Genetic analysis of a child with atypical Hemolytic uremic syndrome and nephrotic-range proteinuria / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical characteristics and genetic etiology for a child with atypical Hemolytic uremic syndrome (aHUS) in conjunct with nephrotic level proteinuria.@*METHODS@#A child patient who had visited the Affiliated Hospital of Qingdao University on June 25, 2020 was selected as the study subject. Clinical data of the patient was collected. Whole exome sequencing (WES) was carried out for the child, and candidate variant was verified by Sanger sequencing of the child and his parents.@*RESULTS@#The child, an 8-month-old male, had presented mainly with edema, oliguria, hematuria, nephrotic level proteinuria, anemia, thrombocytopenia, increased creatinine and urea, hypercholesterolemia but normal complement levels. Genetic testing revealed that he has harbored compound heterozygous variants of the DGKE gene, namely c.12_18dupGAGGCGG (p.P7fs*37) and c.1042G>T (p.D348Y), which were respectively inherited from his father and mother. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variants were classified as likely pathogenic and variant of uncertain significance, respectively. By combining his clinical manifestations and results of genetic testing, the child was diagnosed with aHUS with nephrotic level proteinuria.@*CONCLUSION@#For infants and young children with aHUS in conjunct with nephrotic level proteinuria, variants of the DGKE gene should be screened. Above finding has expanded the mutational spectrum of the DGKE gene.

Clinical characteristics and gene analysis of a family with autosomal dominant Alport syndrome with the onset of nephrotic syndrome / 中华实用儿科临床杂志

The clinical characteristics, mutation analysis results, and family tree of a patient with autosomal dominant Alport syndrome (ADAS), who had nephrotic syndrome as the first manifestation were examined.The proband was a 11-month-old girl who presented with nephrotic syndromes and gross hematuria.During the treatment course, the patient had steroid resistance and a poor response to Cyclosporine and Cyclophosphamide pulse therapy.Renal biopsy was performed 2 years after disease onset.Under the light microscopy, glomerular segmental mesangio-proliferative lesions were observed.The staining of type Ⅳ collagen showed the loss of the α3 chain in the glomerular basement membrane (GBM) and tubular basement membrane, and α5 chain loss in GBM.Electron microscopy showed uneven thickness of GBM, with obviously delaminated and tearing dense basement membrane (BM) layer, showing a typical lace-like change.The segmental BM was loosened and widened.Her father did not develop microscopic hematuria until 10 years later, while her grandmother had asymptomatic hematuria and proteinuria when the proband was diagnosed.A new mutation in the COL4A4 gene was found in the proband, namely c. 1715delG (p.G572Vfs * 81). Her father and grandmother carried the same mutation, but her mother and sister did not have.The clinical manifestation of ADAS is clinically heterogeneous and its incidence may be higher than what we have expected.

Two cases of adenosine deaminase 2 deficiency in children and literature review / 中华实用儿科临床杂志

Objective:To summarize the clinical phenotype and genotype features of 2 children with adenosine deaminase 2 (ADA2) deficiency, and to review the related literature so as to enhance the understanding of this disease.Methods:The phenotype and genotype of 2 cases with ADA2 deficiency who visited the Affiliated Hospital of Qingdao University from March to December 2019 were analyzed.Literature was searched from foreign and domestic databases and studied to summarize clinical and gene mutation characteristics of children with ADA2 deficiency.Results:(1) ADA2 gene mutation was found in both children.One case was characterized by recurrent fever, livedo reticularis, polyarteritis nodosa and immunodeficiency.The mutation site c. 571delC(p.Q191Sfs*5)of the ADA2 gene detected in this case was a homozygous mutation, which was a new mutation point and not reported in China or abroad previously.The other case was characterized by recurrent fever, panniculitis, vasculitis with legs, and immunodeficiency.The mutation site c. 1358A>G(p.Y453C)was a homozygous mutation that was not reported in China previously.(2)There were 171 cases of children diagnosed with ADA2 deficiency in foreign countries, but only 5 cases (3 previously reported cases and 2 cases in this study) were detected in China.The main clinical phenotypes were recurrent fever(5/5 cases), livedo reticularis(4/5 cases), panniculitis(1/5 cases), cutaneous gangrene(1/5 cases), growth retardation(1/5 cases), cerebral infarction(3/5 cases), humoral immunodeficiency(4/5 cases), blood system involvement(3/5 cases), and myalgia(2/5 cases), elevated inflammatory markers(C-reactive protein, erythrocyte sedimentation rate)(5/5 cases). Conclusions:Children with ADA2 deficiency have various clinical phenotypes, and a good understanding of phenotypes can improve the level of clinical diagnosis and treatment.The mutation point of c. 571delC is a novel ADA2 gene mutation type, which further enriches the ADA2 gene spectrum.

Effect of 1,25-dihydroxy vitamin D3 on food allergy in mice / 中华实用儿科临床杂志

Objective To investigate the effects of 1,25-dihydroxy vitamin D3 [1,25 (OH)2D3] on food allergy(FA) in mice and its mechanism.Methods A total of 40 BALB/c mice were randomly divided into 5 groups,8 in each group,including control group (group C) and FA model group (FA group),according to the dose of 1,25 (OH)2 D3 intervention,the mice of the FA group were divided into FA0 group (0),FA1 group [10 μg/(kg · d)],FAm group [50 μg/(kg · d)] and FAh group[100 μg/(kg · d)].Egg albumin was used to establish a food allergy model,with different doses of 1,25 (OH)2D3 for gastric intervention,and the control group was replaced by 9 g/L saline.The serum levels of ovalbumin-immunoglobulin E(OVA-IgE),interleukin (IL)-9 and IL-17 of mice were measured by using enzyme linked immunosorbent assay after the last excitation,and HE staining and histopathological examination were carried out in the small intestine of mice.Results Compared with group C,FAo group and FAh group small intestinal mucosa in mice had different degrees of damage,partial peeling off,structure disorder,villi epithelial cell focal falls peeling off,necrosis,lamina propria edema,congestion,a large number of inflammatory cells infiltration,low but the FA1 group and FAm group had light mucosa damage,intestinal epithelial basically intact,with integrity,no congestion,edema,and inflammatory cells infiltration to a lesser degree.The mean concentrations of serum IgE,IL-9 and IL-17 in different groups were statistically significant (F =40.770,9.530,5.624,all P ＜ 0.05).Compared with the FA0 group [(41.87 ±3.19) ng/L],the OVA-IgE of the FA1 group [(22.71 ±4.77) ng/L] and the FAm group [(16.34 ±2.81) ng/L] were significantly reduced (t =5.533,1 1.835,all P ＜ 0.01),but there was no significant difference between the FAh group [(36.29 ± 6.52) ng/L] (t =1.673,P ＞ 0.05).Compared with the FA0 group [(161.77 ±50.44) ng/L],the IL-9 levels of the FA1 group [(94.29 ± 18.79) ng/L] and the FAm group [(84.45 ± 30.88) ng/L] were significantly lower (t =3.267,3.366,all P ＜ 0.01),while that of the FAh group [(36.29 ±6.52) ng/L] was not significantly lower (t =0.777,P ＞0.05).Compared with FA0 group [(81.55 ±29.37) ng/L],IL-17 levels of FAh group [(133.58 ± 47.05) ng/L] was significantly increased (t =2.653,P ＜0.05),while IL-17 level of FA1 group [(79.41 ± 25.15) ng/L] and FAm group [(58.81 ± 26.00) ng/L] were lower than that of FAo group [(81.55 ±29.37) ng/L],but the difference was not statistically significant (t =0.154,1.640,all P ＞ 0.05).Conclusions Low,medium dose of 1,25 (OH) 2 D3 supplements can inhibit mice food allergies,but high doses of 1,25 (OH)2 D3 improve performance in mice food allergies,and 1,25 (OH)2 D3's influence on the secretion of IL-9 is one that influences mechanism of food allergy.

Effect of 1, 25-dihydroxy vitamin D3 on food allergy in mice / 中华实用儿科临床杂志

Objective@#To investigate the effects of 1, 25-dihydroxy vitamin D3[1, 25(OH)2D3] on food allergy(FA) in mice and its mechanism.@*Methods@#A total of 40 BALB/c mice were randomly divided into 5 groups, 8 in each group, including control group (group C) and FA model group (FA group), according to the dose of 1, 25(OH)2D3 intervention, the mice of the FA group were divided into FA0 group (0), FAl group [10 μg/(kg·d)], FAm group [50 μg/(kg·d)] and FAh group[100 μg/(kg·d)]. Egg albumin was used to establish a food allergy model, with different doses of 1, 25(OH)2D3 for gastric intervention, and the control group was replaced by 9 g/L saline.The serum levels of ovalbumin-immunoglobulin E(OVA-IgE), interleukin(IL)-9 and IL-17 of mice were measured by using enzyme linked immunosorbent assay after the last excitation, and HE staining and histopathological examination were carried out in the small intestine of mice.@*Results@#Compared with group C, FA0 group and FAh group small intestinal mucosa in mice had different degrees of damage, partial peeling off, structure disorder, villi epithelial cell focal falls peeling off, necrosis, lamina propria edema, congestion, a large number of inflammatory cells infiltration, low but the FAl group and FAm group had light mucosa damage, intestinal epithelial basically intact, with integrity, no congestion, edema, and inflammatory cells infiltration to a lesser degree.The mean concentrations of serum IgE, IL-9 and IL-17 in different groups were statistically significant (F=40.770, 9.530, 5.624, all P<0.05). Compared with the FA0 group [(41.87±3.19) ng/L], the OVA-IgE of the FAl group [(22.71±4.77) ng/L] and the FAm group [(16.34±2.81) ng/L] were significantly reduced (t=5.533, 11.835, all P<0.01), but there was no significant difference between the FAh group [(36.29±6.52) ng/L] (t=1.673, P>0.05). Compared with the FA0 group [(161.77±50.44) ng/L], the IL-9 levels of the FAl group [(94.29±18.79) ng/L] and the FAm group[(84.45±30.88) ng/L] were significantly lower (t=3.267, 3.366, all P<0.01), while that of the FAh group [(36.29±6.52) ng/L] was not significantly lower (t=0.777, P>0.05). Compared with FA0 group [(81.55±29.37) ng/L], IL-17 levels of FAh group [(133.58±47.05) ng/L] was significantly increased (t=2.653, P<0.05), while IL-17 level of FAl group [(79.41±25.15) ng/L] and FAm group [(58.81±26.00) ng/L] were lower than that of FA0 group [(81.55±29.37) ng/L], but the difference was not statistically significant (t=0.154, 1.640, all P>0.05).@*Conclusions@#Low, medium dose of 1, 25(OH)2D3 supplements can inhibit mice food allergies, but high doses of 1, 25(OH)2D3 improve performance in mice food allergies, and 1, 25(OH)2D3′s influence on the secretion of IL-9 is one that influences mechanism of food allergy.

Novel mutations of ITGB2 induced leukocyte adhesion defect type 1 / 中华儿科杂志

Objective@#To investigate the pathogenic mechanism of two novel ITGB2 mutations in leukocyte adhesion defect type 1 (LAD1).@*Methods@#The clinical history and blood sample of an 11 years old patient admitted to Affiliated Hospital of Qingdao University in August 2014 were collected. Expression of CD18 (encoded by ITGB2) was analyzed by flow cytometry. Novel ITGB2 mutations were identified by next-generation sequencing technology and confirmed by Sanger sequencing. The functional effect of ITGB2 mutations was detected by PolyPhen2. Expression vectors of both wild type and mutant ITGB2 were constructed and transfected into mammalian cells for analysis of protein stability and subcellular location.@*Results@#The symptoms of the patient (recurrent infections, lowered alveolar ridge and hypodontia) supported the diagnosis of LAD1. Expression of CD18 on the leukocytes was significantly decreased (0.2%) compared with the control samples from the parents (paternal: 99.0%; maternal: 99.1%). The patient was identified to be compound heterozygous for ITBG2 c.954del G (novel mutation) and c.1802C>A (paternal originated). ITGB2 c.954 del G was confirmed to be a harmful frameshift mutation; ITGB2 c.1802C>A was also predicted to be harmful. In terms of protein stability. There was no significant difference between mutant D18 and wild type. However, subcellular location analysis showed the mutant D18 could not locate on cell membrane.@*Conclusion@#The compound heterozygous of ITGB2 mutations (c.954del G and c.1802C>A) decreases the expression and impairs the location of CD18 on leukocytes, which leads to LAD1.

Influence of different doses of vitamin A supplements on the function of T help 17 and regulatory T cells in peripheral blood of asthmatic children / 中华实用儿科临床杂志

Objective To observe the effect of vitamin A(VitA) on T help 17(Th17) and regulatory T cells (Treg) in the peripheral blood in children with asthma and their dose effect relationship,and to investigate the immunoregulation mechanism of VitA.Methods Twenty children with asthma (asthma group) and 16 healthy children (healthy control group) were selected.Peripheral blood mononuclear cells(PBMC) were isolated from venous blood by density gradient centrifugation in the aseptic condition.Different concentrations of VitA [0.0μmol/L (blank control),0.5μmol/L,1.0μmol/L,2.0μmol/L] were added into the cultures in the asthma group.The healthy control group were not interfered with VitA.The supernatant was collected after 72 h.The levels of interleukin 17 (IL-17),interleukin 10(IL-10) and transforming growth factor-β1 (TGF-β1) in the supernatant were determined by enzyme-linked immunosorbent assay.Results (1) IL-17 levels produced by PBMC in the asthma group were significantly higher than those in the healthy control group [(960.53±75.59) ng/L vs (425.07±70.71) ng/L,P＜0.01],and the levels of IL-10 and TGF-β1 were significantly lower than those in the healthy control group [(53.13±6.94)ng/L vs (84.41±6.02) ng/L,(304.51±51.52) ng/L vs (489.45±73.68) ng/L,all P＜0.01].(2) IL-17 levels produced by PBMC in the 0.5μmol/L,1.0μmol/L and 2.0μmol/L VitA concentration of the asthma group [(588.95±44.18)ng/L,(573.13±27.43) ng/L,(686.71±38.98) ng/L] were significantly lower than those in the blank control group[(960.53±75.59) ng/L,all P＜0.01],and IL-17 levels in the 2.0 μmol/L VitA concentration were significantly higher than those in 0.5μmol/L and 1.0μmol/L concentration groop (P＜0.01).(3) IL-10 levels produced by PBMC in the 0.5μmol/L,1.0μmol/L and 2.0μmol/L VitA concentration of the asthma group [(105.35±10.79) ng/L,(111.21±16.11) ng/L,(81.09±6.05) ng/L] were significantly higher than those in the blank control group[(53.13±6.94) ng/L,all P＜0.01],TGF-β1 levels produced by PBMC in the 0.5μmol/L,1.0μmol/L and 2.0μmol/L VitA concentration of the asthma group[(933.01±73.98) ng/L,(1223.31±105.99)ng/L,(776.98±145.44) ng/L] were significantly higher than that in blank control group[(304.51±51.52) ng/L,all P＜0.01],and the levels of IL-10 and TGF-β1 in the 0.5 μmol/L and 1.0μmol/L concentration group were significantly higher than those in the 2.0μmol/L concentration group(all P＜0.01).The level of TGF-β1 in the 1.0μmol/L concentration group were significantly higher than that in the 0.5μmol/L concentration group (P＜0.01).Conclusions The function of Th17 in children with asthma during asthma attack was enhanced,and the function of Treg cells was reduced.The balance disorder of the functions of Th17 and Treg cells occurred.VitA can reduce the function of Th17 in peripheral blood,and enhance the activity of Treg cells in the children with asthma.The physiological level of VitA has the best effect,if high VitA concentration is high its effect is significantly decreased.

Clinical application of serum procalcitonin,high sensitive C reactive protein and interleukin-6 in children with acute respiratory infections / 国际检验医学杂志

Objective To investigate the clinical application of serum procalcitonin(PCT ) ,high sensitive C reactive protein(hs‐CRP)and interleukin‐6(IL‐6)in children with acute respiratory infections .Methods 136 cases of children with acute respiratory in‐fections were selected as observation group ,according to the results of the laboratory diagnosis ,which were divided into bacterial in‐fection group and viral infection group ,at the same time ,at the same period 83 cases of healthy children were selected as control group .Comparison of the level of PCT ,hs‐CRP and IL‐6 between groups .The level of PCT ,hs‐CRP and IL‐6 before and after treat‐ment was studied at the same time .The diagnostic value of PCT ,CRP ,IL‐6 was compared .Results The level of PCT ,hs‐CRP ,IL‐6 in bacterial infection group was significantly higher than the control group ,the difference was statistically significant(P<0 .05) . After treatment ,the level of PCT ,hs‐CRP ,IL‐6 was lower than that before treatment ,the difference was statistically significant (P<0 .05) .The sensitivity ,specificity ,the positive prediction rate ,negative prediction rate and Youden index of PCR was relatively higher compared with the other two index ,the difference was statistically significant(P<0 .05) .Joint detection of sensitivity had improved ,while the specific degree and Youden index was reduced .Conclusion The levels of PCT ,hs‐CRP ,IL‐6 in children with a‐cute respiratory infections have important clinical significance .PCT is better and more real as index for judgement for bacterial in‐fection .Joint detection and comprehensive evaluation of three indicators could significantly improve the sensitivity of the acute re‐spiratory infections ,which is helpful for early diagnosis and prognosis evaluation .

Influence of Azithromycin on peripheral blood helper T lymphocyte 9 cells and helper T lymphocyte 17 cells in ;children with bronchial asthma / 中华实用儿科临床杂志

Objective To observe the influence of Azithromycin on helper T lymphocyte 9 cells ( Th9 ) and Th17 in peripheral blood of children with bronchial asthma,and to investigate the immunomodulating effect of Azithro-mycin. Methods Twenty-six asthmatic children were selected as the experimental group,and 17 healthy children as a control group. Peripheral blood mononuclear cells(PBMC)were isolated from venous blood by density gradient cen-tri-fugation under aseptic conditions. PBMC were split by phytohemagglutinin( PHA) in vitro. Different concentrations of Azithromycin (0,0. 1,1. 0,10. 0 mg/L)were added into the cultures in the experimental group. The control group was not interfered with azithromycin. The supematant was collected after 72 h. The levels of interleukin ( IL)-4, IL-9,IL-17 and IL-23 in the supematant were determined by enzyme-linked immunosorbent assay(ELISA). SPSS 17. 0 software was used to analyze data. Results (1) The levels of IL-4,IL-9,IL-17 and IL-23 produced from PBMC of the experimental group were significantly higher than those in the control group(t=9. 210,3. 040,8. 965,2. 796,P0. 05). The levels of IL-17 and IL-23 of Azithromycin 10 mg/L were lower than those in Azithromycin 0 mg/L and 0. 1 mg/L(P0. 05). (3)IL-9 level was negatively correlated with IL-4 level in the experimental group(r=-0. 255,P>0. 05). The levels of IL-23 and IL-17 secreted by Th17 in asthmatic chil-dren had correlation. The secretion of IL-17 levels rose with the secretion of IL-23 rise(r=0. 64,P<0. 05). Conclu-sions The function of Th9 and Th17 in asthmatic children was enhanced;Azithromycin could restrain the function of Th9 and Th17 at higher tissue concentrations (10. 0 mg/L),the former was unrelated to changes in the secretion of IL-9 levels,and the latter with the secretion of IL-23 levels decreased close. Azithromycin can play a role of immune modulators in higher concentrations,inflammatory cytokines are inhibited in the asthmatic children,and Azithromycin relieves airway inflammation.

Laboratory examination of autoimmune and autoinflammatory diseases / 中华实用儿科临床杂志

Autoimmune disease is an own organization inflammatory lesions,mainly caused by destroying the adaptive immune tolerance mechanism of differentiatingself andnon-self,whose character is appearing the autoantibodies and self-reactive T cells in the body.Autoinflammatory disease is a group of genetic,recurrent and noninvasive inflammatory disease,whose characteristics are fever,rash,joint pain,arthritis,ophthalmic pathological changes and increasing of acute phase proteins,and it can affect many organ systems.These diseases are different in the mode of onset and clinical manifestations,but also can have similar and overlapped symptoms and signs,and often confused with other systemic diseases.Therefore,clinical misdiagnoses or missed diagnoses easily occur.To understand correctly and master the laboratory examination characteristics and its clinical is essential,which has significant value in the clinical diagnosis,differential diagnosis,evaluation and treatment of these diseases.

Significance of the expression of Toll-like receptor and costimulatory molecule of dendritic cells in the peripheral blood in children with Henoch-Sch(o)nlein purpura / 中华实用儿科临床杂志

Objective To observe the expression of Toll-like receptor(TLR) on peripheral blood dendritic cells(DC) in children with Henoch-Schtinlein purpura(HSP),and to investigate the pathogenesis of the abnormal expression of TLR in children with HSP.Methods Twenty hospitalized children with HSP in the Affiliated Hospital of Qingdao University Medical College from Dec.2011 to Jul.2012 were enrolled in the study(HSP group).Twenty agemetched healthy children were selected as a healthy control group.Peripheral venous blood was sampled under aseptic condition,peripheral blood mononuclear cells (PBMC) were isolated from density gradient centrifugation,and DC were generated by recombinat human granulocyte-macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α) in vitro.Expressions of CD83,CD86 and TLR2,TLR3,TLR4 in peripheral blood DC were examined by fluorescent activated cell sorter (FACS).Results 1.No significant distinction was found in the expression of the C Ds3 on peripheral blood DC between HSP group and healthy control group(t =0.80,P ＞ 0.05) ;in HSP group had remarkably increased expression of the CD86 on peripheral blood DC than that of the healthy control group (t =9.56,P ＜ 0.01).2.Expression rates of TLR2,TLR3,TLR4 on peripheral blood DC in the HSP group were higher than those in the healthy control group(t =1 1.79,13.29,9.45,all P ＜ 0.01).3.Expression rates of TLR2,TLR3 and TLR4 in HSP group had positive correlation with expression rates of CD86 (r =0.84,P ＜ 0.01 ; r =0.53,P ＜ 0.05 ; r =0.66,P ＜ 0.05).Conclusions Expressions of TLR2,TLR3 and TLR4 on peripheral blood DC significantly increased and were positively correlated with expression of CD86.This implies that TLR and co-stimulatory molecules might participate in the pathogenesis of HSP by mediating signal transduction,leading to abnormity of cytokines,then inducing Th1/Th2 immune imbalance by showing the advantage of Th2 function.

Expression of Toll-like receptor 2, Toll-like receptor 4 and Toll-like receptor 6 in peripheral blood mononuclear cells and their relationship with Treg immune response in children with Henoch-Schonlein purpura / 中国组织工程研究

BACKGROUND:The influence of Tol-like receptor 2 (TLR2), Tol-like receptor 4 (TLR4), Tol-like receptor 6 (TLR6) signal transduction pathway and active Treg in children with Henoch-Schonlein purpura has been unknown. OBJECTIVE:To investigate the expression of TLR2, TLR4 and TLR6 in peripheral blood mononuclear cells and Treg immune response in patients with Henoch-Schonlein purpura, and to explore the role of TLR2, TLR4, TLR6 and Treg activation in the pathogenesis of Henoch-Schonlein purpura. METHODForty-two hospitalized children with Henoch-Schonlein purpura were enrol ed in this study. Another 15 healthy children were selected as controls. TLR2, TLR4 and TLR6 at protein level in peripheral blood mononuclear cells were detected by flow cytometey;reverse-transcription PCR and real-time PCR were used to evaluate the level of MyD88;the levels of transforming growth factor-βand interleukin-10 were measured by enzyme-linked immunosorbent assay. t-test or t’-test was used to compare the levels of these genes and proteins. Pearson’s correlation test was done for correlation analysis. RESULTS AND CONCLUSION:Compared with the control group, the protein levels of TLR2, TLR4, TLR6 and the relative expression level of MyD88 mRNA were significantly up-regulated (P0.05). The excessive activation of TLR2, TLR4, TLR6 may be involved in the process of Henoch-Schonlein purpura via MyD88-dependent pathway, and the compensatory activation of Treg may participate in protective immunity.

Expression of Toll-like receptor 3 and Toll-like receptor 4 in peripheral blood mononuclear cells from Henoch-Sch(o)nlein purpura nephritis in children and its clinical significance / 中华实用儿科临床杂志

Objective To explore the expressions and clinical significance of Toll-like receptor (TLR) 3 and TLR4 in peripheral blood mononuclear cells in Henoch-Schsnlein purpura nephritis (HSPN) children.Methods According to their 24-hour urinary albumin and whether children with HSP had renal damage or not,105 cases were divided into group A,B and C.Group A were children only with HSP but without renal damage,while group B were children only with HSPN not proteinuria and group C were children with both HSPN and proteinuria.Thirty healthy children were in healthy control group(group N).Flow cytometry (FCM) and real-time PCR detected the mRNA and protein expressions of TLR3 and TLR4 in peripheral blood mononuclear cells.Results 1.The mRNA and protein expressions of TLR4 in peripheral blood mononuclear cells were significantly higher in group A,B,C than those in group N (F =37.33,24.01,all P ＜ 0.05).The mRNA and protein expressions of TLR4 in group C were much higher than those in group A and B (all P ＜ 0.05).Meanwhile,there was no significant difference between group A and B(all P ＞0.05).2.Moreover,there was a positive relationship between protein expression of TLR4 and 24-hour urinary albumin in group C(r =0.69,P ＜ 0.01).3.Expression of TLR3 was of no significant difference in all groups(F =0.86,1.78,all P ＞ 0.05).4.The expression of TLR4 mRNA had a positive correlation with protein expression of TLR4(r =0.61,P ＜ 0.0 1).Conclusions Expressions of TLR4 in peripheral blood mononuclear cells significantly increased and had a positive correlation with urinary protein excretion from HSPN in children.This implied that aberrant activation of TLR4might be relevant to the development of HSPN.

Study on the expression of TLR2 and TLR4 in peripheral blood mononuclear cells and their relation- ship with Th1/Th2 immune response in patients with Henoch-Schonlein purpura / 中华微生物学和免疫学杂志

Objective To explore the role of TLR2 and TLR4 in the pathogenesis of Henoch-Schonlein purpura ( HSP) by investigating their expression at mRNA and protein levels in peripheral blood mononuclear cells ( PBMCs ) and their influences on Th 1/Th2 immune response in children with HSP . Methods 64 hospitalized children with HSP in the Affiliated Hospital of Qingdao University Medical Col -lege from October 2011 to November 2012 were enrolled in the study .They were further divided into non-He-noch-Schonlein purpura nephritis ( NHSPN ) group ( n =36 ) and Henoch-Schonlein purpura nephritis (HSPN) group (n=28).30 age-matched healthy children from Child Health Division of the same hospital were selected as controls .The expression of TLR2 and TLR4 at mRNA level in PBMCs were detected by re-al-time fluorescent polymerase chain reaction .The expression of TLR2 and TLR4 at protein level and T cells subset were detected by flow cytometry .The levels of IFN-γ, IL-4 and IL-6 in plasma were determined by enzyme-linked immunosorbent assay (ELISA).Results (1)Compared with the control group , the expres-sion of TLR2 and TLR4 at mRNA and protein levels were remarkably increased in children with HSP , espe-cially in HSPN group.(2)Compared with the control group, the percentage of CD3+T cells and CD3+CD4+T cells were down-regulated in HSP group , but the percentage of CD 3+CD8+T cells and CD3+HLADR+T cells were up-regulated.(3)The level of IFN-γand the ratio of IFN-γ/IL-4 in plasma from children with HSP were significantly lower than those of the controls , while the level of IL-4 and IL-6 were remarkably higher than those of the controls .(4)The expression of TLR2 and TLR4 at protein level in PBMCs from chil-dren with HSP showed significant positive correlations with the expression of TLR 2 and TLR4 at mRNA level and plasma concentration of IL-4 and IL-6, but a negative correlation with the ratio of IFN-γ/IL-4.Conclu-sions The aberrant activation of TLR 2 and TLR4 might be correlated with the immunological pathogenesis of HSP by enhancing Th2 immune response.The hyper-activation of TLR2 and TLR4 might result in renal injury in patients with HSP .

An in vitro study on influence of intravenous immunoglobulin on function of T cells in cord blood of preterm infants / 中华围产医学杂志

Objective To observe the influence of different doses of intravenous immunoglobulin (IVIG) on function of T cells in cord blood of premature infants and to explore the immunomodulating mechanism of IVIG.Methods Cord blood mononuclear cells (CBMC) were isolated by density gradient centrifugation method from 15 preterm infants born between June 1,2011 to December 31,2011 at 32 34 gestational weeks.Three groups were formed according to the concentrations of IVIG used for CMBC culturing in vitro (Group A,6 mg/ml; Group B,9 mg/ml; Group C,12 mg/ml).After 72 hours in sterile conditions,the levels of IFN 7,IL 4,IL-10 and TGF β1 in the supernatant were determined by enzyme-linked immunosorbent assay.The expression of CD4+ CD25+ Foxp3+ Treg cells was examined by fluorescent activated cell sorter.Differences among groups were compared by one way analysis of variance.For comparison between two groups,LSD test was used.Results (1) The level of IFN γ secreted by CBMC in Group C was 0.03 ± 0.02,which was much lower than that in Group A(0.18±0.08) and Group B(0.13±0.05) (F=5.72,both P＜0.01).The level of IFN-γ in Group B and Group A did not show statistical difference (P＞0.05).Compared with Group A(0.03±0.01),the level of IL4 was much lower in Group B (0.02±0.01) and Group C(0.01±0.01) (F=20.38,both P＜0.01),while no significant difference was shown between Group B and Group C (P＞0.05).(2) No significant difference was found in the expression of CD4+CD25+Foxp3+Treg cells among different groups (F 0.67,P＞0.05).(3) The level of IL 10 in Group C was 3.02 ± 3.79,which was significantly lower than that in Group A (10.78±5.44) and Group B (6.90±4.64)(F=4.68,P＜0.01 and 0.05).The level of IL-10 inGroup B was still lower than that in Group A (P＜0.05).The levels of TGF-β1 in Group C(8.44± 13.71) and Group B(16.15 ±13.94) were significantly lower than that in Group A(30.23 ± 16.32) (F=5.22,P＜0.01,P＜0.05),but there was no significant difference between Group B and Group C (P＞0.05).Conclusions IVIG might inhibit the function of Th cells in CBMC of preterm infants in a dose dependent manner.IVIG could inhibit the function of natural Treg cells by regulating the secretion of cytokines IL-10 and TGF-β1 in a dose dependent effect.

Observation on the Curative Efficacy of Wentong Babu Plaster for Infant Diarrhea / 中国药房

OBJECTIVE:To study the curative efficacy of Wentong babu plaster for infant diarrhea.METHODS:A total of 62 infants with diarrhea were randomly assigned to either treatment group(n=32)or control group(n=30).Both groups were given Dioctahedral smectite,Bifid triple viable,fluid replacement,diet structure adjustment,etc.And the treatment group received additional Wentong babu plaster once daily for 5 days.The curative efficacy and the side effects of the two groups were observed.RESULTS:There are significant difference between the two groups in marked efficacy showing rate and response rate,both higher in treatment group than in control group(P

Effects of astragalus membranaceus on the apoptosis of cord blood lymphocytes in neonates / 中华围产医学杂志

Objective To study the effects and the mechanism of astragalus menbranaceus (AM) on neonatal lymphocyte apoptosis in cord blood. Methods Cord blood mononuclear cells (CBMC) of 30 full-term neonates were cultured with pure phytohemagglutinin (PHA), PHA combined with IL-6 (PHA + IL-6) or AM (PHA + AM) respectively for 48 hours. The apoptosis index (AI) of lymphocytes of CBMC after cultivation were determined by acridine orange-ethidium bromide dying method. The positive expression of CD38 antigen and CD25 antigen were detected by indirect immunofluorescence procedure. The levels of IL-6 in the supernatants of CBMC were detected by ELISA . Results (1) The AI of the PHA + AM group (16. 5?3. 5)% and PHA+IL-6 group (16. 9?4. 0)% was lower than that of the pure PHA group (32.4?2.8)% (P0. 05). (2) The positive expression rates of CD38 of the PHA+AM group and the PHA+IL-6 group were lower than those of the pure PHA group (P0. 05). The positive expression rates of CD25 of the PHA + AM group and the PHA + IL-6 group were higher than those of the pure PHA group (P0. 05). The positive expression of CD38 of CBMC had positive correlation, but CD25 had negative correlation with the AI of CBMC(r=0. 68, -0. 65,P

Preliminary Observation of Apoptotic Feature of T Lymphocyte in Preterm Neonates / 中华围产医学杂志

Objective To study the features and its effect factors of T lymphocyte apoptosis in preterm neonates. Methods The susceptibility to apoptosis of T lymphocytes were determined in 30 full-term neonates and 19 preterm neonates by percentage calculation of apoptosis cell and DNA fragmentation analysis. The expression of CD 25 and CD 38 in peripheral blood mononuclear cells and the volume of IL-2 and IL-6 produced by lymphocytes incubated for 48 h was determined by immunofluorescence procedure and ELISA. Results T lymphocyte apoptosis was easier in preterm(27.2?2.7)%,(34.8?3.9)% than in full-term neonates(21.1?1.4)%,(27.9?2.3)% at incubating 24 h, 48 h(t=10.547,7.789;P