PURPOSE: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8+T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8+CLA+T cells might be underlying mechanism in AD. MATERIALS AND METHODS: CD8+CLA+T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8+CLA+T cells were evaluated. The proliferative responses of CD8+CLA+T cells were assessed by flow cytometry, and the levels of transforming growth factor-beta1 (TGF-beta1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8+CLA+T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8+CLA+T cells in AD. Meanwhile, the levels of TGF-beta1 produced by Tregs were significantly lower in AD, and anti-TGF-beta1 abolished such suppression. CONCLUSION: The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8+CLA+T cells, mediated by TGF-beta1, plays an important role in the pathogenesis of AD.
Adult ; Aged ; CD8-Positive T-Lymphocytes/drug effects/*immunology ; Case-Control Studies ; Cell Proliferation ; Cell Separation ; Dermatitis, Atopic/*immunology/pathology ; Female ; Granzymes/metabolism ; Humans ; Interleukin-10/metabolism ; Lymphocyte Count ; Male ; Perforin/metabolism ; Skin/*immunology/pathology ; T-Lymphocytes, Cytotoxic/drug effects/immunology ; T-Lymphocytes, Regulatory/drug effects/*immunology ; Transforming Growth Factor beta1/pharmacology

OBJECTIVETo study the expression of decoy receptor 3 (DcR3) and its relationship with apoptosis and prognosis in hepatocellular carcinoma (HCC).

METHODSThe expression of DcR3 protein in 43 cases of HCC and 16 cases of non-cancerous liver (including cirrhotic liver tissue and normal liver tissue adjacent to cavernous hemangioma) was studied by immunohistochemistry (using EnVision method). The status of apoptosis was evaluated by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling (TUNEL) technique. Statistic analysis was carried out to assess the correlation between DcR3 expression, apoptotic index (AI) and clinicopathologic parameters.

RESULTSDcR3 protein was expressed in the cytoplasm of HCC cells. The positivity rate of DcR3 in HCC was 74.42% (32/43), which was significantly higher than that in the non-cancerous group (43.75%, P < 0.05). The positivity rate of DcR3 in HCC with metastasis detected within 20 months of diagnosis was 100% (22/22). This was significantly higher than that in HCC without metastasis (52.94%, P < 0.01). The DcR3 expression in HCC also correlated with serum alpha-fetoprotein level (r = 0.444, P < 0.01) and presence of tumor embolus in portal vein (r = 0.414, P < 0.01). However it had no relationship with the patient's age, sex, cirrhotic status, liver capsule invasion, number of tumor nodules and histologic differentiation (P > 0.05). The AI in HCC (0.78 +/- 0.64)% was significantly lower than that in the non-cancerous group [(3.32 +/- 1.81)%, P < 0.01]. The AI in clinical TNM stage I and II tumors (1.03 +/- 0.69)% was significantly higher than that in stage III and IV tumors [(0.52 +/- 0.48)%, P < 0.01]. The AI in HCC without metastasis (1.10 +/- 0.72)% was significantly higher than that in HCC without metastasis [(0.44 +/- 0.27)%, P < 0.05]. The AI correlated with serum alpha-fetoprotein level (r = -0.468, P < 0.01), presence of tumor embolus in portal vein (r = -0.434, P < 0.01) and liver capsule invasion (r = -0.331, P < 0.05). On the other hand, it had no relationship with patient's age, sex, cirrhotic status, number of tumor nodules and histologic differentiation (P > 0.05). The AI in DcR3-positive group (including both HCC and non-cancerous tissues) was significantly lower than that in DcR3-negative group (P < 0.01).

CONCLUSIONSThe expression of DcR3 in HCC correlates with apoptosis of tumor cells and may play a crucial role in tumor pathogenesis and progression. DcR3 protein expression and AI may also serve as important biologic indicators in predicting prognosis of HCC.

Adult ; Aged ; Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Receptors, Tumor Necrosis Factor, Member 6b ; metabolism ; physiology ; Retrospective Studies ; Young Adult

OBJECTIVETo observe the anti-tumor effect of combination TNF-related apoptosis-inducing ligand (TRAIL) with aspirin on liver cancer cell line, SMMC-7721.

METHODSThe survival fraction of SMMC-7721 cells was measured by MTT assay, apoptosis rate and cell cycle was determined by flow cytometry, and the expression of apoptosis-related gene was identified by western blot.

RESULTSThe survival fraction of SMMC-7721 cells treated with 300 ng/ml TRAIL, 3 mmol/L or 10 mmol/L aspirin alone was 82.76%, 81.34% and 71.29% respectively, and the survival fractions of SMMC-7721 cells treated with TRAIL and 3 mmol/L or 10 mmol/L aspirin were 43.54% and 37.8% respectively. The apoptosis rates of SMMC-7721 cells induced by TRAIL and 3 mmol/L or 10 mmol/L aspirin were higher than that induced by TRAIL or aspirin alone (34.76% and 38.56% vs 21.25%, 1.89% and 6.08%), and G0/G1 arrest was observed under TRAIL and aspirin. The expression of Bcl-2 in SMMC-7721 cells treated by 3 mmol/L or 10 mmol/L aspirin decreased markedly, but no effect on Bax.

CONCLUSIONThe cooperative anti-tumor effect of aspirin and TRAIL may be related to the inhibition of the expression of Bcl-2 by aspirin

Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Apoptosis ; Apoptosis Regulatory Proteins ; Aspirin ; pharmacology ; Cell Survival ; drug effects ; Humans ; Membrane Glycoproteins ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; antagonists & inhibitors ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Cells, Cultured ; Tumor Necrosis Factor-alpha ; pharmacology

OBJECTIVETo study the effects of human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) on biological characteristics of hepatocellular carcinoma cell lines HepG2 and SMMC-7721 and on apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

METHODSSmall hairpin hTERT (shTERT) sequence was identified by PCR method; hTERT expressions, morphological features, cell proliferation and replicative senescence were respectively determined using RT-PCR, hematoxylin-eosin (HE) staining, growth curve and beta-galactosidase (b-Gal) staining; cell cycle and apoptosis were identified using flow cytometry after propidium iodide (PI) staining and annexin V/PI double staining.

RESULTSshRNA were found in 6/8 HepG2 and 6/6 SMMC-7721 cell clones transformed by the recombined plasmid pSilencer 3.1-H1 neo-shTERT. The interference rates of hTERT on HepG2 and SMMC-7721 were 100% and 43.3% respectively. Cells in G2-M phases increased from 7.1% to 10.6% and from 6.9% to 7.9% respectively; and the percentage of replicative senenscence cells increased from 0 to 20.4% and from 3.6% to 10.0% respectively. The nucleus/cytoplasm ratios of the cells were obviously decreased after hTERT RNAi treatment. Moreover, apoptosis of hepatocellular carcinoma cells and apoptosis induced by TRAIL were strikingly increased by hTERT RNAi (P < 0.05). For example, apoptosis rates were increased from 3.5% to 5.2% in HepG2 cells and from 4.8% to 7.9% in SMMC-7721 cells after hTERT RNAi treatment. Apoptosis rates were increased from 5.3% to 10.4% in HepG 2 cells and from 13.9% to 77.2% in SMMC-7721 cells after being treated by 100 ng/ml TRAIL for 24 h. However, there were no remarkable changes between control cells and untransformed cells.

CONCLUSIONhTERT RNAi not only has a significant effect on biological characteristics of hepatocellular carcinoma cells, but also obviously can increase cell apoptosis induced by TRAIL.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; pathology ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; RNA, Small Interfering ; genetics ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Telomerase ; genetics ; Tumor Cells, Cultured

OBJECTIVESTo investigate the mechanisms for human telomerase reverse transcriptase (hTERT) RNA interference (RNAi) in increasing hepatocellular carcinoma cell apoptosis induced by TNF-related apoptosis-inducing ligand (TRAIL).

METHODSCell apoptosis was identified by flow cytometry analysis after annexin V/PI double staining. Expression of apoptosis-related proteins, procaspase-8, -9, -3, Bax, Bcl-2 and hTERT, were identified by Western blotting analysis; telomerase activity and telomere length were detected by telomeric repeat amplification protocol (TRAP) and telomere amount and length assay (TALA) methods.

RESULTSHepatocellular carcinoma cell apoptosis induced by TRAIL were all significantly increased by hTERT RNAi (P less than 0.05). For example, apoptosis rates were enhanced from 5.53% (untransformed) to 10.35% (transformed) in HepG 2 cells and from 14.73% to 77.24% in SMMC 7721 cells after being treated by 100 ng/ml TRAIL for 24 h. Moreover, activation of procaspase-8, -9 and -3 in transformed cells after being treated by TRAIL were all significantly raised (P less than 0.05) in a dose-dependent manner. The expression of procaspase-8, -9 and Bcl-2 were effectively augmented (P less than 0.05), but expressions of Bax and hTERT were strikingly decreased (P less than 0.05). Meanwhile, telomerase activity was apparently suppressed and telomere length was markedly shortened (P less than 0.05). There were no remarkable differences in these effects between control cells and the untransformed cells (P more than 0.05).

CONCLUSIONEnhanced cell apoptosis induced by TRAIL through hTERT RNAi may be related to up-regulation of procaspase-8 and -9 expressions. However the down-regulation of hTERT expression, reduced telomerase activity and shortened telomere length may not be related to expressions of Bcl-2 and Bax.

Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; RNA Interference ; TNF-Related Apoptosis-Inducing Ligand ; pharmacology ; Telomerase ; genetics ; metabolism

Aged ; Carcinoma, Papillary ; complications ; diagnosis ; Humans ; Male ; Neoplasm Metastasis ; Skull Base Neoplasms ; secondary ; Thyroid Neoplasms ; complications ; diagnosis

OBJECTIVETo explore the relationship between chronic prostatitis (CP) and prostatic calculus (PC).

METHODSWe used transperineal ultrasonography (TPUS) to detect PC in 500 normal volunteers and 491 CP patients, and divided them into a CP and a CP + PC group according to the ultrasonographic results. Then we analyzed the NIH-CPSI scores, duration of symptoms and white blood cell count in the expressed prostate secretion (ESP).

RESULTSPC was found in 19.8% of the normal controls, 5% (5/100), 12% (12/100), 19% (19/100), 27% (27/100) and 36% (36/100) in the 20-30 yr, 31-40 yr, 41-50 yr, 51-60 yr and > 60 yr groups, respectively. In comparison, PC was detected in 42.2% of the CP patients, 15.8% (12/76), 30.1% (69/215), 55.7% (59/109), 66.2% (43/65) and 82.8% (24/29) in the above five age groups, respectively, with statistically significant differences between the control and CP groups (P < 0.01). The CP and CP + PC groups showed significant differences in the duration of symptoms and white blood cell count in ESP (P < 0.01) but not in CPSI scores (P < 0.05).

CONCLUSIONThe incidence of PC is higher in CP patients than in healthy men, and it is associated with inflammation, aging and symptom duration, but not with CPSI scores.

Adult ; Aged ; Aged, 80 and over ; Bodily Secretions ; Calculi ; pathology ; Case-Control Studies ; Chronic Disease ; Humans ; Inflammation ; Leukocyte Count ; Male ; Middle Aged ; Prostate ; secretion ; Prostatic Diseases ; pathology ; Prostatitis ; pathology ; Young Adult

OBJECTIVETo observe the sensitivity change of SMMC-7721 cells transfected with antisense DNMT1 gene fragment to tumor necrosis factor related apoptosis inducing ligand (TRAIL) and its mechanism.

METHODSCell survival rate was measured by trypan blue, apoptosis rate by TUNEL method and the expression of bcl-2, bax and bad by flow cytometry.

RESULTSCell survival rate of SMMC-7721 cells transfected with antisense DNMT1 gene fragment was markedly lower than that transfected with sense DNMT1 gene fragment or empty vector (P < 0.05 and 0.01), but the apoptosis rate was on the contrary (P < 0.05 or 0.01). The expression of bax and bad (especially the former), but not bcl-2 of SMMC-7721 cells transfected with antisense DNMT1 gene fragment was markedly higher than those of SMMC-7721 cells transfected with sense DNMT1 gene fragment or empty vector.

CONCLUSIONThe sensitivity of SMMC-7721 cells to TRAIL can be enhanced by the transfection of antisense DNMT1 gene fragment, which may be related to the increase of bax and bad expression.

Antisense Elements (Genetics) ; genetics ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; Carrier Proteins ; analysis ; Cell Line, Tumor ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; antagonists & inhibitors ; genetics ; Flow Cytometry ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Membrane Glycoproteins ; pharmacology ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-2 ; TNF-Related Apoptosis-Inducing Ligand ; Tumor Necrosis Factor-alpha ; pharmacology ; bcl-2-Associated X Protein ; bcl-Associated Death Protein

Aged ; Humans ; Laparoscopy ; methods ; Male ; Middle Aged ; Prostatectomy ; methods ; Prostatic Neoplasms ; surgery

OBJECTIVETo propose the clinical classification of Peutz-Jeghers syndrome (PJS).

METHODS AND RESULTSRetrospective analysis of 52 patients with PJS admitted in Nanfang Hospital from 1980 to 2003 was conducted. Twenty-four patients were found to have family history of PJS, who had a mean age of 19 years. In the PJS patients, the incidence of gastric polyps was 64.4%, colorectal polyps 76%, and small bowel polyps 95%. The number of polyps was above 50 in 19 of the 31 patients with gastric polyps, in 18 of the 38 patients with colorectal polyps, and in 8 of the 19 patients with small bowel polyps. The pathology of the majority of the polyps (63/108) was characterized by hamartomas, and the incidence of malignancy was 13.5% in the PJS patients.

CONCLUSIONSPJS can be classified according to family history and location, pathology, and number of the polyps. As most patients with over 50 polyps require surgical intervention, 50 polyps is recommended as the criteria for PJS classification. Endoscopic surgery may suffice for management of patients with fewer polyps (<50), while in patients with more polyps or small bowel polyps, open surgery combined with intraoperative endoscopic surgery is recommended.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Intestinal Polyps ; pathology ; Male ; Peutz-Jeghers Syndrome ; classification ; pathology ; surgery ; Retrospective Studies