By using ~3H—TdR incorporation and CTLL—2 cell line,we observed effects of RU486,anantagonist of type Ⅱ glucocorticoid receptor,on dexamethasone inhibition of lymphocyte prolifer-ation and interleukin—2(IL—2)production in rat spleen.It was shown that RU486 obviouslyantagonized the inhibitory actions of dexamethasone on lymphocyte proliferation and IL—2 pro-duction.By means of dot hybridization and Northern blot analysis,effects of dexamethasone andRU486 on IL—2 gene expression were investigated in lymphocytes of rat spleen.The datademonstrated that dexamethasone markedly decreased IL—2 mRNA production,RU486 alonedidn't affect IL-2 mRNA levels,but obviously reversed dexamethasone-mediated downregala-tion of IL—2 mRNA production in ConA—activated lymphocytes.These results suggest thatthe above effects of dexamethasone may be mediated by glucocorticoid receptor in lymphocytesof rat spleen.

ObjectiveTo explore the effect of estrogen on osteoblast apoptosis induced by serum hungry in vitro.MethodsOsteoblasts of second or third generation from newly born SD rats calvaria were divided randomly into the control group, serum hungry group and serum hungry with estrogen group. Cells of each group were incubated for 24 h, 48 h, 72 h, 5 d, 7 d and 14 d, then labeled using TUNEL staining and examined for morphological characteristics of apoptotic cell under light microscopy after incubated for 72 h. The rates of apoptotic cells of each group were examined with flow cytometry.ResultsThe cells of the control group showed normal appears, the serum hungry group had many cells with purple and blue particles in nuclei, but serum hungry with estrogen group had less such cells. The rate of apoptotic cell significantly increased in serum hungry group and decreased in serum hungry with estrogen group compared with the control group examined with flow cytometry (P<0.05).ConclusionEstrogen can repress osteoblasts apoptosis of rats induced by serum hungry.

Abstract:Objective To investigate the protective effects of blocking CD40/CD40L interactions with human CD40-Ig fusion protein in a murine graft-versus-host disease model.Methods Human CD40 gene extracellular region was inserted into plasmid pIG1, which contains genomic human IgG1 Fc gene. A transient vector containing CD40-Fc fusion gene was transfected into COS-7 cells. The CD40-Ig fusion protein was detected through enzyme-linked immunosorbent assay (ELISA). A constitutive vector was also generated by ligating the CD40-Fc fusion gene into pcDNA3.1 and transfecting it into CHO cells. CD40-Ig was purified by protein A affinity chromatography. SDS-PAGE, Western blot and ligand binding assay were used to identify the qualities of CD40-Ig. Murine acute graft-versus-host disease (GVHD) was induced by intravenous injection of C57BL/6J (H-2b) spleen cells into sub-lethally irradiated BALB/c (H-2d) mice. Protective effects against murine graft-versus-host disease by in vivo administration of CD40-Ig were evaluated.Results Mammalian expression vectors pIG/40Ig and p3.1/40Ig were constructed as described above. Chimeric proteins were expressed in COS-7 and CHO cell culture supernatant and confirmed by ELISA and Western blot. SDS-PAGE showed that fusion proteins had a disulfide-bonded dimeric structure and existed as homodimer. Purified CD40-Ig could bind to CD40L. In vivo administration of CD40-Ig could prevent the development of GVHD and significantly prolong the mean survival time of mice with graft-versus-host disease.Conclusions These results demonstrate that CD40/CD40L interactions play an important role in the pathogenesis of graft-versus-host disease and suggest clinical potential for CD40-Ig in the prevention and treatment of human graft-versus-host disease.

OBJECTIVETo know about content of iodine in foods sold in Tianjing markets presently, and the iodine nutrition conditions in college students. It was also aimed to probe the functions of the iodized salt complement with the dietary iodine intake, and whether the urine iodine could reflect dietary iodine intake.

METHODS278 food samples in markets were collected by a randomly stratified sampling method, while the arsenic-cerium catalytic contact method was used to determine the content in food. The dietary information of students for seven days was recorded, and the urine iodine was determined through the arsenic-cerium catalytic spectrophotometry.

RESULTSThe determination of 47 kinds and 278 food samples indicated that the content of iodine within animal foods (7.8 microg/100 g - 30.8 microg/100 g) was higher than that within plant foods (1.8 microg/100 g - 16.1 microg/100 g). The investigation also showed that students who regarded vegetarian food as principle accounted for 70. 19%. The amount of dietary iodine intake among those students, based on the dietary survey, was (111.67 +/- 53.18) microg/d, while supplementary iodine from iodized salt was about (230.27 +/- 45.55) microg/d. Therefore, the total iodine provided from diet would be (341.95 +/- 89.58) microg/d. Modified by urine creatinine, the median of urine iodine was 271.28 microg/gCr, and the urine iodine and dietary iodine intake was found positively related (r(s) = 0.463, P < 0.01).

CONCLUSIONSRegarding the vegetarian food as the principle, most of students investigated are not rich. The dietary iodine intake is lower than RDA (150 microg), but it can be obtained the iodized salt by 230. 27 microg, which is the possible supplement to the shortage from foods.

China ; Diet Surveys ; Humans ; Iodine ; Nutritional Status ; Sodium Chloride, Dietary ; Students

CD40/CD40L, besides B7/CD28, is an alternative important costimulation signal transduction pathway. It plays a pivotal role in T cell activation. Moreover, it may play a critical role at many levels of sensitization and effector phases of allograft rejection. In order to get the fusion protein of human CD40 extracelluar region and IgG 1 Fc fragment, and investigate the potential role of blocking CD40/CD40L costimulation pathway in immunotherapy, total RNA was extracted from human lymphoma cell line Daudi, and CD40 gene extracelluar region was amplified by RT-PCR. The PCR products were inserted into pGEM T Easy vector, and the cloning vector pGE40 was obtained. The DNA sequence was analyzed by automatic DNA sequencer. After sequencing, the transient expressing vector was constructed by inserting correct fragment into pIG vector, which contains the genomic human IgG1 Fc (hinge, CH2 and CH3) gene. Hence the recombinant fusion expression vector was constructed successfully, and named after pIG/40 Ig. Then, COS-7 cells were transfected through DEAE-Dextran/chloroquine method. The CD40-Ig fusion protein expressed in COS-7 cell culture supernatant was identified by sandwich ELISA and Western blot. Result showed that the CD40-Ig fusion protein can be detected by sandwich ELISA in the cell culture supernatant. Western blot analysis also showed that it could react with McAbs of mouse anti-human CD40 G28-5 and mouse anti-human Ig gamma chain. There is only one obvious band at the position of relative molecular weight 50 kD, and it is equivalent to the expected value. Above all, the recombinant fusion expression vector pIG/40 Ig was constructed, and CD40-Ig fusion protein gene was expressed in COS-7 cells successfully. It could be laid a foundation to investigate the potential role of CD40/CD40L pathway as the target of GVHD prevention and therapy.

Humans ; Tranexamic Acid ; Atrial Fibrillation ; Antifibrinolytic Agents ; Arthroplasty ; Blood Loss, Surgical ; Arthroplasty, Replacement, Hip

OBJECTIVETo investigate the expression of fractalkine (FKN) and its receptor CX3CR1 in cardiac allografts and the effect of Cyclosporin A (CsA).

METHODSThree groups of rats underwent heterotopic cardiac transplantation, 45 cases in each group and 5 cases in control group: SD to SD regarded as isograft group (group A), Wistar to SD divided into CsA untreated allograft group (group B) and CsA treated allograft group (group C), normal SD rats as control group. The FKN mRNA expression was detected by one-step RT-PCR method and the expression of FKN and CX3CR1 protein was detected by standard ABC immunohistochemical technique.

RESULTSThe expression of FKN mRNA and protein was weak in both isografts and normal heart specimens. The changes of FKN mRNA expression were correlated with the process of acute allograft rejection. The peak of FKN mRNA expression (0.8 +/- 0.26) appeared on the seventh day after transplantation, which could be down-regulated by CsA significantly (t = 2.390, P < 0.05). FKN protein was located in endothelia cells and its receptor CX3CR1 was located in infiltrating mononuclear cells in allografts.

CONCLUSIONSUpregulation of FKN and its receptor was significantly correlated with the trafficking of mononuclear cells which play an important role in acute allograft rejection. It may be one of the important mechanisms of CsA to intervene the acute rejection by inhibiting the activation of the FKN-CX3CR1 pathway.

Acute Disease ; Animals ; CX3C Chemokine Receptor 1 ; Chemokine CX3CL1 ; Chemokines, CX3C ; genetics ; metabolism ; Cyclosporine ; pharmacology ; Graft Rejection ; immunology ; pathology ; prevention & control ; Heart Transplantation ; immunology ; pathology ; Immunohistochemistry ; Male ; Membrane Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Receptors, Cytokine ; genetics ; metabolism ; Receptors, HIV ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous

OBJECTIVETo delete IL-11 receptor alpha chain gene from the Bacterial Artificial Chromosome (BAC) chimeric DNA by RecA protein mediated homologous recombination method and establish the transgenic mice model containing whole beta-globin gene cluster.

METHODSTwo 500 bp homologous sequences (A and B) located at the upstream and downstream of IL-11 receptor alpha chain gene respectively were cloned into the Hind III and Xba I sites of pBV vector, then the 1 kb A + B fragment was recovered from the building vector and inserted into the Sal I site of the shuttle vector pSV-RecA. After transforming the shuttle vector into the competent DH10B E. Coli containing BAC DNA, the IL-11 receptor alpha chain gene was finally deleted from the BAC DNA through chloramphenicol positive selection and fusaic acid negative selection. The new BAC clone was characterized by Pulse Field Gel Electrophoresis (PFGE). Then, we microinjected the linearized and purified BAC DNA into the mouse fertilized eggs and prepared the transgenic mice.

RESULTSBy RecA protein mediated homologous recombination method, we deleted the IL-11 receptor alpha chain gene from the BAC DNA containing the complete beta-globin Gene Cluster and established 3 respective transgenic mice lines.

CONCLUSIONHuman beta-globin gene cluster in the transgenic mice mediated by new BAC expresses in a correct mode and level as compared with previous transgenic mice.

Animals ; Chromosomes, Artificial, Bacterial ; genetics ; DNA ; Globins ; biosynthesis ; genetics ; Humans ; Interleukin-11 ; genetics ; Mice ; Mice, Transgenic ; Models, Animal ; Multigene Family ; genetics ; Receptors, Interleukin ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transcription, Genetic

OBJECTIVETo observe the clinical efficacy of acupoint-catgut-embedding therapy combined conventional western medication on chronic obstructive pulmonary disease (COPD) at stable stage due to lung and kidney deficiency.

METHODSThe cases were randomized into observation group and control group, 30 cases in each one. In control group, the conventional western medication was administered. In observation group, on the basis of conventional western medication, the catgut-embedding therapy was applied at Dingchuan (EX-B 1), Feishu (BL 13), Shenshu (BL 23), Fenglong (ST 40) and Zusanli (ST 36). The total attack frequency of acute exacerbation of COPD (AECOPD), the attack frequency of AECOPD at moderate or above moderate stage and TCM syndrome score were compared before treatment and 6 months after treatment in two groups.

RESULTSThe total effective rate was 93.3% (28/30) in observation group and was 90.0% (27/30) in control group, indicating equivalent efficacy between two groups. 6 months after treatment, in two groups, the total attack frequency of AECOPD and the attack frequency of AECOPD at moderate or above moderate stage were reduced remarkably as compared with those before treatment (P < 0.01, P < 0.05). The total attack frequency of AECOPD in observation group was reduced remarkably as compared with that in control group (P < 0.05). The scores of cough, expectoration and chest oppression as well as the total score of TCM syndrome were reduced remarkably after treatment in observation group (all P < 0.01).

CONCLUSIONIntegrated therapy of acupoint-catgut-embedding and conventional medication has similar efficacy as simple medication. But, the combination of acupoint-catgut-embedding therapy and western medication can reduce the attack frequency of AECOPD and improve in cough, chest oppression and other symptoms in patients with COPD at stable stage effectively.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Aged ; Aged, 80 and over ; Catgut ; Combined Modality Therapy ; Female ; Humans ; Kidney ; physiopathology ; Lung ; physiopathology ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; physiopathology ; therapy