Objective To investigate the role of signal transduction of toll-like receptors(TLRs)in immunological pathogenesis in children with acute idiopathic thrombocytopenic purpura(ITP).Methods Thirty children with actue ITP and 30 age-matched healthy children were studied.Real-time fluorescent quantitative PCR was used to evaluate the levels of TLR 1-10 and signal transducing molecules,and cytokines associated with TLRs,such as IL-1?,granulocyte-macrophage colony-stimulating factor(GM-CSF),macrophage colony-stimulating factor(M-CSF)and IFN-?/? mRNA.Expressions of co-stimulatory molecules such as CD40,CD80 and CD86 in mo-nocyte/macrophage(MC)was analyzed by flow cytometry.Results Compared with healthy control group,the mRNA levels of TLR3,7,8 and 9 in ITP group were significantly up-regulated(Pa0.05).Transcription le-vels of MyD88-dependent and-independent pathway molecules such as MD-2,MyD88,IRAK-4,TRAF6,TAK1,TRIF,TRAM,TBK-1 and IFN-? were significantly up-regulated in acute ITP(Pa0.05).Conclusion Aberrant activation of toll-like receptors signaling may be one of the initiating factors of immune dysfunction in children with acute ITP.

OBJECTIVE: To explore the effects of Tanreqing injection, a traditional Chinese herbal preparation for clearing heat and resolving phlegm, in treatment of acute exacerbation of chronic obstructive pulmonary disease (AECOPD) by improving airway inflammation and airway mucus hypersecretion. METHODS: A randomized controlled trial (RCT) was designed. Ninety AECOPD patients were randomly divided into Tanreqing group, ambroxol hydrochloride group and control group. The patients in the three groups were all treated with conventional therapy. Furthermore, intravenous drip infusion of 20 ml Tanreqing injection (once daily) and 15 mg ambroxol hydrochloride injection (twice daily) were administered respectively to the patients in the Tanreqing group and ambroxol hydrochloride group. They were all treated for 10 days. Symptom score of traditional Chinese medicine (TCM), plasma concentrations of interleukin-8 (IL-8), IL-10 and neutrophil elastase (NE) were detected before and after treatment. RESULTS: Cough, sputum amount, expectoration, dyspnea, fever, coated tongue and pulse tracings were improved obviously in Tanreqing group (P<0.05), and the effects of Tanreqing on improving cough, sputum amount and expectoration were better than the conventional therapy (P<0.05), while there was no significant difference between Tanreqing group and ambroxol hydrochloride group (P>0.05). Compared with ambroxol hydrochloride group and the control group, the coated tongue was improved obviously in Tanreqing group (P>0.05). After treatment, plasma concentrations of IL-8, IL-10 and NE were decreased in Tanreqing group and ambroxol hydrochloride group (P<0.05), and the levels of IL-8 and IL-10 in the control group were decreased (P<0.05). The change of IL-8 level before and after treatment in Tanreqing group was greater than that in ambroxol hydrochloride group and the control group. The changes of IL-10 and NE levels in ambroxol hydrochloride group were greater than those in Tanreqing group and the control group, while there was no significant difference in the changes of serum levels of IL-8, IL-10 and NE among the three groups (P>0.05). Total response rates in Tanreqing group and ambroxol hydrochloride group were higher than that in the control group (P<0.05), while there was no significant difference in total response rate between Tanreqing group and ambroxol hydrochloride group (P>0.05). There was no significant difference in total response rate among the three groups (P>0.05). CONCLUSION: Tanreqing injection can improve TCM signs and symptoms in AECOPD patients, and the mechanism maybe due to the decrease of serum levels of IL-8 and NE and improvement of IL-10 level.

OBJECTIVE: To evaluate the safety and efficacy of Chaige Qingre Granule, a traditional Chinese compound herbal medicine, in treating acute upper respiratory tract infection of wind heat syndrome. METHODS: A multi-center, double-blinded, randomized controlled trial was conducted. In the phase II, 60 patients with acute upper respiratory tract infection were randomly divided into the trial group (n=30) and the control group (n=30). In the phase III, 112 patients were randomly divided into the trial group (n=84) and the control group (n=28). The trial group received 6 g Chaige Qingre Granule, and the control group received 6 g Fufang Shuanghua Granule (another traditional Chinese compound herbal medicine). The two groups were all treated for 3 days and four times daily. Clinical symptoms, syndromes, adverse effect, blood, urine and stool test, hepatorenal function and electrocardiogram were examined before and after the treatment. RESULTS: After treatment, the overall obvious response rates of the trial group and the control group were 78.57%, 82.14% (by per-protocol analysis) and 75.86%, 79.31% (by intention-to-treat analysis) respectively, and the overall response rates of the two groups were 96.43%, 100% (by per-protocol analysis) and 93.10%, 96.55% (by intention-to-treat analysis) respectively in phase II. There were no significant differences between the two groups (P>0.05). In the phase III, the overall obvious response rates of the trial group and the control group were 90.54%, 73.08% (by per-protocol analysis) and 88.16%, 70.37% (by intention-to-treat analysis) respectively, and the overall response rates of the two groups were 94.59%, 96.15% (by per-protocol analysis) and 92.11%, 92.59% (by intention-to-treat analysis) respectively. There were no statistical differences between the two groups (P>0.05) too. No adverse effects were found in the trial. CONCLUSION: Chaige Qingre Granule is effective and safe in treating acute upper respiratory tract infection of wind heat syndrome.

Objective To investigate the clinical significance of universal newborn hearing screening for deaf‐ness predisposing genes in newborns .Methods A total of 965 newborns at Subei Hospital in Yangzhou were taken blood samples at heel and received for deafness predisposing genes screening .The most common deafness genes were detected by gene sequencing ,including mt12SrRNA c .1555A > G ,c .1494C > T ,GJB2 35delG ,167delT ,176_191del16 ,235delC ,299_300delAT ,SLC26A4 281C> T ,589G>A ,IVS7 -2A>G ,1174A> T ,1226G> A ,1229C> T ,IVS15+5G> A ,1975G>C ,2027T > A ,2162C> T ,2168A> G ,GJB3538C> T ,547G> A .At the same time ,all infants received hearing screening .Otoacoustic emission(OAE) was used as the first step screening ,and OAE combined with auto-auditory brainstem response(AABR) detection were used as the second step screening . Results Fifty -three cases (5 .49% ) had partial gene mutation ,one case of 12SrRNA gene mutation ,33 cases of GJB2 gene mutation ,18 cases of SCL26A4 gene mutation ,one case of GJB3 gene mutation .Of 965 cases ,28 cases failed to pass hearing screening while 18 cases did not pass rescreening .There were 10 cases taking audiological di‐agnosis at the age of three months .Six cases were confirmed with hearing loss .There were 905 cases passed thehearing screening and genetic screening ,11 failed born hearing and gene screening .Conclusion That the newborn gene screening was added into the hearing screening can be helpful to find out the deafness predisposing genes and drug -induced or late-onset hearing loss .

This study was aimed to investigate the expression, protein localization and function of Ahi-1 gene and its encoding protein in Jurkat cells. The expression of Ahi-1 mRNA and protein were measured by Northern and Western blot respectively. The plasmid containing prototype Ahi-1 was constructed and transfected into Jurkat cells. The Jurkat-A and Jurkat-C cells which either over-expressed the prototype Ahi-1 or not were obtained by selection with G418. The proliferation of the cells was assayed by XTT. The colony formation potential of the leukemia cells was checked by semisolid agarose culture. The results showed that three different transcripts of Ahi-1 (6.5,4.2 and 2 kb) were expressed in peripheral blood lymphocytes (PBLs), Jurkat and HUT78 cells. Ahi-1 protein with 140 kD localized in the cytoplasm majorly while traceless in the nucleus of Jurkat cells and PBLs from normal donor. Ahi-1 protein with 120 kD could be detected in the nucleus fraction of PBLs. Very low level of Ahi-1 protein with 120 kD was expressed in Jurkat cells. Up-regulating expression of Ahi-1 protein with 140 kD in the nucleus was found in Jurkat cells after exposure to meisoindigo, cytarabine, homoharringtonine, methotrexate and etoposide, down-regulating expression of Ahi-1 with 140 kD in the cytoplasm was observed after treatment with meisoindigo. The growth and colony formation potential were inhibited in the Jurkat-A cells, as compared to Jurkat-C cells. Total c-myb and phosphorylated AKT protein were continuously expressed in the Jurkat-C and Jurkat-A cells at similar level, but more phosphorylated c-myb was observed in Jurkat-A cells. It is concluded that three different transcripts of Ahi-1 at 6.5, 4.2 and 2 kb are detected in Jurkat cells; the Ahi-1 protein with 140 kD majorly expresses in the cytoplasm fraction and exposure to multiple chemotherapeutic compounds increased its expression in nucleus fraction. Over-expression of exogenous Ahi-1 can not only inhibit the growth and colony formation potential of Jurkat cells, but also induce the phosphorylation of c-myb in Jurkat cells.
Adaptor Proteins, Signal Transducing ; genetics ; Cell Proliferation ; Gene Expression ; Humans ; Jurkat Cells ; Plasmids ; Proto-Oncogene Proteins c-myb ; metabolism ; RNA, Messenger ; genetics ; Transfection

OBJECTIVE: To evaluate the safety and efficacy of Yiqi Pingchuan Granule in treating acute attack of asthma due to qi deficiency and cold syndrome. METHODS: A randomized controlled trial was conducted. A total of 80 patients with an acute attack of asthma were included. They were allocated into two groups randomly in a ratio of three to one. Sixty patients in the treatment group received Yiqi Pingchuan Granule and 20 patients in the control group received Ruyi Dingchuan Pill. Patients in both groups were treated for 7 days. RESULTS: There were no significant differences in traditional Chinese medicine syndrome, clinical symptoms and lung function between the two groups (P>0.05). After treatment, the forced expiratory volume in 1 second was increased in the treatment group (P<0.05), and the peak expiratory flow was accelerated in the control group (P<0.05). No significant side effects were noted in both groups. CONCLUSION: Yiqi Pingchuan Granule is safe and effective in treating acute attack of asthma due to qi deficiency and cold syndrome.

OBJECTIVETo discuss the mechanism of scar hypertrophy in adenosine receptor A(2A) (A(2A) R) knockout mice.

METHODSAnimal models of hypertrophic scar were established in 12 A(2A) R knockout mice and 12 wild-type mice as control. The thickness and the size of transverse section of the hypertrophic scar were observed by H-E staining. The hydroxyproline (HYP) in the scar was measured colorimetrically. The TGF-beta expression was tested by Western blotting method.

RESULTSThe hypertrophic scar in wild-type mice was more severe than that in knockout mice. Compared with self-control, the increase of the thickness and the size of transverse section of hypertrophic scar was markedly higher in wild-type group than in the knockout group (P < 0.01). There was significant difference in HYP content between the two groups (P < 0.01). Compared with self-control, the increase of TGF-beta expression in wild-type group was much more than that in knockout group (P < 0.01).

CONCLUSIONSThe TGF-beta expression decreases in the A(2A) R knockout mice. The scar hypertrophy is also much less in the A(2A) R knockout mice.

Animals ; Cicatrix ; metabolism ; pathology ; Disease Models, Animal ; Mice ; Mice, Knockout ; Receptor, Adenosine A2A ; genetics ; Transforming Growth Factor beta ; genetics ; metabolism

OBJECTIVETo study the expression of leukocyte-associated Ig-like receptor-1(LAIR-1) in children with immune thrombocytopenia (ITP), in order to explore the possible role of LAIR-1 in the pathogenesis of childhood ITP.

METHODSExpression levels of LAIR-1 on CD4(+) T cells, CD8(+) T cells and CD19(+)CD20(+) B cells of peripheral blood were measured in 40 children with ITP by flow cytometry. Serum level of solubility LAIR-1 (sLAIR-1) was measured using ELISA. Real-time PCR was used to measure LAIR-1 mRNA expression. Thirty-two healthy children served as the control group.

RESULTSThe percentages of CD19(+)CD20(+) B cells in the ITP group were significantly higher than in the control group (P<0.05). In contrast, the percentage of CD4(+) T cells in the ITP group was significantly lower than in the control group (P<0.05). The expression levels of LAIR-1 on CD4(+) T cells and CD8(+) T cells were significantly lower in the ITP group than in the control group (P<0.05). Serum sLAIR-1 level and LAIR-1 mRNA expression in the ITP group significantly increased compared with the control group (P<0.05).

CONCLUSIONSLAIR-1 expression on CD4(+) and CD8(+) T cells decreases and serum sLAIR-1 level increases in children with ITP, suggesting that LAIR-1 may play an important role in immune imbalance in these children.

CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; RNA, Messenger ; analysis ; Receptors, Immunologic ; blood ; genetics ; physiology

OBJECTIVETo obtain the monoclonal antibody against hexon protein of human adenovirus.

METHODSBALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.

RESULTSThe mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.

CONCLUSIONThe monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.

Adenoviruses, Human ; chemistry ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology

Adolescent ; Adult ; Aged ; Bacterial Infections ; drug therapy ; Bone Diseases ; drug therapy ; Child ; Female ; Humans ; Iodophors ; administration & dosage ; Joint Diseases ; drug therapy ; Male ; Middle Aged ; Suppuration ; drug therapy ; Therapeutic Irrigation