JCBI is a glioma eell line established in our department. JCBI cells spontaneously produce a factor inhibiting blastogenic responses of mouse thymocytes and the IL-2 dependent T cell growth of mouse spleen lymphoblast, without inhibiting spontaneous proliferation of mouse thymocytes. The inhibiting activity is high after 72 hours of passage. Physicochemical characterization shows that the factor is nondialyzable. partially inactivated at 37℃ 48 hours and almost inactivated at 56℃ 30 min. But the factor is stable at 4℃ for 2 weeks and in freeze-thawing. The molecular weight of the factor is approximately 10 kd as estimated by gel filtration. The factor may contribute to the impaired immunosurveillance and to the cellular immunodeficiency state in the patients.

Abstract-Eleven mouse monoclonal antibodies (McAbS)specific for human alpha-fetopr-otein(AFP) have been produced by the hybridoma technique. Immunodiffusion was us-ed for determining the immunoglobulin class and subclass of the McAbS. Seven of them are IgG; two are IgM. At least 4 different antigenic determinants on human AFPcan be recognized by using the McAbS and cross-two-site sandwich solid radioimmun-oassay. One of the McAbS can specially react with one of the fragments of AFP dig-ested by pepsin. In addition, the cross-reactivity among the AFPs of different mam-malian species was also tested by using the McAbS.

Objective To introduce the advantages of the endoscopic assistance in primary and secondary face-lift in the frontal and temporal areas.Methods 67 cases were involved in the study,22 of them were secondary facelift cases.The follow-up period ranged from 1 month to 2 years.The patients and doctors satisfactory rate were recorded and the complications were also reported.Results All 67 cases had complete recovery without serious complications.The patient's satisfactory rate was 85％ (57/67),and the doctor's satisfactory rate was 89％ (60/67).Some early minor complications included dimpling at the suture site,asymmetry,overcorrection,transitory paralysis,late edema,scar and fall-off of hair among 80％ or so of patients.4 cases experienced hematoma on the frontal areas.The hematoma disappeared after early aspiration and later fomentation.2 cases had wound ulceration in the temporal 3 months after operation.The anchoring materials were removed and the ulceration tissues were excised 6 months after operation.The wound healed completely.2 patients experienced skin necrosis and depression due to careless electric cauterization on the frontal area.The depression gradually became smooth and inconspicuous after 6 months.All these complications were resolved and became negligible about 6 months after operation.Conclusions Endoscopic assistance is valuable in primary and secondary face-lift in the frontal and temporal areas.It is reliable and worthwhile to introduce the technique for patients aged less than 50-year-old.

Objective To search and analyze the DNA methylation-related genes of ?-thalassemia by oligonucleotide microarray in order to explore a new method for early diagnosis of ?-thalassemia.Methods The cord blood samples of 2 children with ?-thalassemia and 2 health children were detected by a chip containing human DNA methylation 30 178 oligonucleotide probes.The chip results were verified by the methods of methylation-specific PCR (MSP) and real-time PCR.The statistical significance of differentially expressed genes was found on non-supervised clustering (Hierarchical clustering).Results A total of 209 genetic differences (ratio≥2.0 or≤0.5) were showed by 2 groups of chips,and of them 113 genes were up-regulated and 96 genes were down-regulated.The results showed that the methylation-related gene erythroblastic leukemia viral (v-erb-a) was of hypermethylation compared with the normal blood.Conclusion A large number of differentially expressed genes are screened out by the technology of High-throughput DNA methylation of the gene chip in thalassemia.The DNA methylation-related gene of v-erb-a is of hypermethylation in thalassemia.Our methods offers a new idea and approach for prenatal diagnosis for thalassemia by the DNA methylation-related microarray.

OBJECTIVE:To study the rationality of multiple-unit containers of oral drugs.METHODS:The condition of whether the oral western medicines currently used in our hospital were packaged in single-dose container or not were inves?tigated;the information on packaging and distribution of multiple-unit containers was analyzed as well.RESULT:Of the460kinds of oral western medicines investigated,220were in single-dose containers,and138were in multiple-unit containers,of the multiple-unit containers87involved problems in packaging.CONCLUSION:It is suggested that single-dose container should be used and the multiple-unit containers should be improved reasonably.

Objective To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci ( Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods The fusion protein His-CPSIT_p7 was expressed in E.coli BL21 and purified by Ni-NTA affinity chromatography .BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CPSIT_p7 by ELISA.Penicillin sodium was used to establish a model of Cps persistence infection .RT-PCR and Western blot assay were performed to de-tect the expression of CPSIT_p7 at mRNA and protein levels during Cps persistent infection .Results The fusion protein His-CPSIT_p7 was successfully expressed with the use of constructed recombinant expression plasmid pET30 a-CPSIT_p7 and purified .ELISA result showed that the specific antibody titer against CPSIT_p7 reached 1 ∶1 000 000 on the 40th days after immunization .The expression of CPSIT_p7 at mRNA and protein levels were increased in a time-dependent manner in Cps-infected HeLa cells .The peak of mRNA level was reached at the time point of 36 hours after infection , followed by a time-dependent decrease during Cps acute infection .However , the expression of CPSIT_p7 at mRNA and protein levels were not decreased until 60 hours after infection during Cps persistent infection .Conclusion His-CPSIT_p7 protein was suc-cessfully expressed in the prokaryotic expression system and purified , showing an advantage of good immuno-genicity.Highly expressed CPSIT_p7 at mRNA and protein levels were detected during Cps persistent infection.

Objective To observe the optic disc perfusion in anterior ischemic optic neuropathy (AION) patients.Methods Forty eyes of 40 AION patients and 30 eyes of 30 normal subjects were included.The stage of the diseases was defined based on the course of the disease,including acute stage (less than 3 weeks) and recovery stage (more than 3 months).Optic disc blood flow area,outer vascular density and blood flow index were measured by optical coherence tomography angiography in all the subjects.Optic disc perfusion was observed in acute and recovery stage of disease.Results The optic disc blood flow area,outer vascular density and blood flow index were decreased of AION eyes in acute stage compared with the normal subjects,the difference was statistically significant (P＜0.05);while the optic disc blood flow area,outer vascular density and blood flow index of AION eyes in the recovery stage showed no significant difference compared with normal subjects (P＞0.05).Conclusion Disc perfusion is reduced in AION at the acute stage,but recovered at the recovery stage.

Idiopathic parafoveal telangiectasis (IPT) is a retinal vascular disease which is characterized by foveal and parafoveal telangiectasia.The main clinical manifestations are retinal telangiectasis,reduced retinal transparency,retinal venular dilatation,yellow exudation,retinal pigment epithelial lesions,retinal hemorrhage,macular atrophy,macular hole or lamellar hole,subretinal neovascularization and retinal detachment.According to the clinical characteristics and features of fluorescein angiography,IPT can be divided into 3 types and 6 subtypes.Laser photocoagulation,photodynamic therapy,and intravitreal injection of glucocorticoid or anti-vascular endothelial growth factor drugs,can reduce the macular edema and neovascularization.However,due to the unclear etiology of IPT,the existing treatment measures are not specific for its etiology.We need to work hard to understand further the clinical features and pathogenesis of IPT and search the targeted treatments based on its pathogenesis mechanism.

Objective:To analyze the protein expression changes in the retina of non-arteritic anterior ischemic optic neuropathy (NAION) in rats.Methods:The rat NAION (rNAION) model was established by Rose Bengal and laser. Twenty Sprague-Dawley rats were randomly divided into 4 groups, the normal control group, the laser control group, the RB injection control group, and the rNAION model group, with 5 rats in each group. The right eye was used as the experimental eye. The retina was dissected at the third day after modeling. Enzyme digestion method was used for sample preparation and data collection was performed in a non-dependent collection mode. The data were quantitatively analyzed by SWATH quantitative mass spectrometry, searching for differential proteins and performing function and pathway analysis.Results:Compared with the other three control groups, a total of 184 differential proteins were detected in the rNAION group (expression fold greater than 1.5 times and P<0.05), including 99 up-regulated proteins and 85 down-regulated proteins. The expressions of glial fibrillary acidic protein, guanine nucleotide binding protein 4, laminin 1, 14-3-3γ protein YWHAG were increased. Whereas the expressions of Leucine-rich glioma-inactivated protein 1, secretory carrier-associated membrane protein 5, and Clathrin coat assembly protein AP180 were decreased. The differential proteins are mainly involved in biological processes such as nerve growth, energy metabolism, vesicle-mediated transport, the regulation of synaptic plasticity, apoptosis and inflammation. Pathway enrichment analysis showed that PI3K-Akt signaling pathway and complement and thrombin reaction pathway was related to the disease. Conclusion:The protein expressions of energy metabolism, nerve growth, synaptic vesicle transport and PI3K-Akt signaling pathway can regulate the neuronal regeneration and apoptosis in NAION.

Objective To construct a recombinant adenoviral vector carrying and co-expressing human inhibitor of growth 4(ING4) and human interleukin-24(IL-24) mediated by poly( A)-Promoter[Ad-ING4-poly(A)-Promoter-IL-24, referred to as Ad-ING4-IL-24] and explore its effect on the growth of HepG-2 human hepatocellular carcinoma cellsin vitro. Methods The poly(A)-Promoter(hEFl-elF4g) (Sal Ⅰ and Not Ⅰ ), ING4 ( Bgl Ⅱ and Sal Ⅰ ), and IL-24 ( Xho Ⅰ and Xba Ⅰ ) fragments were amplified by PCRusing pORF-mbcl-2α, pcDNA3.0-IL-24, and pcDNA3.0-ING4 plasmids as templates and subcloned into pAdTrack-CMV transfer vector to form pAdTrack-CMV-ING4-poly (A)-Promoter-lL-24, respectively. The pAdTrack-CMV-ING4-poly (A)-Promoter-IL-24 transfer vector linearized with Pme Ⅰ digestion and pAdEasy-1 backbone vector was further cotransformed into the bacteria BJ5183 competent cells for homologous recombination. The resultant pAdEasy-l-pAdTrack-CMV-ING4-poly ( A )-Promoter-IL-24 homologous recombinant plasmids were linearized with Pac Ⅰ digestion and transfected into the human embryonic kidney 293 (QBI-293A) cells by liposome, leading to formation of the recombinant adenoviruses Ad-ING4-IL-24co-expressing ING4 and IL-24. The Ad-ING4-IL-24 were amplified in QBI-293A cells and its titer was up to 3.5 × 109 PFU/ml. Adenovirus-mediated ING4 and IL-24 genes expression in HepG-2 cells was examined by RT-PCR and Western blot. The growth-suppressing and apoptosis-inducingg effect of Ad-ING4-IL-24 coexpressing ING4 and IL-24 on HepG-2 human hepatocellular carcinoma cells was assessed by MTT assay and FCM, respectively. Results DNA sequencing showed that the ING4, poly (A)-Promoter, and IL-24 fragments subcloned into pAdTrack-CMV plasmids were completely identical to those reported in GenBank.ING4 and IL-24 gene mediated by adenovirus could both successfully express in HepG-2 cells. Adenovirusmediated ING4 and IL-24 co-expression significantly suppressed HepG-2 hepatocellular carcinoma cell growth and induced cell apoptosis, and the effect of Ad-ING4-IL-24 group was more significant than AdING4 and Ad-IL-24 group. Conclusion The adenoviral vector co-expressing ING4 and IL-24 mediated by poly(A)-Promoter(Ad-ING4-IL-24) was successfully constructed. Ad-ING4-IL-24 had marked anti-tumor effect in suppressing HepG-2 human hepatocellular carcinoma cell growth and inducing cell apoptosis in vitro. Compared with Ad-ING4 and Ad- IL-24, Ad-ING4-IL-24 enhanced anti-tumor effect.