Objective To explore whether pravastatin protects endothelium against the damage induced by homocysteine (Hcy) in isolated rabbit aorta. Methods Endothelium-dependent relaxation responses to acetylcholine (ACh) of thoracic aortic rings were measured by isometric tension recording before and after aortic rings exposed to Hcy in the absence or presence of pravastatin (PT) to estimate the injury effect of Hcy and the protective effect of pravatatin on rabbit aortic endothelium, respectively. Results Incubation of aortic rings with 1～10mmol/L Hcy for 30min significantly inhibited endothelium-dependent relaxation response to ACh of aortic rings in a concentration-dependent manner. Pre-incubation of aortic rings with 0.3～3mmol/L PT for 15min and co-incubation of aortic rings with 3mmol/L Hcy for another 30min markedly attenuated the inhibition of endothelium-dependent relaxation induced by Hcy in a concentration-dependent manner. Conclusion Pravastatin can improve the impairment of endothelium-dependent relaxation induced by Hcy in isolated rabbit aorta.

Animals ; Helminthiasis ; pathology ; Humans ; Moniliformis ; pathogenicity

The value of snake venom polypeptides in clinical application has drawn extensive attention,and the development of snake polypeptides into new drugs with anti-tumor,anti-inflammatory,antithrombotic,analgesic or antihypertensive properties has become the recent research hotspot.With the rapid development of molecular biology and biotechnology,the mechanisms of snake venom polypeptides are also gradually clarified.Numerous studies have demonstrated that snake venom polypeptides exert their pharmacological effects by regulating ion channels,cell proliferation,apoptosis,intracellular signaling pathway,and expression of cytokine as well as binding to relevant active sites or receptors.

OBJECTIVETo typing of tibial plateau fracture based on spiral CT reconstruction and to explore effect of the typing method for treatment.

METHODSA hundred and twenty-six cases with tibial plateau fracture (male 95, female 31, age from 23 to 58 years old), the fractures were classified based on reconstruction image of spiral CT. Including central compression type in 13 cases, split type in 8, split compression type in 79, comminution type in 26. According to the different typing the suitable incision of operation and fixed method for fracture were select.

RESULTSA hundred and twenty-six cases were followed up for 0.5-4 years with an average of 1.2 years. According to Hohl system score to knee joint function, there were statistical significance in the pain,active movement,active range of motion between before and after operation (P < 0.01) and there were no statistical significance in stability and self-evaluation (P < 0.01).

CONCLUSIONTyping of tibial plateau fracture based on spiral CT reconstruction helpful to choose operative approach, reduction and fixed method and obviously improve clinical effect.

Adult ; Female ; Follow-Up Studies ; Humans ; Image Processing, Computer-Assisted ; methods ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Tibial Fractures ; diagnostic imaging ; surgery ; therapy ; Tomography, Spiral Computed ; Treatment Outcome

Reduced radiosensitivity of lung cancer cells represents a pivotal obstacle in clinical oncology. The hypoxia-inducible factor (HIF)-1α plays a crucial role in radiosensitivity, but the detailed mechanisms remain elusive. A relationship has been suggested to exist between hypoxia and autophagy recently. In the current study, we studied the effect of hypoxia-induced autophagy on radioresistance in lung cancer cell lines. A549 and H1299 cells were cultured under normoxia or hypoxia, followed by irradiation at dosage ranging from 0 to 8 Gy. Clonogenic assay was performed to calculate surviving fraction. EGFP-LC3 plasmid was stably transfected into cells to monitor autophagic processes. Western blotting was used to evaluate the protein expression levels of HIF-1α, c-Jun, phosphorylated c-Jun, Beclin 1, LC3 and p62. The mRNA levels of Beclin 1 were detected by qRT-PCR. We found that under hypoxia, both A549 and H1299 cells were radio-resistant compared with normoxia. Hypoxia-induced elevated HIF-1α protein expression preferentially triggered autophagy, accompanied by LC3 induction, EGFP-LC3 puncta and p62 degradation. In the meantime, HIF-1α increased downstream c-Jun phosphorylation, which in turn upregulated Beclin 1 mRNA and protein expression. The upregulation of Beclin 1 expression, instead of HIF-1α, could be blocked by SP600125 (a specific inhibitor of c-Jun NH2-terminal kinase), followed by suppression of autophagy. Under hypoxia, combined treatment of irradiation and chloroquine (a potent autophagy inhibitor) significantly decreased the survival potential of lung cancer cells in vitro and in vivo. In conclusion, hypoxia-induced autophagy through evaluating Beclin1 expression may be considered as a target to reverse the radioresistance in cancer cells.

This study was aimed to investigate the effects of peripheral blood Th17 cells, IL-17 and IL-21 in the occurrence and development of acute leukemia. 60 patients with acute leukemia (19 patients with ALL, 41 patients with AML) were divided into non-remission group (group A, n=24), remission group (group B, n=36); 25 healthy volunteers were used as control group (group C). In addition to this, these 60 patients were divided into infection group (n=32) and non-infection group (n=28) on the basis of infection status. The concentration of IL-17 and IL-21 in the peripheral blood mononuclear cell culture supernatant after stimulation with anti-CD3 and anti-CD28 mAb were determined with ELISA. The expression of CD4+ IL-17+ cells was determined by flow cytometry. The results showed that (1) the concentrations of IL-17 and IL-21, and proportion of Th17 cells in group A and group B were much lower than those in group C (p<0.05); (2) the expression levels of IL-17 and IL-21, and the proportion of Th17 cells in group A were lower than those in group B (p<0.05); (3) the expression levels of Th17 and IL-17 in infection group were lower than those in non-infection group (p<0.05). It is concluded that Th17 cells may play important roles in the occurrence and development of acute leukemia through secreting IL-17 and IL-21, and their functional level can partially reflect the status of leukemia and can be used to evaluate the risks of infection in patients with leukemia.
Adolescent ; Adult ; CD4-Positive T-Lymphocytes ; Case-Control Studies ; Female ; Humans ; Interleukin-17 ; metabolism ; Interleukins ; metabolism ; Leukemia, Myeloid, Acute ; metabolism ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; metabolism ; Th17 Cells ; secretion ; Young Adult

OBJECTIVETo investigate the expression of HER2/neu, Ki-67 and TK1 protein in meningiomas in correlation with tumor grades and recurrence.

METHODSTwenty cases of each of the following types of meningiomas were selected for the study, namely: the benign non-recurrent, recurrent benign, atypical and malignant, according to the World Health Organization (WHO) histological classification of nervous system, 2007. Immunohistochemistry study for HER2/neu, Ki-67 and TK1 protein was performed. HER2/neu gene amplification was detected using FISH. Cases with HER2 protein overexpression were studied by immunohistochemistry staining. The results of the biomarker assays were also used to study the correlationship with the tumor grades and tumor recurrency.

RESULTSImmunohistochemistry showed that the positive rates of HER2 expression in non-recurrence benign group, recurrence benign group, atypical group and malignant group were 3 cases (15%), 6 cases (30%), 7 cases (35%), and 10 cases (50%), respectively (P < 0.05). A higher tumor grade was correlated with a higher rate of HER2/neu expression. The Ki-67 and TK1 labeling index (LI) in non-recurrence group were lower than those in the atypical or malignant group (P < 0.05), whereas the atypical group had lower LI than that of the malignant group (P < 0.05). Higher levels of LI of Ki-67 and TK1 were correlated with higher tumor grades and recurrence of the benign meningiomas (P < 0.05). Expression of HER2 was positively correlated with Ki-67 and TK1 (r = 0.445, P < 0.01; r = 0.501, P < 0.01, respectively), and there was a positive correlation between Ki-67 and TK1 (r = 0.450, P < 0.01) as well. HER2/neu gene copy amplification in 7 of 26 cases (26.9%) of HER2 immunopositive meningiomas. The rates of HER2/neu gene amplification were 0 in tumors with 1+ immunopositivity, 4/6 in tumor with 2+ immunopositivity and 3/4 in tumor with 3+ immunopositivity. HER2/neu gene amplification in 3+ and 2+ immunopositive cases had no statistical significance (P > 0.05). Aneuploidy of chromosome 17 existed in 9 of 26 of HER2 immunopositive meningiomas (34.6%). However, the rates of chromosome 17 aneuploidy had no significant difference among tumors with variable HER2/neu imumopositivity (P > 0.05).

CONCLUSIONSHigh levels of HER2 and Ki-67 or TK1 expression correlate with the increase of tumor grades and tumor recurrence. HER2/neu gene amplification is seen in a subset of meningiomas with the protein expression (26.9%). A combination of biomarker study including HER2/neu, Ki-67 and TK1 may be useful in predicting the biological behavior of meningiomas.

Aneuploidy ; Chromosomes, Human, Pair 17 ; Female ; Gene Amplification ; Humans ; Immunohistochemistry ; In Situ Hybridization, Fluorescence ; Ki-67 Antigen ; metabolism ; Male ; Meningeal Neoplasms ; genetics ; metabolism ; pathology ; Meningioma ; genetics ; metabolism ; pathology ; Middle Aged ; Neoplasm Recurrence, Local ; Receptor, ErbB-2 ; genetics ; metabolism ; Thymidine Kinase ; metabolism

OBJECTIVETo observe the effect of 1,25(OH)2D3 on thyroid inflammation and Th1/Th2 cells in rats with experimental autoimmune thyroiditis (EAT).

METHODSForty-eight female Wistar rats were randomly divided into 4 groups, namely the prevention group treated with 1, 25-(OH)2D3 from 0 to the 6th week (n=10), treatment group with 1,25(OH)2D3 treatment from the 2nd to the 8th week (n=10) after immune sensitization, positive control group (n=12) and the negative control group (n=16). All the rats were challenged with porcine thyroglobulin for immune sensitization until the 6th or 8th week except for those in the negative control group. In the prevention group and treatment group, the rats received 1,25(OH)2D3 at 5 microg/kg by intraperitoneal injection every other day, while those in the positive and negative control groups were given peanut oil instead. The thyroid pathologies, serum autoantibody level and cytokine levels were examined after the treatments.

RESULTSThe thyroid gland remained structurally intact in the negative control group. In the positive control group, the thyroid showed obvious inflammatory change with structural disruption and even disappearance of the thyroid follicle. The structure of the thyroid gland follicles was intact in the prevention group and treatment group. No significant differences were found in the autoantibody and cytokine levels between the prevention group and negative control group (P>0.05). Compared with the positive control groups, the autoantibody and IFN-gamma and IL-12 levels decreased significantly in the treatment group, but the levels of IL-4 and IL-10 were markedly increased (P<0.05).

CONCLUSION1,25(OH)2D3 given before the establishment of the EAT model helps maintain structural integrity of the thyroid gland and normal levels of the antibodies and cytokines in rats. 1,25(OH)2D3 can ameliorate the pathological changes of the thyroid gland and correct the cytokine disequilibrium in rats with EAT.

Animals ; Autoantibodies ; blood ; Female ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Rats ; Rats, Wistar ; Th1 Cells ; cytology ; Th2 Cells ; cytology ; Thyroiditis, Autoimmune ; drug therapy ; metabolism ; Vitamin D ; analogs & derivatives ; therapeutic use

OBJECTIVETo investigate the expressions of myogenic markers MyoD, myogenin,and desmin in skeletal muscle differentiation of human bone marrow mesenchymal stem cells (hBM-MSCs).

METHODSMyogenic markers MyoD, myogenin,and desmin of hBM-MSCs cultured in vitro were detected by immunofluorescence and RT-PCR. A total of 21 8-to-10 week-old immunosuppressed mdx mice were transplanted with 1x107 passage 5 of hBM-MSCs. The mice were euthanized 2-24 weeks after transplantation,and gastrocnemius muscle were analyzed for human MyoD, myogenin,desmin,and dystrophin (Dys) expressions by immunohistochemistry and RT-PCR.

RESULTSThe numbers of MyoD-,myogenin-,and desmin-positive cells per 100 hBM-MSCs were 23.5∓5.3, 30.7∓6.2, and 28.4∓5.7, respectively. MyoD, myogenin, and desmin mRNA was observed in passage 5 of hBM-MSCs. After two weeks of hBM-MSCs transplantation,a small number of MyoD-and myogenin-positive cells were observed in skeletal muscle of mdx mice,and desmin-positive cells were observed 4 weeks after transplantation. Expressions of MyoD and myogenin were detected in the muscle of mdx mice 2-4 weeks after hBM-MSCs transplantation, which reached a peak 12-16 weeks later. Desmin was expressed in the muscle of mdx mice 4-8 weeks after transplantation,with much more expression after 16 weeks of transplantation. A small number of Dys-positive cell and Dys mRNA expression were presented in the muscle of mdx mice 4 and 8 weeks after hBM-MSCs transplantation,respectively. The expression of Dys in the muscle of mdx mice increased gradually after transplantation.

CONCLUSIONhBM-MSCs have the potential of myogenic differentiation in vitro and contribute to myogenic conversion in xenogeneic animal,during which the up-regulation of MyoD and myogenin expressions may play an important role.

Animals ; Biomarkers ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Desmin ; metabolism ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Mice ; Mice, Inbred mdx ; Muscle, Skeletal ; cytology ; metabolism ; MyoD Protein ; metabolism ; Myogenin ; metabolism ; Up-Regulation

A sensitive, rapid and specific liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method for quantification of gabapentin in human plasma has been developed. After a single plasma protein precipitation with methanol, gabapentin and metformin (internal standard) were chromatographed on a Inertsil ODS-3 column (50 mm x 2.1 mm ID, 3 microm) with mobile phase consisting of methanol-0.2% formic acid aqueous solution (80:20, v/v) at a flow-rate of 0.2 mL x min(-1). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 172 --> m/z 154 and m/z 130 --> m/z 71 were used to quantify gabapentin and metformin, respectively. The run time was 2.2 min. The linear calibration curve was obtained in the concentration range of 40.8-8.16x10(3) ng x mL(-1). The lower limit of quantification was 40.8 ng x mL(-1). The intra- and inter-day precision (RSD) was less than 12%, and the accuracy (RE) was within +/-6.4% calculated from quality control (QC) samples. The method was used to determine the concentration of gabapentin in human plasma after a single oral administration of 600 mg gabapentin capsule to 20 healthy male Chinese volunteers. The method was proved to be selective, sensitive, rapid and suitable for pharmacokinetic study of gabapentin in human plasma.
Administration, Oral ; Amines ; administration & dosage ; blood ; pharmacokinetics ; Anticonvulsants ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Chromatography, High Pressure Liquid ; Cyclohexanecarboxylic Acids ; administration & dosage ; blood ; pharmacokinetics ; Humans ; Male ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; administration & dosage ; blood ; pharmacokinetics