OBJECTIVE: To analyze age-related mandible stress distribution due to midline force. METHODS: Mandibles of children, adults, and elderly individuals were scanned by spiral CT to establish three-dimension imaging models with mesh elements by MIMICS software and HYPERMESH software. The mandible stress distribution was analyzed using ANSIS software. RESULTS: There was no significant difference in mandible stress distribution in various age groups with the greatest stress distribution (Von Mises) present at the mandible angle. Although there was stress present at the mandible neck in adults, no such mandible neck stress was found in children and elderly individuals. CONCLUSION Mandible stress distribution is closely related to the incidence of fracture in various age groups, i.e., more stress and more fracture.
Adolescent ; Adult ; Age Factors ; Biomechanical Phenomena ; Child ; Finite Element Analysis ; Humans ; Image Processing, Computer-Assisted ; Incidence ; Mandible/physiology* ; Mandibular Fractures/etiology* ; Middle Aged ; Models, Anatomic ; Stress, Mechanical ; Tomography, X-Ray Computed/methods* ; Young Adult

This study was purposed to detect the expression level of human endothelial cell tissue factor (hECTF) and concentration of IL-2, and to investigate the alterations of hECTF and IL-2 after using immunosuppressive agent (cyclosporin, CsA) and explore the significance of endothelial cell (EC) lesion and abnormal expression of tissue factor (TF) in GVHD. Human endothelial cells and allogeneic lymphocytes were mixed and cultured as well as were cocultured with CsA for 4-6 hours in vitro, then the expression level of hECTF was detected by flow cytometry and RT-PCR, the concentration of IL-2 in supernatant was assayed by ELISA. The experiment was divided into 4 groups: 1st group--non-mixed-cultured group (negative control group), 2nd group - mixed-cultured group (positive control group), 3rd group - mixed-cocultured group with 1 µg/ml CsA and 4th group--mixed-cocultured group with 2 µg/ml CsA. The results showed that as compared with non-mixed-cultured group (negative control group), the expression level of hECTF and concentration of IL-2 in another 3 groups significantly increased (p<0.01), while as compared with positive control group, the expression level of hECTF and concentration of IL-2 in cocultured groups with CsA both decreased (p<0.01). It is concluded that the lesion of EC and abnormal expression of TF play a crucial role in GVHD, among which the high expression of TF after being stimulated by donor's lymphocytes may be the key step for occurrence and progression of GVHD.
Cells, Cultured ; Coculture Techniques ; Cyclosporine ; pharmacology ; Endothelial Cells ; metabolism ; Graft vs Host Disease ; metabolism ; pathology ; Humans ; Interleukin-2 ; metabolism ; Lymphocytes ; cytology ; Thromboplastin ; metabolism

OBJECTIVETo study the changes in the levels of autophagy-related proteins, Atg1, Atg5, and Beclin1, in organophosphate-induced delayed neuropathy (OPIDN) caused by tri-ortho-cresyl phosphate (TOCP), and to investigate the molecular pathogenic mechanism of OPIDN.

METHODSThirty adult Roman hens were randomly and equally divided into control group and 1, 5, 10, and 21 d intoxication groups. Each hen in the intoxication group was administered TOCP by gavage at a single dose of 750 mg/kg, while each hen in the control group was administered the same volume of corn oil. The hens were killed at the corresponding time points, and their tibial nerves and spinal cords were collected. The levels of Atg1, Atg5, and Beclin1 in the tibial nerves and spinal cords were measured by immunoblotting.

RESULTSCompared with those in the control group, the levels of Atg1 in tibial nerves decreased by 29.8%, 64.4%, 43.5%, and 19.8% at 1, 5, 10, and 21 d, respectively, after intoxication ((P < 0.05); the levels of Atg5 in tibial nerves decreased by 36.8%, 49.6%, 51.2%, and 31.5% at 1, 5, 10, and 21 d, respectively, after intoxication (P < 0.05); the levels of Beclin1 in tibial nerves decreased by 68.5%, 66.3%, and 32.2% at 1, 5, and 10 d, respectively, after intoxication (P < 0.05). Compared with those in the control group, the levels of Atg1 in spinal cords decreased by 23.5%, 48.7%, and 20% at 1, 5, and 10 d, respectively, after intoxication (P < 0.05); the levels of Atg5 in spinal cords decreased by 32.7%, 51.5%, 47.3%, and 39.6% at 1, 5, 10, and 21 d, respectively, after intoxication (P < 0.05); the levels of Beclin1 in spinal cords decreased by 28.9%, 50.2%, 43.2%, and 28.3% at 1, 5, 10, and 21 d, respectively, after intoxication (P < 0.05).

CONCLUSIONThe intoxication of TOCP is associated with the significant changes in the levels of autophagy-related proteins in the nervous tissues of hens, which might be involved in the pathogenesis of OPIDN.

Animals ; Apoptosis Regulatory Proteins ; metabolism ; Autophagy ; drug effects ; Chickens ; Female ; Intracellular Signaling Peptides and Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Nervous System Diseases ; chemically induced ; metabolism ; Neurofilament Proteins ; metabolism ; Spinal Cord ; metabolism ; Tibial Nerve ; metabolism ; Tritolyl Phosphates ; toxicity

This study was aimed to observe the effects of siRNA on Livin expression and function in K562 cells. Livin siRNA were designed and synthesized, then were transfected into K562 cells by using AMAXA nucle transfactor. Expressions of Livin mRNA and protein in transfected K562 cells was detected by RT-PCR and Western blot respectively. Non-transfected cells were used as control. The enhanced green fluorescent protein plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Cell apoptosis was measured by flow cytometry with Annexin V-FITC/PI double staining. The results showed that the transfection efficiency of electroporation method was about 50. The synthesized siRNA inhibited livin expression at both mRNA and protein levels. The rate of K562 cell apoptosis increased from (9.63 ± 0.89) in control group to (12.07 ± 1.39) and (27.41 ± 2.30) at 24 h and 48 h after transfection, respectively (P < 0.05). It is concluded that the siRNA can inhibit anti-apoptosis of livin gene via down-regulating livin gene expression, which may provide the new method for anti-leukemia study.
Adaptor Proteins, Signal Transducing ; genetics ; Apoptosis ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; Neoplasm Proteins ; genetics ; RNA, Small Interfering ; genetics

OBJECTIVETo investigate the effect of 2,5-hexanedione (HD) on degradation of low-molecular-weight neurofilaments (NF-L) in nervous tissue of rats, and to explore the molecular mechanism of n-hexane neuropathy.

METHODSFifty male Wistar rats were randomly divided into one-week poisoning group (n = 10), two-week poisoning group (n = 10), three-week poisoning group (n = 10), four-week poisoning group (n = 10), and control group (n = 10). In the four poisoning groups, a rat model of n-hexane neuropathy was established by intraperitoneal injection of HD (400 mg/kg/d). The change in the sciatic nerve ultrastructure of each rat was observed under an electron microscope. The progression of HD-induced peripheral neuropathy was evaluated using a gait scoring system. The degradation rates of NF-L in the sciatic nerve and spinal cord of each rat were measured by Western Blotting.

RESULTSThe rats showed decrease in muscle strength and abnormal gait after two weeks of HD poisoning and mild or moderate paralysis after four weeks of HD poisoning. The sciatic nerve showed degenerative change, according to electron microscope observation. Compared with the control group, the two-week poisoning group, three-week poisoning group, and four-week poisoning group had the NF-L degradation rates decreased by 25.8%, 70.4%, and 69.7%, respectively, in the supernatant fraction of sciatic nerve, and by 14.7%, 64.6%, and 67.3%, respectively, in the sediment fraction of sciatic nerve, all showing a significant difference (P < 0.01). Compared with the control group, the one-week poisoning group had the NF-L degradation rate decreased by 33.87% in the supernatant fraction of spinal cord, the four-week poisoning group had the NF-L degradation rate increased by 16.2% in the supernatant fraction of spinal cord, and the one-week poisoning group and two-week poisoning group had the NF-L degradation rates decreased by 46.3% and 13.0% in the sediment fraction of spinal cord, all showing a significant difference (P < 0.01).

CONCLUSIONHD poisoning significantly inhibits NF-L degradation in the sciatic nerve, which may be associated with NF degeneration and accumulation in the axons of patients with n-hexane neuropathy.

Animals ; Hexanes ; poisoning ; Hexanones ; pharmacology ; Male ; Nerve Tissue ; drug effects ; metabolism ; physiopathology ; Neurofilament Proteins ; drug effects ; metabolism ; Rats ; Rats, Wistar ; Sciatic Nerve ; drug effects ; metabolism ; physiopathology

Apoptosis ; drug effects ; Arachidonate 12-Lipoxygenase ; analysis ; physiology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Flavanones ; pharmacology ; Humans ; Lipoxygenase Inhibitors ; Multiple Myeloma ; drug therapy ; pathology

OBJECTIVETo investigate the effects of PLK1 gene silence by short hairpin RNA (shRNA) on PLK1 expression and apoptosis in K562 cells, and explore the role of PLK1 in the pathogenesis of leukemia.

METHODSThe shRNA fragment targeting at 1416-1436 bp of PLK1 mRNA was synthesized and cloned into pEGFP-H1 vector, named as pEGFP-H1/PLK1. The empty control, pEGFP-H1 and pEGFP-H1/PLK1 were transfected into K562 cells respectively via electroporation. 24 h or 48 h after transfection, gene and protein expression of PLK1 in the cells were assayed by RT-PCR and Western blot analysis respectively, cells viability by MTT assay, caspase-3 activity by colorimetry, cell cycle and apoptosis by FACS.

RESULTS24 and 48 h after transfection, PLK1 expression in K562 cells was 1.25 +/- 0.07 for control group, 0.52 +/- 0.04 and 0.25 +/- 0.02 for pEGFP-H1/PLK1 group, and 1.24 +/- 0.08 and 1.23 +/- 0.09 for pEGFP-H1 group respectively. The alteration status of PLK1 protein levels were similar to that of PLK mRNA levels. The apoptosis rate was (8.3 +/- 0.6)% in control group, (8.7 +/- 0.7)% in pEGFP-H1 group and (49.7 +/- 3.8)% and (82.3 +/- 6.9)% in pEGFP-H1/PKLK1 group at 24 and 48 h, respectively. In addition, cell fraction at G(2)/M phase was increased obviously compared with control and pEGFP-H1-transfected group.

CONCLUSIONThe constructed shRNA can remarkably inhibit PLK1 expression and transfected K562 cell proliferation, increase apoptosis and block cell-cycle, suggesting that PLK1 play important roles in apoptosis and cell-cycle control of leukemia cells.

Apoptosis ; genetics ; Cell Cycle ; Cell Cycle Proteins ; genetics ; metabolism ; Cell Proliferation ; Genetic Vectors ; Humans ; K562 Cells ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

Objective To explore the conception,mechanism,clinical manifestations,diagnosis,treatment and prognosis of ectopic pituitary adenoma. Methods The clinical data of 3 patients with ectopic pituitary adenoma, admitted to our hospital from October 2010 to March 2011, were retrospectively analyzed and discussed by reviewing the relevant literature. Results Clinical manifestations ofectopic pituitary adenoma were as follows:2 had headache,1 had sexual dysfunction and blurred vision, and 1 had acromegalia combined with psychiatric symptom. Endocrinological examination showed that 1 had obviously increased growth hormone (GH) and 1 increased prolactin (PRL).MRI scan indicated that 2 were located in sphenoid sinus and I was located in the sphenoid sinus and clivus.Contrast-enhanced MRI showed enhanced lesions.Total removal was achieved in 2 patients through the trans-sphenoidal approach; partial removal was achieved in the patient with sphenoid sinus and clivus.Pathology examination demonstrated as pituitary adenomas. Conclusion The patients with ectopic pituitary adenoma have neurological dysfunction or/and endocrinological dysfunction; CT and MRI play valuable role in their diagnosis; radical resection of tumor can achieve satisfactory clinical outcomes.

OBJECTIVETo investigate the inhibition role of anti-Fas hammerhead ribozyme on Fas expression and Fas-mediated apoptosis in mouse cytotoxic T lymphocyte (CTL) cell line--CTLL-2 cells, and explore a novel approach to enhance the ability of T cells against leukemia in donor lymphocytes infusion (DLI).

METHODSA hammerhead ribozyme targeting the Fas mRNA was synthesized and transfected into CTLL-2 cells by electroporation. Fas expression in CTLL-2 cells was detected by using RT-PCR, Western blot and flow cytometry, CTLL-2 cells viability was measured by MTT assay, caspase-3 proteolytic activity by caspase-3 detection kit, and cell apoptosis by flow cytometry. Killing activity of CTLL-2 was detected by LDH releasing assay in vitro.

RESULTSExpression of Fas mRNA and protein in CTLL-2 cells was reduced to 50% after transfection with anti-Fas ribozyme. Being treated with anti-Fas antibody (JO(2)), compared with control and mock-transfected cells, viability of CTLL-2 cells transfected with anti-Fas ribozyme increased by 1-fold, caspase-3 activity and apoptosis rate of ribozyme-transfected cells decreased to 50% and 37%, respectively, and cell killing activity was enhanced by 2-fold.

CONCLUSIONAnti-Fas ribozyme can cleave Fas efficiently and inhibit Fas-mediated apoptosis of CTLL-2 cells, resulting in improvement of their viability.

Animals ; Apoptosis ; Blotting, Western ; Caspase 3 ; metabolism ; Cell Line ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; Cytotoxicity, Immunologic ; immunology ; Flow Cytometry ; Genetic Vectors ; genetics ; Mice ; RNA, Catalytic ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; T-Lymphocytes, Cytotoxic ; cytology ; immunology ; metabolism ; Transfection ; fas Receptor ; genetics ; metabolism