Objective To investigate the effect of IVIG on pre-existing anti-HLA-A2 Ab levels and graft skin survival.Methods C57BL/6 wild type mice were sensitized to HLA-A2 by intraperitoneal injec-tion (IP) of HLA-A2 TgN mouse spleen cells (C57BL/6-TgN [HLA-2.1]1Enge SC) expressing human HLA-A2 at day 0, week 3 and 4.Sensitized mice were respectively treated with human IVIG , albumin, gly-cine, or PBS for 5 days during week 7.Skin transplantation from TgN mice to C57BL/6 wild type mice was performed at week 10 .Efficacy of IVIG DES was assessed by measuring anti-HLA Ab levels by ELISA pre-/post-Rx and graft skin survival was monitored daily post-Tx.Results Changes of HLA-A2-IgG levels:IgG HLA-A2 class I Ab levels in all groups predictably increased from baseline following spleen cell IP and peaked at week 4 ( P <0.01 ) , indicating sensitization .IVIG-treated mice showed significantly lower IgG Ab levels vs.albumin, glycine and PBS-treated at week 9 post-desensitization ( P <0.01).IgG quickly increased to extremely high levels in all groups following skin transplantation ( P <0.01 ) .Graft skin sur-vival between Rx-groups were similar with all rejecting at about 7 days post-transplantation ( P >0.05 ) . Conclusion However, IVIG alone does not inhibit anti-HLA class I Ab production after skin transplanta-tion or prolong SG survival , indicating combination of IVIG with other immunosuppressant or more optimal protocol for desensitization might show more efficacy and should be under explored .

BACKGROUND: Dopaminergic neurons differentiated from neural stem cells have been successfully used in the treatment of Parkinson's disease rats; however, the survival rate of transplanted cells has been low. Most cells die of apoptosis as a result of the formation of oxygen free radical and lipid peroxidation.OBJECTIVE: To observe resveratrol (Res) effects on survival of transplanted cells, transplanted efficacy and dopaminergic differentiation from neural stem cells in a rat model of Parkinson's disease.DESIGN, TIME AND SETTING: Randomized controlled animal experiments were performed at the Animal Experiment Center,Sun Yat-sen University from October 2007 to June 2008.MATERIALS: Thirty-two adult, healthy, male Sprague Dawley rats were equally and randomly assigned to model control, dopaminergic neuron, Res and combination groups. Four healthy Sprague Dawley rat embryos at gastational days 14-15 were selected and fetal rats were used for isolation and culture of neural stem cells. Res (Jingmal Biotech, Shenzhen, China) was used for this study.METHODS: Neural stem cells derived from the mesencephalon of embryonic rats were isolated and cultured in vitro, and passaged in serum-free culture medium containing epidermal growth factor and basic fibroblast growth factor, and then differentiated into dopaminergic neurons in differentia.ion medium. Parkinson's disease rat models were established by the injection of 6-hydroxydopamine in each group. Rats in the dopaminergic neuron group was injected with 3 pL cell suspension (1×10 cells/μL) containing dopaminergic neurons in the corpus striatum. Rats in the Res group received 3 μL of Ras (40 mg/L).Rats in the combination group were subjected to 3 μL of Res (40 mg/L) + 3 μL cell suspension (1×105 calls/μL) containing dopaminergic neurons. Rats in the model control group received 3 ×L of DMEM/F12 culture medium.MAIN OUTCOME MEASURES: The percentage of tyrosine hydroxylase-positive neurons in differentiated cells. The alteration of rotational asymmetry and the survival of tyrosine hydroxylase-positive neurons in graft areas of Parkinson's disease rats after transplantation.RESULTS: Flow cytometry demonstrated that survival rate of tyrosine hydroxytase-positive neurons was (17.8 ±4.2)% at 6 days following differentiation. Compared to the model control group, the rotational asymmetry was significantly improved at 10 days (P < 0.01), was significantly decreased at 20 days following transplantation in the combination group (P < 0.01). At 10-60 days following transplantation, the number of rotational asymmetry was significantly lower in the combination group than in the dopaminergic neuron group (P < 0.01). Tyrosine hydroxylase-positive neurons were not determined in the Res and model control groups. The number of tyrosine hydroxylase-positive neurons was significantly more in the combination group than in the dopaminergic neuron group (P < 0.01).CONCLUSION: Res can increase survival rate of transplanted cells in the corpus striatum, and improve rotational asymmetry in rat models of Parkinson's disease following transplantation of dopaminergic neurons differentiated from neural stem cells.Ke CL, Chen BL, Yu ZH, Huang ZS.Effects of resveratrol on the transplantation of neural stem call-derived dopaminergic neurons in a rat model of Parkinson's disease.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2009;13(27): 5281-5285.[http://www.crter.on http://en.zglckf.com]

Objective To investigate the diagnosis value of serum procalcitonin(PCT)detection combined with antibody de-tection of respiratory pathogens in children with pneumonia.Methods 1 256 cases of pneumonia in children were collected in the hospital from 2013 July to 2014 January were analyzed,detection PCT by chemiluminescence,using enzyme linked immu-nosorbent assay for detection of Mycoplasma pneumoniae ,Chlamydia pneumoniae and common respiratory virus.Results The positive rates of PCT was 30.7%,and MP IgM,CP IgM,RSV IgM,ADV IgM and FLUA IgM were 21.4%,16.8%, 11.8%,8.4%,and 10.9%.PCT in children with different gender in the largest number of positive cases,respectively,male 227 cases and female 158 cases,but the PCT and respiratory pathogen antibody positive in different gender did not exist sta-tistical difference (P >0.05).In different age groups,procalcitonin positive cases in 1 years in the group most,Mycoplasma pneumoniae and Chlamydia pneumoniae IgM positive cases in 1~3 years old were the largest number,the number of respir-atory virus positive patients with different maximum performance in different age groups.PCT and respiratory pathogen an-tibody positive in different age there was no statistical difference (P >0.05).Conclusion The PCT combined with respira-tory pathogen detection of antibodies help the clinic to determine the type of pneumonia in children,to provide help for the diagnosis and treatment of pneumonia in children.

ObjectiveTo compare the efficacy of step-up and top-down infliximab therapy on patients with Crohn's disease (CD).MethodsA prospective and open-label study was performed by the First Affiliated Hospital of SUN Yat-sen University during September 2007 to December 2010.Active CD patients who were refractory to steroid/immunomodulator or who were steroid-dependent were enrolled into step-up group.Active CD patients who had no steroid or immunomodulator therapy before were enrolled into top-down group. All patients were intravenously infused with infliximab of 5 mg/kg body weight in an induction regimen of 3 doses at week 0,2 and 6,followed by maintenance dosing every 8 weeks beginning at week 14.The clinical and endoscopic follow up lasted 30 weeks.Clinical symptoms and mucosal healing status under endoscopy were evaluated by follow-up at week 10 and 30.Results Forty-one CD patients were enrolled,with 24 in step-up group and 17 in top-down group. There were significant differences in disease duration (P =0.006),combination therapy (P ＜ 0.001 ) and severity of disease ( P =0.011 ) in baseline between step-up and top-down groups.At week 10 and 30 during treatment,the clinical remission rates in step-up group were 45.8％ (11/24) and 58.3％ (14/24) respectively; the mucosal healing rates in step-up group were 33.3％ (8/24) and 54.2％ (13/24) respectively; the clinical remission rates in topdown group were 70.6％ ( 12/17)and 82.4％ (14/17) respectively; and the mucosal healing rates in topdown group were 35.3％ (6/17) and 52.9％ (9/17) respectively.No significant differences in clinical remission and mucosal healing rates at both week 10 and 30 were observed between the two groups.The prevalences of adverse events in step-up and top-down group were 41.7％ (10/24) and 29.4％ (5/17)respectively ( P =0.422).ConclusionBoth step-up and top-down infliximab therapy can induce remission in more than half of CD patients,while top-down therapy might be more benefitiary to symptom and endoscopic remission.

Objective To investigate the efficacy and safety of a novel biologies-infliximab in the treatment of patients with Crohn's disease (CD). Methods A prospective study was conducted in 10 patients with CD(8 with active refractory CD and 2 with severe lower gastrointestinal bleeding caused by CD). All patients were intravenously infused with infliximab of 5 mg/kg body weight in an induction regimen of 3 doses at week 0, 2 and 6, followed by maintenance dosing every 8 weeks beginning at week 14. The clinical and endoscopic efficacy of infliximab were evaluated by follow-up of 30 weeks. Results ① Five out of 8 patients with active CD had initial clinical response at week 2. Clinical remission was found in 4 patients at week 30, (3 of them in symptomatic remission without corticosteroids). ② Two patients with severe lower gastrointestinal bleeding caused by CD were in control of bleeding and absence of further recurrence by 30 weeks follow-up. ③ Endoscopy was performed in 6 patients at week 30 to evaluate the healing of mucosal ulceration. Four patients were evaluated for complete or almost mucosal healing. ④ Adverse events were seen in 7 out of 10 patients with infliximab treatment, among whom 2 cases had severe side effect including pneumonia in one and delayed hypersensitirity reaction in the other. Conclusion Infliximab has efficacy for the induction and maintenance of remission in part of patients with active CD. Some patients achieved mucosal healing with infliximab therapy and low incidence of serious adverse events.

AIM: To observe the relationship between aortic atherosclerosis plaque formation and telomerase activation in vascular smooth muscle cells (VSMC). METHODS:Iliac artery atherosclerosis plaque from 20 renal failure patients and five cases of normal iliac arteries were obtained and confirmed by histological examination. The endothelial cells and the connective tissue on surface were lightly scraped and removed. Telomerase activity of the residual tissue was detected by telomeric repeat sequence amplification protocol. RESULTS:Five cases of normal iliac arteries showed negative telomerase activity. Eight of 20 cases athorosclerosis plaque showed positive telomerase activity, the other 12 cases showed negative telomerase activity. CONCLUSION: Telomerase activation may play a certain role in VSMC proliferation and atherosclerosis formation.

Objective:To investigate the phenotype and function of the intestinal γδT lymphocytes in post-infectious irritable bowel syndrome mouse model. Methods:The mouse model for post-infectious irritable bowel syndrome was established by the infection with trichinella spiralis. The intestinal inflammation,abdominal withdrawal reflex( AWR) and colon transportation test were observed. 2 and 8 weeks later,the animals were sacrificed and the lymphocytes in the intestinal lymph nodes and spleen were collected,from which the γδT lymphocytes were isolated and purified by monoclonal antibody-immuno-microbeads method. The functions of the purified γδT lymphocytes were evaluated,including proliferation by 3 HTdR;CD69,CD62L molecule staining by flow cytometry. Furthermore,the con-centration of cytokine IL-17 and IFN-γ in the supernatant of the cultured γδT lymphocytes were detected by ELISA. Results: At 2nd weeks after infection,significant intestinal inflammation was observed,with increasingγδT lymphocytes,proliferating and activating with increasing production of IL-17. At 8th weeks after infection, the intestinal inflammation disappeared, whereas the number of γδT lymphocytes remained increasing,also with proliferating and activating with increasing production of IL-17. Meanwhile,the mice show higher AWR score and Bristol score. Conclusion: γδT lymphocytes could participate in the pathogenesis of PI-IBS via their proliferation,activation and production of IL-17.

Objective To investigate the expression change of cytokines in peripheral blood and intestinal mucosa in the patients with diarrhea post-infectious irritable bowel syndrome(PI-IBS) and its relation with clinical symptoms scores.Methods Thirty outpatients and inpatients with diarrhea PI-IBS(observation group) and contemporaneous 30 individuals undergoing physical examination(control group) in the Hainan Provincial People's Hospital from January to December 2013 were selected.The peripheral blood mononuclear cells(PBMC) were separated and cultured.Then the levels of IFN-y and IL-10 in peripheral blood and cell culture supernatant fluid were detected by ELISA.The colonic mucosal tissue was taken by coloscopy.Then colonic mucosal IFN-γ and IL-10 protein expression was detected by immunohistochemistry staining.Furthermore,the correlationship between the level change of IFN-γ and IL-10 with clinical symptom score was analyzed by using the Spearman correlation method.Results Peripheral blod IL-10 and IFN-γ levels had no statistical difference between the two groups(P＞0.05).Compared with the control group,in PBMC seperation and cuture,the IFN-γ level in the observation group was increased and IL-10 level was decreased,the difference was statistically signifieant(P＜0.01).The intestinal main symptom score in the observation group had the positive correlation with IFN-γ expression level of PBMC culture supernatant fluid and colonic mucosal IFN-γ expression level(r=0.45,0.94,P＜0.01),and had the negative correlation with IL-10 expression level(r=-0.52,-0.79,P＜0.01).Conclusion The unbalance of IFN-γ and IL-10 level could be involved in the pathogenesis of diarrhea PI-IBS,which can serve as the observation indicators of disease activity.