OBJECTIVE To study the effect of optimal decontamination methods on the management of implantation instruments of orthopedics.METHODS To clean the instruments by the Campany(control group) and by the nurse after hospital training(management group),and compare the decontamination quality of the two methods by visual inspection using magnifying glass.RESULTS The qualification rate of instruments of the management group was more than control group(P

Objective To characterize the clinical features and outcome of massive lower gastrointestinal hemorrhage in patients with Crohn's disease.Methods The clinical data form 13 patients Who identified as Crohn's disease accompanied by massive lower gastrointestinal hemorrhage between 1998 and 2007 were retrospectively analyzed.Results The ratio of men to women was 3.3 : 1.Bleeding was the first symptom of disease in foor patients.The lesion was involved in small howel in 10 cases.The origin of bleeding was identified in 8 cases.Four patients received surgical operation and none of them died,while 9 patients received medical treatment and 3 of them died.Totally 10 patients were followed up for 6 to 94 months.Eight patients had recurrent massive hemorrhage,and two of them subsequently required surgery.One patient died after surgery.All patients who had recurrent massive bleeding and still survival were characterized by recurrent bleeding as only or major clinical feature,and they were all with small bowel involvement.Conclusions Diagnostic and therapeutic approach of massive lower gastrointestinal hemorrhage in Crohn's disease is very challenging.It is possible that there exists a particular phenotype of Crohn's disease that presents a tendency of bleeding recurrence,whose bleeding probably related to small bowel lesions.

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 .

Objective To analyze the clinical features,clinical prognosis and predictive factors of ulcerative colitis (UC) complicated with cytomegalovirus (CMV) infection.Methods From May 2004 to November 2014,120 hospitalized patients diagnosed as UC and screened for CMV infection were enrolled.A total of 31 patients with moderate to severe UC accompanied by CMV infection were screened out.Demographics,clinical features,endoscopic appearance and treatment of patients with UC complicated with CMV infection were analyzed,and compared with 60 moderate to severe UC patients without CMV infection at the same period of hospitalization.Mann Whitney U test was performed for statistical analysis.Logistic regression analysis was used to analyze risk factors of UC complicated with CMV infection.Results Among 120 patients with UC,29 were mild or in remission period,whose CMV screening tests were all negative.Ninety-one were moderate to severe,31 patients (34.1 ％) of them had CMV infection,and 20 of 31 patients were steroid-refractory.Among the 31 patients with UC complicated with CMV infection,median age was 39 years (22 years,51 years),median disease duration was 24.0 months (6.0 months,42.0 months) which was shorter than that of patients without CMV infection (36.0 months (13.5 months,84.0 months)),and the difference was statistically significant (U=639.5,P=0.015).A total of 23 patients (74.2％) had extensive colitis and 26 patients (83.9％) had history of severe colitis.A total of 29 patients (93.5％) had history of corticosteroids treatment,12 patients (38.7％) had history of immunosuppressive agents treatment,and six patients (19.4 ％) had history of infliximab treatment.Compared with UC patients without CMV infection,fever,abdominal pain and weight loss were more common in UC patients with CMV infection.Five CMV-infected patients had mild liver dysfunction.Endoscopic appearance was longitudinal ulceration,irregular ulceration,large deep ulceration,punchedout ulceration and worm-like ulceration.Among the 25 CMV-infected patients who were treated with corticosteroids,11 patients (44.0％) had no response.Among the 39 CMV-negative patients who were treated with corticosteroids,eight patients (20.5％) had no response.The rate of patients who needed rescue therapy of the former was higher than that of the latter,and the difference was statistically significant (x2 =4.026,P=0.045).The results of multivariate Logistic regression analysis showed that hemoglobin over 100 g/L(OR=0.144,95％ confidence interval (CI) 0.040 to 0.516,P=0.003) was a protective factor of CMV infection,however corticosteroids use within a month before the onset (OR=8.946,95％oCI 2.459 to 32.541,P=0.001) was a risk factor.Conclusions UC patients treated with corticosteroids and immunomodulator therapy may predispose UC patients to CMV infection,on the other hand,CMV infection can exacerbate the severity of UC.CMV infection should be screened and monitored in UC patients,and anti viral therapy should be taken in time in case of CMV infection.

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]

BACKGROUND: Recently, several scientists have found that differentiation of neural stem cells (NSCs) towards dopaminergic neurons may be increased in vitro by combination of some special cytokines. They have also found that dopaminergic neurons differentiated from NSCs can be used for the treatment of Parkinsn's disease. To improve the therapeutic effects of/n vitro transplantation, we should further study the biologic characteristics of NSCs at the induction and differentiation.OBJECTIVE: To explore the differentiation of NSCs which were incubated in differentiation solution for different time towards dopaminergic neurons in vitro.DESIGN: Single sample observation.SETTING: Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University.MATERIALS: This study was performed at the Department of Ncurosurgery, First Affiliated Hospital of Sun Yat-sen University from May to October 2007. Six healthy Sprague Dawley (SD) rats, gestational age 14 days, of clean grade, weighing 350-400 g,were provided by the Laboratory Animal Center, Sun Yat-sen University[permission No. SCXK (yue)2007-0034]. The protocol was performed in accordance with ethical guidelines for the use and care of animals.METHODS: NSCs derived from rat embryonic mesencephalon were cultured in serum-free culture medium containing epidermal growth factors and basic fibroblast growth factors. After passage, the NSCs were induced to differentiate towards dopaminergic neurons in the differentiation medium supplemented with interleukin 1o, interleukinl 1, human leukaemia inhibitory factors, and glial cell line-derived neurotrophic factors. The percentage of tyrosine hydroxylase positive neurons in differentiated ceils was detected with flow cytometer when NSCs were cultured in differentiation solution for 2, 4, 6, 8 and 10 days, respectively.MAIN OUTCOME MEASURES: Cellular morphological alteration of rat NSCs after differentiation. The percentage of tyrosine hydroxylase positive neurons in differentiated cells derived from NSCs.RESULTS: In differentiation medium, NSC spheres attached the bottom of plates and began to collapse. Cells inside the spheres grew out gradually and became irregular in shape. Six days later, most of the cells had I or 2 long processes and a few short processes. The percentage of tyrosine hydroxylase positive neurons in differentiated cells was respectively (3.2_+_0.9)%, (6.8 +1.6)%, (16.7-+2.6)%, (14.8_+1.8)% and (12.2_+2.5)% after culture for 2, 4, 6, 8 and 10 days, with significant differences (F =26.449, P ＜ 0.05).CONCLUSION: Induction time influences the differentiation of NSCs towards dopaminergic neurons in vitro. The percentage of dopaminergic neurons is the highest in differentiated cells derived from NSCs which are cultured in differentiation solution for 6 days.

BACKGROUND: Differentiation inducing factors and gestational age influence the differentiation potential of embryonic neuralstem cells.OBJECTIVE: This study was designed to observe the differentiation potential of rat mesencephalic neural stem cells at differentgestational ages towards dopaminergic neurons.DESIGN: A randomized controlled observation.SETTING: Department of Neurosurgery, First Affiliated Hospital of Sun Yat-sen University, Guangzhou City, GuangdongProvince, China.MATERIALS: This study was performed at the Laboratory of the First Affiliated Hospital of Sun Yat-sen University between Marchand September 2007. Thirty adult gestational SD rats, weighing 350 400 g, were provided by the Laboratory Animal Center of SunYat-sen University (Permission No. 2007-0034). The protocol was performed in accordance with ethical guidelines stated in Guide forthe use and care of laboratory animals, approved by the Committee on the Care and Use of Laboratory Animals of the Institute ofLaboratory Animal Resources Commission on Life Sciences, National Research Council, China (1985). DMEM/F12 serum-free medium,B27 additives, epidermal growth factor, basic fibmblast growth factor, and fetal bovine serum (volume fraction:0, 1) were purchased fromGibco Company, British; Interleukin lα, interleukin 11, and glial cell-derived neurotrophic factors were purchased from R&D Company,USA; In addition, leukaemia inhibitory factor (Perpotech, British), tyrosine hydroxylase(Santa Cruz, USA), nidogen antibody,microtubule-associated protein 2 antibody, and glial fibrillary acidic protein antibody(Chemicon, USA) were also used.METHODS: Six rats were randomly selected at each time point (on days 10,12,14,16, and 18 after gestation). After anesthesia, therats were sacrificed. Under the aseptic condition, fetal rat was harvested. Rat mesencephalic ventral brain tissue was isolated forculture of neural stem cells. Different gestational ages of rat brain-derived neural stem cells were separately cultured in theserum-free medium containing epidermal growth factors and basic fibroblast growth factors. After passage and amplification, theneural stem cells were induced to differentiate towards dopaminergic neurons in the medium containing interleukin lu, interleukin11, leukaemia inhibitory factors, glial cell-derived leukaemia inhibitory factors. On day 6 after induction and differentiation, thedopaminergic neurons were observed and identified by immunocytochemistry. After labeled by tyrosine hydroxylase, thedifferentiated dopaminergic neuron proportion was detected by a flow cytometer.MAIN OUTCOME MEASURES: The growth state of differentiated rat neural stem cells at different gestational ages and theimmunocytochemistry results. The tyrosine hydroxylase staining-positive neural stem cell proportion after induction anddifferentiation.RESULTS: Rat mesencephalic neural stem cell spheres on days 10,12, 14, 16, and 18 after gestation adhesively grew in thedifferentiation-inducing medium. The neural stem cells in the spheres gradually grew in radial tendency. On day 6 afterdifferentiation, most of the neural stem cells exhibited 1-2 long processes or several short processes. After nidogenimmunocytochemical staining, most of neural stem cells exhibited cytoplasm-positive. After culture for 6 days in the differentiationinducing medium, rat mesencephalic neural stem cells at gestational 10,12, 14, 16, and 18 days were detected by a flow cytometer.Results demonstrated that the proportion of tyrosine hydroxylase-positive cells was (10.3±2.5)%, (21.6±3.4)%, (16.7±2.8)%,(14.2±3.2)%, and (8.9±1.8)%, respectively. There was a significant difference in the proportion of tyrosine hydroxylase-positivecells among the cells at different gestational days (P < 0.05). Rat neural stem cells at gestational 12 days could be induced todifferentiate into dopaminergic neurons at the highest proportion.CONCLUSION: Mesencephalic neural stem cells of rats at different gestational days have different capabilities to differentiatetowards dopaminergic neurons. The proportion of dopaminergic neurons is the highest when mesencephalic neural stem cells ofrats at gestational 12 days.

[Objective] To investigate the change of liver histology, proliferation of BDEC, and expression of TGF-β in different stages of liver chelestusis. [Methods] ①Rat cholestatic livers were induced by common bile duct ligation (BDL) and separated into 3 groups, namely control group (DO), 7 days after BDL group (DT), and 18 days after BDL group (D18). ② Histological changes of livers in different groups were evaluated based on Knodell HAI score. ③ Real-time PCR was used to detect the expression of TGF-β in liver tissue and isolated BDEC in different groups. ④ Statistically analyzing the correlation between Knedell HAl score and the levels of TGF-β_1 mRNA. ⑤ In vitro study was performed to investigate the effect of TGF-β_1 on an immortalized mouse intrahepatic bililary epithelial cell line (mIBEC). [Results] (I) Knedell HAI score and the proliferation of intrahepatic bile ducts increased as the liver cholestasis aggravated. ② The levels of TGF-β_1, TGF-β_2, and TGF-β_3 mRNA were significantly up-regulated in liver tissues and BDEC as the liver cholestasis aggravated. ③ Positive correlation was found between Knedell fibrotic score and the levels of TGF-β_1 mRNA in liver tissues and BDEC(r=0.9376, P<0.05 and r=0.9682, P < 0.01). ④ In vitro study showed that TGF-β_1 inhibited the proliferation of mIBEC. [Conclusion] ① Liver injury, biliary proliferation, and the levels of TGF-β mRNA expression increased as liver cholestasis aggravated. ② The interaction of TGF-β_1 and BDEC plays an important role in the pathogenesis of BDL induced cholestatic liver disease. ④ Up-regulated expression of TGF-β_1 mRNA in the proliferated BDEC participates in the formation of BDL induced cholestatic liver fibrosis.

Objective To inspect the efficacy and mucosa healing condition of infliximab with azathioprine combination therapy in Crohn's disease (CD) and its correlation with prognosis.Methods A total of 20 active CD patients who received infliximab and azathioprine combination therapy at The First Affiliated Hospital of Sun Yat-sen University were objects of this study.The clinical efficacy was evaluated at 10 weeks,30 weeks,54 weeks and 2 years respectively according to CD activity index.The efficacy was evaluated under endoscopy at 10 weeks,30 weeks,54 weeks and 2 years respectively according to mucosal response situation under endoscopy.The data were analyzed by analysis of variance or Fisher's exact test between two groups.The factor affecred mucosal healing was analyzed by Logistic regression analysis.Results The clinical remission rate of patients without steroid at week 10,30,54 and 2 year were 12/20,16/20,15/20 and 15/20 respectively.Mucosal healing rate at week 10,30 and 54 weeks were 8/20,12/20 and 10/20 respectively.Logistic regression analysis indicated that age was the only factor affected mucosal healing at 30 weeks (OR =0.774,95％ CI:0.630 to 0.950).There was significant differences in clinical remission between mucosa response patients and invalid under endoscopy at 30 weeks and 2 years without steroid (at 30 weeks:14/14 vs 2/6; at 2 years:14/14 vs 1/6; all P＜0.01).Infliximab were withdrawn in 4 of 16 patients who was in non-steroid clinical remission at 30 weeks,and the other 12 patients were continued with infliximab therapy.There was no significant difference in non-steroid clinical remission rate (4/4 vs 11/12) and mucosa healing rate (2/4 vs 7/12) between withdrawal and continue of infliximab therapy (all P＞0.05).Conclusions Infliximab with azathioprine combination therapy can effectively promote and maintain mucosa healing in CD.The mucosa response patients can maintain long time non-steroid clinical remission.

Objective To investigate the evaluation standard and proper time point of anti-tuberculosis trial for differential diagnosis between intestinal tuberculosis (ITB) and Crohn's disease (CD). Methods Clinical data and endoscopic changes of 28 patients with confirmed ITB and 11 with confirmed CD,who underwent anti-tuberculosis trail, were retrospectively analyzed. Results No significant difference could be found in clinical characteristics of ITB and CD patients on baseline, such as active pulmonary tuberculosis, strong positive skin test and anal fistula/perianal abscess. Clinical symptoms were relieved in both groups right after anti-tuberculosis treatment. After 3 months of treatment, the no-improvement rate in ITB group was 0, whereas that of CD group was 27.3% (P =0. 004). The disappearance rate plus improvement rate of ulcer in ITB group was 90. 9% (20/22) plus 9. 1% (2/22) and 100% ( 28/28 ) plus 0 at 3 and 6 months of treatment, respectively. The disappearance rate plus improvement rate of nodular lesion was 58. 8% (10/17) plus 41.2% (7/17) and 76. 5% (13/17) plus 23.5% (4/17), respectively. There was no obvious improvement of active ulcer or nodular transformation in CD group at any time point ( P ＜ 0. 01 ).Conclusion With deficiency of special index for differential diagnosis of ITB and CD, some cases hard to differentiate still have to accept anti-tuberculosis treatment. Three months of anti-tuberculosis treatment is a proper time point to evaluate the efficacy. Disappearance of active ulcer and nodular transformation, together with cure or obvious improvement in clinic are taken as effective for treatment trail.