Abstract Objective: To study the effect of Trichosanthin on cytokines production and ion channel of human PBMC, to demonstrate its immune regulatory mechanism.Methods: The levels of IL-2 and IL-6 produced by PBMC under stimulation of PHA-M were measured by radio-immunoassay after cultured with Trichosanthin. The current intensity of voltage-dependent potassium channel [K(V) channel] were recorded by patch-clamp technique after added Trichosanthin.Results: Both IL-2 and IL-6 production were enhanced by Trichosanthin, and the current in-tensity of K(v) channel were markedly increased from (210.2 ?53.6)PA to (2 223.0 ?310.7)PA.Conclusion: Trichosanthin can enhance the production of both IL-2 and IL-6 of human PBMC and increase the current intensity of K( v) channel on T lymphocyte membrane, which im-prove cell-mediated immunity and humoral immunity.

Objective To compare the changes of serum hepcidin,hemoglobin and other indexes of pure hemodialysis and hemodialysis combined with hemodiafiltration in treatment of maintenance hemodialysis patients,and to investigate the effect of hemodialysis combined with hemodiafiltration therapy on serum hepcidin clearance,the improvement of anemia and the relationship between them of maintenance hemodialysis patients.Methods Twenty-two patients with maintenance hemodialysis patients were collected from Shengjing Hosphal Affiliated to China Medical University from January 2015 to February 2017.This study was self-controlled study,before enrollment all patients were given only regular hemodialysis,after enrollment all patients were given hemodialysis combined with hemodiafiltration therapy for 3 months,compared the changes of serum hepcidin,hemoglobin,reticulocyte percentage,ferritin,serum iron,transferrin saturation and EPO of pure hemodialysis(before treatment) and hemodialysis combined with hemodiafiltration treatment for 3 months of patients,and analyzed the correlationship between serum hepcidin and other indexes.Results After hemodialysis combined with hemodiafiltration therapy for 3 months,compared with pure hemodialysis serum hepcidin levels were significantly lower((77.61±3.31) μg/L vs.(89.49±7.87) μg/L,t=9.660,P<0.01),hemoglobin levels were increased((112.73±11.02) g/L vs.(88.32±11.83) g/L,t=-11.957,P<0.01).The correlations between serum hepcidin and other indexes show that serum hepcidin and hemoglobin and ferritin were related,serum hepcidin was negatively correlated with hemoglobin(r=-0.851,P<0.01)and positively correlated with ferritin(r=0.385,P<0.05),and found no correlations between hepcidin and reticulocyte percentage(r=-0.151,P=0.328),serum iron(r=0.123,P=0.427),transferrin saturation(r=0.211,P=0.170) and EPO(r=-0.173,P=0.261).Conclusion To compare with pure hemodialysis,hemodialysis combined with hemodiafiltration therapy can more effectively remove hepcidin and improve anemia,and they are related.

Objective To explore the expression of Excision repair cross -completion 1 in non-small cell lung cancer and to analyze its clinical significance .Methods ERCC1 protein expression was detected by IHC method in 160 cases of non-small cell lung cancer tissue .We analyzed relationship between the ERCC 1 ex-pression and clinicopathological parameters of non -small cell lung cancer ,as well as its correlation between ER-CC1 expression and the prognosis .At the same time ,160 cases of intraoperative specimens were primarily cul-tured.The relationship between ERCC1 expression and the drug sensitivity of DDP was analyzed .Results ER-CC1 protein was mainly expressed in the nucleus .ERCC1 protein expression in lung cancer tissue showed no rela-tionship with age,gender,tumor size and pathological types .ERCC1 protein expression was associated with TNM stages and lymph node metastases .High expression of ERCC1was significantly correlated with poor prognosis in pa-tients,indicating that ERCC1 is the independent factor of NSCLC patients .MTT results showed that the positive ex-pression rate of ERCC1 in DDP sensitive groups was higher than in DDP resistant groups .Conclusion ERCC1 is an independent factor influencing the prognosis of non -small cell lung cancer ,and associated with DDP resistance .

Objective To analyze the pathogenesis, clinical features, treatment and prognosis of pulmonary hypertension (PHT) in systemic lupus erythematosus (SLE) patients. Methods Thirty-eight patients admitted during Jan 2001 to Aug 2008 were diagnosed as SLE associated with PHT and were analyzed retrospectively. Results All cases were female with the mean age of 35.7 yds. The mean period from PHT to the diagnosis of SLE was 2.3 years. Three cases were severe PHT with remarkable heart failure. Twenty-seven eases demonstrated Raynaund's phenomenon. Antinuclear antibody (ANA) was positive in all 38 cases and the titer was 1:320. Anti-DNA antibody, anti-Sin antibody, anti-U1RNP antibody and anti-SSA, anti-SSB were positive in 21 cases, 16 cases, 15 cases, 8 cases respectively. Antiphospholipid antibody and rheumatoid factor (RF) were elevated in 4 cases and 13 cases respectively. Seven cases were involved in lung fibrosis and pulmonary function test showed mild to moderate restrictive abnormality in these 7 cases. After treated with the combination of steroid and immunosuppressive agents, one case died on the 10th day and 2 cases died in 2 to 3 years after the diagnosis was established. The other 35 cases stayed stable. Conclusion PHT is one of the severe complications of SLE and often accompanied by poor outcome. Therefore early diagnosis and early combined therapy is very important.

Objective To investigate the efficacy and the adverse effect of mycophenolate mofetil(MMF)in HBV associated glomerulonephritis.Methods Twenty patients with HBV-GN diagnosed by renal biopsy were treated with MMF.The initial dosage of MMF was 1.5 g/d.The dosage was reduced to 1.0 g/d when 24 h excretion of urinary protein decreased to 0.5 g/d.Blood routine test,hepatic and renal function,plasma albumin,serum triglyceride and cholesterol,24 h urinary protein excretion,HBV DNA were performed before and after 2,3,6,9 months' treatment.Results Twenty-four-hour urinary protein excretion was significantly reduced by the treatment with mycophenolate mofetil(P

BACKGROUND:Erythropoietin and progenitor cel transplantation both have therapeutic effects on lower extremity arterial occlusive disease.
OBJECTIVE:To investigate the erythropoietin modification effect and magnetic resonance imaging feasibility of superparamagnetic iron oxide (SPIO)-labeled endothelial progenitor cel s in vitro.
METHODS:Rat bone marrow-derived endothelial progenitor cel s at logarithmic growth phase were randomized into four groups:endothelial progenitor cel group, SPIO labeled transfection group (pcDNA3-EPO transfection fol owed by SPIO labeling), SPIO labeled empty vector group (empty plasmid transfection fol owed by SPIO labeling), and SPIO labeling group (only SPIO labeling). 4.7T MRI was used to observe SPIO-labeled endothelial progenitor cel s. Cel proliferation, cel cycle distribution, and expression of erythropoietin protein in the four groups were measured.
RESULTS AND CONCLUSION:MRI findings showed with the increasing cel number, gradual y lowered signal intensity on T1-weighted imaging (T1WI), T2WI and T2*WI was seen, and the reduction in the signal intensity was the maximum on T2*WI sequence and the minimum on T1WI sequence. For T1WI, T2WI and T2*WI sequences, the minimum number of cel s was 2×104, 1×104 and 0.5×104, respectively. Cel proliferation and cel cycle distribution showed no significant difference among three SPIO labeling groups. In addition, the expression of erythropoietin protein was only found in the SPIO-labeled transfection group. These findings showed that under SPIO labeling, erythropoietin gene-modified endothelial progenitor cel s show no changes in cel proliferation and cel cycle, and the 4.7T MR is capable of imaging SPIO-labeled erythropoietin gene-modified endothelial progenitor cel s in vitro.

Objective To investigate the effect of aliskiren on type 2 diabetic nephropathy in db/db mice.Methods Eight-week old db/db and db/m mice were subjected to right nephrectomy to hasten the development of diabetic nephropathy.At age of 16 weeks,they were divided into four groups:db/m group (normal control),db/db group ( diabetic control),db/db+ A3 group (db/db mice treated with aliskiren 3 mg·kg-1d-1) and db/db+A25 group (db/db mice treated with aliskiren 25 mg kg-1 d-1).Body weight,blood glucose,glycosylated hemoglobin,proteinuria and systolic blood pressure were measured before and after treatment.After treatment,renal histological examination by PAS staining,TGF-β1 and PAI-1 protein by ELISA,expression of ColⅣ and FN protein by immunofluorescence,mRNA expression of TGF-β1,PAI-1,ColⅣ,FN and renin by real time PCR,renin activity and angiotensin Ⅱ level by radioimmunoassay were performed.Results Treatment for 4 weeks of aliskiren at a dose of 25 mg kg-1 d-1 markedly decreased urinary albumin excretion,glomerulosclerosis and suppressed synthesis of TGF-β1,PAI-1,Col Ⅳ,FN without inducing significantl change in systolic blood pressure,compared to those of db/db group (all P＜0.05).Aliskiren also suppressed renin activity and angiotensin Ⅱ level in renal cortex (all P＜0.05).Conclusion Aliskiren protects against type 2 diabetic nephropathy.

Objective To investigate the correlation of serum anti-M-type phospholipase A2 receptor (PLA2R) antibody with laboratory parameters of idiopathic membranous nephropathy (IMN) in adult patients with membranous nephropathy (MN),and to explore the role of anti-PLA2R antibody in the pathogenesis of IMN. Methods Forty-six adult patients with biopsy-proved glomerular diseases were involved in this study,including 20 cases with IMN,7 cases with IgA nephropathy (IgAN),6 cases with hepatitis B-associated membranous nephropathy (HBV-MN),6 cases with minimal change nephropathy (MCN),4 cases with focal segmental glomerulosclerosis (FSGS) and 3 cases with class Ⅴ lupus nephritis.Total RNA was extracted from human glomeruli and was reversely transcribed to the first-strand cDNA.The full-length human M-type PLA2R was amplified by PCR and the 605 bp product was subcloned into eukaryotic expression vector containing CMV promoter.The recombinant human M-type PLA2R plasmid vector was transiently transfected into human embryonic kidney (HEK) 239T cell line using the FuGene6 transfection reagent.Western blotting was used to detect serum anti-PLA2R antibodies.Correlations of antiPLA2R antibody level with laboratory parameters,including serum albumin,total cholesterol,Scr and 24-hour urine protein,of IMN patients were evaluated. Results Among 20 cases with IMN,15 cases showed positive anti-PLA2R antibodies (positive rate 75％).Of 7 cases with IgAN and 6 cases with HBV-MN,only 1 case showed positive anti-PLA2R antibody respectively (positive rate 14.29％ and 16.67％ respectively).Anti-PLA2R antibody was negative in other patients.The positive rate of anti-PLA2R antibody in IMN patients was significantly higher than that in patients with secondary MN and other types of glomerlonephritis (all P＜0.01).Furthermore,anti-PLA2R antibody level was positively correlated with 24-hour urine protein(r=0.803,P＜0.01) and negatively correlated with serum albumin in IMN patients (r=-0.816,P＜0.01). Conclusions The high positive ratio of anti-PLA2R antibody may indicate that it is the specific autoantibody in IMN.AntiPLA2R antibody is correlated with IMN disease severity,which indicates that it may be the pathogenic autoantibody in IMN.

BACKGROUND:Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials were developed in our previous studies. OBJECTIVE:To observe the capability of gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials in the repair of rabbit radius defects. METHODS: Thirty healthy adult New Zealand rabbits were selected to make critical-size lacunar bone defects of the upper radius (15 mm×6 mm). Then, the rabbit models were randomized into experimental group (n=15), autogenous bone graft group (n=10) and blank group (n=5). Gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite materials were implanted into radial bone defects in the experimental group. Bone defect in blank group was implanted without any materials; in the autogenous bone graft group, the contralateral radius with same length was taken and implanted into the defect. General observation, histological observation and X-ray observation were performed respectively at 2, 4, 8, 12 weeks postoperatively. RESULTS AND CONCLUSION:At 12 weeks after operation, the experimental group showed obvious new blood vessels at the defect region, complete bony union and disappearance of the composite implant, but lamelar bone structure appeared, smal blood vessels were visible, the edge of new bone was connected to the original bone edge, exhibiting a continuity of bone, the bone density was slightly lowered, and the defect region became unobvious. In the autogenous bone graft group, bony union and trabecular bone reconstruction were distinct, the lamelar bone became mature, the medulary cavity was recanalized, the fracture line disappeared completely, and the bone density was completely consistent with that of the original bone. In the blank group, there was no obvious bone formation, which led to bone nonunion, and there were a great amount of fiber tissues and inflammatory cel infiltrated. To sum up, the gelatin-chitosan-hydroxyapatite-minocycline biomimetic nanocomposite material can obviously promote the repair of critical-size bone defects, and the repairing effect is basicaly the same with that of autologous bone grafting.

Objective:To evaluate the curative efficacy of tacrolimus (FK506)combined with corticosteroids in the treatment of idiopathic membranous nephropathy (IMN ), and to analyze the response of anti-M-type phospholipase A2 receptor (PLA2R)antibody to the treatment.Methods:Sixty-one adult IMN patients were divided into FK506 group (FK506 combined with corticosteroids,n = 24)and CTX group (cyclophosphomide combined with corticosteroids,n=37)according to their willing to the acceptance of different immunosuppressives. The remission rates of the patients at 4,8,12 and 24 weeks after treatment in two groups were analyzed.The ELISA method was used for the detection of serum anti-PLA2R antibodies of the patients before and after treatment.The antibody level changes were observed and the difference of remission rates was compared between the antibody positive patients and the negative patients in FK506 group.Results:The total remission rates of the patients in FK506 and CTX groups 24 weeks after treatment were 91.7% and 64.9% respectively and the total remission rate of the patients in FK506 group was significantly higher than that in CTX group (P < 0.05).The total remission rates at 8,12,24 week after treatment and the partial remission rates at 12 and 24 weeks in FK506 group were significantly higher than those in CTX group (P <0.05).The anti-PLA2R antibody positive rates in FK506 group and CTX group were 75.0% and 70.3%,respectively;the negative conversion rate of anti-PLA2R antibody at 24 weeks after treatment in two groups were 61.6% and 57.7%,respectively;there were no significant differences between two groups (P >0.05).The total remission rates of the antibody positive patients and negative patients at baseline in FK506 group 24 weeks after treatment were 88.9% and 100.0%,respectively;there was no significant difference (P > 0.05).Except the characteristic side effects of corticosteroids,the patients in FK506 group presented no hyperglycemia,tubulointerstitial damage,hepatic lesion or neurotoxicity. But the patients in CTX group showed mild hepatic lesion or gastrointestinal symptoms.Conclusion:The remission rate of FK506 combined with corticosteroids is higher than that of CTX combined with corticosteroids in the treatment of IMN patients.FK506 treatment has relatively rapid effect and less side effects as well.The negative conversion of serum anti-PLA2R antibody follows the remission after treatment;however,the negative conversion rate has no significant difference between FK506 group and CTX group.The remission rate of IMN patients treated with FK56 combined with hormone has no relationship with the baseline anti-PLA2R antibody.