1.Current problems and solution for urinary albumin measurements
Chinese Journal of Laboratory Medicine 2015;(9):586-588
Measurement of urine albumin plays a vital part in the early detection and appropriate management of chronic kidney disease. However, urinary albumin measurements have yet not been standardized due to lack of a reference method and its reference material.It is very important for laboratory technicians to be aware of biochemical characteristics of urinary albumins, laboratory techniques, urine sampling methods, reference interval setting, and units of reporting.
2.Biomarkers of acute kidney injury in clinical laboratory
Chinese Journal of Laboratory Medicine 2014;(6):410-414
Acute kidney injury ( AKI) is a common and serious clinical problem.Serum creatinine (SCr) and urine output are still used as the most common biomarkers for diagnosis of AKI.However, neither of them is an ideal indicator for diagnosis of AKI.In recent years , a number of potential biomarkers of AKI with more favorable test characteristics than SCr have been identified and studied in a variety of experimental and clinical settings.This review describes the novel biomarkers of AKI , including NGAL , IL-18, KIM-1, Cys C and L-FABP, which have now progressed to the clinical phase.
3.Yictims'Serum electrolyte disturbances in sufferers during the Wenchuan earthquake in China
International Journal of Laboratory Medicine 2009;30(5):442-445
Objective To study the electrolyte disturbances in the sufferers during the Wen-chuan earthquake in China,and investigate their possible pathomechanism. Methods Serum concentra-tions of potassium (K), sodium (Na), chlorine (Cl), calcium (Ca), magnesium (Mg) and inorganic phosphate (P) were determined in four hundred sufferers. Their values were analyzed via statistical method. Results Significant differences of all determined six electrolytes were observed among various groups. Pearson correlation analysis showed that correlative changes among electrolytes were similar in various groups except the correlation coefficients of Na-Mg in dialysis group (r= 0.066, P = 0.546) and non-dialysis group (r=-0.433, P= 0.044). Multivariate logistic analysis revealed the highest risk factor was K+,which led to crush injury (OR= 28.037, P= 0.000) and crush syndrome (OR=6.434,P=0.000). The receiver operating characteristic (ROC) curve revealed that K+ was the most specific (specificity: 98.5%; cutoff value: 5.8 mmol/L),Ca2+ was the most sensitive (sensitivity: 81.1%; cutoff value: 1.75 mmol/L) for crush injury diagnosis. The specificity of serum K+ , Mg2+ amd P was comparative in diagnosis of crush syndrome. The sensitivity of Ca2+ was the highest,ac-counting for 75.3%. Conclusion Electrolyte disturbances of suffers occur during a natural disaster; serum electrolyte measurement contributes to correct diagnosis and severity assessment.
4.Correlation between oxidative stress and trace elements in blood of patients with cancer
Chinese Journal of Tissue Engineering Research 2006;10(4):187-190
BACKGROUND: The occurrence and development of tumor may result from the reaction of a series of factors. Up to now, earlier detection, monitoring treatment, and prognosis assessment for tumor are very difficult. It can provide the theoretical foundation of rehabilitation and survival for cancer patients to study the mechanisms of occurrence and development of tumors, or investigate the changes of blood level and tissue distribution.OBJECTIVE: To investigate the changes of levels of some trace elements and serous prooxidant-antioxidant system of cancer patients.DESIGN: A controlled observation trialSETTING: Department of Laboratory Medicine, Mianyang CentralHospital.PARTICIPANTS: From September 1999 to December 2000,111 cancer patients were selected from the Third People's Hospital of Zigong, Yibin First People's Hospital of Sichuan Province, and Longchang County people's Hospital of Neijiang City, Sichuan Province.Of them, 21 cases were of liver cancer, 16 of gastric carcinoma, 15 of colorectal cancer, 11 of breast cancer, 13 of lung cancer, 13 of esophageal cancer, 7 of brain cancer and 15 of other cancers. At the same time; 36 control subjects were recruited from those who came to the hospital for health examination.METHODS :Blood samples of subjects after overnight fasting were pre pared by collecting venous blood in 5-mL Vacutainer tubes (Becton Dickinson, American). After 2-3 hours, the blood samples were centrifuged at 3 000 r/min for 15 minutes. 2.0 to 3.0 mL of serum was separated. The sera were stored in the refrigerator until analysis. Serum glutathione(GSH)level was determined by dithiobis-2-nitrobenzoic acid colorimetry. Xanthine oxidase (XOD) concentration was determined with xanthine-nitrotetrazole colorimetry. Glutahione oxidase (GSHPx) activity was determined with dinitrobenzoic acid colorimetry. Malondialdehyde(MDA) concentration in serum was measured with thiobarbituric acid reaction. Vitamin C (Vit C),vitamin E(Vit E) and total antioxidant power(TAOP) levels were measured with phenanthroline colorimetry. Albumin concentration was determined with bromcresol purple. Serum transferrin (Trf) and ceruloplasmin (CER)concentrations were determined with immunonephelometry. The concentrations of Cu, Zn, Fe, and Se were measured with flame atomic absorption spectroscopy.MAIN OUTCOME MEASURES: MDA (as lipid peroxidation marker),trace elements copper, zinc, iron, and selenium and their transport proteins (albumin, transferrin and ceruloplasmin); antioxidation status markers: the concentrations of XOD, GSH, GSHPx, vitamin C, and vitamin E, and total antioxidant power levels.RESULTS: It was shown that lipid peroxidation ( measured as MDA)was significantly higher in cancer patients than in healthy controls [(5.21±1.05) nmol/L vs (4.04±0.68) nmol/L,P < 0.001], and the TAOP level was significantly decreased in cancer patients than in the health controls[(4.34±0.980) U/L vs (5.87±0.93) U/L,P < 0.001]. There were not obvious changes of antioxidation components (XOD, GSH, GSHPx, vitamin C,and vitamin E). Serum albumin concentration was found to be significantly lower in the cancer patients than in the health controls [(34.19±6.94)g/L vs (42.34±4.89) g/L ,P < 0.001], and serum ceruloplasmin concentration was found to be significantly higher in the cancer patients than in the health controls [(0.371 ±0.031) g/L vs (0.346±0.026) g/L,P < 0.05] butserum transferrin concentration remained unaltered (P > 0.05). As com pared with the healthy controls, serum copper level was significantly increased[(19.27±4.74) μmol/L vs (14.29±2.71) μmol/L, P < 0.001], serum selenium levels was significantly decreased[(1.175±0.333 0 μmol/L vs (1.413±0.446) μmol/L,P < 0.001)]. However, the concentrations of zinc and iron remained unchanged. Correlation was observed between copper and MDA levels (r=0.281, P=0.003) in the cancer patients but not in the healthy controls. Moreover, a correlation was also observed between serum iron and MDA levels in the patients with liver cancer (r=0.680,P=0.001).CONCLUSION: The presence of an association between oxidative stress and some trace elements was found in cancer patients; however, the results are possibly inconsisteut because of different cancer types, cancer grades,or other characteristics of the patients engaged in the test.
5.Preparation,characterization and preliminary application of monoclonal antibody against cystatin C
Tingmei CHEN ; Jiafu FENG ; Zhiguang TU
Chinese Journal of Immunology 2000;0(09):-
Objective:To prepare monoclonal antibodies(McAb)against cystatin C(Cys C)and to establish the particle enhanced turbidimetric immunoassay(PETIA)for determining human serum Cys C.Methods:The prokaryotic expression vector pET32a(+)/Cys C was constructed and Cys C expression was induced.McAbs against Cys C were prepared with the hybridoma technique after mice were immunized with the purified recombinant protein.Then the McAbs were covalently attached to uniform microparticles,PETIA method for determination of human serum Cys C was established,and primary evaluation tests of methodology were performed.Results:Three hybridoma cell lines were obtained successfully,the secreted antibodies were isotype of IgG1,and Western blot confirmed that the antibodies reacted specifically to the Cys C protein.After one of the hybridoma cell lines was injected into mice abdominal cavity,the ascites abundant for McAb was obtained.The titer of the McAb against the purified protein was 1∶4?106.With the self-made McAb,PETIA for human serum Cys C was established.The primary evaluation tests of methodology revealed that self-established PETIA method had a satisfactory performance,which was equal to the import kit.Conclusion:The prepared McAb against Cys C is prepared,which could be used to establish PETIA for determining human serum Cys C.
6.Alteration of serum Cystatin C concentrations in patients with hepatopathy infected with hepatitis B or C virus
Jiafu FENG ; Tingmei CHEN ; Zhiguang TU
Chinese Journal of Laboratory Medicine 2003;0(11):-
Objective To investigate the clinical application of Cystatin C as a biological marker for monitoring hepatic pathological change in patients with virus hepatitis.Methods Two hundred and seven patients infected with hepatitis B or C virus(HBV, HCV)were divided into cirrhosis group(group A),chronic HBV group(group B),chronic HCV group(group C),and liver cancer group(group D). 32 healthy controls(group H) were recruited . The serum TIMP-1,TIMP-2,and Cystatin C as well as some traditional markers for monitoring liver function and renal function including creatinine, creatinine clearance rate, alanine transaminase, and aspartate transaminase were determined.Results In these groups, serum Cystatin C(F=28.334, P
7.Effectiveness analysis of determining the content of notopterol and isoimperatorin in notopterygium
Jiang YU ; Jiafu FENG ; Bangguo ZENG
Chongqing Medicine 2014;(35):4769-4770,4773
Objective To establish the effect of determining the content of notopterol and isoimperatorin in notopterygium from different habitat and harvest time by HPLC method .Methods HPLC instrument was used ,Waters symmetry C18(4 .6 mm × 250 mm ,5 μm) was adopted with a mobile phase of acetonitrile‐water (45∶55) ,the separation was carried out at a flow rate of 1 mL/min ,column temperature was 30 ℃ and the detection wavelength was 310 nm .Results Notopterol and isoimperatorin showed good linearity in the ranges of 0 .240 8-1 .505 0 μg and 0 .117 6-0 .735 0 μg with average recoveries rate (n=6) of 0 .98% and 1 .42%respectively .The content of notopterol and isoimperatorin in crude drugs of notopterygium from different habitat marked differ‐ence ,and harvest period and growth years made effect to content of two components .Conclusion HPLC method is easy and fast , and can be used as a basis for confirming the best habitat and harvest period of notopterygium .
8.Effects of quercetin on proliferation and expression of P38MAPK and HMGB1 protein in neonatal rat cardiac fibroblasts induced by TNF-α
Chun ZHANG ; Jian FENG ; Jiafu LI
Chongqing Medicine 2017;46(23):3189-3191
Objective To study the effect of quercetin on proliferation and expression of P38MAPK and HMGB1 protein in neonatal rat cardiac fibroblasts induced by TNF-α.Methods Purified cardiac fibroblasts were obtained by trypsin digestion and differential adherence method.The proliferation of cardiac fibroblasts was detected by MTT assay.The expression of P38MAPK and HMGB1 protein was detected by Western blot.Results Quercetin had no effect on the proliferation of cardiac fibroblasts in the basal state but inhibited the proliferation of cardiac fibroblasts induced by TNF-α,and the A value of each group was increased with the increase of quereetin concentration(P<0.05).The exspression of P38MAPK、HMGB1 were decresed with the icrease of quercetin.Conclusion Quercetin may inhibit the proliferation of cardiac fibroblasts induced by TNF-α.The mechanism may be inhibit the expression of HMGB1 through P38MAPK signaling pathway.
9.Establishment of enzymatic kinetic assay of serum argininosuccinate lyase activity and its preliminary clinical application
Jiafu FENG ; Tingmei CHEN ; Xiaolin YU ; Jian WANG ; Zhiguang TU
Chinese Journal of Laboratory Medicine 2009;32(2):217-221
Objective To establish a continuous monitoring assay of serum argininosuccinate lyase (ASL) activity with automatic biochemistry analyzer, and perform methodology validation and preliminary clinical application.Methods According to the chemical reaction catalyzed by ASL and the working characteristics of automatic biochemistry analyzer, an enzyme coupled reaction system with high specificity was set up, and the methodology validation was performed.Three hundred and nine patients with various liver diseases, 269 non-liver disease patients and 40 healthy controls were enrolled in this study.Serum ASL, ALT, and AST level were determined in all subjects.Results A new kinetics assay of ASL activity was set up with automatic biochemistry analyzer.The methodological validation demonstratod that inter-assay and intra-assay coefficient of variation were 4.0% and 5.9% respectively and the mean recovery was 100.5%.The linear range was 0-167.7 U/L.The lowest detection limit was approximately 0 U/L.The interference test showed that there is no significant interferences while the concentration of bilirubin is less than 342 μmoL/L or commonly used anticoagulants is employed at their routine concentrations.However,interference was significant when Hb level is more than 0.06 g/L.Preliminary study of clinical application showed that there was no significant difference of serum ASL level between non-liver disease group and healthy group ( q = 0.027, P = 0.979 ), but there was significant differences for both serum ALT and AST levels (ALT:q =6.461,P =0.000;AST:q =6.481,P =0.000).Conclusions A continuous monitoring assay for the determination of serum ASL activity is successfully established. Serum ASL may be a good biomarker for liver injury.
10.Determination of Sulfurous Anhydride in Rhizoma Dioscoreae by Ion Chromatography-Direct Extraction
Xiaolin WANG ; Minling YU ; Wenqiu CHEN ; Jiafu FENG
China Pharmacist 2015;(1):166-168
Objective:To establish a quantitative analysis method to determine the content of sulfurous anhydride in Rhizoma di-oscoreae by ion chromatography-direct extraction. Methods:Sulfurous anhydride was extracted by KOH solution (25 mmol·L-1). An IonPac? AS11-HC column(250 mm × 4 mm, 9. 0 μm) was used. The column temperature was 20℃, the eluent was KOH solution (20 mmol·L-1 ) at flow rate of 1. 00 ml·min-1 and the conductivity temperature was 20℃. Results:There was a good linear rela-tionship between the injection quantity (1.160-29.100 μg)and the peak area of sulfite(r =0.999 9). The average recovery was 98. 9%(RSD=0. 6%, n=9). The quantitation limit was 1. 38 ng·ml-1. Conclusion: The method is simple, accurate and rapid, which is appropriate for the quantitative analysis of sulfite anhydride in Rhizoma dioscoreae.