Objective To explore the expression of TLR4/MyD88/NF-κB signaling and its correlation with the progression of acute intestinal graft-versus-host disease (iGVHD) after allogeneic bone marrow transplantation in mice.Method Recipient BALB/c female mice were lethally irradiated and were reconstituted within 4-6 h with a transplant of bone marrow cells (1 × 107) and different amounts of splenocytes (1 × 107,n =12 or 2 × 107,n =12) from MHC-mismatched C57BL/6 donors to induce iGVHD,and 6 healthy BALB/c mice served as controls.A globe survey observation of GVHD by survival,clinical manifestation,and histological detection was performed.RT-PCR,immunohistochemistry,and Western blotting technology were used to detect the mRNA and protein levels of TLR4,MyD88 and NF-κB p65 in the small intestine tissue.Result The exacerbation of iGVHD was associated with the increasing dose of allogeneic spleen lymphocytes.The mRNA expression of TLR4,MyD88 and NF-κBp65 was increased as the iGVHD progressed.All of them in severe iGVHD model were significantly increased as compared with the healthy controls (P＜0.05).The expression of corresponding proteins had the same tendency as mRNA.All of the three genes expression was not only positively correlated with each other,but also with the clinical GVHD score:TLR4 (R =0.814,P＜0.001),MyD88 (R=0.828,P＜0.001),and NF-κB p65 (R=0.568,P =0.034).Conclusion Excessive activation of TLR4/MyD88/NF-κB signaling pathway does exist in iGVHD,and the enhanced levels of gene transcription and translation are positively correlated with the deterioration of iGVHD.

In 2013 Shanghai took the lead to carry out the specialist standardized training which is common in the international medical education. This paper first gives a picture of the general condition of the standardized training on specialists of obstetrics and gynecology in Shanghai. Then from the perspectives of strategic deployment, department management, clinical skill training and the training of examiners, the paper explores the assessment model in which both the assessment of train-ing process and the scores of the final examination are considered whereas the process assessment is given more weight. This paper is aimed to provide experience and suggestions for the further advance of the specialists standardized training in the field of obstetrics and gynecology.

0.05).However,neither CpG-ODN nor hTNF-? failed in improving the expression rates of CD1a.Conclusions CpG-ODN,like hTNF-?,has remarkable im- munostimulatory effect on the differentiation and maturation of monocyte-derived DC from patients with chronic hepatitis B.

Objective To explore the effects of deoxynivalenol (DON) on apoptosis of human gastric carcinoma cell line SNU in vitro. Methods SNU cells were treated with DON at different concentrations (50, 100, 1000, 2000 μg/L) for 12 hours, and then cells were harvested for cell apoptosis by flow cytometric (FCM) DNA analysis and the expression of Bax, Bcl-2 and Caspase-3 at protein level with FCM and Western blotting. Results FCM results showed that the apoptosis rates of SNU cells in DON treatment groups were all higher than that in control, especially in DON 1000 μg/L and 2000 μg/L groups (P<0.05). In the concentration range from 50 to 2000 μg/L, a significant concentration-depended response correlation could be found between apoptosis rate and DON concentration (r =0.940, P <0.01). FCM and Western blotting showed Bax and Caspase-3 expression in often SNU cells DON treatment for 12 hours were up-regulated while that of Bcl-2 was down-regulated. Conclusion DON can induce apoptosis of SNU cells in vitro in dose-dependent manner, and possible mechanisms of apoptosis induction effects may be up-regulation of the expression of Bax and down-regulation of that of Bcl-2 and activation of the key enzyme of apoptosis Caspase-3.

Objective To evaluate the applied value of functional immunity measured by the ImmunKnow assay in the diagnosis of post-transplant infection in Chinese 1iver recipients.Methods Thirty-eight normal adults and 68 adult liver transplant recipients were under investigation.Whole blood samples from either normal volunteers(each sample for one person)or the liver recipients(one or more samples for one person)were collected freshly and cultured within 6h.The CD4+T cells were selected and their ATP value was assayed the next day.The liver recipients were grouped in stable status(n=52)or infection(n=64)according tO their clinieal manifestation.Results The average ATP value in the recipients with infection after liver transplantation was 165.7±100 μg/L,significantly lower(P＜0.05)than that in stable recipients(309±126 μg/L)or normal volunteers (292±83 μg/L).The low ATP levels in post-transplant recipients had fair good correlation to infection clinically(RR=0.5021,P＜0.01).Infectious risk was high when ATP value was less than 165μg/L(OR=11,95%CI 3.9-32.2,P＜0.01).Specificity and sensitivity of low ATP value in post-transplant infection were 86.53%and 73.81% respectively,Conclusion ImmuKnow assay provides a new tool in monitoring immune status in post-transplant recipients,and call helpfully predict and diagnose clinical infection.

Objective: To observe the tolerability of Chinese melanoma patients to four-week high-dose interferon alfa-2b(INTRON A(R)，Schering-Plough)therapy. Methods:A total of 29 patients with high risk melanoma[American Joint Committee on Cancer Staging(AJCC)ⅡB-ⅢC]who received adjuvant interferon therapy in our hospital between September 2007 and May 2009 were retrospectively reviewed.Patients received 4 hours of intravenous infusion of interferon alfa-2b fdose range，22.00 million international unit(MIU)to 33.75 MIU]Ⅳ 5 days/week for 4 weeks.The adverse events were evaluated with National Cancer Institute Common Toxicity Criteria(NCI 2.0 version). Results: The average daily dose was 17.63 MIU/(m~2·d).The therapy was ended in two patients because of poor wound healing or intolerability to severe fatigue.The most common adverse events were myelosuppression.Grade 3/4 neutropenia was observed in 69% (20/29)patients and was rapidly reversed after conventional support interventions.Grade 1/2 abnormal hepatic function occurred in 18 cases(62%).Twenty-six patients were followed up for 3 to 22 months.Five patients developed early progression:one with local recurrence，two with regional lymph node metastasis one with in-transit metastasis in the affected limb,and one with distant metastasis. Conclusion: High-dose interferon alfa-2b regimen can be well tolerated by Chinese patients but cannot effectively inhibit subclinical lesions.

The aim of this study is to clarify whether edaravone postconditioning had protective effect against renal ischemia/reperfusion injury and to compare the protective effect between ischemic postconditioning and edaravone postconditioning. Rats were subjected to 45 min ischemia followed by 24 h reperfusion. The rats were randomly assigned to seven groups: a sham-operated control group, an ischemia/reperfusion group, an ischemic postconditioning group, a normal saline vehicle postconditioning group and an edaravone postconditioning (1, 3, and 6 mg x kg(-1)) group. Renal function was assessed by serum creatinine and BUN concentration, while histological damage of renal tissue was assessed with HE staining. MDA content and SOD activity of renal tissue were determined. TUNEL staining was performed to analyze the apoptosis of the tubular epithelial cells, the protein level of Bcl-2 and Bax in renal tissue was examined by Western blotting. Compared to the ischemia/reperfusion group, edaravone postconditioning significantly decreased serum creatinine and BUN concentration, and ameliorated histological damage of renal tissue. MDA was less after 24 h reperfusion in the edaravone postconditioning group than that in the ischemia/reperfusion group, consistent with an increase in SOD activity. In addition, edaravone postconditioning decreased TUNEL-positive cells and Bax expression, and increased Bcl-2 expression. Results detected in the edaravone postconditioning group showed no significant difference from the ischemic postconditioning group. Edaravone administered during the last 3 min of ischemia, prior to reperfusion induces a pharmacological postconditioning in vivo against renal ischemia/reperfusion injury in rats. This protection is similar to that observed with ischemic postconditioning.
Animals ; Antipyrine ; analogs & derivatives ; therapeutic use ; Apoptosis ; drug effects ; Blood Urea Nitrogen ; Creatinine ; blood ; Free Radical Scavengers ; therapeutic use ; Ischemic Postconditioning ; Kidney ; blood supply ; pathology ; Male ; Malondialdehyde ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Superoxide Dismutase ; metabolism ; bcl-2-Associated X Protein ; metabolism

Background Oxygen-induced retinal neovascularization is the main pathological basis for many retinal vascular diseases.Research showed that light exposure at night can suppress retinal neovascularization in oxygen-induced retinopathy(OIR),but there were few reports discussing its effect on ROP.Objective This study aimed to observe the effect of light exposure at night on retinal neovascularization in an OIR mouse model.Methods Sixty-four newborn C57 BL/6J mice were randomly divided into four groups,with 16 mice for each group.OIR models were established by rearing the newborn C57BL/6J mice with their mothers in a(75±2)％ oxygen environment from postnatal day 7(P7)to Pl2,and then transferred to room air.In the OIR model group,the environmental illumination level was the same as the normal control group,and the model mice were exposed to 100 lx light at night in the OIR+ light exposure group.In the simple light exposure group,normal mice were reared in room air and were exposed to light at night from P12 to P17.All the mice were sacrificed on P17,and retinal flat mounts were prepared to assess the oxygen-induced changes of retinal vessels using the adenosine diphosphatase(ADPase)histochemical technique.The amount of proliferative neovascularization was quantified by counting the number of endotheliocyte nuclei in new vessels extending from the retinal inner limiting membrane into the vitreous in ocular cross-sections.The expression of the vascular endothelial growth factor(VEGF)protein was detected by immunohistochemistry.Real-time PCR analysis was performed to examine the expression of VEGF mRNA.The rearing and usage of the animals complied with the Statement of ARVO.Results Less free-vascular areas and new blood vessels were seen in the OIR+light exposure group compared with the OIR model group.On day 17 of the mouse life,the number of the endotheliocyte nuclei in new vessels extending from retinal inner limiting membrane were 0.97±0.83,1.00±0.72,38.57±5.01 and 16.92±3.39 in the normal group,simple light exposure group,OIR model group and OIR+light exposure group,respectively,showing significant differences among them(F =78.767,P =0.000).The number of nuclei in the OIR+light exposure group were less than that of the OIR model group(t=20.446,P＜0.01).Immunochemistry showed that the expression of VEGF in retina was weaker in the OIR+light exposure group than the OIR model group.The relative expression values of VEGF mRNA were 1.00±0.00,0.94±0.07,2.08±0.50 and 1.43±0.21 in the normal group,simple light exposure group,OIR model group and OIR+light exposure group,respectively,showing a significant difference (F=11.268,P =0.003),where the VEGF mRNA level in the OIR+light exposure group was lower than that of the OIR model group(t =20.163,P＜0.05).Conclusions Light exposure at night can weaken retinal neovascularization in OIR mice

Background The incidence of posterior capsular opacification (PCO) is increasing with the growing of cataract surgery rate.Recent researches provend that disintegrin has inhibitory effect on PCO,and echistatin is one of the disintegrin prime families.Objective This study was to investigate the effects of disintegrin and echistatin on proliferation,adhestion and migration in human lens epithelial cells (LECs) line (SRA01/04).Methods Human LECs line at logarithmic growth phase was used in the study.Cells were cocultured with medium and different concentrations of echistatin (0,2.5,5.0,7.5,10.0,15.0,20.0 mg/L) for different time.The proliferative inhibitory rates of LECs were detected by MTT method 24,48 and 72 hours after cultured.Anti-adhesion effect of echistatin were analyzed by the same assay in 90 minutes.Cell scratching test was performed to evaluate the migration ability of LECs.The width of the scratch was recorded in the culture plate covered with cells under an inverted microscope.After being cultured for 24 hours and 48 hours with echistatin,cell migration distances was examined.Results Compared with the 0 mg/L echistatin group,cells proliferation was obviously inhibited.After cultured with 2.5,5.0,7.5,10.0,15.0,20.0 mg/L echistatin,the proliferation inhibitory rate was 2.6％,15.4％,21.2％,34.7％,46.1％,58.2％ at 24 hours;6.6％,21.9％,38.2％,50.0％,60.7％,76.9％ at 48 hours and 9.8％,29.0％,46.6％,63.4％,69.1％,92.4％ at 72 hours,respectively.The absorbance value (A) in the 5.0,7.5,10.0,15.0,20.0 mg/L groups were significantly lower than that in the 0 mg/L group (P＜ 0.05).With the prolongation of acting time of Ecs,the A value of the cells was gradually reduced,with statistically significant difference (P＜0.05).The adhesion inhibitory rate was 2.6％,15.0％,26.1％,35.3％,45.2％ and 54.5％ in the 2.5,5.0,7.5,10.0,15.0,20.0 mg/L group,respectively.Compared with the result in the 0 mg/L group,the A value in the 5.0,7.5,10.0,15.0,20.0 mg/L group was statistically significant (P＜0.05).After cultured for 24 hours and 48 hours,cell migration distance shortened in the 5.0,7.5,10.0,15.0,20.0 mg/L group,showing a statistically significant difference among them (P＜0.05).Cell migration distance was gradually shortened with the lapse of action time of Ecs with the significant difference (P ＜ 0.05).Conclusions echistatin has inhibitory effects on proliferation,adhestion and migration for human LECs in vitro in time-and dose-dependent manner.It is inferred that echistatin may play a role in the prevention and treatment of PCO.

In order to investigate the role of the PTEN expression in carcinogenesis and development of endometrial carcinoma and clarify whether and how PTEN and PI3K/Akt pathway relate to endometrial carcinoma, the expression of PTEN and phospho-Akt was detected by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) methods and Western-blot from 24 cases of endometrial carcinoma, 10 cases of endometrial atypical hyperplasia, 10 cases of endometrial hyperplasia, and 10 cases of normal endometrium. SP immunohistochemical methods were used to measure levels of PTEN protein expression in following 5 study groups: 31 cases of endometrium in proliferative phase, 30 cases of endometrium in secretory phase, 71 cases of endometrial hyperplasia, 25 cases of atypical hyperplasia and 73 cases of endometrial carcinoma. Immunostaining score of PTEN was 3.39+/-0.15 in proliferative phase, 1.90+/-0.21 in secretory phase, 3.34+/-0.29 in endometrial hyperplasia, 0.62+/-0.11 in atypical hyperplasia, and 0.74+/-0.19 in endometrial carcinoma, respectively. PTEN mRNA relative value in normal endometrium, endometrial hyperplasia, endometrial atypical hyperplasia, and endometrial carcinoma was 2.45+/-0.51, 2.32+/-0.32, 0.46+/-0.11, and 0.35+/-0.13 respectively. The expression levels of PTEN mRNA and protein in patients with endometrial carcinoma and atypical hyperplasia were significantly lower than in those of proliferative phase and with endometrial hyperplasia. The level of PTEN expression in patients with endometrial carcinoma was significantly related to tissue type (P<0.005), differentiation (P<0.05) and clinical stage (P<0.05), but not to depth of myometrium invasion (P>0.05). Western blot analysis revealed that Phospho-Akt level in PTEN negative cases was significantly higher, and there was a negative correlation between PTEN and phospho-Akt (r=-0.8973, P<0.0001). It was suggested that loss of PTEN expression was an early event in endometrial tumorigenesis. The phosphorylation of Akt induced by the loss of PTEN took part in the tumorigenesis and development of endometrial carcinoma.