.Method DNA samples of 18 children with viral hepatitis/ G6PD deficiency were studied for the 3 common mutations by using the natural primers or mismatched primers mediated PCR followed by restriction enzyme analysis and their clinical manifestation was analyzed.Results Among 18 cases,8 cases were G1388A,and 4 cases were G1376T,and 1 case was A95G.The 3 mutations of G1388A,G1376T and A95G added up to 72.22%.There were no significant difference between the patients with viral hepatitis/ G6PD deficiency and the children only with G6PD deficiency.The incidence rates of acute hemolysis and acute renal failure in the viral hepatitis/G6PD deficiency patients were higher than those in children with G6PD deficiency only.Conclusions It is supposed that the G6PD deficiency is primary in the patients of viral hepatitis with G6PD deficiency.The children of viral hepatitis with G6PD deficiency have more severe pathogenetic conditions and more complications than the children with G6PD deficiency only.

Identification of G6PD mutations in Guangxi by combination of denaturing high performance liquid chromatography, DNA sequencing and restriction endonuclease analysis / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore new genotypes of glucose-6-phosphate dehydrogenase (G6PD) deficiency in Guangxi.</p><p><b>METHODS</b>G6PD mutations were identified by combination of denaturing high performance liquid chromatography (DHPLC), DNA sequencing and restriction endonuclease assay.</p><p><b>RESULTS</b>Three cases (10.0%) of G6PD Viangchan (871G-->A, 1311C-->T) and one case (3.3%) of G6PD Union (1360C -->T) were first identified in China mainland. G6PD Ganton (1376G-->T, 30.0%) was the commonest mutation, followed by Kaiping (1388G-->A, 26.7%), and Gaohe (95A-->G, 23.3%).</p><p><b>CONCLUSION</b>Besides G6PD Ganton, Kaiping and Gaohe, there are G6PD Viangchan and Union in Guangxi. G6PD Viangchan and Union is reported for the first time in China mainland.</p>

The application of the dissociate bone flap in treatment of the macrosis depressed skull fracture on the frontal orbit part / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the reconstructive effect of the dissociate bone flap to repair the macrosis depressed skull fracture on the frontal and orbit part.</p><p><b>METHODS</b>The coronal scalp flap was elevated and dissociate bone flap was expanding to the 2cm width beside the edge of depressed skull fracture. The first step was to extract the dissociate bone flap and make there is an area for operating . Then extract free bone fragments, and elevate the depressed orbital lamina and use the biological glue to stick it to its position. The free fragments extracted were stacked into a whole one and it to its position in use of the biological glue on the dissociate bone flap. The uneven inner table should was smoother with bon-wax. The prosthetic dissociate bone flap was put back on its position and fixation.</p><p><b>RESULTS</b>From January 2000 to December 2004, 17 cases of the macrosis depressed skull fracture on the frontal and orbit part undertaken plastic surgery by the dissociate bone flap to treat the macrosis depressed skull fracture and obtained excellent curative effect.</p><p><b>CONCLUSIONS</b>Using dissociate bone flap to treat the mocrosis depressed skull fracture on the frontal and orbit part can avoid the complication of the traditional operation, and make the method become a plastic surgical operation.</p>

Distribution of Vitamin D Receptor BsmⅠ and Tru9I Site Gene Polymorphism in Children in Guangxi Region / 实用儿科临床杂志

Objective To investigate the distribution of vitamin D receptor(VDR) gene BsmI and Tru9I site gene polymorphism in children in Guangxi region.Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method and DNA sequencing technology were used to analyze the VDR genotype in 268 healthy children [including 143 boys and 125 girls which aged (4.15?0.63) years] in Guangxi region.Hardy-Weinberg balance analysis was used to check up the samples.Results There were 246 cases which were bb genotgpes in BsmI site and 21 cases which were Bb genotypes and 1 case which was BB genotypes in 268 cases.The frequency of bb,Bb and BB in BsmI site were 91.79%,7.84% and 0.37%,respectively.Frequencies of b and B allelic gene were 95.71% and 4.29%;There were 12 cases which were tt genotypes in Tru9I site and 85 cases which were Tt genotypes and 171 cases which were TT genotypes in all 268 cases.tt,Tt and TT in Tru9I site were 4.48%,31.72% and 63.80%,respectively.Frequencies of t and T allelic genes were 20.34% and 79.66%.The result of PCR-RFLP was according to the result of DNA sequencing.The samples had group representation through Hardy-Weinberg balance analysis.Conclusions The polymorphism frequency and distribution of VDR gene BsmI and Tru9I site gene polymorphism of children in Guangxi region of China exhibit its own characteristics.BB and tt genotypes minority.

The influence of acupuncture on Nestin expression in the cortex of hypoxia-ischemic neonatal rats / 中华物理医学与康复杂志

Objective To investigate the influence of acupuncture on Nestin expression in the cerebral cor- tex of hypoxia-ischemic neonatal rats.Methods A total of 72 Sprague-Dawley(SD)rats aged 7 days were random- ly divided into three groups:A normal group(n=24),the hypoxic-ischemic encepbalopathy(HIE)group(n=24), and an acupuncture+HIE group(n=24).The rats were sacrificed at time points including 3d,7d,14d and 28d af- ter insult,their brains were removed and sections were made.Any nerve stem cells(NSCs)in the cerebral cortex were detected using immunohistochemical staining of Nestin.Results Nestin-positive cells were found in the cere- bral cortex in the normal neonatal SD rats early after birth.After HI insult,the Nestin-positive cells had increased in the injured areas of the cortex and in contralateral mirror areas,especially around the injured areas.Peak Nestin ex- pression was 3 days after insult.In the acupuncture+HIE group,Nestin positive cells were significantly increased when compared with that in the HIE group,the peak was postponed,and the peak was longer than in the HIE group. The Nestin positive cells decreased,were seldom seen,and were not different among three groups 28 days after the HI insult.Conclusions Endogenous neural stem cells in the cerebral cortex were increased by acupuncture in hy- poxia-ischemic in neonatal rats,and acupuncture made the peak of Nestin expression longer.

Study Progress of B Lymphocyte Abnormal in Idiopathic Thrombocytopenic Purpura / 实用儿科临床杂志

Idiopathic thrombocytopenic purpura(ITP) has been confirmed a clinical syndrome because platelets are much destroyed by immunologic mechanism,it is also called immunethrombocytopenic purpura.The pathogenesis of that is very multiplicity,and at present it isn′t understood.This review article is about study progress of B lymphocyte abnormal and the possible pathogenesis of ITP in children.