Animals ; Apoptosis ; Bradykinin ; metabolism ; Bradykinin Receptor Antagonists ; Ischemia ; metabolism ; Ischemic Preconditioning ; Muscle, Skeletal ; blood supply ; metabolism ; pathology ; Myocardial Infarction ; metabolism ; Myocardium ; pathology ; Swine ; Swine, Miniature

OBJECTIVETo study the short- and long-term effects of multi-glycosides of tripterygium wilfordii (GTWon) the histological structures of testes in pubertal rats and possible mechanisms.

METHODSForty-eight 5-week-old male Sprague-Dawley rats were randomly intragastrically administered with low-does GTW(6 g/kg daily)and high-does GTW (12 mg/kg daily) or 1% sodium carboxymethylcellulose (6 mL/kg, control group) for four weeks. The testes were sampled for detecting histological structures and c-kit expression by immunohistochemistry 24 hrs and four weeks after drug discontinuance.

RESULTSThe number of spermatogenic cells and the expression of c-kit in testes were reduced in the two GTW treatment groups 24 hrs and 4 weeks after drug discontinuance compared with those in the control group(P<0.05). Four weeks after drug discontinuance atrophy and interstitial edema of seminiferous epitheliumin in testes were observed, and the testis weight and the expression of c-kit in testes were reduced in the high-does GTW group compared with those in the control group (P<0.01). There were no significant differences in the parameters observed between the low-dose GTW and the control group 4 weeks after drug discontinuance.

CONCLUSIONSGTW has adverse effects on testes in a dose-dependent manner in puberty rats. Low-dose GTW may cause reversible short-term injuries to testis tissues. The damage of the interstitial tissue of testes induced by high-dose GTW may be one of the causes of long-term injuries of testes.

Animals ; Dose-Response Relationship, Drug ; Glycosides ; pharmacology ; Male ; Organ Size ; drug effects ; Proto-Oncogene Proteins c-kit ; analysis ; Rats ; Rats, Sprague-Dawley ; Sperm Count ; Testis ; chemistry ; drug effects ; pathology ; Tripterygium ; chemistry

Objective:To study the efficacy and safety of natural calcined bone repair material(NCBM)in repairing bone defect af-ter tooth extraction.Methods:A randemized,double-blinded,parallel,positive control(Bio-Oss)and multi-center clinical trial was employed.Imaging examination was used as the main efficacy evaluation index,surgical wound healing,rejection reaction,bone me-tabolic changes,bone infection signs were the subordinate efficacy evaluation indexes,the incidence of adverse reactions was observed for safety evaluation.Results:280 cases were included,269 cases completed the trial.In NCBMand Bio-Oss group the effective rate of imaging examination was 93.08% and 93.70%(P >0.05)respectively.The wound healing time of the 2 groups was less than 7 days,no rejection reaction,bone metabolic change and bone infection sign were observed.The incidence of adverse events in NCBM and the Bio-Oss group was 0.72% and 2.14%(P >0.05)respectively.Conclusion:The efficacy and safety between natural cal-cined bone repair material is not inferior to Bio-Oss in repairing bone defect after tooth extraction.

Objective:To investigate effects of hydrogen sulfide (H2S) on inducible nitric oxide synthase (iNOS) in kidneys of Type 1 diabetic rats.Methods:Thirty-two male SD rats were randomly divided into four groups:A normal control (NC) group,a diabetes mellitus (DM) group,a NaHS (NaHS+DM) group,and a NaHS control (NaHS) group (n=8 per group).Type 1 diabetes was induced by a single intraperitoneal injection of streptozotocin (55 mg/kg).After successful establishment of models,the rats in NaHS+DM and NaHS groups were injected with NaHS solution (56 μmol/kg) intraperitoneally.Eight weeks later,the activities of total nitric oxide synthase (T-NOS) and iNOS,as well as the level of nitric oxide (NO) were detected in serum and renal tissues,respectively,The activity of glutathione peroxidase (GSH-Px) was determined in renal tissues,The ultrastructures of renal tissues were observed by transmission electron microscope,The protein expression ofiNOS in renal tissues was detected by Western blot.Results:Compared with the NC group,there was no significant difference in the various indexes in the NaHS group (P＞0.05).However,in the DM group,the activities of T-NOS and iNOS,and the level of NO were all increased significantly in serum and renal tissues,while the activity of GSH-Px was decreased in renal tissues.Under the electronic microscope,the thickening of the glomerular capillary basement membrane,the proliferation of mesangial matrix,and the foot fusion were observed.The protein expression ofiNOS was increased obviously in renal tissues in the DM group (P＜0.01).Compared with the DM group,the activities of T-NOS and iNOS and the level of NO were all decreased in serum and renal tissues,while the activity of GSH-Px was increased in renal tissues in the NaHS+DM group (P＜0.01).The renal ultrastructural damages were ameliorated obviously.The protein expression ofiNOS was decreased significantly (P＜0.01).Conclusion:H2S exerts a protective effect on kidney injury in type 1 diabetic rats.The mechanism might be related to inhibition of iNOS activity and protein expression,in turn leading to reduction of NO content in renal tissues.

Background There are a lot of studies about the carrier of corneal endothelial transplantation,but the best carrier has not been defined.Objective This study was to investigate the biocompatibility of chitosan carrier with rabbit corneal endothelium in vivo.Methods Fresh eye-balls were obtained from 10 New Zealand white rabbits.Rabbit corneal endothelial cells (CECs) were isolated and cultured on chitosan carrier in vitro.The morphology and density of rabbits CECs were observed every day,and the expressions of fibronectin (FN),collagen-1 (Coil-I) and Zonula occludens 1 (ZO-1) were detected by immunoinfluorescence.The morphology and ultrastructure of CECs were observed under the scanning and transmission electron microscope.Chitosan carrier with CECs was implanted into the anterior chamber of the left eyes in ten healthy New Zealand white rabbits,and only paracentesis of anterior chamber was performed in the right eyes as controls.The inflammation of ocular anterior segment was examined under the slit lamp microscope,and corneal thickness was measured 1 week,4 and 8 weeks after operation.Corneal endothelium cell density and morphology were examined under the corneal endothelial microscope at postoperative 2 weeks.Corneal samples were collected for the regular histopathological examination to observe the inflammatory reaction at postoperative 1 month and 3 months.Paired t test was used for statistical analyses between the control group (left eyes) and the experimental group (right eyes).The use and care of the animals followed the Statement of ARVO.Results CECs formed an intact monolayer of cells with the uniform shape and size on the chitosan membrane after incubated for 5 days.The cells reached confluence of 90％ 7 days after cultured with the 40％ hexagon cells.Under the scanning electron nicroscope,rabbit CECs showed the round or polygon in the shape with the microvillus on the cell surface.The cells connected closely by desmosome.The processes,pseudopodiums and microvillus on the cellular surface,vacuole in the cytoplasm,expanded endoplasmic reticulum with ribosome and abundant chromatin were exhibited under the transmission electron microscope.The immunofluorescence examination revealed the positive expressions of FN,Coll-Ⅰ and ZO-1 in the CECs on the chitosan carrier.In the in vivo experiment,the exudation in the anterior chamber and corneal edema were seen under the slit lamp microscope 3 days after implantation of chitosan carrier with CECs.However,the inflammation was gone 14 days after operation.The differences of the corneal thickness were no significant between the experimental group and the control group 1 week and 4,8 weeks after operation (t =1.377,P=0.265;t =1.795,P=0.165 ; t =0.390,P =0.760).In addition,no significant differences were found in the CECs density and the hexagon cells rate between the two groups(P =0.365,0.062).The histopathological examination showed that the inflammatory cells around the chitosan membrane were disappeared 3 months after operation and showed a good corneal structure.Conclusions Chitosan carrier has a good biocompatibility with rabbit CECs and anterior chamber,and it may be a potentially good carrier for CECs transplantation.

Biomarkers ; blood ; Child ; Chorionic Gonadotropin ; blood ; Follicle Stimulating Hormone ; blood ; Growth Disorders ; etiology ; Humans ; Klinefelter Syndrome ; complications ; diagnosis ; genetics ; Magnetic Resonance Imaging ; Male ; Mediastinal Neoplasms ; complications ; diagnosis ; surgery ; Puberty, Precocious ; diagnosis ; etiology ; Teratoma ; complications ; diagnosis ; surgery ; Testis ; pathology

Objective:To study the expression of MMP1 and TIMP1 in simple and radiation-combined wound healing and their effects on the healing process and tissue remodeling. Methods: A rat model of radiation-combined wound healing was used. Immunohistochemistry and in situ hybridization were performed which enabled the detection of MMP1 and TIMP1 expression in the healing process. Ultrastructural changes were observed with transmission EM. Results: The wound healing process was impaired and delayed. In rats receiving 25 Gy of gamma ray locally the irradiated wounds healed 6 days later than non-irradiated controls. The following changes in MMP1 and TIMP1 expression were found: (1) In the early inflammatory phase and in the period of granulation tissue formation, MMP1 expression in the newly-formed epidermis of irradiated wounds approximated that in the controls. Later, the epidermal expression of MMP1 in radiation wounds was comparatively increased with the delay of the healing process.On days 3 to 14 after wounding, TIMP1 was weakly positive in the proliferating keratinocytes of control wounds and became negative after epidermal covering, whereas no or only slight epidermal expression was detected in radiation wounds before epidermal covering.(2)MMP1 and TIMP　1 expression in radiation wounds was markedlydecreased in fibroblasts　, endotheliocytes and macrophages as compared with the controls. The expression phase was prolonged due to the delay of the healing process.Conclusions:The reduced expression of MMP1 and TIMP1 in granulation tissue retards such important processes as cell migration, angiogenesis and tissue remodeling, thus retarding the healing process. The expression of MMP1 in the newly-formed epidermis may help the process of reepithelialization,but in the late healing period, overexpression of MMP1 and decreased expression of TIMP1 in the epidermis may hinder the establishment of basal membrane and the formation of granulation tissue, and thus affect the matrix remodeling process.

Objective To investigate the effect of autophagy on the apoptosis of H 9C2 cardiomyocytes.Methods H9C2 cardiomyocytes were incubated with different concentrations of high glucose and high lipids ( HGHL ) for different time.The surface area of cardiomyocytes was measured after HE staining .The cell viability and apoptotic rate were measured by flow cytometry.Western blot was used to detect the expression levels of LC3Ⅱ, p62, Beclin-1,LAMP2 and cleaved caspase-3.Results After treatment with HGHL, the cells appeared hypertrophy in a concentration-and time-dependent manner , the cell hypertrophy was most obvious under the condition of HGHL(500 μmol/L,36 h)(P<0.001).Cell apoptosis increased in a concentration-and time-dependent manner, the apoptotic rate was nearly 50%under the condition of HGHL ( 500 μmol/L,36 h) ( P<0.001 ) .After treatment with HGHL for 24 h, compared with the control group , the expression of LC3Ⅱ was very significantly increased ( P<0.01) , the expression of Beclin-1 and LAMP 2 were significantly decreased ( P<0.05) , but the expression of p 62 was significantly increased ( P<0.01 ) .Compared with the control group and the intervention group , the expression of cleaved caspase-3 was significantly increased ( P<0.01 ) after 1 h of chloroquine pretreatment .Conclusions HGHL induced H9C2 cardiomyocytes hypertrophy promote H 9C2 cardiomyocytes apoptosis in a concentration-and time-dependent manner .HGHL may inhibit autophagy formation and degradation of H 9C2 cardiomyocytes at the same time , leading to abnormal autophagy accumulation of cardiomyocytes , and promote apoptosis , suggesting that inhibition of autophagy may be an important cause for promote apoptosis .

BACKGROUND:Clinical treatment of hemivertebrae-induced congenital scoliosis is a complex medical problem. OBJECTIVE:To find the optimal treatment for hemivertebrae accompanied by congenital scoliosis.
METHODS:Total y 142 hemivertebrae patients who had received surgical treatment in the Department of Orthopedics, the 306 Hospital of Chinese PLA, China from 2010 to 2012 were enrol ed. The main surgical treatment was hemivertebrae resection and bone fusion with internal fixation, apical osteotomy for severe scoliosis and spinal shortening with internal fixation, one-stage posterior thoracolumbar osteotomy with internal fixation, spinal decompression with internal fixation.
RESULTS AND CONCLUSION:After treatment, the average correction rate was 70.9%for scoliosis and 71.7%for kyphosis. The fol ow-up period was 14-35 months, with an average of 23.4 months. By the end of the final fol ow-up, the loss rate for Cobb’s angle was 7.3%for scoliosis and 7.7%for kyphosis. Fol ow-up X-ray films showed bone fusion and internal fixation without loosening, fracture, and decompensation. Implementation of one-stage posterior thoracolumbar osteotomy with internal fixation can effectively correct hemivertebrae-induced kyphoscoliosis to obtain a satisfactory spinal sagittal and coronal balance.

OBJECTIVETo investigate the effects of rabbit limbs ischemia/reperfusion on myocardial necrosis and apoptosis in vivo.

METHODSThirty-six healthy new zealand rabbits were randomly divided into 3 groups: (1) Sham group; (2) I/R(Ischemia/reperfusion) group; (3) RPostC (remote postconditioning) group. The activity of blood serum creatine kinase (CK) and lactate dehydrogenase (LDH) were measured at baseline, the end of ischemia after 60 min and 120 min of reperfusion respectively. The extent of myocardial ischemia and the scope of myocardial infarction were assessed by evans blue and Triphenyl tetrazolium chloride (TTC). The myocardial cell's apoptosis at the area of myocardial ischemia was estimated by Tunel. Protein expression of caspase-3, Bcl-2 and Bax in myocardial ischemic area were analyzed with the method of immunohistochemistry.

RESULTSCompared with I/R group, the myocardial infarct size and the CK activity were significantly reduced in RPostC group. The Tunel positive index of RPostC group in ischemic myocardium was significantly lower than that in I/R group (21.79% +/- 1.07% vs 35.81% +/- 1.10%, P < 0.05). Caspase-3 positive cells index was calculated with randomly selected five regions in each slide and then the positive cells in per hundred cells were calculated. The RPostC group of caspase-3 positive cells was significantly lower than that in I/ R group(25.03% +/- 1.16% as 39% +/- 2.43%, P < 0.05). Compared with the sham group, the Bax protein expression index and the Bcl-2 protein expression index of I/R group and RPostC group were increased. The Bax/Bcl-2 ratio of RPostC group decreased, while it was increased in I/R. Compared with the I/R group, the Bax protein expression and Bax/Bcl-2 ratio of RPostC group significantly reduced, but the expression index of Bcl-2 ratio was significantly increased, the differences were statistically significant.

CONCLUSIONLimbs ischemia/postconditioning could significantly reduce necrosis and apoptosis of ischemia/reperfusion myocardium. The mechanism of reducing the myocardial cell apoptosis may have relation to inhibiting the activation of pro-apoptotic gene caspase-3 and increased expression of Bcl-2.

Animals ; Apoptosis ; Caspase 3 ; metabolism ; Creatine Kinase ; blood ; Ischemic Postconditioning ; L-Lactate Dehydrogenase ; blood ; Lower Extremity ; Male ; Muscle, Skeletal ; blood supply ; Myocardial Reperfusion Injury ; pathology ; Necrosis ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Rabbits ; bcl-2-Associated X Protein ; metabolism