BACKGROUND: Thawed allografts are usually discarded for various reasons. Whether these discarded allografts can be refrozen for later use and their histological changes in vivo have not been reported. OBJECTIVE: To investigate the histological characteristics of anterior cruciate ligament (ACL) reconstructed with the repetitive freeze-thawing allogenic Achilles tendon in New Zealand white rabbits. METHODS: Allogenic Achilles tendons were harvested from adult male New Zealand white rabbits, and were stored at -8 ℃ and thawed at 20 ℃ for 1, 2, 3 and 10 times, respectively, after sealed package and 60Co irradiation. Twenty-four adult male New Zealand white rabbits were enrolled: the left and right knees of 12 rabbits were respectively reconstructed with 1 (control group) and 2 times of freeze-thawing allogenic Achilles tendon, and another 12 rabbits underwent reconstruction with 3 and 10 times of freeze-thawing allogenic Achilles tendon, respectively. Three specimens from each group were evaluated with modified histology grading scores at 6, 12 and 24 weeks to assess the cell morphology, cell quantity, matrix staining intensity, fibrocartilage formation, new bone formation, tendon healing and cartilage injury. RESULTS AND CONCLUSION: The cell morphology, matrix staining intensity and total scores of the 10 times group were significantly higher than those of the other groups at 6 weeks (P < 0.05), but other parameters showed no significant differences among groups (P > 0.05). ACL reconstructed with 10 times of repetitive freeze-thawing allogenic Achilles tendons had higher histological scores at 6 weeks after modeling, but no significant differences were shown at 12 and 24 weeks after modeling. To conclude, our study only testifies better histological scores on the multiple times of freezethawing Achilles tendon than the less times of freeze-thawing Achilles tendon at the early period after operation.

Objectives:To investigate the association and the prognostic significance of the expression of telomerase activity (TA) and apoptosis (APO) in tissues of primary breast cancer.Methods:TRAP-PCR and ELISA were developed to detect the expression of TA,and make quantitative and qualitative analysis respectively.TUNEL was also used to evaluate apoptosis cells in cancer lesions and count apoptosis index (AI).The expression levels of proteins of oncogene Bcl-2 and p53 were measured by means of S-P immunohistochemistry.Results:The mean TA positive expression rate was 89.8%,and its mean optic density was (A) 0.63±0.29,which were significantly higher than those of normal tissues and benign breast lesions (P<0.05).The 5 years survival rate of patients with positive TA expression was lower (60.3%) than those of negative expression (77.4%).There were no relations between TA expression and patients ages,TNM staging,lymph nodes metastasis,size,locations and pathological types of cancer.But TA was positively correlated with APO (r=0.733 1,P<0.05) and the protein expression of oncogene Bcl-2 (r=0.781 4,P<0.05) and negatively correlated with that of p53 (r=-0.625 5,P<0.05).Moreover,in patients receiving preoperative chemotherapy,the levels of TA expression were much lower than those without chemotherapy (64.7% vs 85.1%),and the former had higher AI than the latter (7.19±3.75% vs 3.23±2.04%),intraarterial chemotherapy group were superior significantly to adjuvant group (P<0.05).Multivariate Cox Regression Analysis showed that only TA and AI influenced the survival time of primary breast cancer patients,and patients with higher positive TA expression and AI>4.0% had shorter survival time than those of negative TA expression and AI<4.0% patients.Conclusions:There was a significant correlation between TA and APO and its related oncogene Bcl-2,p53.TA and APO are of great diagnostic values,therapeutic helps and prognostic significance in the formation and the development of primary breast cancer.

Acupuncture Therapy ; Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Respiration Disorders ; physiopathology ; therapy ; Yawning ; Young Adult

Background The rejection following keratoplasty still is a leading cause of corneal transplantation failure.Studies showed that the interleukin-22 (IL-22) ,one of the effector molecules of T helper cell 17 (Th17) participated on the rejection after heart,liver and bone marrow transplantation.However,the effect of IL-22 on corneal graft rejection is not well understood.Objective This study was to investigate the expression of IL-22 mRNA in the corneal grafts and the role of IL-22 in the immune rejection after corneal transplantation in rats.Methods Seventy-two Wistar rats were randomized into autologous keratoplasty group,allograft keratoplasty group and anti-rejection group,and other 4 normal Wistar rats served as normal control group.Autologous keratoplasty was operated on the Wistar rats of the autologous keratoplasty group,and allograft keratoplasty were carried out with the 24 SD rats as donors and 48 Wistar rats as recipients.Tobramycin and dexamethasone eye drops were topically administrated after autologous keratoplasty for 2 weeks in the anti-rejection group.The experimental eyes were examined by slit lamp microscope after surgery and graft survival was evaluated based on the rejection scoring criteria of Larkin.Intergroup accumulated survival rates of grafts were compared using Kaplan-Meier analysis.Histopathological examination of grafts was carried out in 5 and 14 days after operation respectively,and the related expression levels of IL-22 mRNA and aryl hydrocar-bon receptor (AhR) mRNA were carried out by real-time fluorescence quantitative PCR.The feeding and use of the experimental animals followed the Guangdong provincial regulations on the management of experimental animals.The experimental design was approved by the ethics committee of Southern Medical University.Results The median survival time of grafts in the allograft keratoplasty group was 10 days,and that in the anti-rejection group was 17 days,showing a significant survival extention in the anti-rejection group (x2=16.442,P =0.000).Significant differences were found among the 4 groups in the related expression levels of IL-22 mRNA in both 5 days and 14 days after surgery (postoperative 5 days : F=2.44,P =0.00;postoperative 14 days: F=267.92, P =0.00), and the related expression levels of IL-22 mRNA were remarkably higher in the allograft keratoplasty group than those in the anti-rejection group at different time points (postoperative 5 days :9.70±0.35 vs.0.46±0.21;postoperative 14 days : 23.12 ± 1.89 vs.3.14±0.94) (both at P＜0.05).The related expression levels of AhR mRNA in the grafts were considerably different among the 4 groups (postoperative 5 days : F =395.73, P =0.00;postoperative 14 days : F =942.37, P =0.00) , and the expression levels were significantly elevated in the allograft keratoplasty group compared with the anti-rejection group at various time points (postoperative 5 days:2.52±0.32 vs.1.89±0.10;postoperative 14 days:7.20±0.25 vs.2.60±0.17) (both at P＜0.05).Conclusions The expression level of IL-22 RNA up-regulates in the grafts with immuno-rejection.Topical administration of tobramycin and dexamethasone eye drops inhibits the rejection after keratoplasty.AhR plays a regulative role to the expression of IL-22 in rats after keratoplasty.

Objective To investigate the expressions of cell phenotypes CD_8 and CD_(28) of peripheral blood lymphocytes in patients with colorectal cancer. Method CD_8 and CD_(28) of cell phenotypes were measured by flow cytometry in 50 patients with colorectal cancer (cancer group) and 30 nontumorous patients (control group). Results The expression of CD_8 in cancer group was significantly higher than that in control group [(32.24±8.38)% vs (22.18±7.55)%](P < 0.01). CD_(28) and CD_8~+/CD_(28)~+ were much lower than those in control group [(52.03±10.94)% vs (60.60±7.98)%,12.18±4.28 vs 16.38±4.94](P<0.01). CD_8~+/CD_(28)~+ in Dukes D stage patients were significantly lower than that in Dukes B stage patients (P<0.05). CD_(28) and CD_8~+/CD_(28)~+ in lymph node metastasis patients were much lower than those in lymph node negative patients (P < 0.01). CD_(28) and CD_8~+/CD_(28)~+ in serosa involved patients were lower than those in no serosa involved patients (P < 0.01). Conclusions The expressions of cell phenotypes CD_8 and CD_(28) of peripheral blood lymphocytes in patients with colorectal cancer are closely related to biological characteristics of the tumor. The assays of cell phenotypes CD_8 and CD_(28) might be useful for evaluating the immunal statement and prognosis of patients with colorectal cancer.

Objective Some features, as numbers, morphology and distribution, of the Langerhans cells (LC) with various surface marker infiltrating in the lesion of basal cell carcinoma (BCC) were explored in order to apply some experimental data for assessment of the roles of LCs in the cutaneous tumor immunity.Methods Expression of CD1a, S-100 and HLA-DR on the surface of infiltrating LC of 30 cases of BCC and 15 cases of normal skin tissues were investigated by immunohistochemistry. The expression qualities were studied with computer image analysis system. Results It was observed that CD1a+ LC distributed mainly in tumor nest compared with the control group, CD1a+ LC in BCC revealed morphological changes: the dendrites shortened or disappeared. The density of CD1a+ LC, detected by cell number counting as well as staining strength scanning, showed a statistic difference between the control group and BCC (P ＜0.05). S-100(+)LC spread evenly throughout BCC lesions. The number of cell dendrites as well as its length increased markedly.The cell number as well as the staining strength in BCC lesions increased markedly compared with the control group (P ＜0.05). HLA-DR(+)LC infiltrated mainly around the located tumor. This kind of LC in the lesion did not show significant differences compared with the controls (P ＞0.05). Conclusion LC showed differences between BCC lesion and normal skin tissues. The alterations in numbers, morphology and distribution of LC with various markers in the lesion of BCC supplied special local cellular immunity status, resulting in disable metastasis and good prognosis.

BACKGROUND: Epidemiologic studies have shown an association between higher insulin levels and coronary artery disease, and metabolic studies have associated insulin resistance and compensatory hyperinsulinemia with non-insulin-dependent diabetes mellitus, hypertension, obesity,and lipid disorders.OBJECTIVE: To investigate the relationship between insulin resistance (IR) and erythrocyte insulin receptors (EIRs) in the patients with cerebral infarction (CI).DESIGN: Case-control trial.SETTING: Department of Neurology, China-Japan Union Hospital of Jilin University.PARTICIPANTS: From January 2004 to October 2004, 40 patients with CI, who were in-patients in China-Japan Union Hospital of Jilin University,were selected for the study. Meanwhile, 30 healthy doctors or nurses were recruited as normal controls.METHODS: The levels of blood glucose and serum insulin under fasting and 2-hour after oral glucose tolerance test (OGTT) were detected in the 40 patients with CI and 30 healthy doctors or nurses. Fasting blood glucose multiplied by fasting serum insulin was insulin resistance index (IRI). The number of insulin receptors and their binding affinity on every erythrocyte were determined using modified Gambhir's method. The correlation between the number of EIRs and IRI was analyzed.MAIN OUTCOME MEASURES: Comparison of CI group with controlRESULTS: Data of 40 patients with CI and 30 controls were analyzed, and under fasting and 2-hour after OGTT: The level of serum insulin under fasting and blood glucose, serum insulin at 2-hour after OGTT in CI group were higher than those in normal group [(13.30±5.15), (9.85±4.36) mU/L,(8.27±1.65), (6.32±1.37) mmol/L, (75.21±21.12), (28.26±6.31)mU/L,P ＜ 0.01,EIRs: The number of insulin receptors with high and low affinity and maximum specific binding rate in the patients with CI were significantly less than those in normal group [20.30±4.50, 23.80±4.10; 2 223.80±509.30,2 610.10±435.10; (10.62±3.55)%, (13.21±2.94)%, P ＜ 0.01]. Multiple linear regression analysis and correlation analysis showed the numbers of two types of EIRs in the patients with CI were negatively correlated with IRI (r=-0.458, -0.439, P ＜ 0.01, both).CONCLUSION: Insulin resistance can lead to cerebral infarction. Decrease in insulin receptors may play an importance role in cerebral infarction induced by insulin resistance.

Animals ; Apoptosis ; Cell Hypoxia ; Cell Proliferation ; Cells, Cultured ; Hydrogen-Ion Concentration ; Muscle, Smooth, Vascular ; cytology ; Myocytes, Smooth Muscle ; cytology ; Oxygen ; metabolism ; Rats

Objective To explore the effects of simvastatin on the protein expressions of urokinase-typeplasminogen activator (uPA) and plasminogen activator inhibitor-1 (PAI-1). Methods Male Sprague–Dawley rats were divided into saline group , LPS group and LPS plus simvastatin group , and were then pretreated with simvastatin (1 mg/kg) for 30 minutes before addition of LPS (8 mg/kg). Changes in left ventricular pressure were recorded. Ninety minutes after LPS injection, whole blood was collected from the inferior vena cava, and neutrophils were separated. The neutrophils were then lysed to detect levels of uPA and PAI-1. Results Left ventricular systolic pressure (LVSP: mmHg), maximal differential of left ventricular pressure (+dp/dtmax:mmHg/s), and heart rate (beats/min) were markedly decreased at different time points after administration of LPS, and maximal differential of left ventricular pressure increased in the rats receiving LPS as compared with those receiving saline, although the differences between the control and LPS groups were not statistically significant. LPS caused a great decline in uPA content and an elevation in PAI-1 content in neutrophils, but simvastatin diminished the impact of LPS on neutrophils. Conclusion Simvastatin plays a role in protection of cardiac function in rats with LPS-induced septic shock , and controls expressions of uPA and PAI-1 in neutrophils.