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.

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.

Cardiovascular System ; GATA4 Transcription Factor ; metabolism ; Humans

Objective To clarify the role of B7-H1 in the immune privilege after corneal transplantation in homogeneity variant mice.Methods We established the experimental animal model of allograft mice by using C57BL/6 mouse as donor and Balb/c mouse as recipient.We allocaated the mice with long time survival (＞50 days) corneal graft into survival group,mice with rejection occurring in 50 days into rejection group,and normal C57BL/6 mice into control group.The transplanted corneal grafts were obtained for future reference at the 8th week after transplantation in survival group,and the time of rejection in rejection group.The expression of B7-H1 mRNA was detected by using immunohistochemistry and real time quantitative PCR (RT-PCR),and the relationship between B7-H1 and the immune privilege after corneal transplantation was analyzed. Results The B7 H1 mRNA was highly expressed in epithelium and endothelium of corneal grafts both in survival and control group,in comparison to an obviously lower expression in rejection group.The relative expression level of B7-H1 mRNA was 200.0 ± 11.5 in survival group,44.7 ± 10.8 in control group,and 6.9 ± 12.0 in rejection group,respectively. There were statistically significant differences among the three groups (F=241.164,P＜0.01 ).The was a significant correlation between the level of B7-H1 mRNA and occurrence rate of rejection in corneal graft (P＜0.01 ).Conclusion The results suggest that the immune privilege after corneal transplantation might be mediated by B7-H1,which plays an important role in maintaining the state of corneal immune privilege.

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 To evaluate the differences of the lower extremity atherosclerosis between patients with and without type 2 diabetes using dual-source CT angiography.Methods Dual-source CT angiography of lower extremity was performed in 87 patients with (n=30)or without (n= 57 )diabetes.Extent of luminal stenosis,and the type,distribution and range of the plaques were compared.Results 342 plaques in 540 segments (63.3%)in diabetic patients,and 500 plaques in 1 026 segments (48.7%)in non-diabetic ones were detected respectively.Compared with non-diabetic patients,the diabetic ones had a higher overall incidence of plaques (P <0.05).Calcified plaques were the most common in both kinds of patients,and the incidence of mixed plaques was high-er in diabetic patients than that in non-diabetic ones (35.6 % vs.28.4%,P <0.05).Light to moderate stenosis occurred in most diabetic patients,and fewer occlusion was found compared with non-diabetic ones (9.1% vs.1 7.0%,P <0.05).The most common sites of the plaques in diabetic patients were located at distal small arteries below the knee.However,those were located at proximal arteries above the knee for non-diabetic ones.The involvement of atherosclerosis in diabetic patients was more diffused,and the de-gree of Ⅳ (75%-100%)was higher than that in non-diabetic ones (P <0.05).Conclusion Atherosclerosis in lower extremity on dual-source CT angiography is very common in diabetic patients with multi-segmental,diffused,non-obstructive involvement of dis-tal small arteries below the knee.

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

Adult ; Benzene ; poisoning ; Cord Blood Stem Cell Transplantation ; Female ; Humans ; Infant, Newborn ; Leukemia ; etiology ; surgery ; Male ; Treatment Outcome ; Young Adult