Objective To investigate the expression of human glucocorticoid-induced tumor necrosis factor receptor(hGITR)in synovial and cartilage tissue of patients with rheumatoid arthritis(RA),and analyze their relationships with the severity of synovial inflammation in RA.Methods Expression of hGITR in synovial and cartilage biopsy material of 16 RA patients,9 osteoarthritis(OA)patients,4 contmIs were detected in situ by immunohistochemical technique and the results were analyze then.Results hGITR was strongly expressed in the vascular endothelial cells and inflammatory cells in RA and hGITR positive cells could be detected in approximately 69%of RA synovial cells.Significant differenee Was foand in hGITR expression between RA patients and control group(P<0.01).hGITR was less expressed in the cartilage than the synovial in RA.No significant difference was found in expression in the cartilage tissue expression between RA patients and the control group.Significant difference Was found in hGITR expression between synovial and cartilage in RA.Furthermore.the hGITR positive cells in synovial tissues were also found to correlate significandy with the severity of synovial inflammation in RA patients(r=0.895,P<0.01).Conclusion These results suggest that the abnormal expression of hGITR in the synovial my bean important mechanism leading to the synovial destruction found in RA.

Objective To evaluate the preliminary outcome of postoperative three-dimensional conformal radiation therapy(3DCRT)as compared with conventional radiation therapy(CRT)in stageⅢA non-small cell lung cancer(NSCLC).Methods From December 1994 to February 2002,114 such patients received postoperative radiotherapy in M.D.Anderson Cancer Center,with 63 patients in CRT group and 51 in 3DCRT group.The median radiation dose was 53.5 Gy/27f/5-6w and 52.8 Gy/26f/5-6w in CRT and 3DCRT group, respectively.Results The median follow-up time was 28 and 13 months in CRT and 3DCRT group.The 1-,2- year overaLl survival were 80%,62% and 93%,70%.The 1-,2-year disease free survival was 62%,44% and 58%,32% in CRT group and in 3DCRT group,respectively.There was no significant difference between CRT group and 3DCRT group in terms of overall survival,disease-free survival,local-regional control,distant metastasis-free survival or treatment-related complication.There was significant difference in the death cause between two group,with 4 patients having died of cardiac complication in the CRT group and none in the 3DCRT group.Conclusions Postoperative three-dimensional conformal radiotherapy can reach the same immediate results of conventional radiotherapy in stageⅢA NSCLC,with a lower incidence of cardiac complication. However,the long-term results need further follow-up.

Objective To investigate the expression levels of telomeric protein TPP1 and POT1mRNA in the peripheral blood mononuclear cells (PBMCs) of patients with systemic lupus erythematosus (SLE), and the relations between these gene expression levels and disease activity are explored. Methods TPP1 and POT1 genes were measured using real-time polymerase chain reaction (RT-PCR) in PBMCs. The expression levels of TPP1 and POT1 genes in PBMCs were compared between 48 SLE patients and 30 healthy indivi-duals. Results The expression levels of TPP1 and POT1 in the PBMCs of SLE patients significantly decreased compared to healthy individuals (P<0.01). The expression levels of TPP1 and POT1 were much lower in SLE patients with lupus nephritis than those in patients without lupus nephritis (P<0.01), and in LN patients, the levels of TPP1 and POT1 were negatively correlated with proteinuria. The expression levels of POT1 in active SLE patients was lower than that in inactive SLE patients (P<0.05). The expression levels of POT1 was negatively correlated with serum IgG and SLEDAI scores, but positively correlated with complement C3, but no correlation between ESR, CRP, C4,ANA and the expression levels of TPP1, POT1, SLEDAI scores. Conclusion TPP1 and POT1 genes play a key role in the etiology of SLE, and they are involved in the pathogenesis of lupus nephritis. POT1 is helpful in evaluating SLE disease activity and severity.

Objective To evaluate the safety and efficacy of endovascular intervention to revise peripheral bypass problems through prosthetic approach.Methods Among 17 cases undergoing graft bypass anastomotic stenosis and graft thrombosis was identifled in 16 cases(inflow or outflow obstructive lesions in 10),inflow obstructive lessions in 1(without anastomotic and graft thrombosis).All revision procedures were taken under local anesthesia,16 patients were treated by means of surgical thrombectomy followed by endovascular intervention through prosthesis itself in addition to one who had no thrombectomy.The graft patency and clinical outcome were observed.Resuits Thirteen stents were implanted in 13patients with distal anastomotic stenosis and 1 with proximal anastomotic stenosis including 10 stentings/PTAs in iliac popliteal,posterior tibial or anterior tibial arteries.One stent was implanted in 1 patient with common femoral stenosis.Stenting were not used(abandoned)in 2 patients,of which one underwent a foot amputation and calf gangrene occurred a week later,and the other had a redo of grafting.Follow-up time is 1-35 months.with an average of 12±4 months.One had a below-knee amputation two months after intervention,the other had symptoms recurred and treated with a redo 3 months afterwards.the third died of myocardial infarction six months later.Grafts remained patent in the rest 13 patients at follow-up.Conclusions Endovascular intervention through prosthesis is a safe and effective method,which offers an alternative means to treat anastomotic stenosis.inflow or outflow obstructive lesions.

Increasing evidence has revealed that maternal cytomegalovirus (CMV) infection may be associated with neurodevelopmental disorders in offspring.Potential relevance between the placental inflammation and CMV-related autism has been reported by clinical observation.Meanwhile,abnormal expression of Toll-like receptor 2 (TLR2) and TLR4 in placenta of patients with chorioamnionitis was observed in multiple studies.IL-6 and IL-10 are two important maternal inflammatory mediators involved in neurodevelopmental disorders.To investigate whether murine CMV (MCMV) infection causes alterations in placental IL-6/10 and TLR2/4 levels,we analyzed the dynamic changes in gene expression of TLR2/4 and IL-6/10 in placentas following acute MCMV infection.Mouse model of acute MCMV infection during pregnancy was created,and pre-pregnant MCMV infected,lipopolysaccharide (LPS)-treated and uninfected mice were used as controls.At E13.5,E14.5 and E18.5,placentas and fetal brains were harvested and mRNA expression levels of placental TLR2/4 and IL-6/10 were analyzed.The results showed that after acute MCMV infection,the expression levels of placental TLR2/4 and IL-6 were elevated at E13.5,accompanied by obvious placental inflammation and reduction of placenta and fetal brain weights.However,LPS 50 μg/kg could decrease the IL-6 expression at E13.5 and E14.5.This suggests that acute MCMV infection during pregnancy could up-regulate the gene expression of TLR2/4 in placental trophoblasts and activate them to produce more proinflammatory cytokine IL-6.High dose of LPS stimulation (50 tg/kg) during pregnancy can lead to down-regulation of IL-6 levels in the late stage.Imbalance ofIL-6 expression in placenta might be associated with the neurodevelopmental disorders in progeny.

Objective To analyze the constituent of the 32 kD protein band and its expression in schizophrenia se?rum. Methods Sixty schizophrenia patients and 58 health controls were recruited. The serum samples were collected and precipitated with 7%PEG. The sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to ob?tain the abnormal 32 kD proteins band in patients. This protein band was cut and then analyzed using mass spectrometric technique. Results The 32 kD protein band was present in 38 schizophrenia patients but not in control and positive rate was 63.33%. The mass spectrometric analysis showed that 32 kD protein band contained 14 proteins ranging from 30 kD to 35 kD, including 6 high-frequency proteins (cDNA coded protein 1 and 2, Apolin protein A-1, Isoform 2 of ficolin-2, Complement factor H and clusterin) and 8 low-frequency proteins (IgG H chain, zinc-alphg-2-glycoprotein, fermitin,family apolin protein L-1, isoform 10 of collectin-1, purine nucleoside, anne xin and cDNA coded protein 3). Three cD?NA coded unknown proteins were highly similar to complement C4-B, β2-glycoprotein and erythrocyte band 7 integral membrane protein. Conclusion There is a unknown specific 32 kD protein that is consisted mainly of fourteen proteins in serum of schizophrenia.

Objective To investigate the initiation pathway of corneal endothelial cell apoptosis induced by high-pression.Methods Primary rabbit corneal endothelial cells were identified by immunohistochemistry and cultured under high pressure 50 mmHg (1 kPa =7.5 mmHg) for 1 h,2 h,24 h,respectively,while cells cultured under the normal pressure 15 mmHg served as the normal pression group.In addition,the first generation of rabbits corneal endothelial ceils with 70％ to 80％ fusion were pretreated with 10-6 mol · L-1 anti-Caspase 8 and anti-Caspase 9 for lh,followed by 50 mmHg pression for the treatment of the cells;while cells cultured with no inhibitor in the same pression served as the control group.Then the expression of P53 and Bcl-2 protein was detected by Western blot,and cytochrome C in rabbit corneal endothelial cells was determined by immunofluorescence staining in all groups.Results The expression levels of P53 in the 50 mmHg group were 0.651 +0.007,0.805 ±0.006 and 0.839 ±0.011 after 1 h,2 h,24 h high-pression respectively,which were significantly higher than those in the normal pressure group (0.033 ± 0.004),and the difference approached statistical significance (all P ＜ 0.01).The expression of P53 protein in corneal endothelial cells gradually increased as time went on,and the difference was statistically significant between each two time-points (all P ＜ 0.01).Moreover,the expression of Bcl-2 in the 50 mmHg pressure group was 0.590 ± 0.009,0.724 ± 0.005 and 0.34 ± 0.016,respectively,which was higher than that in the normal pressure group (0.081 ±0.013),with signifi cant difference (all P ＜ 0.01),and the difference approached statistical significance between each two time points in this group (all P ＜ 0.01).The expression level of P53 in anti-Caspase 9 and anti-Caspase 8 group was 0.535 ± 0.007 and 0.703 ± 0.010,respectively,which was significantly lower than that in the control group (0.727 ± 0.021),and the difference was statistically significant (all P ＜ 0.01).The expression of Bcl2 was 0.312 ± 0.003 and 0.442 ± 0.011,respectively,which were significantly lower than that in the control group (0.501 ± 0.011),with statistical difference (P ＜ 0.01).Finally,the expression of P53 and Bcl-2 in anti-Caspase 9 group was lower than that of anti-Caspase 8 group (P ＜ 0.01),indicating that anti-Caspase 9 had more enhanced inhibitory effect on the apoptosis of corneal endothelial cells than anti-Caspase 8.Conclusion AntiCaspase 9 inhibitor could effectively block the corneal endothelial cell apoptosis induced by high pressure.And the damage from high pressure on corneal endothelial cells mainly triggers the release of cytochrome C from chondriosome to activate the endogenous enzyme linked apoptotic pathway in which Caspase 9 involves.

To explore a rapid and easy method to detect labile iron of pool (LIP) in cells, HL-60 and K562 cells were cultured at a concentration 1 x 10(6)/ml in RPMI 1640 containing 10% heat-inactivated fetal bovine serum. The iron deprivation was induced by adding desferrioxamine (DFO) 10 - 100 micromol/L for 0 - 48 hours. The intracellular LIP was measured by probe calcein-AM. Calcein fluorescence was monitored in 1420 multilabel counter. The results indicated that when HL-60 and K562 cells were incubated with different concentrations of DFO, the calcein fluorescence intensity was higher than that of control group at 12, 24 and 48 hours (P < 0.05). Fluorescence value of representing LIP in DFO groups was lower than that in the control group. In conclusion, DFO can decrease LIP in leukemia cells. The approach used in this study may provide a simple and reliable method for detection of intracellular iron homeostasis.
Cation Transport Proteins ; antagonists & inhibitors ; biosynthesis ; metabolism ; Deferoxamine ; pharmacology ; Fluoresceins ; Fluorescent Dyes ; HL-60 Cells ; Humans ; Iron ; metabolism ; Iron Chelating Agents ; analysis ; metabolism ; Iron-Regulatory Proteins ; metabolism ; K562 Cells

OBJECTIVE12-0-tetradecanoylphorbol-13 acetate (TPA) plays an important role in precipitating cell differentiation for various tumor cells, especially leukemic cells. Changes of many genes may be involved in this process. The purpose of this study was to observe the relationship between the EGR1mRNA expression and cell differentiation during TPA-induced K562 cell differentiation.

METHODSIncubation of human K562 cells in vitro was applied to cultivate K562 cells. The cells were treated in two different ways. K562 cells of experiment group were treated with TPA and those of control group were treated without TPA. Using morphology (Wright's staining and NSE staining) and flow cytometry (FCM), the investigators observed the differentiation characteristics of K562 cells, cell-cycle and the differentiation antigen expressions of CD33 and CD14 on cell membranes. RT-PCR was carried out to assay EGR1 mRNA expression.

RESULTSAfter treated with TPA for 7 d, the morphology of K562 cells obviously tended to mature differentiation, like monocytes. The differentiation rate of induced K562 cells was up to 95% in experiment group and 4.5% in control group, respectively. Using SPSS software, the above result showed statistical significance (P < 0.01). Using NSE staining, K562 cells showed positive reaction. Some of them were densely stained. The positive rate was up to 86%. More than half of the positive cells could be inhibited by NaF. The inhibiting rate of NaF was up to 58.72%, showing statistical difference when compared with that of control group. FCM analysis showed that most of K562 cells stimulated by TPA underwent G1/S phase cell-cycle arrest. The composing rate of cell-cycle in TPA-treated group showed that (53.7 +/- 1.25)% of cells were at G0 + G1 phase and (44.3 +/- 1.32)% were at S phase (P < 0.05). The level of CD33 expression on cell membranes was mildly decreased from 0.997% to 0.893% (P > 0.05). However, the level of CD14 expression was significantly increased from 0.049% to 0.387% (P < 0.05).

CONCLUSIONK562 cells could express EGR1mRNA during TPA-induced differentiation, which suggested that EGR1mRNA might participate in the process of K562 cells differentiating into monocyte/macrophages, and might play an important role in precipitating and maintaining cell differentiation for leukemic cells.

Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Carcinogens ; pharmacology ; Cell Cycle ; drug effects ; genetics ; Cell Differentiation ; drug effects ; genetics ; Cell Division ; drug effects ; genetics ; Cell Membrane ; chemistry ; drug effects ; DNA-Binding Proteins ; genetics ; Early Growth Response Protein 1 ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Immediate-Early Proteins ; genetics ; K562 Cells ; Lipopolysaccharide Receptors ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Sialic Acid Binding Ig-like Lectin 3 ; Tetradecanoylphorbol Acetate ; pharmacology ; Transcription Factors ; genetics

OBJECTIVETo investigate the changes of hair dermal papilla and its regulatory role in the growth cycle of the hair follicles.

METHODSSingle hair follicles were isolated from surgical specimens of human scalp and cultured in Williams E medium. The growth of the hair follicle was measured and the morphology and structure of the dermal papilla in the different growth cycles were observed continuously.

RESULTSThe hair follicle could grow in the medium for 12 days at the average growth rate of 0.2-0.3 mm/day. The flat and round dermal papilla lay at the bottom of the hair bulb in the telogen and anagen stages. In the hair follicle with accelerated growth, the dermal papilla became elongated, loosened, and closely adhered to the hair matrix. In the catagen stage the dermal papilla shrunk, and became separated from the hair matrix. A new hair bulb was regenerated when the hair follicle was transected at a low level. The hair follicle stopped growing after transection at a higher position.

CONCLUSIONThe hair dermal papilla is the essential for hair follicle growth, and plays an important role in regulating the hair growth cycle.

Dermis ; cytology ; growth & development ; Hair ; growth & development ; Hair Follicle ; cytology ; growth & development ; Humans ; Tissue Culture Techniques