Simultaneous determination of 8 bioactive isoflavonoids in Astragali Radix by UHPLC equipped with core-shell column / 中国中药杂志

This research aims to develop an UHPLC method, based on core-shell column(i.e. superficially porous particles), for simultaneous determination of eight isoflavonoids including formononetin,(6αR,11αR)-3-hydroxy-9,10-dimethoxypterocarpan, calycosin-7-O-β-D-glucopyranoside,(3R)-7,2-dihydroxy-3,4-dimethoxyisoflavone, calycosin, ononin,(6αR,11αR)-9,10-dimethoxypterocarpan-3-O-β-D-glucopyranoside, and(3R)-7,2-dihydroxy-3,4-dimethoxyisoflavan-7-O-β-D-glucopyranoside in Astragali Radix. The analysis was performed on an Agilent Poroshell EC-C_(18 )column(2.1 mm×100 mm, 2.7 μm) with 0.2% formic acid solution(A)-acetonitrile(B) as mobile phase for gradient elution. The flow rate was 0.5 mL·min~(-1), with column temperature of 40 ℃ and the wavelengths were set at 260 and 280 nm. According to the results, all calibration curves showed good linearity(R~2>0.999 8) within the tested concentration ranges. Both the intra-and inter-day precisions for 8 isoflavonoids were less than 0.80%, with the mean recovery at the range of 94.71%-104.6%. Thus, the newly developed UHPLC method using core-shell column owned the advantages in terms of rapid analysis, low column pressure and less solvent consumption, thus enabling the usage of conventional HPLC systems. Meanwhile, quantitative evaluation was carried out for 22 batches of commercial Astragali Radix. It has been found that great variations occurred for the content of the individual isoflavonoids among different batches; in contrast, the total content of total 8 isoflavonoids(>0.1%) was stable in most samples, indicating that it was reasonable to involve all isoflavonoids as the chemical markers for the quality control of Astragali Radix.

Study on effect of tetramethylpyrazine on proliferation and apoptosis of leukemic U937 cells and its mechanism / 中国中药杂志

<p><b>OBJECTIVE</b>To study the proliferation and apoptosis of tetramethylpyrazine (TMP) on leukemic U937 cells and its possible mechanism.</p><p><b>METHOD</b>The inhibitory effect of TMP on the proliferation of U937 cells was detected by CCK-8 assay. The cell apoptosis and cycle distribution were examined by the flow cytometry. The mRNA expressions of bcl-2 and P27 were determined by the Real-time PCR. Western blot was carried out to detect bcl-2, caspase-3, cyclin E1, CDK2 and P27 expressions.</p><p><b>RESULT</b>TMP inhibited the proliferation of U937 cells in a dose-and-time dependent manner, with IC50 value of 160 mg x L(-1) at 48 h. In addition, TMP could induce the apoptosis of U937 cells and block the cell cycle in G0/G1 phase. According to the results of Real-time PCR and Western blot, TMP could down-regulate the expression of apoptosis-related molecule bcl-2, cycle-related protein cyclin E1 and CDK2 and up-regulate caspase-3 and P27.</p><p><b>CONCLUSION</b>TMP shows the effects in inhibiting the proliferation of leukemic U937 cells and inducing the apoptosis. Its mechanism may be related to the impacts on the cell cycle distribution, down-regulation of the bcl-2 expression, which finally activates caspase-3, starts the apoptosis path and causes the cell apoptosis.</p>

Clinical significance of HA117, mdr1, mrp1, lrp and bcrp resistance genes expressions in children with acute leukemia / 中华实用儿科临床杂志

Objective To explore the expressions of the HA117,mdr1,mrp1,lrp and bcrp genes in bone marrow mononuclear cells(BMMNC) with acute leukemia (AL) and the clinical significance in AL.Methods Expressions of HA117,mdr1,mrp1,lrp and bcrp genes in 81 children with AL and 14 children with idiopathic thrombocytopenic purpura (ITP) were tested using semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR) technique.Results 1.The expressions of HA117,mdr1,mrp1,lrp and bcrp genes in AL patients were higher than that in ITP children.And the expressions of these genes in refractory-relapsed patients also higher than that in the initially diagnosed patients or remission patients.2.The first complete remission (CR1) rate in HA117 and lrp positive cases was lower than that in negative cases(all P ＜ 0.05),but the CR1 rates in mdr1,mrp1 and bcrp positive cases had no significant difference from it in negative cases(all P ＞ 0.05).3.Correlation analysis showed:there was no correlation between the expression of HA117 and mdr1 or HA117 and rnrp1 or HA117 and 1rp or HA117 and bcrp in childhood AL(r =0.031,0.319,0.203,0.176,11.178,all P ＞ 0.05).mdr1 and mrp1 or mdr1 and bcrp or lrp and bcrp had obviously positive correlation (r =0.260,0.308,0.317,all P ＜ 0.05).In refractory-relapsed group,HA117 and rndr1 or rnrp1 or lrp or bcrp had obviously perfect positive correlation.Conclusions The gene HA117 and mdr1,mrp1,lrp,bcrp may be associated with muhidrug resistance of AL in children,therefore the detection of HA117 expression is helpful in management of AL patients.Comparing to the mdr1 or mrp1 or bcrp genes,the genes of HA117 and lrp can better predict the multidrug resistance in childhood AL.

Effect of HOXA10 gene silenced by shRNA on proliferation and apoptosis of U937cell line / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effects of lentivirus-mediated RNA interference targeting HOXA10 gene on the proliferation, apoptosis and morphology of leukemic cell line U937.</p><p><b>METHODS</b>Four different shRNA plasmids were designed and built to interfere with HOXA10 gene. The four interference plasmids were transfected into 293T cells with the HOXA10 over expression plasmid and then the RNAi efficiency of the four interference plasmids was determined by Western blot. The best one was chosen to transfect 293T cells with lentiviral helping plasmids to produce packaged lentivirus (lenti-shHOXA10). U937 cells were divided into interference group (lenti-shHOXA10), negative control group and untreated group. After infection with the packaged lentivirus, infection efficiency of lentivirus for U937 was detected by flow cytometry, and the expression of HOXA10 gene mRNA and protein was detected by real-time PCR and Western blot. Cell survival was determined by MTT assay. Apoptosis rate was detected by flow cytometry.</p><p><b>RESULTS</b>Lentiviral-shRNA vector of HOXA10 gene was successfully constructed. Compared with the negative control and untreated groups, mRNA level of HOXA10 decreased by (92.3±1.3)%, protein levels decreased by 91.1%, and the inhibition rate of U937 cells [(43.9±0.7)%] increased in the interference group (P<0.05). Wright's staining showed that the ratio of karyon to cytoplasm was reduced and mitotic phase was rare in the interference group. Apoptosis rate in the interference group [(27.1±1.4)%] was significantly higher than in the negative [(19.4±1.9)%] and untreated groups [(5.5±1.3)%] (P<0.05).</p><p><b>CONCLUSIONS</b>Lentivirus mediated RNAi can reduce the expression level of HOXA10, effectively inhibit proliferation and promote apoptosis of U937 cells. HOXA10 gene is expected to become a new target for the treatment of leukemia at gene level.</p>

Analysis of thirteen cases with secondary coagulation disorder caused by raticide exposure / 中华儿科杂志

<p><b>OBJECTIVE</b>To summarize the clinical characteristics of secondary coagulation disorders caused by exposure to poison (raticide) in children and to investigate the diagnosis and corresponding treatment.</p><p><b>METHOD</b>The process of diagnosis, clinical characteristics, response to treatment and the prognosis were analyzed.</p><p><b>RESULTS</b>The main clinical manifestation was mucosal bleeding (66.6%), including epistaxis, gingival bleeding, hematomas and so on. All these children were previously well and had no history of bleeding. Activated partial thromboplastin time (APTT) and prothrombin time (PT) were prolonged, factor II was undetectable and the levels of factors VII, IX, and X were lower. The fibrinogen was normal. A raticide was detected in blood and urine of 13 children although 12 of the patients had no definite history of raticide ingestion. Prothrombin complex, fresh frozen plasma and vitamin K(1) were effective in these cases. However, 2 - 3 weeks later, 6 patients presented with recurrent bleeding.</p><p><b>CONCLUSION</b>For children with secondary coagulation disorders of unknown cause, intoxication of raticide should be considered. The administration of blood coagulation factors and vitamin K(1) are effective in early treatment, and the treatment period should be more than 2 months. The PT and APTT should be followed up. Vitamin K(1) should be stopped when PT and APTT are normal.</p>

Effects of Yinian Jiangya Decoction containing serum on cytokines secretion of vascular endothelium of spontaneously hypertensive rats / 中国结合医学杂志

<p><b>OBJECTIVE</b>To observe the effects of Yinian Jiangya Decoction (YNJYD) on cytokine secretion in spontaneoulsy hypertensive rats (SHRs) vascular endothelium.</p><p><b>METHODS</b>Aortic endothelial cells (ECs) were primarily cultured from SHRs; male SD rats were treated with different doses (high, medium, and low doses) of YNJYD, the blood was collected on the 21st day, and then, the serum was separated. ECs were cocultured with the serum for different time courses, and the culture supernatant concentrations of endothelin (ET)-1, nitric oxide (NO), tissue-type plasminogen activator (t-PA), and plasminogen activator inhibitor (PAI-1) were determined by ABC-ELISA methods.</p><p><b>RESULTS</b>ET-1, NO, t-PA, and PAI-1 levels in endothelial cell culture supernatant were increased in a time-dependent manner; YNJYD could significantly elevate NO and t-PA expressions in ECs, while ET-1 and PAI-1 expressions were dramatically decreased; these effects of YNJYD were in a concentration-dependent manner.</p><p><b>CONCLUSION</b>The therapeutic effect of YNJYD on hypertension is attributed to its effect on regulating vessel dilation and blood coagulation, in which ET-1/NO and PAI-1/t-PA are two pairs of pivotal mediators.</p>

Clinical study on childhood acute lymphoblastic leukemia diagnosed and treated with 04 Protocol in Chongqing, China / 中华儿科杂志

<p><b>OBJECTIVE</b>To analyze the clinical and laboratory data from acute lymphoblastic leukemia (ALL) patients and the results of treatment using 04 Protocol (suggested by the Pediatric Hematology Group of Chinese Medical Association in 2004).</p><p><b>METHODS</b>This study included 88 children with ALL below the age of 18 years during the period from October 1, 2004 to June 30, 2007. Minimal inhibitory concentration (MIC) and clinical risk classification were done and the new chemotherapy regimen was used according to the protocol. Patients were stratified into low-risk (LR), medium-risk (MR), and high-risk (HR) groups. Life table method was used to estimate survival rate and statistical analysis was done by using software SPSS for Windows.</p><p><b>RESULTS</b>From October 2004 to June 2007, 88 childhood ALL patients were treated with the 04 Protocol. Sixty-three (91.30%) patients attained complete remission (CR) and 17 patients lost to follow up. The overall 4-year-event-free survival (EFS) rate (+/- SE) was (59.73 +/- 7.22)%. EFS was (75.60 +/- 9.71)% in the LR (n = 30), (65.50 +/- 11.69)% in the MR (n = 20) and (44.03 +/- 12.36)% in the HR. Relapse occurred in 18.18% of patients. Seven (7.95%) of 88 patients with ALL died during he induction therapy. Infection was the most common cause of death.</p><p><b>CONCLUSION</b>The outcome of patients treated with the 04 Protocol was favorable. Clinical risk classification and the leukemia cells of D19 are independent predictors of prognosis of ALL. High dose methotrexate played an important role in prevention and treatment of central nervous system leukemia. The mortality rate of this chemotherapeutic protocol during induction therapy was high.</p>

Effects of Tanshinone IIa on cytokines and platelets in immune vasculitis and its mechanism / 中国实验血液学杂志

The objective of this study was to investigate the effect of Tanshinone IIa on IL-1beta, IL-6 and TNF-alpha cytokines in immune vasculitis and platelets, as well as their relationship. The model of immune vasculitis of rabbits were established by intravenous injection of bovine serum albumin twice. Experiment was divided into 4 groups: control group, model group, tanshinone IIa-treated group and aspirin-treated group. The platelet count, platelet aggregation of peripheral blood were determined. The levels of serum IL-1beta, IL-6, TNF-alpha were detected by ELISA. The pathological changes of immune vasculitis were analyzed by hematoxylin & eosin staining, elastic fibers staining and electron microscopy. The results showed that the levels of IL-1beta and TNF-alpha in model group were significantly higher than those in normal group (p < 0.05), while the level of IL-6 was not significantly different between various groups. The serum level of IL-1beta was correlated with platelet number, while serum levels IL-1beta and TNF-alpha were both correlated with the platelet aggregation. The treatment with tanshinone IIa could significantly decrease the serum levels of IL-1beta, TNF-alpha and platelet number, and the efficacy of tanshinone IIa was same as aspirin. The tanshinone IIa and aspirin both could alleviate the vessels damage in patients with immune vasculitis. It is concluded that the tanshinone IIa may diminish the inflammation damage of vessels in patients with immune vasculitis through the inhibition of cytokines and platelets.

Successful treatment of a patient with Wiskott-Aldrich syndrome using hematopoietic stem cell transplantation--case report and literature review / 中华儿科杂志

<p><b>OBJECTIVE</b>Wiskott-Aldrich syndrome (WAS) is a primary immunodeficiency diseases. The patients with classical WAS have poor prognosis. The hematopoietic stem cell transplantation is the most effective method to cure WAS at present. In this report, a patient with WAS was cured with HLA identical sibling bone marrow transplantation (BMT).</p><p><b>METHODS</b>Wiskott-Aldrich syndrome protein (WASP) was detected using flow cytometry and WASP were analyzed for the diagnosis. The bone marrow was collected from the elder sister who was the HLA identical sibling donor. A total of 4.38x10(8)/kg mononuclear cell (MNC) and 3.78x10(6)/kg CD34+ cells were collected and transfused into the patient after the conditioning regimen with busulfan/cyclophosphamide. Cyclosporine only was used for graft-versus-host disease prophylaxis. WASP and short tandem repeats (STR) were detected as the evidence of engraftment.</p><p><b>RESULTS</b>The diagnosis was WAS: WASP (-IVS9+2T>C, WASP-negative). The patient received busulfan/cyclophosphamide 9 days before the transplantation. WBC decreased to 0.1x10(9)/L in d+4; The absolute number of neutrophils (ANC) was 0.8x10(9)/L in d+13, and exceeded 1.0x10(9)/L later on. From d(-9)-d+14 the patient was dependent on platelet transfusion. From d+15 the patient's PLT>50x10(9)/L and returned to normal after d+30. In d+9-d+10 mild GVHD (I degree) occurred but subsided after the steroid treatment. From d+50, WASP was detected positive and STR showed full donor DNA chimera. Follow-up for 510 d post-transplant, the patient suffered only from mild cold twice, no eczema, no bleeding occurred. The PLT is normal and no chronic GVHD occurred. The levels of IgG, IgM and IgA of the patient were approximately normal.</p><p><b>CONCLUSION</b>The HLA-identical sibling's BMT seems to be the periorit treatment of choice for the WAS patient.</p>

Effect of siRNA targeting VEGF on cell apoptosis and the expression of survivin in K562 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of adenovirus vector-mediated siRNA targeting vascular endothelial growth factor(VEGF) on apoptosis and the expression of survivin in K562 cells.</p><p><b>METHODS</b>K562 cells were infected with recombinant adenovirus Ad5-VEGFsi for 72 hours as experimental group (K562/Ad5-VEGFsi), and empty vector group (K562/Ad5) and blank control group (K562) as controls. VEGF mRNA and survivin mRNA expression were determined by RT-PCR. The protein levels of VEGF and survivin were measured by ELISA and Western blot, respectively. The apoptosis of K562 cells was detected by flow cytometry.</p><p><b>RESULTS</b>The levels of VEGF and survivin mRNA expression in experimental group cells were significantly decreased (P < 0.01). The protein concentration of VEGF in experimental group supernatant was (1121 +/- 15) pg/ml, being lower than that in empty vector group \[(1290 +/- 28) pg/ml\] and black control group \[(1303 +/- 28) pg/ml\] (P < 0.01), and the level of survivin protein in experimental group (0.26 +/- 0.11) was significantly reduced compared with that in blank control group (0.74 +/- 0.10) (P < 0.01). The apoptosis rate of K562/Ad5-VEGFsi cells (16.45 +/- 0.14)% was higher than those of K562/Ad5 cells (3.54 +/- 0.17)% and K562 cells (2.56 +/- 0.20)% (P < 0.01).</p><p><b>CONCLUSIONS</b>VEGF can up-regulate the expression of survivin. After inhibition of VEGF by RNAi, the expression of survivin is decreased subsequently and the rates of cell apoptosis are increased.</p>

Hematopoiesis support of mesenchymal stem cells in children with aplastic anemia / 中国当代儿科杂志

<p><b>OBJECTIVE</b>The abnormality of hemopoietic inductive microenvironment (HIM) is involved in the pathophysiology of aplastic anemia (AA). Mesenchymal stem cells (MSC) are main source of bone marrow stromal cells which constitute the bone marrow HIM. Thus, the bone marrow failure in AA may be related to the function of MSC. The aim of the study was to investigate the hematopoiesis support function of MSC in children with AA in vitro.</p><p><b>METHODS</b>Bone marrow samples were collected from 24 children with AA at diagnosis and 19 children with idiopathic thrombocytopenic purpura (ITP), infectious mononucleosis or lymphadenitis (controls). MSCs from bone marrow samples were isolated, cultured and expanded. Morphology, proliferation activity and colony forming unit-fibroblast (CFU-F) were measured. The ability of bone marrow MSC to adhere hemopoietic cells was assayed by MTT. The concentration of stem cell factor (SCF) released from MSC was tested using ELISA. Mononuclear cells (MNC) of bone marrow were plated onto a feeder layer formed by MSC. Cells count and BFU-E, CFU-GM, CFU-GMME productions were measured.</p><p><b>RESULTS</b>The first and third passage time of MSC in children with AA was longer than that in the controls. The number of CFU-F in children with AA (15.70+/-5.78) was less than that in the controls (21.73+/-5.74) (P<0.05). The concentration of SCF in MSC supernatants in children with AA (30.69+/-16.82 pg/mL) was significantly lower than the controls (50.74+/-14.83 pg/mL) (P<0.01). The total MNC count and the number of BFU-E, CFU-GM and CFU-GMME colonies in the support of MSC in children with AA were significantly lower than those in the controls (P<0.01).</p><p><b>CONCLUSIONS</b>The hematopoiesis support function of MSC was significantly reduced in children with AA in vitro. The decreased hematopoiesis support function of MSC may be related its decreased proliferation capacity and SCF release activity.</p>

Effects of matrine on invasion and metastasis of leukemia cell line Jurkat / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the inhibitory effects of matrine, in different concentrations, on invasion and metastasis of human acute lymphocytic leukemia cell line Jurkat.</p><p><b>METHODS</b>In vitro cultured Jurkat cells were treated by matrine in concentration of 0 g/L, 0.1 g/L, 0.15 g/L and 0.2 g/L, respectively. Then cell adhesion assay, cell migration assay and matrigel invasion assay were used respectively to observe the effects of matrine on adhesion, migration and invasive capacity of Jurkat cells. Meantime, RT-PCR was performed to detect the matrix metalloproteinase (MMP)-2 and MMP-9 mRNA expression levels. Comparison of measurement data among groups was analyzed by variance analysis.</p><p><b>RESULTS</b>As compared with the control group, the adhesion of Jurkat to fibronectin (FN) was significantly inhibited by 0.15 g/L and 0.2 g/L of matrine (P < 0.05); the cell migration and invasive capacity were significantly lowered by 0.1 g/L, 0.15 g/L and 0.2 g/L matrine (P < 0.01). High mRNA expression of MMP-9 presented but that of MMP-2 was expressed insignificantly in Jurkat cells, matrine at 0.1 g/L, 0.15 g/L and 0.2 g/L showed obvious effect in down-regulating MMP-9 mRNA expression (P < 0.01). Besides, MMP-9 mRNA expression was found to be positively correlated with the invasive capacity of Jurkat cells (r = 0.940, P < 0.01).</p><p><b>CONCLUSIONS</b>Matrine is a good drug for antagonizing the invasion and metastasis of leukemia cells, it may roundly inhibit the adhesion, migration and invasive capacity of Jurkat cells, the mechanism might be related with the down-regulation of MMP-9 mRNA expression.</p>

How to Cultivate the Pediatrics Interns' Clinical Work Ability / 中华医学教育探索杂志

The internship is the transition period of a medico becoming a doctor,the cultivation of clinical work ability of interns is a comprehensive ability cultivation which includes the foundation theories' consolidation and use,the practical operative train- ing,the cultivation of clinical thought ability and communication between doctors and patients,etc.To educate pediatrics intern has its characteristics.

Differentiation of K562 cells induced by antisense oligonucleotide targeting survivin / 中华儿科杂志

<p><b>OBJECTIVE</b>Gene therapy of leukemia is a new and effective method. It is known to all that the pathogenesis and development of leukemia are related to a variety of genes. Survivin is a member of inhibitors of apoptotic proteins (IAP). Its cDNA was cloned from target cell protease receptor-1 (EPR-1). It is expressed in common tumors, but there is no expression in normal and mature tissues. High expression of survivin was detected in leukemic cells. The present study was conducted to examine the role of survivin in the differentiation of leukemic cells by using antisense-oligonucleotides.</p><p><b>METHODS</b>Human leukemic cell K562 was used as the model for the study. K562 cells were divided into 4 groups randomly: antisense oligonucleuotide (ASON) group, nonsense oligonucleotide (NSON) group, lipofectin group and control group. There were 5 samples in each group, and the experiment was repeated for three times. ASON was designed with the reference to targeting survivin mRNA. K562 cells were cultured in RPMI1640 contained fetal cattle serum at a concentration of about 10 percent. Cell transfection was induced by lipofectin. Forty-eight hours after thansfection, the morphology and ultrastructure were observed. Twenty-four hours and 48 hours after thansfection, the function of K562 cells was detected by benzidine staining, POX staining and NBT staining, respectively. The mean fluorescence intensity of CD33 was determined by flow cytometry. The method of immunohistochemistry was used to examine the protein level of survivin.</p><p><b>RESULTS</b>After thansfection with ASON, the size of K562 cells was reduced, but the cytoplasm was increased. The metarubricyte, segment granulocyte, apoptotic cells could be found. Morphologically and ultrastructurally, erythroid and myelocytic differentiation was observed. The positive level of benzidine staining in ASON group (11.90 +/- 2.30 at 24 h and 18.20 +/- 2.93 at 48 h) was higher than that of NSON group, lipofectin group and control group, respectively. The positive level of POX staining in ASON group (17.40 +/- 3.54 at 24 h and 29.40 +/- 3.70 at 48 h) was also higher than that of any other groups. The positive level of NBT staining in ASON group (7.50 +/- 2.26 at 24 h and 12.10 +/- 2.63 at 48 h) was significantly higher than that of NSON group, lipofectin group and control group, respectively (P < 0.01). In ASON group, the mean fluorescence intensity of CD33 (21.43 +/- 1.61 at 24 h and 14.86 +/- 1.20 at 48 h) was significantly lower than that of any other groups (P < 0.01). After thansfection for 24 h, the protein level of survivin in ASON group was decreased significantly compared to that of control group. There was no difference in survivin protein level amongst ASON group, NSON group and lipofectin group at 24 h (P > 0.05). Forty-eight hours after thansfection, the protein level of survivin was decreased significantly.</p><p><b>CONCLUSIONS</b>ASON targeting survivin can induce K562 to erythroid and myelocytic differentiation. Survivin is related to differentiation of K562 cells, and it can be a target of gene therapy for leukemia.</p>

Study Progress on Pathogenesis of Idiopathic Thrombocytopenic Purpura / 实用儿科临床杂志

Idiopathic thrombocytopenic purpura(ITP) as an autoimmune disease,is the most common clinical hemorrhagic disease.But,its pathogenesis is not completely clear yet.Low platelet count can easily repeate in chronic ITP patients,there are poor efficacy in refractory ITP patients,too.In recent years,a series of important progress on the pathogenesis of ITP had been made.In the humoral mechanism,there were significant progress on the emergence of autoantibodies;the theory on anomaly of the megakaryocyte quantity and quality caused by autoantibody were put forward.In the cellular mechanisms,the theory on reduction in the number of Tr cells,Co-stimulative signal and platelets directly dissolved by cytotoxicity T cells were put forward also.This review will be made to sum up the study progress on the pathogenesis of ITP.

Identification of two novel WASP gene mutations in 3 boys with Wiskott-Aldrich syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>The Wiskott-Aldrich syndrome (WAS) is a rare X-linked primary immunodeficiency caused by mutations in the WAS protein (WASP) gene. The disease is characterized by recurrent infections, eczema, and thrombocytopenia with small platelets, and it is known to be associated with extensive clinical variability, and mutation studies indicated that genotypes are also highly variant among WAS patients. The present study was conducted to identify the mutation types of Wiskott-Aldrich syndrome protein (WASP) gene in 3 boys suffering from Wiskott-Aldrich syndrome.</p><p><b>METHODS</b>Based on the typical clinical manifestations of Wiskott-Aldrich syndrome including thrombocytopenia, eczema, and recurrent infections and scanning electron micrographs, 3 patients were suspected of having WAS. The WASP gene of the 3 patients and their mothers were detected by PCR-direct sequencing analysis.</p><p><b>RESULTS</b>By sequence analysis using sense and antisense primer separately, the authors found two novel WASP gene mutations. For the twin brothers, a C deletion at nucleotide 984 was detected in exon 10 of WASP gene (984delC). The consequence of the C deletion involved frameshift mutation after H317 and premature stop at 444 (H317fsX444). Their mother was a carrier of the mutated WASP gene. For another WAS patient, a nonsense mutation with nucleotide substitution of G to T at position 1388 (1388G-->T) in exon 11 of WASP gene, led to premature translational termination at amino acid position 452 (E452X). His mother had not been found to have WASP gene mutation.</p><p><b>CONCLUSION</b>Genetic analysis is useful in definite diagnosis of Wiskott-Aldrich syndrome patients and in carrier detection and prenatal diagnosis, especially of atypical or sporadic WAS patients.</p>

Significance of Changes of Serum B Cell Activating Factor Belonging to Tumor Necrosing Factor Family in Children with Idiopathic Thrombocytopenic Purpura / 实用儿科临床杂志

Objective To explore the significance of the changes of serum B cell activating factor(BAFF) in children with idiopathic thrombocytopenic purpura(ITP).Methods The concentrations of BAFF were detected by ELISA in 42 children with ITP(test group) before and after treatment in the Children′s Hospital Affiliated to Chongqing Medical University from Apr.to Nov.2008,the blood platelet count(BPC) was detected also.And 40 children who were operated selectively were selected as control group.The variation of concentrations of BAFF were analyzed among children with ITP before,after treatment and control group.Moreover,the relationship between serum BAFF expression and BPC were analyzed by the Pearson test.Results The concentrations of serum BAFF were higher in children with ITP before treatment compared with that in control group [(0.943 3?0.583 5) ?g?L-1 vs(0.538 9?0.234 7) ?g?L-1,P0.05).There was negative correlation between serum BAFF expression and BPC(r=-0.305,P

Expression and Clinical Significance of Cyclin Kinase Inhibitor P21~(WAF1) and P27~(KIP1) in Children with Acute Leukemia / 实用儿科临床杂志

Objective To investigate the expression of cyclin kinase inhibitor P21~(WAF1) and P27~(KIP1)in children with acute leukemia and its clinical significance.Methods A total of 32 hospitalized children with acute leukemia(AL) were included in this study.Their bone marrow samples were collected before chemotherapy and individual patient was detected after complete remission(CR).The method of immunocytochemistry was used to estimate the expression of P21~(WAF1) and P27~(KIP1).Both positive percentage and intensity of the cells were counted.Results Findings showed that the positive ratios of P21~(WAF1) and P27~(KIP1) in total samples,ALL samples and AML samples were lower than the control group(P