Effect of combined treatment with cyclophosphamidum and allicin on neuroblastoma-bearing mice

Objective:To evaluate the efficacy of allicin combined with cyclophosphamide on neuroblastoma(NB)-bearing mice and explore the immunological mechanism in it.Methods:A total of 30NB-bearing mice were equally randomized into model group,cyclophosphamide group and combined therapy group,10 nudemice were set as normal saline(NS) group.Cyclophosphamide group and combined therapy group were weekly injected with 60 mg/kg cyclophosphamide for four weeks;besides,combined therapy group was given with allicin(10 mg/kg/d) by gastric perfusion for 4 weeks;model group and NS group were given with the same volume of NS.Serum VEGF content was detected by ELISA pre-treating(0 d) and on the 3rd d.14 th d and 28 th d;on29th d,all mice were sacrificed and the tumor,liver,spleen and thymic tissues were weighted.Tumors were made into paraffin section for detecting tumor cell apoptosis and proliferation by TUNEI.and BrdU method,respectively.Survival curves were drawn by Kaplan-Meier method.Results:After treatment,both treatment groups relieved on viscera indexes,VEGF level,T cell subsets distribution and tumor growth and each index of combined therapy group was better than cyclophosphamide group(P【0.05 or 0.01);only combined therapy group could significantly increase the lifetime of NB-bearing mice(χ~2=5.667,P=0.017).Conclusions:Allicin can improve T cell subsets distribution and inhibit VEGF expression through its immunomodulatory activity,thereby improve the efficiency on NB in coordination with cyclophosphamide.

Effect of orthodontic force on inflammatory periodontal tissue remodeling and expression of IL-6 and IL-8 in rats

Objective:To investigate effect of orthodontic force on inflammatory periodontal tissue remodeling and expression of IL-6 and IL-8 in rats.Methods:Eighty SD rats were randomly divided into 4 groups,blank control group(group A)with 5 rats,treatment normal group(group B)with 25 rats,inflammation control group(group(group C)with 25 rats,inflammation treatment group(group D)with 25 rats.Immunohistochemistry and histomorphometric analysis was performed to measure the expression of IL-6,IL-8 and the firet molar to the recent movement in the distance.Results:The expression of IL-8 reached a maximum on day 5 and declined thereafter in group B;the expression of IL-6 reached a maximum on day 5 in group B.The expression of IL-6 and IL-8 was gradually weakened with time in group C.The expression of IL-6 and IL-8 were high,and reached a maximum on day 5 and declined thereafter in group D.AD of positive cells in group D were higher than group B at each time point(P<0.05).The time which 0.49 N orthodontic force was loaded was longer,orthodontic tooth movement distance was greater.Movement distance in group D were longer than group B(P<0.05).Conclusions:Orthodontic force as well as inflammatory stimulus can evoke the expression of IL-6 and IL-8.Under the combined effects of inflammation and orthodontic force,the expression of IL-6,IL-8will increase.

Causes of immune dysfunction in hyperbilirubinemia model rats

Objective:To explore the causes of immune dysfunction in neonatal rats with hyperbilirubinemia.Methods:A total of 60 newborn SD rats were equally randomized into normal saline(NS) group,LPS control group,bilirubin control group,low-dose group and high-dose group.After anesthesia,0.1 mL NS was given to the NS and LPS control group and different doses of bilirubin for the other groups;1 h later,the NS and bilirubin control group received the intraperitoneal injection of 0.05 mL NS and 1mg/kg LPS for the other groups.After 5 or 24 hours of model establishment,spleens were collected for detecting the expression levels of MyD88 and p-TAK1 protein and the spleen cells apoptosis by immunohistochemmistry and TUNEL method.After 24 hours of model establishment,scrum inflammatory factors levels and T cell subsets distribution were determined by ELISA and flow cytometry.Results:In contrast to low-dose bilirubin,high-dose bilirubin could induce spleen cells apoptosis in coordination with LPS.After 5 hours of model establishment,compared with NS group.MyD88 expression level in low-dose group elevated while p-TAK1 level in high-dose group reduced(P<0.05).In high-dose group,inflammotory factors levels and CD8^+T cells percentage were all higher than LPS control and NS group(P<0.05),while CD4^+ T cells percentage was lower than NS group(P<0.05).Conclusions:High-concentration plasma bilirubin in coordination with LPS could inhibit NF- κB signal pathways activation and aggravate inflammatory reaction,thus caused immunosuppression with inflammation cascade,which resulted in the immune dysfunction.

Study on the influence of curcumin on chemosensitivity of nephroblastoma cells

Objective:To study the influence of curcumin on chemosensitivity of nephroblastoma cells.Methods:Human nephroblastoma cells line SK-NEP-1 was transplanted to the nude mice subcutaneously to establish the implantation tumor model of human nephroblastoma cells.A total of 30 tumor-bearing mice were divided into three groups of ten randomly.The routine chemotherapy group was given vincristine(0.05 mg/mL·0.2 mL/d) and actinomycin D(15 ng/mL·0.2 mL/d) combined chemotherapy regime.The curcumin chemotherapy group was given the same combined chemotherapy regimens and curcumin(30 mg/kg/d) by intraperitoneal injection.The control group was given normal saline(NS) of the same volume by intraperitoneal injection.Continuous administration would be kept for 4 weeks and 3 days a week.The volumetric changes of every group were recorded.The serum of every group in different time was collected and the VEGF content was detected by ELISA.All mice were cercrificed and the tumor tissues were stripped and weighed after 4 weeks’ treatment.The tumor inhibition rate was calculated.The cell proliferation activity and apoptosis rate were detected by MTT and flow cytometry method.All data were statistically analyzed by SPSS 19.0.Results:The tumor volume,serum VEGF content,tumor inhibition rate,cell proliferation activity and apoptosis rate of routine chemotherapy group and curcumin chemotherapy group had significant differences comparing with the control group(P<0.05) after 4-week’s treatment.The cancer growth of curcumin chemotherapy group was obviously decreased and even tended to shrink comparing with routine chemotherapy group(χ2=15.732,P=0.007).The cell proliferation activity was significantly reduced and the apoptosis rate was significantly higher,(χ~2=9.427,P=0.012)which showing the effect of chemotherapy was enhanced.Conclusions:The chemosensitivity of nephroblastoma cells could be improved by curcumin,then the effect of preoperative adjuvant chemotherapy scheme would be enhanced,the growth of nephroblastoma cells would be inhibited and the surgical risk of nephroblastoma would be reduced.

ntravitreal Ranibizumab for Aggressive Posterior Retinopathy of Prematurity

Retinopathy ofprematurity （ROP） is a proliferative disease that affects infants of young gestational age （GA） and low birth weight （BW）. Aggressive posterior ROP （AP-ROP） is a rapidly progressing and severe presentation of ROP. It is characterized as posterior location （zone 1 or posterior zone I1）, Stage 3, and with plus disease （arterial tortuosity and venous dilation）. It can quickly lead to retinal detachment and blindness if not treated in time.

ASSOCIATION ANALYSIS OF POLYMORPHISMS IN SIX GENES WITHIN THE GH/IGF-1 AXIS IN PATIENTS WITH IDIOPATHIC SHORT STATURE IN THE CHINESE HAN POPULATION

Objective To investigate relationships of polymorphisms in six genes（GHR,IGF-1, IGF-1R,IGFBP-3,JAK2,and STAT5b）in the growth hormone（GH）/insulin-like growth factor-1 （IGF-1）axis with idiopathic short stature（ISS）in the Chinese Han population.Methods A casecontrol study was carried out on a cohort of 198 ISS patients and 306 healthy controls.A total of 106 tagging single nucleotide polymorphisms（tagSNPs）from the six genes were selected from the HapMap（haplotype map of the human genome）Han Chinese in the Beijing subset.Results of genotyping conducted by highthroughput Illumina GoldenGate?Assay were analyzed by statistical software.Results Both individual tagSNPs and haplotypes showed an association with ISS in the Han Chinese population（P <0.05）.For each single test,both allele and genotype were tested.By allele frequency analysis,six positive SNP sites （rsNo.l,rsNo.2,rsNo.3,rsNo.4,rsNo.5,and rsNo.6）of 3 genes（JAK2,IGF-1 R,and GHR）were found having associations with ISS.By genotype frequency analysis,there were significant differences between the patient and control groups in the following SNP sites;4 sites in JAK2 gene（rsNo.1,rsNo.2, rsNo.3,and rsNo.4）and 1 site in GHR gene（rsNo.6）.The risk which affected ISS was found related to the JAK2 gene in 4 sites（increase in rsNo.1 and decrease in rsNo.2,rsNo.3,and rsNo.4）and to the GHR gene in 1 site（decrease in rsNo.6）.They were four haplotypes in gene of IGF-1 R as "TGC","CGCT", "TA",and " CA",one haplotype in IGFBP-3 as "TA",and one haplotype in JAK2 as " CTG",which revealed high significance for risks of affecting ISS.At last,multivariate logistic regression analysis of specific site rsNo.6 of the GHR gene revealed that the serum IGF-1 was related to genotypes AA and AC, with genotype CC as the reference（P=0.015）.Conclusion Genetic variances in six genes within the GH/IGF-1 axis may be important etiological factors for ISS in the Chinese Han population.

Study on the Methodology of Developing Evidence-Based Clinical Practice Guidelines of Chinese Medicine

At present, evidence-based clinical practice guideline （EBCPG） is the main mode of developing clinical practice guidelines （CPGs） in the world, but in China, most of CPGs of Chinese medicine （CM） are still guidelines based on expert consensus. The objective of this study is to construct initially the methodology of developing EBCPGs of CM and to promote the development of standardization of CM. Based on the development of ＂Guideline for Diagnosis and Treatment of Common Pediatric Diseases in CM＂, the methodology of developing EBCPG of CM was explored by analyzing the pertinent literature and considering the characteristics of CM. In this study, the key problem was to put forward the suggestion and strategies. However, due to the methodology study of developing EBCPG of CM is still in the initial stage, there are still some problems which need further study.

Delayed-onset adenosine deaminase deficiency with a novel synonymous mutation and a case series from China

Background Adenosine deaminase(ADA)is a key enzyme in the purine salvage pathway.Genetic defects of the ADA gene can cause a subtype of severe combined immunodeficiency.To date,few Chinese cases have been reported.Methods We retrospectively reviewed the medical records of patients diagnosed with ADA deficiency in Beijing Children's Hospital and summarized the previously published ADA deficiency cases from China in the literature.Results Nine patients were identified with two novel mutations(W272X and Q202=).Early-onset infection,thymic abnor-malities and failure to thrive were the most common manifestations of Chinese ADA-deficient patients.The ADA genotype has a major effect on the clinical phenotype.Notably,a novel synonymous mutation(c.606G>A,p.Q202=)was identified in a delayed-onset patient,which affected pre-mRNA splicing leading to a frameshift and premature truncation of the protein.Furthermore,the patient showed γδT cells expansion with an increased effect or phenotype,which may be associated with the delayed onset of disease.In addition,we reported cerebral aneurysm and intracranial artery stenosis for the first time in ADA deficiency.Five patients died with a median age of four months,while two patients received stem cell transplantation and are alive.Conclusions This study described the first case series of Chinese ADA-deficient patients.Early-onset infection,thymic abnormalities and failure to thrive were the most common manifestations in our patients.We identified a synonymous muta-tion that affected pre-mRNA splicing in the ADA gene,which had never been reported in ADA deficiency.Furthermore,we reported cerebral aneurysm in a delayed-onset patient for the first time.Further study is warranted to investigate the underlying mechanisms.