1.Time resolved fluoroimmunoassay for Glypican 3 and its preliminary application
Dan, LI ; Jing, ZHANG ; Xin, BAI ; Biao, HUANG ; Yi, ZHANG ; Zhong-wei, L(U) ; Hong, TU
Chinese Journal of Nuclear Medicine 2011;31(3):201-204
Objective To establish a time resolved fluoroimmunoassay (TRFIA) method for detecting Glypican 3 (GPC3) and to explore the diagnostic value of serum GPC3 for hepatic carcinoma (HCC). Methods Microplate coated with anti-GPC3 monoclonal antibody 7C8 and GP9 labeled with Eu3+ were used to establish TRFIA kit. The serum concentrations of GPC3 in 41 HCC patients and 44 chronic hepatitis (CH) patients were quantitatively analyzed. AFP was detected by with lowest limit of 2.06 μg/L. The CV of inter and intra assay were 12.25% and 12.91%, respectively. The average serum concentration of GPC3 in HCC patients was (86.68±110.39) μg/L (median: 56.98 μg/L). But in CH patients it was only (14.77±29.48) μg/L, which was significantly lower than that in HCC (Wilcoxon W=1335.00, Z=-4.99, P<0.001). With diagnostic cut-off value set at 42.94 μg/L, the diagnostic sensitivity and specificity of TRFIA GPC3 for HCC were 58.5% (24/41) and 95.5%(42/44) respectively. The diagnostic sensitivity of AFP was 46.3% (19/41) in 41 HCC patients, and was raised to 78.0% (32/41) when combined with GPC3. Conclusions Serum GPC3 assay by TRFIA is established and it could increase the diagnostic sensitivity for HCC when combined with AFP.
2.Investigation of iodine nutritional status and thyroid function among pregnant and lactating women from rural areas of Jinlin province in 2009
Li-fen, YANG ; Hui-xin, CHEN ; Wei, LI ; Min, WANG ; Dan-ni, YANG ; Hong-yan, L(U) ; Yan-hua, LUO ; Jing-shen, ZHAO ; Li-xia, ZHU
Chinese Journal of Endemiology 2011;30(2):162-164
Objective To investigate the iodine nutritional level and thyroid function of pregnant and lactating women in rural areas of Jilin province. Methods The investigation sites were selected from rural areas of three towns (Baoshan, Mingcheng and Yantongshan of Panshi county, Jilin province) in 2009. The pregnant and lactating women were selected as subjects in these three towns. The blood samples were collected and the thyroid function (including serum TT3, TT4, FT3, FT4) were measured with chemiluminescence, and serum thyroglobulin antibodies(TgAb), thyromicrosome antibody(TMAb), and thyroglobulin(Tg) were measured with radioimmunoassay (RIA). The urine samples were collected three times within one month and were measured for iodine concentration by As-Ce catalytic spectrophotometry method. Results In the pregnant women, serum TT3 was higher than that of healthy pregnant women, accounted for 14.3%(8/56), while serum TT4, TT3, FT4 were lower than those of healthy pregnant women, accounted for 3.6%(2/56),5.4% (3/56), and 1.8%(1/56), respectively. In the lactating women,serum TT3 was higher than that of healthy lactating women, accounted for 3.6%(2/56), while serum TT4, FT4 were lower than those of healthy lactating women, accounted for 1.8%(1/56), respectively. Five per cent to 20% of the pregnant and lactating women had higher TgAb and TMAb. Conclusions Existing salt iodine level is appropriate for pregnant women and lactating women, but there was a tendency towards hypothyroid in some women. Routine monitoring of urinary iodine and thyroid function should be carried out among pregnant and lactating women.
3.Gene express and antibody preparation of Neurocan protein
Mou-Xuan DU ; Xiao-Dan JIANG ; Ru-Xiang XU ; Zhen-Zhou CHEN ; Jun L(U)
Chinese Journal of Neuromedicine 2009;8(2):109-113
Objective To offered some prophase works by preparing Neurocan protein, antiserum, and assaying their characteristics, in order to construct the isopropy-β-D-thiogalactoside (IPTG)-participated DNA vaccine, which can neutralize the inhibitors in the injured CNS following the immune administration and then promote the nerve regeneration. Methods Neurocan gene was syntbetized with HisTag label in beginning and enzyme-cut sites at amphi- of the sequence. The prokaryotic expression plasmid, PET30a-Neurocan, was constructed as usual, converted into the Escherichia coli, and induced by IPTG to express positively. The interest protein was identified by SDS-PAGE and Western blot respectively. The preimmune serum was as the negative control during the ELISA assay of antiserum valency. The immune serum was as the first antibody, and the goat-anti-rabbit labeling with alkaline phosphatase (AP) was employed as the second one. Coloration was with NBT/BCIP method. Results The correct sequence of the synthetic Neurocan gene was clearly showed by identification with enzyme-cut, PCR and sequencing. The Neurocan protein expressed by prokaryotic showed its molecular weigh as 55 000 following the SDS-PAGE identification, and it could specifically bind with anti-HisTag, which implied the interesting protein just as the expression product of Neurocan gene. The valency of antiserum was shown by ELISA as 1:1 000 000, the purpose strap of which was confirmed by Western blot. Conclusions Neurocan protein could be successfully expressed in prokaryotic, the antibody of which could be specifically obtained by immune administration to the rabbit. The Neuroncan antibody could bind with the Neurocan protein specifically.
4.Clinical and MR imaging characteristics of patients with delayed encephalopathy after acute carbon monoxide poisoning
Shu-Yan CONG ; Dan L(U) ; Jian-Fei NAO ; Xiu-Lan ZHAO
Chinese Journal of Neuromedicine 2012;11(5):512-515
Objective To discuss the clinical and MR imaging characteristics of patients with delayed encephalopathy after acute carbon monoxide (CO) poisoning (DEACMP). Methods The clinical and imaging data of 23 patients with DEACMP,admitted to our hospital from January 2007 to July 2011,were retrospectively analyzed. Results The dominant symptoms of DEACMP included mentally falling in 21 patients (91.3%),extrapyramidal impairments in 15 patients (65.2%),and mental and behavioral abnormalities in 12 patients (52.2%).Cranial MRI features included bilateral subcortical white matter and/or periventricular white matter damage in 11 patients, bilateral basal ganglia (mainly bilateral globus pallidus) damage in 4,and damage in both sub-cortical white matter and basal ganglia in 8.The prognosis of patients with simple globus pallidus damage was better.All patients were given high pressure oxygen,medications improving the cerebral blood circulation,brain nutritional medicine and symptomatic treatment with total efficiency reaching 86.9%. Conclusion Cranial MRI features has important values in early diagnosis,treatment efficacy evaluation and prognosis estimation of patients with acute DEACMP; full-time course of hyperbaric oxygen therapy is very important and effective.
5.Feasibility of local LINGO-1 polyclonal antibody treatment for spinal cord injury in adult rats
Jun L(U) ; Ru-Xiang XU ; Zhi-Qiang FA ; Xiao-Dan JIANG ; Xin LU ; Yi-Quan KE ; Ying-Qian CAI ; Mou-Xuan DU ; Yu-Xi ZOU ; Ling-Sha QIN
Chinese Journal of Neuromedicine 2009;8(5):476-478,483
Objective To analyze the feasibility of local LINGO-1 polyclonal antibody administration for treatment of spinal cord injury in adult rats. Methods Twenty-four adult female SD rats were randomized into sham-operated group, rabbit IgG group and LINGO-1 antibody group. In the latter two groups, partial transaction of the T9 segment of the spinal cord was performed to completely sever the dorsal eorticospinal tract, followed immediately by administration of rabbit IgG and anti-LINGO polyclonal antibody via a mini-osmotic pump, respectively. At 3 and 28 days after the operation, the T8~10 segments of the spinal cord were harvested to prepare cryosections, and immunofluorescence staining was used to analyze the penetration of LINGO-1 polyclonal antibody into the spinal cord tissue and its specific binding to LINGO-1 molecules. Results In LINGO-1 antibody group, the presence of rabbit antibodies was detected at the injured sites of the spinal cord at 3 and 28 days after the operation. The mean immunofluorescence density was significantly lower in L1NGO-1 antibody group than in rabbit IgG group at 3 days after the operation (P<0.05). In rabbit IgG group, the mean immunofluorescence density for LINGO-1 in the crysections pre-treated with LINGO-1 polyclonal antibody was significantly lower than that in sections pre-treated with rabbit IgG(P<0.05). Conclusion Locally administered LINGO-1 polyclonal antibody can penetrate into the injured sites in the spinal cord in a wide time window and recognizes LINGO-1 molecule specifically, suggesting the feasibility of passive immunotherapy for spinal cord injury.