1.Evaluation of the effects of calibration modes and sample reconstitution on the analytic precision of 26 clinical chemistry analytes using system measurement procedure
Huiying SUN ; Yan SHAO ; Shuming LIU ; Bin HU ; Baorong CHEN
Chinese Journal of Laboratory Medicine 2018;41(2):149-154
Objective To evaluate the system measurement procedure effects on the analytic precision of clinical chemistry analytes.Methods In June 2009, June 2010 and September 2010 respectively,the National Center for Clinical Laboratories of China and the Organization of Five Hospitals in Fukuoka Japan organized comparison activities of 26 clinical chemistry analytes which were ALT,AST,GGT, ALP,CK,LDH,AMY,ChE,TG,TC,HDL-C,LDL-C,Glu,Cr,BUN,UA,K,Na,Cl,Ca,TP,Alb,TBil,DBil, P,Fe.In this paper, we investigated 26 analytes of three sets in Beijing Aerospace General Hospital as follows.(1)The precision of different reconstitution methods was observed by using three kinds of pipetting tools, such as measuring pipette, pipette and dispenser.(2)The experiments were carried out in three stages by testing the dried powder control samples of two concentration levels(101-Ⅰ,101-Ⅱ)provided by Hitachi Japan.They were measured on 28 consecutive days at each stage in order to observe the precision of 26 clinical chemistry analytes.In the first stage,we used the former measurement procedure to measure the control samples;in the second stage we added three conditions of the measurement procedure.The first was two calibration modes,which were once-a--day calibration and twice-a--day calibration.The second was the calibration standard and the last was the conditions of the freeze-thaw samples.In the third stage, we used the twice-a-day calibration only for GGT,ALP,ChE,TG,Cr,Na,K,CL,ALB.(3)JSCC and Health Industry Standard quality objectives were implemented to evaluate whether the precision of the improved measurement procedure met the requirements.(4)Paired T test were used to compare the precision of measurement between the second stage and the first stage, and between the third stage and the second stage of the measurement procedure.Results (1)The precision of three kinds of pipetting tools were 0.56%,0.10%, 0.01%.(2)The ranges of precision of ALT,AST,GGT,ALP,CK,LDH,AMY,ChE,TG,TC,HDL-C,LDL-C,Glu,Cr,BUN,UA,K,Na,Cl,Ca,TP,Alb,TBil,DBil,P,Fe were 0.99%-10.5% about 101-Ⅰ and 0.91%-7.03%about 101-Ⅱin the first stage.The ranges of precision were 0.66%-8.81%of 101-Ⅰand 0.66%-4.28%of 101-Ⅱin the second stage.The ranges of precisions were 0.60%-3.91%of 101-Ⅰand 0.73%-3.39%of 101-Ⅱin the third stage.(3)73%/80%of the samples met the standard of JSCC about 101-Ⅰand 101-Ⅱand 80%/88%of the samples met the standard of Health Industry Standard in the first stage.88%/100% of the samples met the standard of JSCC about 101-Ⅰand 101-Ⅱ and 100%/100%samples met the standard of Health Industry Standard in the second stage.The ratio of samples meeting the standard of JSCC about 101-Ⅰand 101-Ⅱwere 96%/100% and that of Health Industry Standard were 100%/100%in the third stage.(4)Precision of 101-Ⅰand 101-Ⅱwas statistically significant between the measurement procedures of second stage and the first stage,and there was no significant difference between the third stage and the second stage.Conclusion (1)The precision of samples using dispenser to reconstitute is higher than that of the other two pipetting methods.(2)Improving the calibration mode and reconstitution of samples increase the precision of 26 clinical chemistry analytes by over 50%.
2.Therapeutic window of whole blood rapamycin concentration in recipients of renal transplantation
Changxi WANG ; Wenjun SHANG ; Lizhong CHEN ; Jiguang FEI ; Bin REN ; Shuxia LI ; Keli ZHENG ; Xiaoda TANG ; Yu FAN ; Zhilian MIN ; Juan Qi ; Zhihong LIU ; Shuming JI ; Leishi LI ;
Chinese Journal of Nephrology 1997;0(05):-
Objective To study the therapeutic window of rapamycin(RPM) concentration in primary recipients of renal transplantation. Methods An open label, multi center study was performed. One hundred primary renal allograft recipients with cadaveric donors were enrolled from 4 transplantation centers in China. The immunosuppressive regimen was triple therapy,i.e.RPM combined with CsA and steroid. A loading dose of RPM 6 mg/d was administered within 48 hours after transplantation, then a maintaining dose of 2 mg/d was administered. The whole blood concentration of RPM was measured by HPLC method. Results The whole blood concentration of RPM in this group was (6.65?2.75)ng/ml, the 10th and 90th percentile for RPM concentration was 3 2 ng/ml and 10 26 ng/ml,respectively.9 5%(8/84)patients suffered from acute rejection during the 6 month period after transplantation in this study, and the concentration of RPM in these was lower than that in non rejection patients(P=0.001). Hyperlipidemia and liver dysfunction were the most frequently adverse events, and RPM concentration was significantly associated with the concentration of triglyceride. Conclusions 4~8 ng/ml is a suitable level for RPM concentration. Regular drug monitoring and reasonable dose modulation may increase the validity and security of RPM.
3.Protective effect of herba artemisiae scopariae aqueous extract on neonatal parenteral nutrition-associated cholestasis induced by multidrug resistance protein 3 gene mutation
Xiufang YANG ; Shuming BIN ; Dan LI ; Huiying LIANG ; Kang CHEN ; Kaijun ZHENG ; Juncai DING ; Qiaowei ZHU ; Shangwen SHI ; Guiling CHEN
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(3):308-314
Objective To investigate the protective effect of herba artemisiae scopariae extract on multidrug resistance protein 3(MDR3)gene mutation-induced neonatal parenteral nutrition-associated cholestasis(PNAC)and its possible mechanism.Methods ①Human primary hepatocytes were treated with cell culture in vitro,CRISPR/Cas9 lentivirus infection and MDR3 mutant gene lead-in.The levels of hepatic and biliary biochemical indexes[alanine transaminase(ALT),aspartate transaminase(AST),total bilirubin(TBil),direct bilirubin(DBil),indirect bilirubin(IBil),total bile acid(TBA)]in the supernatant of hepatocytes before and after 16,32,48 hours were compared to determine the time required for fatty acid induction of PNAC hepatocyte model with MDR3 gene mutation.② Human primary hepatocytes were divided into blank control group,MDR3 gene wild type group,MDR3 gene mutation group,and herba artemisiae scopariae extract intervention group according to random number table method.The blank control group was treated with culture medium only,the MDR3 gene wild type group was infected with lentivirus and mixed with wild type MDR3 gene and culture medium,the MDR3 gene mutation group was infected with lentivirus and cultured in culture medium with the mutant genes lead-in of LV-MDR3KI(c.485T>A,c.2793insA,c.1031G>A,c.3347G>A)mutation,while the MDR3 mutant gene was lead-in by lentivirus infection and cultured in culture medium,and then pretreated with 100 g/L herba artemisiae scopariae extract in the herba artemisiae scopariae extract intervention group,then the four groups of hepatocytes were induced with 1%fat emulsion,and the treatment time was the time needed to construct the PNAC hepatocytes model with MDR3 gene mutation.The levels of ALT,AST,TBil,DBil,IBil and TBA in the supernatant of hepatocytes were measured by enzyme-linked immunosorbent assay(ELISA).The mRNA expression abundance of adenosine triphosphate binding cassette proteins(ABCB4,ABCB11,ABCC2,ABCC3,ABCC4)encoding MDR3,bile salt export pump(BSEP),multidrug resistance associated protein(MRP)2-4,and tumor necrosis factor-α(TNF-α)genes were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results Compared to the blank control group and MDR3 gene wild type group,there was no significant difference in the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 gene mutant group before and 16 hours after induction with 1%fat emulsion,however after treated with 1%fat emulsion for 32 hours and 48 hours,the levels of ALT,AST,TBil,DBil,IBil,TBA in the supernatant of MDR3 mutant hepatocytes were significantly increased(P<0.05),consequently the time required for fatty acid induction of PNAC hepatocyte model was 32 hours.At 32 hours after treatment with fat emulsion,the levels of ALT,AST,TBil,DBil,TBA in the supernatant of hepatocytes in the herba artemisiae scopariae extract intervention group were significantly decreased[ALT(ng/L):148.3±2.3 vs.164.9±7.0,AST(ng/L):2767.4±78.8 vs.3239.4±107.1,TBil(μmol/L):7.6±0.2 vs.13.6±0.3,DBil(μmol/L):1.8±0.1 vs.5.7±0.2,TBA(μmol/L):3.4±0.2 vs.6.7±0.1,all P<0.05].The ABCB4,ABCC2,ABCC3,ABCC4 mRNA expression of MDR3,MRP2,MRP3,MRP4 in the blank control group,MDR3 wild type group,MDR3 gene mutation group and the herba artemisiae scopariae extract intervention group had no significant difference.The expression of TNF gene mRNA was highly expressed in MDR3 gene mutation group(2-??Ct:1.258±0.200 vs.1.001±0.052),and was low expressed in the herba artemisiae scopariae extract intervention group(2-??Ct:0.387±0.247 vs.1.258±0.200),and there was a significant difference between the two groups(both P<0.05).Compared to the MDR3 gene mutation group,the ABCB11 gene encoding BSEP mRNA expression in the herba artemisiae scopariae extract intervention group was significantly increased(2-??Ct:2.955±0.479 vs.1.333±0.529,P<0.05).Conclusion The herba artemisiae scopariae extract has a protective effect on PNAC induced by MDR3 gene mutation,which may be related to antagonizing inflammatory reaction,decreasing the expression of TNF mRNA and improving the expression of ABCB11 gene encoding BSEP.
4.Application of allogeneic platelet gel in surgical operation for giant aortic aneurysms, one case
Jianjun WU ; Bin JIANG ; Renxue XIANG ; Ronghui SHI ; Kang YANG ; Wei LIU ; Xin CUI ; Shuming ZHAO
Chinese Journal of Blood Transfusion 2021;34(7):698-701
【Objective】 To explore the effect of allogeneic platelet gel on hemostasis and repair of vascular anastomoses and wounds in patients with giant aortic aneurysms during surgery. 【Methods】 One adult dose of allogeneic platelets, applied as platelet rich plasma (PRP), was prepared as platelet gel (PG) (about 220 mL) for spraying or smearing at the vascular anastomosis and thoracic wound during the surgery of giant aortic aneurysms. 【Results】 The gel formation was presented about 30 seconds after spraying or smearing with PG on the anastomotic and wound surface. The hemostatic effect is good, with less postoperative drainage fluid from pericardium and mediastinum than usual, and the ICU stay was 4 days. 【Conclusion】 Allogeneic platelets as a source of PRP to prepare PG may be applied to obtain the clotting and healing during surgical operation.