Objective To investigate the protective effect of mucopolysaccharide polysulfate cream(MPC) and micro needle array drug delivery technologarray(MNADDT) on the blood vessel of arteriovenous fistula.Methods Eighty cases maintenance hemodialysis patients with autogenous arteriovenous fistula were randomly divided into control group(with the treatment of MPC) and study group(with the treatment of MPC plus MNADDT),40 patients in each group.Both groups were studied for the duration of 12 months.The vascular complications of puncture pain sense,puncture failure times,phlebitis,hardening of the arteries,internal fistula stenosis or embolism,pseudoaneurysm,swollen hands comprehensive syndrome of two groups were observed,and the changes of arteriovenous fistula blood flow,blood vessel diameter by Doppler ultrasound were observed in the two groups for 12 months.Results (1)The incidence of puncture pain(2.71±0.11 vs.2.76±0.14,t=2.21,P<0.05),puncture failure times(38 times vs.73 times,χ2=11.425,P<0.05),phlebitis(2 cases vs.8 cases,χ2=4.341,P<0.05) and blood vessel sclerosis(2 cases vs.9 cases,χ2=5.446,P<0.05) of the treatment group were significantly lower than that of the control group during the study(P<0.05).(2)Blood flow of arteriovenous fistulain of control group before and after treatment were (859.7±148.3) ml/min and (946.5±169.2) ml/min respectively,and the difference was significant(t=2.356,P<0.05).While blood flow of arteriovenous fistulain,cephalic vein diameter,head of venous flow velocity of study group before and after treatment were (824.6±171.5) ml/min and (1 218.1±241.9) ml/min,(5.59±0.74) mm and (6.02±0.57) mm,(94.23±27.35) cm/s and (115.85±36.63) cm/s,respectively,and the differences were significant(t=8.212,2.382,2.877,P<0.05),there were no significantly changes in the above indexes in the study group after treatment(t=5.3612,2.152,2.281,P<0.05).Conclusion MPC plus MNADDT can reduce the vascular complications of arteriovenous fistula,improve blood vessel diameter and increase blood flow.

ObjectiveTo investigate the outcome and risk factors for kidney involvement by analyzing 64 patients with anti-neutrophil cytoplasmic antibody(ANCA)-associated vasculitis.MethodsData analyzed including the demographic information,survival status,renal survival status and laboratory parameters such asserum albumin level,serum creatinine level,urinary protein excretion level,hematuria,high sensitivity C-reactive protein(CRP),ANCA titer,and the Birmingham vasculitis activity score (BVAS).Logistic regression analysis,Cox regression analysis and ROC curve were used to evaluate the risk factors of patients with renal involvement and all-event survival.ResultsTotally 64 patients were enrolled [24 females with the average age of (59.9±2.0) years] and followed up for a median of (38±16) months.The morality rate was 14％,and the prevalence of end stage renal disease was 39％.Compared with those who had better outcomes,patients who died or with end stage renal disease had higher serum creatinine level [ (624±246),(245±127 ) μ mol/L,respectively,t=7.17,P＝0.005 ] and erythrocyte sediment rate [ (112±24),(76±48) mm/1 h,respectively,t=3.74,P＜0.01 ],but lower serum albumin level [(294±31 ),(316±42) g/L,respectively,t=-2.27,P=0.01 ] and hemoglobin level [ (79±13),(99±33) g/L,respectively,t=-3.23,P＜0.01 ] at baseline.Logistic regression analysis found that serum creatinine level and erythrocyte sediment rate at baseline were associated with poor outcome and Cox regression analysis further confirmed this result[Scrβ=1.004,95％CI1.002～1.006,P＜0.01; ESR β=l.018,95％CI 1.000～1.037,P=0.046].ROC curve analysis showed that serum creatinine and erythrocyte sediment rate were predictors for AAV patients' prognosis and their AUC were 0.95 and 0.80,the sensitivity of these parameters was both 94％,and the specificity was 93％ and 70％respectively.ConclusionThe intensity of initial treatment should be based on disease severity and activity in order to improve the prognosis of those with ANCA-associated vasculitis with renal involvement.Increased serum creatinine and erythrocyte sediment rate may serve as predictors for poor prognosis in this patient cohort.

Objective To elucidate the prevalence and risk factors of cardiovascular disease (CVD) in end-stage renal disease (ESRD) patients on peritoneal dialysis (PD), and to investigate the associated problems in treatment. Methods A total of 254 PD patients in our division were enrolled in this study. CVD history, laboratory measurements, examinations of carotid atherosclerosis and left ventricular hypertrophy by ultrasonography were collected and associated factors were analyzed. The median follow-up time was 49 months. Results The overall prevalence of CVD was 37% (93/254). Diabetes, longer dialysis duration, hypertfiglyceridemia, hypoalbuminemia, hypoprealbuminemia were commonly found in the patients with new CVD event. The patients without pre-existing CVD had the higher Ccr, Kt/V, D/Pr, nPCR, serum albumin level. In those with pre-existing CVD, the hypertriglyceridemia and the duration of dialysis were independent predictors of progression of CVD. Differences of LAD, LVST, LVMI and IMT were significant between with and without pre-existing CVD groups. Kaplan-Meier curves showed that the presence of CVD was the independent risk factor of survival. Alb<330 g/L, LAD>39.6 mm and peritonitis were risk factors of CVD. Conclusion The prevalence of CVD in PD patients is quite high. CVD history should be realized, dialysis adequacy should be maintained, and peritonitis should be prevented.

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α(+) dendritic cells (DCs) in vitro was investigated in this study. Immature CD8α(+) DCs were prepared from C57BL/6 (H-2(b)) bone marrow cells by using a cytokine cocktail. On the 3rd day of culture, CD8α(+) DCs were pulsed by allogeneic (Balb/c, H-2(d)) EL9611 leukemia antigen, or RM-1 syngeneic prostate cancer antigen, with the concentration series of 0, 2.5, 5.0, 10.0, 20.0 μg/mL, respectively, then antigen-loaded immature CD8α(+) DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1:1, 2:1 and 4:1. T cell proliferation was measured by MTT assay. Cytokines including interferon gamma (IFN-γ) and interleukin-10 (IL-10) in CD8α(+) DCs and T co-culture supernatant were detected by using ELISA. Cytotoxic effect of antigen-specific T cells was tested by LDH release assay. Conventional mature DCs (mDCs) induced from C57BL/6 (H-2(b)) bone marrow cells by using granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control. The results showed that the proliferative activity of T cells stimulated by CD8α(+) DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α(+) DC/T ratio increased (P<0.05). When antigen concentration ≤ 5 μg/mL and CD8α(+) DC/T ratio ≤ 2:1, the ability of CD8α(+) DCs to stimulate T cell proliferation was higher than mDC control in allogeneic tumor antigen-pulsed groups (P<0.05), but not in syngeneic tumor antigen-pulsed groups (P>0.05). The level of IFN-γ and IL-10 in CD8α(+) DCs and T cell co-culture supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P<0.05), and the cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when the CD8α(+) DC/T was 1:1 or 2:1 (P<0.05). There existed a negative correlation between the level of IL-10 and T cell proliferation. T cell cytotoxicity assay showed that when CD8α(+) DCs were pulsed with allogeneic tumor antigen, the maximal T cell killing efficiency could reach (100±7.7)%, whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)%. It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α(+) DCs could stimulate T cells to exert the GVT effect in vitro, and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen. The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α(+) DC/T ratio (1:1 and 2:1).

The graft-versus-tumor (GVT) effect of T cells induced by tumor antigen-pulsed CD8α+dendritic cells (DCs) in vitro was investigated in this study.Immature CD8α+ DCs were prepared from C57BL/6 (H-2b) bone marrow cells by using a cytokine cocktail.On the 3rd day of culture,CD8α- DCs were pulsed by allogeneic (Balb/c,H-2d) EL9611 leukemia antigen,or RM-1 syngeneic prostate cancer antigen,with the concentration series of 0,2.5,5.0,10.0,20.0 μg/mL,respectively,then antigen-loaded immature CD8α+ DCs were co-cultured with syngeneic T cells according to the DC/T ratio of 1∶1,2∶1and 4∶1.T cell proliferation was measured by MTT assay.Cytokines including interferon gamma (IFN-γ)and interleukin-10 (IL-10) in CD8α+ DCs and T co-culture supernatant were detected by using ELISA.Cytotoxic effect of antigen-specific T cells was tested by LDH release assay.Conventional mature DCs　(mDCs) induced from C57BL/6 (H-2b) bone marrow cells by using granulocyte-macrophage colony　stimulating factor (GM-CSF) and interleukin-4 (IL-4) served as a control.The results showed that the　proliferative activity of T cells stimulated by CD8α+ DCs loaded with allogeneic or syngeneic tumor antigen was augmented with the CD8α+ DC/T ratio increased (P＜0.05).When antigen concentration ≤ 5μg/mL and CD8α+ DC/T ratio ≤ 2∶1,the ability of CD8α+ DCs to stimulate T cell proliferation was　higher than mDC control in allogeneic tumor antigen-pulsed groups (P＜0.05),but not in syngeneic tumor antigen-pulsed groups (P＞0.05).The level of IFN-γ and IL-10 in CD8α+DCs and T cell co-culture　supernatant were increased in both allogeneic and syngeneic antigen-pulsed groups (P＜0.05),and the　cytokine level was higher in allogeneic antigen-pulsed groups than in syngeneic antigen groups when　the CD8α+DC/T was 1∶1 or 2∶1 (P＜0.05).There existed a negative correlation between the level of　IL-10 and T cell proliferation.T cell cytotoxicity assay showed that when CD8α+ DCs were pulsed with　allogeneic tumor antigen,the maximal T cell killing efficiency could reach (100±7.7)％,whereas syngeneic tumor antigen-pulsed group had only (65.0±3.4)％.It was concluded that syngeneic and allogeneic tumor antigen-pulsed immature CD8α+ DCs could stimulate T cells to exert the GVT effect in vitro,and the GVT effect was more obvious with allogeneic tumor antigen than with syngeneic tumor antigen.The optimal condition was low allogeneic tumor antigen pulsation (≤ 5 μg/mL) and low CD8α+ DC/T　ratio (1∶1 and 2∶1).

Objective:To establish autoverification rules for coagulation tests in multicenter cooperative units, in order to reduce workload for manual review of suspected results and shorten turnaround time (TAT) of test reports, while ensure the accuracy of results.Methods:A total of 14 394 blood samples were collected from fourteen hospitals during December 2019 to March 2020. These samples included: Rules Establishment Group 11 230 cases, including 1 182 cases for Delta check rules; Rules Validation Group 3 164 cases, including 487cases for Delta check; Clinical Application Trial Group 77 269 cases. Samples were analyzed for coagulation tests using Sysmex CS series automatic coagulation analyzers, and the clinical information, instrument parameters, test results, clinical diagnosis, medication history of anticoagulant and other relative results such as HCT, TG, TBIL, DBIL were summarized; on the basis of historical data, the 2.5 and 97.5 percentile of all data arranged from low to high were initially accumulated; on the basis of clinical suggestions, critical values and specific drug use as well as relative guidelines, autoverification rules and limits were established.The rules were then input into middleware, in which Stage I/Stage II validation was done. Positive coincidence, negative coincidence, false negative, false positive, autoverification pass rate, passing accuracy (coincidence of autoverification and manual verification) were calculated. Autoverification rules underwent trial application in coagulation results reports.Results:(1) The autoverification algorisms involve 33 rules regarding PT/INR, APTT, FBG, D-dimer, FDP,Delta check, reaction curve and sample abnormalities; (2)Autoverification Establishment Group showed autoverification pass rate was 68.42% (7 684/11 230), the false negative rate was 0%(0/11230), coincidence of autoverification and manual verification was 98.51%(11 063/11 230), in which positive coincidence and negative coincidence were respectively 30.09% (3 379/11 230) and 68.42%(7 684/11 230); Autoverification Validation Group showed autoverification pass rate was 60.37%(1 910/3 164), the false negative rate was 0%(0/11 230), coincidence of autoverification and manual verification was 97.79%(3 094/3 164), in which positive coincidence and negative coincidence were respectively 37.42%(1 184/3 164) and 60.37%(1 910/3 164); (3) Trialed implementation of these autoverification rules on 77 269 coagulation samples showed that the average TAT shortened by 8.5 min-83.1 min.Conclusions:This study established 33 autoverification rules in coagulation tests. Validation showedthese rules could ensure test quality while shortening TAT and lighten manual workload.

Objective : To investigate whether the application of melatonin (MT) in embryo culture in vitro can improve the treatment effect of in vitro fertilization embryo transfer(IVF⁃ET) in patients with decreasing ovarian reserve (DOR) . Methods : 128 DOR patients receiving assisted reproductive therapy were collected. All patients were treated with an antagonist scheme of super⁃ovulation. Patients were divided into melatonin group (n = 56) and control group (n = 72) according to whether melatonin ( melatonin concentration 10 - 9 mol/L) was added into embryo culture medium. Results : There was no statistically significant difference in oocytes fertilization rate and cleavage rate between the two groups during later embryo culture , but blastocyst formation rate ( 65. 22% vs 56. 16% ) and high⁃quality blastocyst rate (52. 96% vs 40. 94% ) in the melatonin group were higher than those in the control group , and the differences were statistically significant ( P < 0. 05 ) . There were no significant differences in the implantation rate (50. 00% vs 38. 67% ) and clinical pregnancy rate (48. 39% vs 46. 00% ) of blastocysts after freezing⁃thawing between the two groups , but the cycle number of high⁃quality blastocysts obtained in the melatonin group was higher than that in the control group (85. 71% vs 69. 44% ) , and the difference was statistically significant (P < 0. 05) . Conclusion In a way , the application of melatonin in the in vitro culture of early embryos can promote the development of oocytes in patients with DOR , improve the quality of embryos , and finally substantially improve the therapeutic effect of such patients.

Cell Cycle Checkpoints/genetics* ; Checkpoint Kinase 1/metabolism* ; Heterozygote ; Humans ; Mitosis/genetics* ; Mutation ; Zygote/metabolism*