Objective Establish quality control methods for critical quality attribute of bispecific antibody against programmed cell death protein 1(PD-1)/cytotoxic T-lymphocyte-associated protein 4(CTLA-4).Methods The biological activity of PD-1 target was determined by reporter gene assay,and the competitive binding activity of CTLA-4 target was determined by flow cytometry;The antibody molecular size variants were controlled by reducing/non-reducing capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS) and size exclusion chromatography-high performance liquid chromatography(SEC-HPLC);Charge heterogeneity was determined by imaging capillary isoelectric focusing electrophoresis(iCIEF);Bispecific anti-PD-1/CTLA-4 antibody was identified by peptide map analysis;Glycosylation was analysed by high performance liquid chromatography(HPLC)Results The concentration for 50% of maximal effect(EC_(50)) of PD-1target was(6.91±0.78) nmol/L,and the relative biological potency to the reference was(103.50±13.08)% with the RSD of 12.64%;The EC_(50) of CTLA-4 target activity was(0.35±0.28) nmol/L,and the relative biological potency was(99.30±9.15)% with the RSD of 8.32%.The percentage of peak area of light chain and heavy chain of reducing CE-SDS was(98.86±0.02)%.The main peak area percentage of non-reducing CE-SDS was(93.07±0.13)%,fragment percentage was(4.44±0.13)%,and polymer percentage was(2.49±0.15)%.The peak area percentage of SEC-HPLC monomer and polymer were(97.20±0.01)% and(2.68±0.01)%,respectively.The area percentage of peak A group,peak B group,peak C group and peak D group were(38.43±0.54)%,(43.26±0.32)%,(11.31±0.14)% and(7.00±0.17)%,respectively.Peptide mapping showed the specific spectrum of the bispecific anti-PD-1/CTLA-4 antibody,which could be adopted for identification test.The highest proportion of glycotype was GOF,with a content of(41.06±0.11)%,There were three types of glycan containing sialic acid,namely G2F+G1F-NANA,G2F-NANA and G2F-2NANA,with the content of(12.44±0.12)%,(12.00±0.05)% and(5.37±0.05)%,respectively.The total content of glycan containing sialic acid was(29.80±0.20)%.Conclusion The critical quality attributes of bispecific anti-PD-1/CTLA-4 antibody were studied and the corresponding quality control methods were established to ensure its safety,effectiveness and quality control,which provides a reference for the quality control methods and strategies of this type of monoclonal antibody products.

Autoimmune diseases are characterized by the presence of various autoantibodies and may cause injuries to multiple organs,with kidney as the most common and important organ involved.Autoantibodies are of great importance in the diagnosis,treatment and prognosis of autoimmune renal diseases.Lupus nephritis,anti-neutrophil cytoplasmic antibody (ANCA) associated vasculitis and antiglomerular basement membrane (GBM) disease are the most common autoimmune renal diseases.Anti-C1q antibody,ANCA and anti-GBM antibody play important roles in those diseases,respectively.Appropriate and steady detecting methods are crucial to clinicians,and the results should also be interpreted with great cautions.

Objective To describe the work experience and needs of enterostomal therapist in Shanghai.Methods Phenomenological methodology was adopted in this study.In-depth interview was conducted on 9 enterostomal therapists.Data were analyzed by inductive codes and coding methods.Results Five themes were extracted,including responsibility and mission,pride,accomplishment,helplessness,and satisfaction.When they began their new jobs,they confronted many challenges,but they conquered them with responsibility and mission.Finally,they felt growing up and harvested a lot during this time.Conclusions Hospital managers should pay attention to problems they encountered in their work and provide necessary supports for their training,work and development.

Objective To study the effect of short-term inte ns ive insulin treatment on the pancreatic B-cell function in type 2 diabetic mell itus (T2DM).Methods Insulin and C-peptide serum levels were measured befor e and after intensive insulin treatment in T2DM by chemical luminescence immunoa ssay.Results The levels of insulin and C-peptide were increased at 0、30、60、120、180min in OGTT in T2DM after insulin intensive treatment.Conclusion Short-term intensive insulin treatment can partiall y improve the B-cell function of T2DM with lower function and made it rest.

Objective To study the levels of serum resistin and sICAM-1 in diffferent glucose tolerance status.Methods Fasting serum resistin and sICAM-1 concentration were measured by enzyme immunoassay in 52 cases suffering from T2DM,18 cases with glucose tolerance impaired and 30 cases with normal glucose tolerance.Meanwhile,fasting blood glucose,insulin and lipids were determined.HOMA-IR was calculated.Results Fasting serum resistin concentration (ng/mL) was higher in IGT than that in those patients with T2DM and NGT (P

Objective To study the clinicopathological features of C3 glomerulonephritis and the associations between plasma complement fragments level and clinical manifestations.Methods The clinical and pathological data of the 12 patients with C3 glomerulonephritis in our division from January 1999 to June 2010 were analyzed retrospectively.Concentrations of plasma factor B,Ba,C3,C3a,C4a and C5a were detected by commercial available sandwich ELISA kits on the day of renal biopsy.Results Ten of the 12 patients were C3 glomerulonephritis without MPGN,and the rest 2 were C3 glomerulonephritis with MPGN.All the patients presented proteinuria.Two of the 10 C3 glomerulonephritis patients without MPGN presented nephrotic range proteinuria,6 with microhematuria,1 with gross hematuria,and 2 with renal insufficiency.One of the 2 C3 glomerulonephritis patients with MPGN presented nephrotic range proteinuria,accompanied by microhematuria,hypertension and renal insufficiency.The other patient showed moderate proteinuria with normal renal function.Most of C3 glomerulonephritis patients without MPGN showed mild mesangial proliferative glomerulonephritis,and 4/10 patients had various degree cresentic formation.One C3 glomerulonephritis patient with MPGN had 47.1％ cresentic formation.The concentration of plasma C3 in C3 glomerulonephritis patients was normal,while the plasma factor B was significantly decreased,and the concentrations of plasma Ba,C3a,C4a and C5a were significantly elevated.The concentration of plasma Ba was positively correlated with the proteinuria level,while the concentrations of plasma C3a,C4a and C5a were not correlated with the levels of proteinuria or plasma creatinine.Conclusions Majority of these 12 patients were C3 glomerulonephritis without MPGN,manifests as nephritic syndrome clinically and mild mesangial proliferative glomerulonephritis histopathologically.Complement activation via alternative pathway may play an important role in the pathogenesis of C3 glomerulonephritis.

Objective To explore the relationship between different airway pressure and expression of related cytokines to airway remodeling.Methods Fourty-two chronic obstructive pulmonary disease cases (COPD group) and thirty-three control cases were collected.The above cases underwent mechanical ventilation in the period of general anesthesia.According to different levels of peak inspiratory pressure(PIP), above two groups, randomly and respectively, were divided into high PIP (HPIP, 24 cmH2O) group, moderate PIP (MPIP, 22 cmH2O) group, low PIP (LPIP, 20 cmH2O) group.All positive end expiratory pressure (PEEP) was set to 5 cmH2O.Then the collection of BALF was performed before and 3 hours after applying ventilator.The related factors to airway remodeling, such as fibroblast growth factor 2 (FGF-2), transforming growth factor β1 (TGF-β1) and matrix metalloproteinase-9 (MMP-9), were measured by enzyme-linked immunosorbent assay (ELISA) and Western blot.Results 1)Beforemechanical ventilation, the levels of FGF-2, TGF-β1 and MMP-9 in COPD group were higher than control group (P<0.01).2)3 hours after mechanical ventilation, we saw significant upregulated expression of FGF-2, TGF-β1, MMP-9 in HPIP subgroup in control group (P<0.05) and the 3 factors levels of COPD group were all increased (P<0.05).Moreover, HPIP subgroup was significantly higher than MPIP and LPIP subgroup in COPD group (P<0.05).3)The expression of FGF-2, TGF-β1, MMP-9 in BALF had a positive correlation with the airway pressure levels in COPD group(P<0.01).Conclusions Under mechanical ventilation, sustained high airway pressure may enhance the expression of FGF-2, TGF-β1 and MMP-9 which may result in airway remodeling by mechanosensitive cation channel in bronchial epithelial cells, especially in COPD patients.

Objective To study the influence of couple-centered mental intervention on marital quality and subjective happiness among young-and middle-aged breast cancer patients undergoing modified radical mastectomy.Methods A total of 100 postoperative young-and middle-aged breast cancer patients were divided into observation group (n=50) and control group (n=50) using random digit number table according to the order of hospitalization.The patients in the observation group were given the couple-based mental intervention,and the control were given the routine mental intervention only.The marital quality and subjective happiness between the 2 groups were evaluated and compared by Olson marital quality questionnaire and Memorial University of Newfoundland scale of happiness (MUNSH) respectively before intervention and 6 months after intervention.Result After intervention,both the score of marital quality and subjective happiness in the observation group were significantly higher than those in the control group (P＜ 0.05).Conclusion The enforcement of couple-centered mental intervention among the young-and middle-aged breast cancer patients undergoing modified radical mastectomy can significantly improve the marital quality and subjective happiness.

At present,reports at home and abroad suggest a low probability of successful preservation of ovarian endocrine function after ovarian shift radiotherapy.After radiotherapy for cervical carcinoma,the ovarian function is associated with various factors,such as radiotherapy dose and method,patient's age,shift position,and concurrent chemotherapy drugs.Therefore,as for each patient,the dose to the ovarian tissue should be controlled within the individual dose limit to effectively preserve the ovarian function.