1.Laboratory study on reproducibility of no-albumin human plasma by rivanol precipitation Ⅰ.Purification or isolation of protein C and protein S
Yonghui TAO ; Jian JIN ; Bocheng WANG ; Manda ZHANG ; Changgeng RUAN
Chinese Journal of Biochemical Pharmaceutics 2001;22(3):121-124
Purpose The aim is to isolate and purify Protein C(PC) and Protein S(PS) from no-albumin human plasma by rivanol precipitation. Methods The isolated and purified steps included adsorption onto and elution from barium, salting-out, ion-exchange chromatography, affinity chromatography,preparative isoelectric focusing and so on. Results The molecular weights of the obtained PC and PS were (61±0.9)kD and (83±0.8)kD, respectively, the isoelectric point, 4.70±0.03 and 5.20±0.03,and the yield, 28.3% and 12.6%. The purified PC and PS were shown to be highly homogeneous by capillary zone-electrophoresis(CZE), and rich in Glu, Leu and Gly or Asp, Glu and Leu respectively. Conclusion The methods could be used for large-scale isolation and purification of PC and PS from no-albumin human plasma.
2.Purification of human urine erythropoietin and thrombomodulin
Yonghui TAO ; Jian JIN ; Rongjun ZHANG ; Yang HE ; Manda ZHANG ; Changgeng RUAN
Chinese Journal of Biochemical Pharmaceutics 2001;22(2):57-60
Purpose The aim is to isolate and purify the erythropoietin(EPO)and thrombomodulin(TM) from human urine.Methods Purifying and isolating human urine with means of filtration and concentration,ion-exchange chromatography, affinity chromatography and so on.Results 4.47 mg EPO and 9.92 mg TM were obtained from 2 000 kg human urine.Their molecular weights were(38.6±1.0)kD and (60±1.4)kD erespectively, the isoelectric point, 3.60±0.02 and 3.70±0.02, yield, 31.9% and 25.0%.The purified EPO and TM was shown to be highly homogeneous by capillary zone-electrophoresis(CZE),abundant Glu,Ala and Gly include on TM. Conclusion The high purity EPO and TM was obtained.
3.Cross-species Transmission of Avian Leukosis Virus Subgroup J.
Yanwei SHEN ; Menglian HE ; Ji ZHANG ; Manda ZHAO ; Guihua WANG ; Ziqiang CHENG
Chinese Journal of Virology 2016;32(1):46-55
Avian leukosis virus subgroup J (ALV-J) is an avian retrovirus that can induce myelocytomas. A high-frequency mutation in gene envelope endows ALV-J with the potential for cross-species transmission. We wished to ascertain if the ALV-J can spread across species under selection pressure in susceptible and resistant hosts. First, we inoculated (in turn) two susceptible host birds (specific pathogen-free (SPF) chickens and turkeys). Then, we inoculated three resistant hosts (pheasants, quails and ducks) to detect the viral shedding, pathologic changes, and genetic evolution of different isolates. We found that pheasants and quails were infected under the selective pressure that accumulates stepwise in different hosts, and that ducks were not infected. Infection rates for SPF chickens and turkeys were 100% (16/16), whereas those for pheasants and quails were 37.5% (6/16) and 11.1% (3/27). Infected hosts showed immune tolerance, and inflammation and tissue damage could be seen in the liver, spleen, kidneys and cardiovascular system. Non-synonymous mutation and synonymous ratio (NS/S) analyses revealed the NS/S in hypervariable region (hr) 2 of pheasants and quails was 2.5. That finding suggested that mutation of isolates in pheasants and quails was induced by selective pressure from the resistant host, and that the hr2 region is a critical domain in cross-species transmission of ALV-J. Sequencing showed that ALV-J isolates from turkeys, pheasants and quails had moved away from the original virus, and were closer to the ALV-J prototype strain HPRS-103. However, the HPRS-103 strain cannot infect pheasants and quails, so further studies are needed.
Amino Acid Sequence
;
Animals
;
Avian Leukosis
;
transmission
;
virology
;
Avian Leukosis Virus
;
classification
;
genetics
;
physiology
;
Chickens
;
Ducks
;
virology
;
Galliformes
;
virology
;
Host Specificity
;
Molecular Sequence Data
;
Poultry Diseases
;
transmission
;
virology
;
Quail
;
virology
;
Sequence Alignment
;
Turkeys
;
virology
;
Viral Envelope Proteins
;
chemistry
;
genetics
;
metabolism
4.Radioimmunoassay of human cardiac acidic isoferritin: a new index for hepatic cancer
Jian JIN ; Manda ZHANG ; Bocheng WANG ; Hualong XIAO ; Junming SUN
Chinese Medical Journal 1998;111(2):150-153
Objective To investigate human cardiac acidic isoferritin as a specific index of hepatic cancer.Methods Acidic isoferritin was isolated and purified from human heart muscle. A radioimmunoassay for the acidic isoferritin in human serum has been developed, on the equilibrium method. The antiserum was obtained from rabbits immunized with purified acidic isoferritin. The 125 I-acidic isoferritin was prepared by the chloramine-T method. The data were processed using the automated smoothed spline function data processing program.Results The intra- and inter-assay CV of acidic isoferritin RIA were 1.65% and 9.71%, respectively, and the recovery rate was 102%. The antiserum provided a linear response from 7.0 to 369.6 μg/L with ED50 of 27.50 μg/L. The cross reactivity with AFP, CEA, lactoferrin and transferrin was negligible, and that with ferritin was 1.74%. The serum acidic isoferritin concentration showed a considerable variation in different sex and age groups. The serum acidic isoferritin was measured in liver diseases including hepatic cancer, hepatic cirrhosis and acute and chronic hepatitis. Its sensitivity for diagnosis of hepatic cancer was 73.05%, independent from the severity of hepatic injury. In 8 malignant tumors studied, acidic isoferritin appeared the most valuable in the diagnosis of hepatic cancer, with its positive, negative, false positive and false negative rates all being ideal.Conclusions Acidic isoferritin may turn to be a rather specific index of hepatic cancer. Combination of monitoring both acidic isoferritin and AFP would raise the positive detection and specificity in the diagnosis of hepatic cancer.