Objective To investigate a clinically perspective way of inducing transplantation tolerance. Methods Kidney transplantation model was established using Lewis (RT1 l) rats as recipients and DA (RT1 a) rats as donor. Before transplantation the recipient rats were conditioned with anti lymphothyte serum (ALS) injected for three times, followed by 10 8 donor bone marrow cells transfusion (BMT) and injection of 100 mg/kg cyclophosphamide (CP), survival time of kidney allografts, mixed lymphocyte reaction (MLR) and delayed type hypersensitivity (DTH) of recipient to donor spleen cells were observed and studied. Results 5 of the 7 kidney allografts survived 73 to 90 days without sign of rejection. The donor specific MLR and DTH were suppressed. Conclusions Induction of transplantation tolerance is a prospective way for the prevention of allograft rejection.

Objective:To investigate the antitumor activity of the compound HS-4 and the action mechanism.Methods:MTT method was used to testin vitroantitumor activity of the compound HS-4. Orthotopic xenotransplantation tumor model of liver cancer was established in nude mice, and,in vivoantitumor activity of compound HS-4 was tested with a small animal in-vivo imaging system. Sequencing of small RNA library and RNA library was performed in HS-4 treated tumor cell group and control group to investigate the anti-cancer mechanism of HS-4 at level of functional genomics, using high-throughput sequencing technology. Results:HS-4 was found to have relatively highin-vitro antitumor activity against liver cancer cells, gastric cancer cells, renal cancer cells, lung cancer cells, breast cancer cells and colon cancer cells. The IC50 values against SMMC-7721 and Bel-7402 of liver cancer cells were 0.14 and 0.13 nmol/L respectively, while the IC50 values against MGC-803 and SGC-7901 of gastric cancer cells were 0.19 and 0.21 nmol/L, respectively. It was demonstrated that HS- 4 possessed a better therapeutic effect in liver cancer.Conclusions: A new reliable orthotopic xenotransplantation tumor model of liver cancer in nude mice is established. The new compounds HS-4 was found to possess relatively highin vivo andin vitroantitumor activity against liver cancer cells.

Objective To analyze the significance of levels and combined detection of plasma homocysteine(Hcy),high-sensitivity C reactive protein(hs-CRP) and lipoprotein-associated phospholipase A2(Lp-PLA2) in patients with ischemic cerebral stroke(ICS).Methods A total of 140 patients with ICS were selected into ICS group,and 100 healthy individuals were recruited in the control group.The plasma levels of Hcy,hs-CRP,Lp-PLA2 were examined,then the positive rates of them were statistically analyzed.Analyzed the value of the levels and joint detection of Hcy,hs-CRP and Lp-PLA2.Results The plasma levels and the positive rates of Hcy,hs-CRP and Lp-PLA2 in ICS group were statistically higher than those in the control group(P<0.05).The area under the receiver-operating characteristic curve of combined detection of Hcy,hs-CRP and Lp-PLA2 in diagnosis of ICS was 0.949,the sensitivity was 0.850,and the specificity was 0.807,which were higher than Hcy,hs-CRP and Lp-PLA2 alone.Conclusion The plasma levels of Hcy,hs-CRP and Lp-PLA2 increase in ICS patients,and the combined detection of the three indicators might improve the early diagnostic value of ICS.

Objective:To validate an enzyme linked immunosorbent assay (ELISA) method for the quantification of rhCNB in long-tailed macaque sera.Methods: The linear,sensitivity,accuracy,precision and recovery were determined using ELISA.Results:The present ELISA method had high linearity within 0.195 ng/ml-12.5 ng/ml,the working curve of rhCNB was Y=15.1X-0.26, R2=0.996 8 , the method showed good sensitivity of 0.195 ng/ml, the accuracy were in the range of 91.9%-108.8%, and the Coefficient of variation ( CV) for inter-assay were 3.55%,1.39%and 4.71%,the intra-assay were 1.59%,3.2%and 3.8%,all less than 10%, the recoveries were in the range of 88.5% -108.3%, <110% .Thus the method was coincidence with requirement.Conclusion:Double antibody sandwich ELISA assay of rhCNB in long-tailed macaque sera has good sensitivity ,accuracy, precision and recovery and it can be used to measure rhCNB concentration in biological samples .

Objective To investigate the role and significance of neuropilin-2(NRP2)for regulating the angiogenesis of pancreatic neuroendocrine tumors(PNETs).Methods The NRP2 expression in pancreatic neuroendocrine tumer BON-1 cell line was intevened.The BON-1 cells cultural supernatants in the control group and interference group were used to treat human umbilical vein endothelial cells(HUVEC).CCK-8 was used to detect the cell proliferation,Transwell was used to detected the cell migration and the tubule formation test was used detect the pro-angiogenesis.Results The CCK-8 detection showed that there was no statistically significant difference in the supernatant treated HUVEC proliferations between the interference group and control group medium(P>0.05):the absorbancy in the control group was 0.35±0.04,while which in the interference group was 0.32±0.04.The Transwell test showed that the invasion ability of HUVEC treated with cultural supernatants in the interference group was weakened compared with the control group,the control group was(203±13)/hole,while the interference group was(100±10)/hole(P＜0.01);the tubule formation test showed that HUVEC tubular formation treated by cultural supernatant in the interference group was decreased,the control group was 40±5,while the interference group was 24±3(P＜0.01).Conclusion Interfering NRP2 expression of BON-1 cells can inhibit the vessel formation ability of co-cultured HUVEC,suggesting that NRP2 may have the pro-angiogenesis effect of PNETs,and may be a potential new target for the treatment of PNETs.

OBJECTIVE:To establish a method for determining recombinant human calmodulin B subunit(rhCNB)in rat plas-ma,and study its pharmacokinetics characteristics. METHODS:ELISA double-antibody sandwich method was adopted. 1 μg/ml rhCNB monoclonal antibody mAb was wrapped,added to the to-be-test sample,rhCNB polyclonal antibody pAb(dilution ratio of 1∶5 000)and HRP-labeled conjugate of anti-IgG(dilution ratio of 1∶10 000)were added. Using tetramethylbenzidine for develop-ing,microplate reader was conducted in wavelength of 450 nm to determine the absorbance value(OD value)and plasma concen-tration of 6 rats after 2,15,30,60,120,240,480,720 min of iv 2.5 mg/kg rhCNB,and the pharmacokinetic parameters were calculated by BAPP 3.0 software. RESULTS:The linear range of rhCNB were 0.195-12.5 ng/ml(r2=0.995 0),lower limit of quan-titation was 0.195 ng/ml,accuracy were 97.300%-103.622%(RSD<7.5%,n=6);RSDs of within-batch,inter-batch,freezing and thawing 3 times were no higher than 8.5%(n=6,18,15). rhCNB pharmacokinetics characteristics in rat fitted to two-com-partment model,AUC0-720 min was 173.038 mg·min/L and t1/2 was 94.62 min. CONCLUSIONS:The established method has high specificity and sensitivity,good accuracy and precision,which can be used for rhCNB quantitative detection and pharmacokinetics study in biological samples.

Objective: To explore the related factors of subependymal hemorrhage (SEH) and cerebral hemodynamic changes. Methods: From October 2012 to October 2017, 200 cases of children with subependymal hemorrhage diagnosed by ultrasound in our department of pediatrics were selected as the observation group , and a total of 150 children who were admitted to the Department of Pediatrics in the same period due to craniocerebral diseases and other serious diseases were selected as control group. The independent risk factors of the children in the observation group were analyzed, and the difference of the maximum systolic blood flow velocity (SV), the diastolic maximum flow velocity (DV), the systolic and diastolic velocity ratio (S/D), the resistance index (RI), and the pulsatile index (PI) were compared between the two groups. Results: Neonatal asphyxia, preterm birth, acidosis, neonatal respiratory distress syndrome (NRDS), patent ductus arteriosus and coagulation dysfunction were independent risk factors for subependymal hemorrhage. The bleeding side SV and DV of the observation group were higher than those of the control group, with statistically significant difference (P<0.001). In the observation group, the bleeding side SV and DV were higher than those of the healthy side, with statistically significant difference (P<0.001). There was no significant difference in bleeding side SV, DV, S/D, RI and PI in 110 cases of single side ependymal hemorrhage (P>0.05). Conclusions Children with ependymal hemorrhage can observe the hemodynamic indexes of anterior cerebral artery (ACA) dynamically by craniocerebral ultrasound, and judge the therapeutic effect by evaluating the systolic and diastolic blood flow velocity, so as to prevent the further aggravation of subependymal hemorrhage

Objective This study aimed to analyze the knowledge, attitude, behavior, and practice of pneumococcal vaccination in type 2 diabetes mellitus patients. Methods From January to March 2018, 3000 patients with type 2 diabetes mellitus were randomly selected from patients with type 2 diabetes registered and managed in community health service centers to participate in a questionnaire survey, using multi-stage cluster random sampling. The questionnaire included basic information, pneumococcal vaccine awareness, and pneumococcal vaccination status and inoculation intention, 2896 valid questionnaires were collected. The chi-squared test and multivariate logistic regression analysis were used to analyze the knowledge, attitude, behavior, and practice and influencing factors of pneumococcal vaccination. Results Of all the patients, 1267 (43.75％) patients with type 2 diabetes were willing to receive pneumococcal vaccination, and 23 (0.79％) patients with type 2 diabetes were vaccinated against pneumococcal, 1295(44.72％) patients with type 2 diabetes believed that patients with chronic diseases were susceptible to pneumococcal. Eight hundred seventy-seven (30.38％) patients believed that pneumococcal vaccination for chronic disease patients could reduce the risk of outpatient, hospitalization and death. Seven hundred ninety-nine (27.59％) patients believed that diabetic patients were the primary recipients of the pneumococcal vaccine. Multivariate logistic regression analysis showed that retired patients; those who thought that patients with chronic diseases were susceptible to pneumonia; those who thought that pneumococcal vaccination could reduce the risk of needing outpatient care, hospitalization, and death;and those who thought that diabetes was the priority of pneumococcal vaccination were more willing to receive pneumococcal vaccination (OR=1.442, 0.764, 1.494, 1.713, P all<0.05). Conclusion Patients with type 2 diabetes in Shenzhen have lower pneumococcal vaccine awareness, inoculation intention, and vaccination rates. Health education on the prevention of diabetes complications and pneumococcal vaccination should be promoted.

Objective To analyze the clinical characteristics of community-acquired influenza virus pneumonia in hospitalized children and improve the clinicians' understanding level of this disease.Methods Data of 70 cases with community-acquired influenza virus pneumonia admitted to the Respiratory Department and Infectious Disease,Beijing Children's Hospital,Capital Medical University,from November 2009 to April 2018 were collected and the clinical characteristics were analyzed.Results Of the 70 cases,61 cases(89.7％) were discharged after improvement.The median age was 3.5 years old,and 50 cases(71.4％) were 0 to 5 years old.There were 29 cases with severe influenza pneumonia,41 cases with mild influenza pneumonia,3 cases died,and 19 cases (27.1％) had underlying diseases.Sixty-four cases (91.4％) were hospitalized in winter and spring.The first symptoms were mainly fever in 64 cases (91.4％) and cough in 65 cases (92.9％),and temperatures were mostly from 39.1 ℃ to 41.0 ℃.Lung auscultation was dominated by moist rales (30 cases,58.8％) and wheezing (8 cases,15.7％).There were many complications of influenza virus pneumonia,including 19 cases with myocardial injury,11 cases with liver function injury,4 cases with toxic encephalopathy,3 cases with electrolyte disturbance,2 cases with multiple organ failure,2 cases with hemophagocytic syndrome,and 1 case with septic shock.Chest radiographic results reveal bilateral inflammation in 40 children (57.1％),prodominatly in lower lobe lesions (39 cases).The common changes were patchy shadow,interstitial parenchymal lesion,ground glass shadow,and pleural effusion.Forty-seven children (67.1％) were infected by influenza A,and 23 children(32.9％) were co-infected.The percentage of severe cases with underlying diseases (68.4％) was significantly higher than that in children without chronic diseases (31.4％),the difference was statistically significant (x2 =7.830,P =0.005).The increase rate of C reaction protein (CRP) in severe cases (54.3％) was significantly higher than that in mild cases (28.6％),the difference was statistically significant (x2 =4.769,P =0.029).Conclusions Community-acquired influenza virus pneumonia in children mainly occurs in winter and spring.It is more common seen in children under 5 years of age.The main clinical manifestations of community-acquired influenza virus pneumonia are high fever and cough,extrapulmonary complications are more common.Most children have moist rales and showed bilateral inflammation and lower lobe lesions in chest radiography.Children with underlying diseases are more likely to develop severe influenza virus pneumonia.Elevated CRP is associated with severe influenza virus pneumonia.Most patients have a good prognosis,but there are still cases of death.

Objective To detect the serum level of 3-carboxy-4-methyl-5-propyl-2-furanpropanoic acid (CMPF),a significant metabolite offish oil,in subjects with normal glucose tolerance (NGT) in local communities,and to investigate the association of CMPF with fatty acid metabolism.Methods A total of 272 NGT participants from screening for diabetes in Shanghai in 2013 were enrolled.Anthropometric measurements,biochemical evaluation,and questionnaire interview were performed for all the participants.The participants were divided into normal weight group [body mass index (BMI) ≤23.9 kg/m2,n =143] and overweight/obesity group (BMI ≥ 24 kg/m2,n =129).The serum CMPF concentrations were determined using an enzyme-linked immunosorbent assay.Results Serum CMPF level in overweight/obesity group was lower than that in normal weight group [96.50 (46.11,169.56) μmol/L vs 153.20 (83.16,282.97) μmol/L,P＜0.05].The serum CMPF level was negatively correlated with BMI (r =-0.256,P＜0.01),triglycerides (r =-0.175,P =0.004),and free fatty acid (r =-0.126,P =0.041) according to bivariate correlation analyses.A multivariate stepwise linear regression analysis showed that the serum CMPF level was independently associated with BMI,triglycerides,free fatty acid,and HbA1C.A logistic regression analysis showed that the CMPF was a protective factor against obesity (OR =0.324,95％ CI 0.158,0.664).Conclusion Serum CMPF level is reduced in overweight/obese subjects.CMPF is beneficial to lipid metabolism.