Objective To construct a regulatable plasmid containing single chain fusion gene of murine interleukin-12 (mIL-12) which was regulated with mifepristone (RU486) and explore its expression in vitro. Methods The p40 and p35 subunit sequence of mIL-20 were respectively obtained from the plasmid GCp35Ep40PN by polymerase chain reaction (PCR) and they were cloned into pCA14 plasmid after introducing a linker by overlap PCR. The single chain mIL-12 gene was comfirmed by sequencing and subcloned into pRS-17 vector which contains RU486 regulator cassette. The positive clone named pRS-RUmIL-12 was identified by restriction endonuclease digestion and PCR. Lipofectamine 2000 was used to transfect the pRS-RUmIL-12 to HEK293 cells followed by manufacturer’s recommendations. The protein concentration of mIL-12 induced with RU486 in supernatant of the transfected HEK293 cells was measured by ELISA. Results The sequence of single chain mIL-12 what we obtained was the same as the expected result. The results of restriction endonuclease digestion and PCR showed that the RU486-inducible regulatory vector (pRS-RUmIL-12) was successfully constructed. No significant mIL-12 protein concentration in supernatant of HEK293 cells activation was measured without the inducer RU486, whereas higher concentration of the mIL-12 protein was observed in the presence of RU486. The relationship of concentration of the mIL-12 protein and RU486 was positive correlated under definite range. Conclusion A regulatable eukaryotic expression plasmid of mIL-12 single chain fusion gene was constructed, which could be used in the further research of gene regulation and gene therapy.
Objective To determine the concentration of int erleukin-12(IL-12),interleukin-2(IL-2) and tumor necrosis factor(TNF)and the irpossible role in the pathogenesis of proliferative vitreoretinopathy(PVR) ． Methods Patients were divided into 3 groups:18 with PVR,7 with simples retinal detachment caused by macular hole and 4 samples from normal eyes were used as control.Sample s of vitreous were obtained by aspiration through pars plana before cryotherapy ,vitrectomy and gas injection and stored in liquid nitrogen at －70℃ within 30 minites for ELISA. Results ①The levels of IL-12,IL-2，and TNF in the vitreous of PVR were positively correlated with the degree of severity of disease.②The levels of IL-12, IL-2,and TNF in the PVR were higher than those in simple retinal detachment caused by macular hole and those in control group(Plt;0.01 ).③The levels of IL-12,IL-2，and TNF in retinal detachment caused by macular hole were also higher than those in the control group(Plt;0.01). Conclusion IL-12,IL-2，and TNF may play a role at lease to some extent in the pathogenesis of PVR. (Chin J Ocul Fundus Dis,1999,15:75-77)
Objective To explore the changes and interrelationship of serum interleukin-12 (IL-12) and T lymphocyte subset in patients with primary hepatic carcinoma (PHC). Methods Serum IL-12 level was determined by ELISA in 36 patients with PHC. The peripheral blood T lymphocyte subset was assessed with flow cytometry. The distribution and changes of T lymphocyte subset in the tumor tissue were detected by immunohistochemistry analysis. Results The numbers of the CD＋4 T cell were reduced and of the CD＋8 T cell increased either in peripheral blood or tumor tissue, and showed the trend of the ratio (T4/T8) declined progressively with the aggravation of the state with PHC. IL-12 and T4/T8 had significant interrelationship.Conclusion IL-12 is an important antitumor factor of the patients with PHC. T lymphocyte subset plays a great role in the process of antitumor.
ObjectiveTo investigate the overall accuracy of interleukin-12 (IL-12) for diagnosis of tuberculous pleurisy. MethodsWe searched in PubMed, Embase, Web of Science, China National Knowledge Infrastructure databases, WanFang Data, and VIP Information for qualified studies that reported diagnostic accuracy of IL-12 for tuberculous pleurisy up to February 2014. The methodological quality of each study was evaluated by Quality assessment of diagnostic accuracy studies. Statistical analyses were performed by Meta-Disc 1.4 software and the pooled sensitivity, specificity and other diagnostic indexes. Meta-analysis of the reported accuracy of each study and summary receiver operating characteristic (SROC) curve were also performed. ResultsEight studies met the inclusion criteria for the analysis. The summary estimates for IL-12 in the diagnosis of tuberculous pleurisy were:sensitivity 0.80 [95% CI (0.76, 0.84)], specificity 0.76 [95% CI (0.71, 0.81)], positive likelihood ratio 3.23 [95% CI (2.26, 4.60)], negative likelihood ratio 0.30 [95% CI (0.20, 0.45)], diagnostic odds ratio 13.57 [95% CI (6.66, 27.64)], and the area under the curve of SROC was 0.86. ConclusionIL-12 plays a valuable role in the diagnosis of tuberculous pleurisy, and IL-12 may be a useful diagnostic marker for tuberculous pleurisy.