BACKGROUND: Donor antigen presenting cells immigrated into the recipient can induce the immunological tolerance of recipient T cells to donor, leading to a final acceptance to grafts, lnterleukin-10 (IL-10) modification maintains dendritic cells at a desirable differentiating state, which is an effective method to promote the protection to kidney in the simultaneous liver-kidney transplantation.OBJECTIVE: To observe the immigration of IL-10-modified dendritic cells in rats subjected to simultaneous liver-kidney transplantation and to investigate the mechanism of action.DESIGN, TIME AND SETTING: A randomized, controlled, animal experiment was performed in the Medical College of Sun Yat-sen University between June 2004 and February 2006.MATERIALS: Male DA donor rats (n=60) and Lewis recipient rats (n=60), both were adult and of clean grade, were included.Sixty Lewis rats were randomly and evenly divided into 3 groups: IL-10-modified cell group, simple cell group, and model control group.METHODS: Donor rat liver and kidney were harvested by simultaneous liver-kidney transplantation. Recipient rats in each group were subjected to orthotopic liver and left kidney transplantation to establish models of immunological rejection. Under sterile condition, donor rat femur and tibia were harvested. Dnlbecco's modified eagle's medium (DMEM) was used to wash out the bone marrow. After removal of red cells, dendritic cells were isolated and cultured by adherent method. After modified with 20 μg/L IL-10 for 72 hours, dendritic cells were intravenously transfused into rat bodies in the IL-10-modified group, 2×10(7) cells/rat, In the simple cell group, rats were treated with donor dendritic cells without modification with IL-10. Rats in the model control group received no any interventions.MAIN OUTCOME MEASURES: [1]Dynamic changes of vital sign, urine volume, liver and renal function in recipient tissues;[2] Pathohistological detection results;[3]Distribution of donor dendritic cells in the recipient rats by in situ hybridization.RESULTS: In the simple cell and model control groups, urinary volume was reduced to 0 mL 5-6 days after transplantation. In addition, both groups presented with severe acute rejection. In the IL-10-modified cell group, urinary volume maintained at 6-12mL within 2 weeks after transplantation. The acute rejection of liver and kidney transplantation was obviously inhibited, surviving for(20.0±2.6) days on average, which was significantly longer than that in the simple cell group and model control group. A probability value of less than 0.05 in the Log Rank test was considered statistically significant. There were many Y chromosome-labeled dendritic cells immigrated into the mesenteric lymph node in the recipient rats.CONCLUSION: IL-10-modified dendritic cells play an immunoprotective effect on the liver and kidney transplanted simultaneously. Donor immature dendritic cells immigrated into recipient tissue could reduce acute rejections and prolong the survival time of liver and kidney grafts and recipients.

The experiment was conducted to examine the effect of zinc toxicity on the peripheral blood T-lymphocyte by the method of flow cytometry(FCM) and ANAE.200 one-day-old Avian broilers were divided into four groups,and fed with the diets as follows:①control group(Zn 100 mg/kg diet) and ② zinc toxic groups(Zn 1 500 mg/kg diet,zinc toxic group Ⅰ;Zn 2 000 mg/kg diet,zinc toxic group Ⅱ;Zn 2 500 mg/kg diet,zinc toxic group Ⅲ) for seven weeks.The ANAE positive ratios of the peripheral blood T-lymphocytes were much lower in the three zinc toxic groups than that in control group at 7 weeks of age.CD4+ T cell numbers were reduced from 2 to 7 weekds of age in zinc toxic group Ⅲ and from 6 to 7 weeks of age in zinc toxic group Ⅱ,as compared with that of control group.The numbers of CD8+ T cell decreased at 2 and 4 weeks of age in zinc toxic groups Ⅱ and Ⅲ,and at 7 weeks of age in zinc grups Ⅰ and Ⅲ.CD4+/CD8+ ratio was higher at 2 and 4 weeks age,and lower at 6 and 7 weeks of age in zinc toxic groups Ⅰ,Ⅱ and Ⅲ than in control group.The results showed that zinc toxicity would suppress the development of T-lymphocytes and reduce the peripheral blood T-lymphocyte populations.Potential mechanism underlying these observations are also discussed.

The experiment was conducted to examine the effect of copper toxicity on the peripheral blood T-lymphocyte using the flow cytometry(FCM) and ANAE.180 one-day-old Avian broilers were divided randomly into three groups,and fed diets as follows:(1)Controls(Cu 11.97 mg/kg diet) and (2)copper toxic(Cu 650 mg/kg diet,copper toxic group Ⅰ;Cu 850 mg/kg diet,copper toxic group Ⅱ) for six weeks.The ANAE positive ratios of the peripheral blood T-lymphocytes were much lower in the two copper toxic groups than in control group from 1 to 6 weeks of age(P＜0.05 or P＜0.01).Also,there was significant difference between copper toxic groups Ⅰ and Ⅱ at 1,3,5 and 6 weeks of age(P＜0.05 or P＜0.01).CD+4 T cell numbers reduced from 2 to 6 weeks of age in both copper toxic group Ⅰ and copper toxic group Ⅱ as compared with those of control group(P＜0.05 or P＜0.01).At the same time,there was significant difference between copper toxic groups Ⅰ and Ⅱ at 6 weeks of age(P＜0.05).But the numbers of CD+8 T cell were not varied from 2 to 6 weeks of age in copper toxic groups Ⅰ and Ⅱ in comparison with those of control group(P＞0.05).The CD+4/CD+8 ratio was lower from 2 to 6 weeks of age in copper toxic groups Ⅰ and Ⅱ than in control group.The results showed that copper toxicity could suppress the development of T-lymphocytes and reduce the peripheral blood T-lymphocyte populations.Potential mechanisms underlying these observations are also discussed.

150-day-old Tianfu ducklings were divided into three groups,and fed with diets as follows :Zn deficient (22. 9 mgZn per kg diet),controls(100 mg Zn per kg diet) and Zn toxic (1 300 mg Zn per kg diet) for seven weeks (Zn deficiency,ZD)or four weeks (Zn toxieity,ZT). The ANAE+ positive ratios of the peripheral blood T-lymphocytes were much lower (P＜0.01 ) in Zn deficient and toxic groups than in the control group. The results showed that Zn deficiency and toxicity would suppress the development of T-lymphocytes and reduce the peripheral blood T-lymphocyte populations. Potential mechanisms underlying these observations are also discussed.

AIM:To study whether the pulmonary infection of Escherichia coli (E.coli) interferes the glu-colipid metabolism in high-fat diet-induced obese mice.METHODS:High-fat diet-induced obese mice (n=48) and nor-mal chow-fed control mice ( n=48) were intranasally infused with 40 μL fluid containing 4 ×109 CFUs E.coli.The ser-um, periepididymal adipose tissue and liver were obtained at 0 d, 1 d, 2 d, 3 d and 4 d after infection.The body mass, periepididymal adipose tissue and liver were weighed , and the levels of fasting blood glucose (FBG), fasting blood insulin ( FINS) , free fatty acid ( FFA) and very-low-density lipoprotein ( VLDL) were measured by ELISA .The serum total cho-lesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol ( LDL-C) , and hepatic TG contents were detected , and the hepatic steatosis was observed under microscope with oil red O staining.RESULTS:Compared with day 0, the body mass, fat mass and fat index were decreased significantly from day 1 to day 4 after infection (P<0.05).The levels of FBG, FINS and HOMA-IR were apparently raised from day 2 to day 4 af-ter infection (P<0.05).The contents of serum FFA, TG and VLDL were increased markedly from day 1 to day 4 after in-fection (P<0.05).However, the concentrations of serum TC, LDL-C and HDL-C were decreased obviously from day 1 to day 3 ( P<0.05 ) .The liver mass , liver index and TG content were significantly increased from day 1 to day 4 ( P<0.05 ) .Consistently , the lipid droplet accumulation in the liver cells was increased obviously at day 2 and day 4 after infec-tion.Compared with control group , except the levels of serum TC , LDL-C and HDL-C in obese group substantially de-creased, the other indexes were increased by different degrees during the whole experiment period (P<0.05).CONCLU-SION:Pulmonary infection of Escherichia coli exacerbates the disorder of glucose and lipid metabolism in high-fat diet-in-duced obese mice , which contributes the development of insulin resistance and hepatic steatosis .

Objective To analyze the pathologically confirmed pulmonary Actinomycosis in the 11 patients in focusing on clinical features and mis-diagnostic reasons so as to improve physicians' awareness of this rare disease and reduce the misdiagnosis.Methods We retrospectively reviewed the medical records of 11 cases with pathologically confirmed pulmonary Actinomycosis during January 2003-August 2015.The clinical data and main causes of misdiagnosis in these cases were collected and analyzed.Results The study included 11 patients with a mean age of(53.0 ± 11.6.0)years.Among the 11 cases,8 (72.7 ％) patients had complications,6 (54.5 ％) were current or ex-smokers.Main clinical manifestations of 11 cases were cough(11/11,100.0 ％),sputum(11/11,100.0 ％),hemoptysis (7/11,63.6％),chest pain(6/11,54.5％)and fever(3/11,27.3％).Ten patients presented with one lobe of lung lesions,including 4 patients in the lower lobe and 3 in the upper lobe of the left lung,2 in the upper lobe and 1 in the lower lobe of the right lung.While,the remained one case presented with lesion locating in right main bronchus.Iconography often presented as pulmonary mass shadow,consolidation shadow,spicule sign,lobulation sign,hilar and/or mediastinal lymphadenopathy and pleural effusion.Vacuolar lesions were observed in some of the focuses.Flexible bronchoscopy was performed in 8 (72.7％)patients.Among them,7 patients showed mucosal swelling and congestion,luminal occlusion with purulence secretion,2 cases with polypoid neoplasm.Initial misdiagnosis rate were 100％ (11/11),among which 7 cases were misdiagnosed as lung cancer,2 cases as fungus infection,and 1 case as pulmonary tuberculosis and 1 case as pneumonia,respectively.All patients were definitely diagnosed by biopsy finding an evidence of hyphae of Actinomycosis in lung tissue specimens.The definitive diagnosis was made by CT-guided percutaneous lung biopsy in 4 cases,by transbronchial lung biopsy (TBLB)in 5 cases and by thoracotomy or video-assisted thoracoscopic surgery(VATS) in 1 case respectively.Actinomycosis in most patients was cured with high-dose penicillin administration over a prolonged period.Conclusions The diagnosis of pulmonary Actinomycosis remains challenging via its non-specific clinical symptoms and iconography features,and the presence of comorbidity may further increase the difficulty and complexity of diagnosis,leading to delaying-or mistaking-diagnosis.Obtaining positively pathological specimens is diagnostic key.Transbronchial lung biopsy through a bronchoscope and CT-guided percutaneous needle biopsy are the priority methods.

Antibody-drug conjugates(ADCs)are commonly heterogeneous and require extensive assessment of exposure-efficacy and exposure-safety relationships in preclinical and clinical studies.In this study,we report the generation of a monoclonal antibody against monomethyl auristatin E(MMAE)and the development,validation,and application of sensitive and high-throughput enzyme-linked immunosor-bent assays(ELISA)to measure the concentrations of MMAE-conjugated ADCs and total antibodies(tAb,antibodies in ADC plus unconjugated antibodies)in cynomolgus monkey sera.These assays were suc-cessfully applied to in vitro plasma stability and pharmacokinetic(PK)studies of SMADC001,an MMAE-conjugated ADC against trophoblast cell surface antigen 2(TROP-2).The plasma stability of SMADC001 was better than that of similar ADCs coupled with PEG4-Val-Cit,Lys(m-dPEG24)-Cit,and Val-Cit linkers.The developed ELISA methods for the calibration standards of ADC and tAb revealed a correlation be-tween serum concentrations and the OD450 values,with R2 at 1.000,and the dynamic range was 0.3-35.0 ng/mL and 0.2-22.0 ng/mL,respectively;the intra-and inter-assay accuracy bias％ranged from-12.2％to-5.2％,precision ranged from-12.4％to-1.4％,and the relative standard deviation(RSD)was less than 6.6％and 8.7％,respectively.The total error was less than 20.4％.The development and validation steps of these two assays met the acceptance criteria for all addressed validation parameters,which suggested that these can be applied to quantify MMAE-conjugated ADCs,as well as in PK studies.Furthermore,these assays can be easily adopted for development of other similar immunoassays.