OBJECTIVETo investigate the apoptosis effect of diffuse large B-cell lymphoma cell line (DLBCL) SUDHL-4 induced by IL-21 and its related mechanism.

METHODSSUDHL-4 cells were treated with IL-21 at different concentration (1000 ng/ml, 100 ng/ml, 10 ng/ml, 1 ng/ml) for 24 h, 48 h, 72 h, respectively. The inhibitory rate of cell proliferation was detected by CCK-8 assay. The cell growth curves were drawn and half inhibitory concentration (IC(50)) values were calculated. The cell apoptosis were detected by flow cytometry (FCM), the expression of the caspase-9, caspase-3, cleaved caspase-3, Bcl-2, Bcl-XL, Bid, Bax and c-myc protein in SUDHL-4 cells treated with IL-21 by western blot, the mRNA expression of Bcl-2, Bcl-XL, Bid, Bax, c-myc by Survivin gene with RT-PCR.

RESULTSIL-21 markedly inhibited SUDHL-4 cell growth in a time- and dose-dependent manner. The 48 hIC(50) was 140.9ng/ml; The FCM showed that the apoptosis proportion of SUDHL-4 cells treated with 100 ng/ml of IL-21 apoptosis (AnnexinV-FITC(+) positive cells) gradually increased (48 h: 19.7 ± 2.3%). The protein expression of caspase-9, caspase-3, Bcl-2 and Bcl-XL decreased in a time-dependent manner. The Bax and c-myc protein markedly increased, but the Bid protein level did not change. IL-21 up regulated c-myc and Bax gene expression, however down regulated Bcl-2 and BCL-XL gene expression, but the gene expression of Bid and Survivin hadn't been changed significantly.

CONCLUSIONSIL-21 can inhibit proliferation and induce apoptosis of SUDHL-4 cell. The mechanism may involve in endogenous mitochondrial pathway mediated by the c-myc and the Bcl-2 genes.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Humans ; Interleukins ; administration & dosage ; pharmacology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-X Protein ; metabolism

This study was aimed to investigate the impact of specific siRNA on survivin gene in transfected lymphoma cell line and provide experimental evidences for future treatment of mantle cell lymphoma. The small interfering RNA (siRNA) targeted survivin mRNA was synthesized in vitro and was transfected into Jeko-1 that showed high survivin expression in mRNA level. The levels of survivin mRNA and protein expression were detected by quantitative reverse transcription polymerase chain reaction (RT-PCR) and Western blot respectively. The apoptosis effect was examined by calculating the ratio of Annexin V-FITC/PI positive cells using flow cytometry. The inhibition of cell proliferation was assayed with CCK-8 reagent after transfection. The results showed that expression of survivin mRNA was markedly suppressed by the siRNA. The relative expression levels were 0.49 ± 0.03, 0.38 ± 0.02 and 0.17 ± 0.02 at time points of 24, 48 and 72 h respectively, compared with the control group; the inhibitive rates of cell proliferation were (31.2 ± 2.1)%, (43.3 ± 3.4)% and (52.6 ± 2.5)%; the apoptotic rates of cells were (6.3 ± 0.5)%, (13.5 ± 1.1)% and (23.6 ± 1.6)% respectively; survivin protein expression levels were gradually reduced. It is concluded that the siRNA targeting survivin down-regulates the expressions of survivin mRNA and protein evidently. The siRNA of survivin displays the potent ability to inhibit the proliferation of lymphoma cell line Jeko-1; survivin may become a potential molecular target for the therapy of lymphoma in the future.
Cell Line, Tumor ; Cell Proliferation ; Gene Silencing ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Transfection

Objective To investigate the effects of high spermatic vessel dissection on testicular morphological alteration of SD rats in prepuberty,puberty and sexual maturity phases.Methods Thirty-day-old SD rats were divided into 2 groups underwent sham operation and left high spermatic vessel dissection as a simulation of Palomo′s maneuver.Detailed morphological investigations were made at 3 different postoperative intervals among the 3rd day,30th day and 56th day.Results High spermatic vessel dissection in prepubertal rats induced acute testicular ischemia in the operated testes on the 3rd day.Most of the operated testes on the 30th day showed testicular atrophy.And all the operated testes showed testicular atrophy and sperm disappearance in epididymis on the 56th day.Conclusion High dissection of spermatic vessel in prepubertal rats induced testicular ischemia in prepuberty and testicular growth failure in puberty,testicular atrophy completely and sperm production losing in sexual maturity phase.

OBJECTIVETo investigate the association between non alcoholic fatty liver disease (NAFLD) and metabolic syndrome (MS).

METHODSA cross-sectional multiple-stage stratified survey was performed. A total of 2190 civil servants of Chongqing city were invited to participate in the survey covering physical examination, serum biochemistry-profile and ultrasonographic examination of liver.

RESULTSOf 2176 valid questionnaires, altogether 455 cases were diagnosed as NAFLD and 231 individuals were diagnosed as MS. The prevalence of obesity, hyperglycemia, blood lipid disturbance, primary hypertension, NAFLD and MS was 38.3%, 5.5%, 31.7%, 29.9%, 20.9% and 10.6% respectively, which was increased along with aging (chi2 = 31.775, P = 0.000; chi2 = 25.985, P = 0.000; chi2 = 44.818, P = 0.000; chi2 =149.802, P = 0.000; chi2 = 61.302, P = 0.000; chi2 = 43.508, P = 0.000 a partly). The prevalence of obesity, hyperglycemia, dyslipidemia, primary hypertension, metabolic syndrome was significantly higher than those in control group (chi2 = 384.554, P = 0.000; chi2 = 25.597, P = 0.000; chi2 = 370.849, P = 0.000; chi2 = 40.252, P = 0.000; chi2 = 215.077, P = 0.000 separately), and the level of body mass index (BMI), fasting plasma glucose (FPG), triglyceride (TG), systolic blood pressure (SBP), diastolic blood pressure (DBP) in NAFLD group was remarkably higher than those in control group (t = 26.308, P = 0.000; t = 6.055, P = 0.000; t = 15.980, P = 0.000; t = 10.550, P = 0.000; t = 13.628, P = 0.000 respectively), while the level of high-density lipoprotein cholesterol (HDL-C) was on the opposite (t = 20.067, P = 0.000). Compared with the control group, odds risk for NAFLD was 22.82 folds (95% CI: 12.64-41.19) in obesity, 20.97 folds (95% CI: 11.21-39.24) in hyperglycemia, 24.40 folds (95% CI: 13.51-44.07) in dyslipidemia, 15.73 folds (95% CI: 8.66-28.60) in primary hypertension, while the risk for NAFLD was the highest in MS (OR = 31.06, 95% CI: 17.12-56.35). There were simple or multiple metabolic disorders in 455 individuals diagnosed as NAFLD, and 21 case (4.6%) with obesity, hyperglycemia, dyslipidemia and primary hypertension.

CONCLUSIONNAFLD is closely related with MS, which may be considered as a feature of MS.

Adult ; China ; epidemiology ; Fatty Liver ; epidemiology ; Female ; Humans ; Male ; Metabolic Syndrome ; epidemiology ; Middle Aged ; Prevalence

AIMTo investigate the impact of 8-(N,N-diethylamino)-n-octyl-3,4,5-trimethoxybenzoate (TMB-8) on the change of cerebral blood flow(CBF) induced by 5-HT or KCl in rats.

METHODSThe CBF in rats was measured by a Laser-Doppler flowmeter. The cranial window field was superfused with artificial cerebral spinal fluid containing TMB-8, 5-HT or KCl.

RESULTS12.5, 25 and 50 mumol.L-1 TMB-8 showed no significant effect on rest CBF in rats. 12.5, 25 and 50 mumol.L-1 TMB-8 apparently inhibited the decline of CBF evoked by 1 or 2 mumol.L-1 5-HT. When persistent reduction of CBF were evoked by 1 mumol.L-1 5-HT, TMB-8 markedly increased the CBF in a concentration-dependent manner. The reduction of CBF induced by 20 or 40 mmol.L-1 KCl was also suppressed by 12.5, 25 and 50 mumol.L-1 TMB-8. While persistent reduction of CBF was evoked by 20 mmol.L-1 KCl. TMB-8 markedly increased the CBF in a concentration-dependent manner.

CONCLUSIONThese results indicate that TMB-8 is effective in preventing and treating the reduction of CBF induced by 5-HT or KCl, and improved the supply of blood in rat brain during ischemia.

Animals ; Brain ; blood supply ; drug effects ; Calcium Channel Blockers ; pharmacology ; Drug Interactions ; Gallic Acid ; analogs & derivatives ; pharmacology ; Potassium Chloride ; antagonists & inhibitors ; Rats ; Rats, Sprague-Dawley ; Regional Blood Flow ; drug effects ; Serotonin Antagonists ; pharmacology

The sigma-1 receptor is a molecular chaperone protein highly enriched in the brain. Recent studies linked it to many diseases, such as drug addition, Alzheimer's disease, stroke, depression, and even cancer. Sigma-1 receptor is enriched in lipid rafts, which are membrane microdomains essential in signaling processes. One of those signaling processes is ADAM17- and ADAM10-dependent ectodomain shedding. By using an alkaline phosphatase tagged substrate reporter system, we have shown that ADAM10-dependent BTC shedding was very sensitive to both membrane lipid component change and sigma-1 receptor agonist DHEAS treatment while ADAM17-dependent HB-EGF shedding was not; and overexpression of sigma-1 receptor diminished ADAM17- and ADAM10-dependent shedding. Our results indicate that sigma-1 receptor plays an important role in modifying the function of transmembrane proteases.
ADAM Proteins ; metabolism ; ADAM10 Protein ; ADAM17 Protein ; Amyloid Precursor Protein Secretases ; metabolism ; Animals ; Betacellulin ; COS Cells ; Cercopithecus aethiops ; Gene Expression ; HEK293 Cells ; Heparin-binding EGF-like Growth Factor ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Membrane Microdomains ; metabolism ; Membrane Proteins ; metabolism ; Receptors, sigma ; agonists ; metabolism

OBJECTIVETo investigate angiogenesis of collagen-chitosan porous scaffold, and to study survive of skin grafts on the scaffold after bilayer dermal equivalent (BDE) was transplanted on wounds with full thickness skin defects.

METHODSThe full thickness skin defects were made on 10 Bama miniature pigs and the BDE composed of collagen-chitosan porous scaffold and silicone membrane was transplanted on wound. Angiogenesis in dermal equivalent, wound healing, and healing and survive of skin grafts on dermal equivalent were observed in 1, 2, and 3 weeks after the BDE transplantation. At the same time, CD34 positive signals (neo-forming micro-vessels) were detected by immunohistochemical staining.

RESULTSInflammatory cells and fibroblasts infiltrated into dermal equivalent and a few new micro-vessels had been formed in 1 week after the BDE transplantation; neo-forming micro-vessels perpendicular to wound bed had increased significantly in 2 weeks after the BDE transplantation; neo-forming micro-vessels could be observed in almost all dermal equivalents in 3 weeks after the BDE transplantation. CD34 positive signals (neo-forming micro-vessels) in 3 weeks after the BDE transplantation was much more than those in 2 weeks after the BDE transplantation, and CD34 positive signals in 2 weeks after the BDE transplantation was much more than those in 1 week after the BDE transplantation. Survival rate of intermediate split thickness skin graft were 10%, 70% and 100% respectively after the skin grafts had been grafted for 2 weeks on surface of the scaffold which had been transplanted for 1, 2 and 3 weeks. Epidermis which had been grafted on surface of the scaffold for 1 or 2 weeks could perfectly survive after BDE had been transplanted for 1 or 2 weeks.

CONCLUSIONSCollagen-chitosan porous scaffold plays a very important role in wound healing of full thickness skin defect and can induce fibroblast infiltration and new micro-vessel formation. Epidermis grafted on surface of collagen-chitosan porous scaffold can perfectly repair wounds, and it has brilliant applied prospects in repairing skin defect.

Animals ; Chitosan ; Collagen ; Disease Models, Animal ; Female ; Graft Survival ; Neovascularization, Physiologic ; Silicones ; Skin ; injuries ; Skin Transplantation ; Swine ; Swine, Miniature ; Tissue Scaffolds ; Wound Healing

The aim was to study the mechanisms of HL-60 cell apoptosis induced by nimodipine (NMDP) and cytarabine (Ara-C). The DNA fragment was detected by agarose gel electrophoresis. The expressions of bcl-2 and bax gene proteins related with apoptosis were investigated by immunohistochemistry. The results showed that HL-60 cell apoptosis rate had been increasing in the experimental groups compared with the control group since culturing 8 hours. The expression of Bcl-2 protein was lower and the expression of Bax protein was higher in the experimental groups than that in the control group, while ratio of bcl-2/bax was lower in the experimental groups than that in the control group. It is concluded that NMDP and Ara-C induce apoptosis of HL-60 cells, and the mechanism of apoptosis induced by them may down-regulate the expression of bcl-2 gene and up-regulate the expression of bax gene. The mechanism of HL-60 cell apoptosis induced by Ara-C and NMDP is probably associated with the down-regulation of Bcl-2 protein expression.
Antimetabolites, Antineoplastic ; pharmacology ; Apoptosis ; drug effects ; Cytarabine ; pharmacology ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Gene Expression Regulation, Neoplastic ; HL-60 Cells ; Humans ; Nimodipine ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; bcl-X Protein ; biosynthesis ; genetics

OBJECTIVETo investigate the mechanism of apoptosis of EC9706 tumor-bearing nude mice induced by all-trans retinoic acid (ATRA).

METHODSHuman esophageal carcinoma cell line EC9706 cells were inoculated into nude mice to establish the solid tumor model. The tumor models were divided into the following groups: ATRA group, fluorouracil group, the two-drugs combination group, and with an equal volume fraction of solvent as the control group. The nude mice were sacrificed after 10 days of medication. TUNEL staining was used to detect cell apoptosis. RT-PCR was used to detect the expression level of mRNA and immunohistochemistry was used to detect the expression level of protein of caspase-3 and survivin, the apoptosis-related genes in the tumor tissue.

RESULTSThe apoptosis rates of the ATRA group, 5-Fu group and ATRA + 5-Fu group were 44.3%, 39.7% and 91.0%, respectively. There was a significant difference in comparison with the control group (0.7%), and the ATRA group had no significant difference compared with that of the fluorouracil group (P > 0.05), but the apoptosis rate of the two-drugs combination group was significantly higher than that in the two single-drug groups (P < 0.05). The average gray value of caspase-3 protein expressed in the control group was 46.12 ± 0.33 and the relative expression of caspase-3 mRNA was 0.14 ± 0.03, both were significantly lower than that in the ATRA group, 5-Fu group and the two-drugs combination group (P < 0.05). The average gray value of survivin protein expressed in the control group was 96.07 ± 0.13 and the relative expression of survivin mRNA was 0.84 ± 0.04, both were significantly higher than those of other groups (P < 0.05). The ATRA group had no significant difference compared with the fluorouracil group (P > 0.05), but the two-drugs combination group was significantly different compared with the single-drug groups (P < 0.05).

CONCLUSIONApoptosis in the EC9706 tumor cells in nude mice can be induced by ATRA. The mechanism may be related with down-regulation of the level of survivin gene expression and up-regulation of the level of caspase-3 gene expression.

Animals ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; genetics ; metabolism ; Cell Line, Tumor ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Fluorouracil ; pharmacology ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Nude ; Neoplasm Transplantation ; RNA, Messenger ; metabolism ; Tretinoin ; pharmacology

OBJECTIVETo investigate biosynthetic and apoptotic mechanisms in repair of full thickness skin defect with collagen-chitosan porous scaffold transplantation, and to determinate differences between wound repair with the scaffold transplantation and scar healing without the scaffold transplantation.

METHODSThe full thickness skin defects were made on 10 Bama miniature pigs and the bilayer dermal equivalent (BDE) composed of collagen-chitosan porous scaffold and silicone membrane was transplanted on wounds. Surfaces of wounds were observed at 1, 2, and 3 weeks after the BDE transplantation, and so were done the wound repairs after epidermis had been grafted for 2 weeks on surface of the scaffold which had been transplanted on skin defect wounds for 2 weeks. At the same time, TGF-beta1 expressions, apoptosis and self collagen replacement of scaffolds in wounds were detected in situ by immunohistochemical staining, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL) and picrosirius red polarized light. Wounds without scaffold transplantation were studied as control.

RESULTS1) Wounds with the scaffold transplantation were different from granulation tissue. 2) The peak of TGF-beta1 expression in the scaffold wounds was from 1 to 2 weeks after BDE transplantation, and TGF-beta1 expressions decreased continuously from 3 to 4 weeks. TGF-beta1 expressions increased continuously in the control wounds from 1 to 3 weeks and decreased on 4 weeks. TGF-beta1 expressions in the scaffold wounds on 1st and 2nd week were significantly higher than those in the corresponding control wounds, whereas, TGF-beta1 expressions in the scaffold wounds on 3rd and 4th week were significantly lower than those in the corresponding control wounds. 3) Apoptosis increased continuously in the scaffold wounds from 2 to 4 weeks after BDE transplantation, and so did in the control wounds from 3 to 4 weeks. However, apoptosis signals in the scaffold wounds on 2nd, 3rd, and 4th week after BDE transplantation were significantly more than those in the corresponding control wounds, and there was no difference between apoptosis signals in the scaffold wounds on 1st week after BDE transplantation and those in the corresponding control wounds. 4) Observation by picrosirius red polarized light method: self collagen began to synthesize in the scaffold wounds on 1st week after BDE transplantation, and scaffolds had been replaced by self collagen from 2 to 3 weeks after BDE transplantation.

CONCLUSIONSCollagen-chitosan porous scaffold plays a very important role in wound healing of full thickness skin defect. The mechanisms of wound repair by dermal scaffold are different from those by granulation and scar healing. It has a good future in repairing skin defect.

Animals ; Apoptosis ; Chitosan ; metabolism ; Collagen ; biosynthesis ; metabolism ; Dermis ; Extracellular Matrix ; Female ; Skin Irritancy Tests ; Skin, Artificial ; Stents ; Swine ; Swine, Miniature ; Tissue Engineering ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing