Objective To evaluate the application of Cystatin C(Cys C)international primary reference material and national secondary reference materials for quantity transfer in seven conventional systems.Methods Seven Cys C reagents from different manufacturers commonly used in clinical laboratories were used to simultaneously measure two standard substances[ERM-DA471/IFCC,GBW(E)091173～6]and 42 individual serum samples.The theoretical and measured values of the standard substances were linearly fitted,and the measured values of the single serum samples were substituted into the fitting equation to obtain the calibrated values,and the measured values and calibrated values were compared with CLSI EP9-A3 file.Results The theoretical and measured values of ERM-DA471/IFCC and GBW(E)091173～6 were linearly fitted,and the regression equations were Y=0.941X+0.159,Y=0.963X+0.162,respectively,with correlation coefficients of 1.000(taking system B as an example).All system analysis models were analyzed using Passing-Bablok regression analysis.The comparison bias between the calibration values and measurement values of two standards in seven systems were-20.38%～10.58%and-16.76%～9.90%,respectively,with the same bias trend.The comparison bias of the calibrated values of the two standard substances was-4.18%～2.31%.The calibration of the standards had a significant improvement for the bias between calibration value and measurement value exceeding±4%.The bias of the measured values in the pairwise combinations of each system at each medical decision level was-17.87%～14.97%and the bias range of GBW(E)091173～6 calibration values at each medical decision level was-4.96%～4.51%,with most values being less than 3%.The bias of ERM-DA471/IFCC calibration values was-3.92%～6.30%,with most values being less than 4%.Among them,the most common situation was that the bias of ERM-DA471/IFCC calibration value was less than that of measurement value,and the bias of GBW(E)091173～6 calibration value was less than that of ERM-DA471/IFCC calibration value,with 51.19%(43/84)of the comparison bias bata at the level of medical decisionmaking.Conclusion Both reference materials can improve the alignment bias of conventional system,and GBW(E)091173～6 calibration effect is better than ERM-DA471/IFCC.

Objective To evaluate the application of Cystatin C(Cys C)international primary reference material and national secondary reference materials for quantity transfer in seven conventional systems.Methods Seven Cys C reagents from different manufacturers commonly used in clinical laboratories were used to simultaneously measure two standard substances[ERM-DA471/IFCC,GBW(E)091173～6]and 42 individual serum samples.The theoretical and measured values of the standard substances were linearly fitted,and the measured values of the single serum samples were substituted into the fitting equation to obtain the calibrated values,and the measured values and calibrated values were compared with CLSI EP9-A3 file.Results The theoretical and measured values of ERM-DA471/IFCC and GBW(E)091173～6 were linearly fitted,and the regression equations were Y=0.941X+0.159,Y=0.963X+0.162,respectively,with correlation coefficients of 1.000(taking system B as an example).All system analysis models were analyzed using Passing-Bablok regression analysis.The comparison bias between the calibration values and measurement values of two standards in seven systems were-20.38%～10.58%and-16.76%～9.90%,respectively,with the same bias trend.The comparison bias of the calibrated values of the two standard substances was-4.18%～2.31%.The calibration of the standards had a significant improvement for the bias between calibration value and measurement value exceeding±4%.The bias of the measured values in the pairwise combinations of each system at each medical decision level was-17.87%～14.97%and the bias range of GBW(E)091173～6 calibration values at each medical decision level was-4.96%～4.51%,with most values being less than 3%.The bias of ERM-DA471/IFCC calibration values was-3.92%～6.30%,with most values being less than 4%.Among them,the most common situation was that the bias of ERM-DA471/IFCC calibration value was less than that of measurement value,and the bias of GBW(E)091173～6 calibration value was less than that of ERM-DA471/IFCC calibration value,with 51.19%(43/84)of the comparison bias bata at the level of medical decisionmaking.Conclusion Both reference materials can improve the alignment bias of conventional system,and GBW(E)091173～6 calibration effect is better than ERM-DA471/IFCC.

Objective:To explore the clinicopathological characteristics and prognosis of pancreatic gastrointestinal interstitial tumors(pGISTs).Methods:Three cases of pGISTs diagnosed in the Affiliated Tumor Hospital of Guangxi Medical University from August 2015 to October 2019 were analyzed retrospectively. Relevant literatures at home and abroad were searched and reviewed through PubMed, China knowledge Network, Wanfang and VIP databases. The sex, age, tumor size, tumor location, cystic or solid tumor, mode of treatment, mitosis, gene mutation, survival status and survival time were recorded, and the correlation between tumor cystic-solid characteristics and clinicopathological parameters was analyzed. Kaplan-Meier′s method was used to calculate the overall survival (OS) rate and disease-free survival (DFS) rate. Univariate and multivariate Cox regression models were used to analyze the independent risk factors affecting the prognosis of pGISTs.Results:In this group, 3 cases were combined with 71 cases reported in the literature, and a total of 74 cases of pGISTs were included. Among them, 36 cases were male and 38 were female, the age of onset was 55(19-84) years, and the diameter of the tumor was 8 cm(2-35 cm). The tumor location of 71 patients was recorded by literature; 30 cases (42.3%) were located in the head of the pancreas. The solid-cystic characteristics of tumor in 63 patients were recorded by literature, and 33 cases (52.4%) were solid. The mode of treatment of 74 patients was recorded, and 60 cases (81.1%) underwent radical resection. The mitosis figures of 59 patients were recorded, and 33 cases (55.9%) were <5/50 high power field of vision (HPF). The gene mutation of 14 patients was recorded, and 11 cases (78.6%) were c-kit exon gene mutation. Correlation analysis showed that the cystic-solid characteristics of the tumor were significantly correlated with tumor location, tumor diameter and mitosis figures, but not with age, sex, histological type, Ki-67 index and modification National Institutes of Health(mNIH) classification. The 5-year OS rate of 51 patients after radical resection was 88.8%, and the 5-year DFS rate was 60.3%. The 1-year OS rate of patients receiving palliative treatment was 51.9%, and the 1-year fatality rate was 33.3%. Univariate Cox regression analysis showed that male ( P=0.083), mitosis figures >5/50 HPF ( P=0.008)and CD 34 negative ( P=0.055)were risk factors for postoperative recurrence of pGISTs, while multivariate Cox regression analysis showed that mitosis figures >5/50 HPF ( P=0.023)was an independent risk factor for the prognosis of pGISTs. Kaplan-Meier survival analysis showed that patients with mitosis figures ≤5/50 HPF had a higher survival rate ( P=0.0003), but there was no significant difference on prognosis between patients with 10/50 HPF and >10/50 HPF( P=0.3075). Conclusions:pGISTs usually occured in the head of pancreas, and the tumor volume was usually found to be large. The main treatment was radical operation, and the main mutation type was exon mutation of c-kit gene. Nuclear fission image figures >5/50HPF was an independent risk factor for postoperative recurrence.

Objective　To investigate the protective effect of complement C5a receptor 1 (C5aR1) antagonist on cerebral ischemia-reperfusion （CIR） in mice.Method　Mice were randomly divided into sham operation group,cerebral ischemia-reperfusion group (model group) and C5aR1 antagonist （PMX53）group.At 3 h before,24 h after and 48 h after cerebral reperfusion timepoint,the PMX53 group was given with PMX53,the sham group and the model group were given same volume of saline by intraperitoneall injection.At 72 h after cerebral reperfusion timepoint,neurological deficits score of mice were evaluated by the modified Longa method,the infarcted brain volume was calculated after TTC staining,the cerebral tissue water content of the ischemic hemisphere was calculated by dry and wet weight method,the mRNA expression of inflammatory factors in the ischemic hemisphere were detected by real-time PCR,and the relative expression of ZO-1 in cerebral tissue of the ischemic hemisphere was calculated by Western blot.Results　At 72 h after cerebral reperfusion,compared with the model group,neurological deficits function score,cerebral water content,cerebral infarction volume and inflammatory cytokines (IL-1β,TNF-α) were significantly decreased in the PMX53 group (all P<0.05),ZO-1 expression was significantly higher in the PMX53 group (P<0.05).Conclusion　C5aR1 antagonist can improve the neurological function score after CIR,reduce the volume of cerebral infarction,reduce the degree of cerebral edema and inflammatory response,and has a protective effect on the blood-brain barrier.

OBJECTIVE:To study clinical effect of argatroban for progressive stroke patients of different cerebral ischemic ranges. METHODS:A total of 116 progressive stroke patients selected from neurology department of our hospital during Feb. 2015-May 2016 were divided into anterior circulation (ischemia) group (n=60),posterior circulation (ischemia) group (n=50) and lacunar(cerebral infarction)group(n=6)according to cerebral ischemic ranges. They all received routine treatment combined with argatroban,and given continuous intravenous infusion of argatroban 60 mg/d on the day and 2nd day of disease aggravation, and then continuous intravenous infusion of argatroban 5th day after relieving,3 h/time,bid,7 d as a treatment course. NIHSS scores,modified RANKIN (mRS) scores,APTT and ADR were compared among 3 groups. RESULTS:Forteen days after treat-ment,NIHSS scores and mRS scores of 3 groups were decreased significantly compared to before treatment,with statistical signifi-cance (P<0.05). There was statistical significance between anterior circulation group and posterior circulation group (P<0.05). Two hours after treatment,APTT of 3 groups were prolonged significantly,especially in lacunar group,with statistical significance (P<0.05). Forty hours after treatment,APTT of 3 groups were recover to normal,without statistical significance (P>0.05). No obvious ADR was found in 3 groups. CONCLUSIONS:Argatroban shows significant therapeutic efficacy for progressive stroke of different ischemic ranges with good safety;especially for the patients with anterior circulation ischemics stroke,the effect is quick and anticoagulant effect is significant.

Objective Few reports are seen on the methods of establishing the rabbit model of pancreatic cancer .This study was to compare the effect of Panc-1 cell suspension orthotopic implantation with that of VX-2 tissue orthotopic implantation in construc-ting the rabbit model of pancreatic cancer . Methods Using the random number table method , we divided 30 healthy rabbits into a tissue suspension group ( n=15) and a cell suspension group ( n=15) , VX-2 tissue suspension employed for in-situ implanting in the former group and panc-1 cell suspension utilized in the latter .Then we evaluated the two modeling methods by B-ultrasonography , 3.0T MRI, and CT. Results In the third week after modeling , transpla-ntive metastasis of lots of tumor tissues was observed in the duode-num, colon, appendix, and peritoneal wall in 5 rabbits of the tissue suspension group , but only in the greater omentum of 3 rabbits in the cell suspension group , with high signals of MR T 2 in the posterior gastric body .One case of duodenal metastasis was seen in the cell suspension group , with slightly high signals of MR LAVA in the posterior gastric body .The model of pancreatic cancer was successfully established in all the 15 rabbits of the tissue suspension group , but only in 3 of the cell suspension group .The success rate of tumor im-planting at 3 and 4 weeks was significantly higher in the former ( 46.66%and 100%) than in the latter group ( 6.67%and 20.00%) (P<0.05). Conclusion VX-2 tissue orthotopic implantation is a more feasible and convenient method than Panc -1 cell suspension orthotopic implantation for establishing the rabbit model of pancreatic cancer .

Objective To explore the application value of magnetic resonance molecular functional imaging diffusion weighted imaging(DWI) in the identification of pancreatic carcinoma and mass-type pancreatitis of animal model.Methods Each 8 cases of laboratory pancreatic head transplantation tumor model,chronic mass-type pancreatitis model and normal rabbits were selected and performed the MR DWI molecular functional imaging,the b values were 333,667,1 000 s/mm2 respectively.The apparent diffusion coefficients(ADC) of pancreatic carcinoma model,mass-type pancreatitis model and normal pancreas under different b values were observed.Then the change situation of ADC values of pancreatic carcinoma model,mass-type pancreatitis model and normal pancreas under different b values and difference of ADC(DADC) was analyzed.Moreover the differences in molecular diffusion,tissue perfusion among various groups were observed.Results Throughout the study period,the mortality rate of pancreatic head transplantation tumor model was 50%;the mass-type pancreatitis model and 8 normal rabbits were normally survival.The ADC value of pancreatic carcinoma under the same b value was significantly lower than that of chronic inflammation and normal pancreas area.The ADC value in each group was decreased with the increase of b value,and there was significant difference in ADC value when the b value was 333 s/mm2(F=6.662,P=0.014),in the pairwise comparison among groups,the difference between pancreatic cancer and pancreatitis (t=6.773,P=0.003) and between pancreatic cancer and normal pancreas(t=5.883,P=0.016) had statistical significance (P<0.05).The b value was increased,DADC was smaller,the difference change of DADC between pancreatic cancer area and chronic pancreatitis mass area,between pancreatic cancer area and normal pancreatic head area had statistical significance (P<0.05).Conclusion Rationally selecting the molecular functional imaging DWI technology of b value can better distinguish pancreatic cancer from mass-type pancreatitis,which may be promoted and applied in the evaluation of animal pancreatic head cancer model.

OBJECTIVETo explore the molecular mechanism in the formation of unstable plaques.

METHODSThe cDNA microarray E-MTAB-2055 was downloaded from ArrayExpress database to screen the differentially expressed genes in 24 ruptured plaques against 24 stable plaques. Functional enrichment analysis was conducted to define the biological processes and pathways involved in disease progression. The protein-protein interaction network was constructed to identify the risk modules with close interactions. Five pairs of carotid specimens were used to validate 3 differentially expressed genes of the risk modules by real-time PCR.

RESULTSA total of 439 genes showed differential expression in our analysis, including 232 up-regulated and 207 down-regulated genes according to the data filter criteria. Immune-related biological processes and pathways were greatly enriched. The protein-protein interaction network and module analysis suggested that TYROBP, VCL and CXCR4 might play critical roles in the development of unstable plaques, and differential expressions of CXCR4 and TYROBP in carotid plaques were confirmed by real-time PCR.

CONCLUSIONOur study shows the differential gene expression profile, potential biological processes and signaling pathways involved in the process of plaque rupture. TYROBP may be a new candidate disease gene in the pathogenesis of unstable plaques.

Adaptor Proteins, Signal Transducing ; genetics ; Disease Progression ; Down-Regulation ; Gene Expression Profiling ; Humans ; Membrane Proteins ; genetics ; Oligonucleotide Array Sequence Analysis ; Plaque, Atherosclerotic ; genetics ; Protein Interaction Maps ; Real-Time Polymerase Chain Reaction ; Receptors, CXCR4 ; genetics ; Transcriptome ; Up-Regulation ; Vinculin ; genetics

Objective To explore the molecular mechanism in the formation of unstable plaques. Methods The cDNA microarray E-MTAB-2055 was downloaded from ArrayExpress database to screen the differentially expressed genes in 24 ruptured plaques against 24 stable plaques. Functional enrichment analysis was conducted to define the biological processes and pathways involved in disease progression. The protein-protein interaction network was constructed to identify the risk modules with close interactions. Five pairs of carotid specimens were used to validate 3 differentially expressed genes of the risk modules by real-time PCR. Results A total of 439 genes showed differential expression in our analysis, including 232 up-regulated and 207 down-regulated genes according to the data filter criteria. Immune-related biological processes and pathways were greatly enriched. The protein-protein interaction network and module analysis suggested that TYROBP, VCL and CXCR4 might play critical roles in the development of unstable plaques, and differential expressions of CXCR4 and TYROBP in carotid plaques were confirmed by real-time PCR. Conclusion Our study shows the differential gene expression profile, potential biological processes and signaling pathways involved in the process of plaque rupture. TYROBP may be a new candidate disease gene in the pathogenesis of unstable plaques.

Objective To explore the molecular mechanism in the formation of unstable plaques. Methods The cDNA microarray E-MTAB-2055 was downloaded from ArrayExpress database to screen the differentially expressed genes in 24 ruptured plaques against 24 stable plaques. Functional enrichment analysis was conducted to define the biological processes and pathways involved in disease progression. The protein-protein interaction network was constructed to identify the risk modules with close interactions. Five pairs of carotid specimens were used to validate 3 differentially expressed genes of the risk modules by real-time PCR. Results A total of 439 genes showed differential expression in our analysis, including 232 up-regulated and 207 down-regulated genes according to the data filter criteria. Immune-related biological processes and pathways were greatly enriched. The protein-protein interaction network and module analysis suggested that TYROBP, VCL and CXCR4 might play critical roles in the development of unstable plaques, and differential expressions of CXCR4 and TYROBP in carotid plaques were confirmed by real-time PCR. Conclusion Our study shows the differential gene expression profile, potential biological processes and signaling pathways involved in the process of plaque rupture. TYROBP may be a new candidate disease gene in the pathogenesis of unstable plaques.