Objective To observe the distribution of circum-patella nerve fibers in the soft tissue to provide experimental evidence,which is significant in denervation for Total Knee Arthroplasty (TKA).Methods Patella specimens were collected from 4 cadavers (2 cases of diabetic foot,lcase of lower extremity arterial occlusive,and 1case of car accident),all 4 of which were resected soft tissue with a dimension of 0.5cm × 0.5cm and full depth thickness around patella more than 0.5cm for histology and morphology observation.The nerve fibers histology and morphology were observed in all resected specimens with HE staining and silver-gilt glycine staining in the same field of microscopic vision.Results Anatomy found that the vascular network form skins directly involved in the patella nourish hole area and in the 10,2,4,7 clock point have found that blood vessels into the patella.There have a large number of nerve fibers near to the patella under the microscope,but there were no significant difference in the nerve fibers region distribution of all specimens.There were some into patella nerve fiber paths in side of patella soft tissue,which lied in 7,11 and 13 clock point,but outside no this phenomenon.The distribution of circum-patellar nerve fibers were described as distribution of regional concentration,which lied in much more 5,6,7 clock points and 10,11,12,1,2,clock points,in which the quadriceps tendon and patellar tendon have more than the others.In the 13 clock point,the fascia and periosteum of nourish hole area were also found in a large number of nerve fibers,and there were laminar distribution in different soft tissue layers,which were collected much more in synovial layer,fat pad,tendon near to patella.Conclusion There are much more nerve fibers near to the patella and some into patella nerve fiber paths in the medial side and nourish hole area.Nerve fibers distribution of circum-patella can be described as laminar distribution and regional concentration ,which is more in the centre,bottom more than top,outside more than inside,the bipolar more than the others.The patella denervation operation by reducing the number of peripheral nociceptors to achieve desensitization is feasible in TKA.

Objective To evaluate the efficacy of non-Hodgkin's lymphoma(NHL)treated by high does MTX,autologous peripheral stem cell transplantation and biotherapy for 67 cases.Methods Sixty-seven NHL patients from June,2003 to March,2007 were treated by three times HD-MTX,APBSCT and biotherapy of IL-2.Results There were 36 cases(87.8%)in complete relase(CR)period;5 cases(12.2%)in relapse(RE)period and 1 patient(2.4%)died in CR group;in PR group,there were 15 cases(57.7%)in CR period;11 cases(42.3%)in RE period and 5 patients(19.2%)died.Conclusion These preliminary results suggest that the therapy can be performed safely.It is an efficacious therapeutic measure for the patients with non-Hodgkin's lymphoma.

Objective To evaluate clinical effect of autologous peripheral blood stem cell transplantation(APBSCT)on the treatment of hematologic malignancies.Methods Totally 231 patients(ALL in 45 cases,AML in 34 cases,NHL in 100 cases,HD in 31 cases,MM in 21 cases)with hematologic malignancies received APBSCT from March 2001 to February 2007.Therapeutic effect and complication were oberserved.Results Totally 230 patients obtained hematopoietic reconstitution quickly,one case failed.ALL CR1(first time CR):13 in DFS,4 alive with disease(LWD),11 in death;ALL CR2(second time CR):3 in DFS,4 in LWD,10 in death;AML CR1:12 in DFS,3 in LWD,6 in death;AML CR2:6 in DFS,2 in LWD,6 in death;NHL CR:43 in DFS,7 in LWD,9 in death;NHL CR2:18 in DFS,5 in LWD,7 in death;NHL NR:2 in DFS,4 in LWD,5 in death;HD CR1:10 in DFS;HD PR:12 in DFS,3 in LWD;HD RE:3 in DFS,2 in LWD,1 in death;MM:7 in DFS,6 in LWD,8 in death.Conclusion APBSCT is a safe and effective therapy method for treating hematologic malignancies.

BACKGROUND:Different structures of matrix models, such as grating, holes and pil ars make different effects on the differentiation of neural stem cel s. OBJECTIVE:To explore the effects of the diameter and spacing, known as physical signals of micro pil ars on neural stem cel differentiation. METHODS:Micro pil ars with different diameters and spacing, both of which had four dimensions of 2.5, 5, 10 and 20μm, were fabricated on silicon substrates by photolithographic method. Purified primary neural stem cel s were incubated on the each micro pil ar for 7 days in vitro. Then the differentiation of neural stem cel s into neuron-like cel s was observed using immunofluorescence staining and quantitative real-time PCR. RESULTS AND CONCLUSION:When the diameters of the micro pil ars were constant and the spacing of micro pil ars varied in the range of 2.5-10μm, the differentiation rate of neural stem cel s increased with the spacing increase. When the spacing was invariable and the diameters changed in the range of 2.5-20μm, the differentiation rate of neural stem cel s declined with the diameter increase. Especial y, the micro pil ars with 2.5μm diameter and 10μm spacing significantly promoted the differentiation of neural stem cel s into neuron-like cel s. These results show that specific micro pil ars with smal diameters and large spacing facilitate the differentiation of neural stem cel s, thus providing guidance for developing tissue-engineered scaffolds.

Objective To investigate the optimal condition for culturing human umbilical cord blood-derived stromal cells(hUCBDSCs),and to observe their biological behaviors.Methods The umbilical cord blood was obtained from the Department of Obstetrics of the authors' hospital.The influence on the growth of hUCBDSCs was determined by the isolation method,the medium and the time of renewal of first medium were analyzed.The status of cell growth was observed under inverted microscope and the morphological characters were studied with the cells stained by Wright's staining.The hUCBDSCs were identified by cytochemistry and immunocytochemistry methods.Results The gelatin precipitation was better than other techniques for isolation.The optimal time for first medium renewal was the fourth day of culturing,and the improved Dexter-type cultural system was better than the classical Dexter-type cultural system in primary culture.The colonies of adherent cells began to form in 9-14 days(with a median of 12.1 days),and the number of colonies reached it maximum in 15-21 days(mean 19.4 days).On day 28,adherent cells spread all over the bottom of dish.On day 28 of culturing,these cells were found under light microscopy to have differentiated into three kinds of cells: fibroblast-like cells,macrophage-like cells and small-round cells.Cytochemistry assay revealed that the positive rate reached 100% with non-specific esterase(NSE) and saccharogen(PAS) staining.26% of the hUCBDSCs were positive with alkaline phosphatase(ALP) staining,but negative with peroxydase(POX) staining.Immunocytochemistry staining revealed that the positive rates of hUCBDSCs for CD31,CD68 and Fn were 96% and 95%,and 94% respectively,and for CD45 was 0%.Conclusion The hUCBDSCs could be successfully cultured in vitro,and it sets a foundation for further study on the clinical application of hUCBDSCs.

Objective To investigate the effects of thrombopoietin (TPO) on the proliferation of megakaryocytic line-HEL cells co-cultured with human umbilical cord blood-derived stromal cells (hUCBDSCs), which is supposed to further elucidate the mechanism of TPO-mediated functions. Methods HEL cells were co-cultured with hUCBDSCs, and with human bone marrow stromal cells (hBMSCs), and suspended HEL was used as control. HEL The concentration of TPO in supernatant of hUCBDSCs and hBMSCs was detected by ELISA assay. The expression of c-Mpl membrane-bound protein of HEL cells was detected by laser scanning confocal microscopy and flow cytometry, and the expression of c-Mpl mRNA was detected by RT-PCR. Results It was revealed by ELISA assay that the concentration of TPO secreted by hUCBDSCs was higher than that by hBMSCs, even though the passage was done. The secretion peak in hUCBDSCs group appeared at the 8th day of culturing, somewhat delayed to that in hBMSCs group which the peak appeared at the 6th day. The expression of C-mpl protein displayed uniform green fluorescence on the surface of HEL cell line as determined by laser confocal microscopy, and mean fluorescence intensity detected by flow cytometry was 22.19?2.15, 29.65?0.82 and 37.43?1.69 in HEL suspended culture group, HEL/hBMSCs co-culture group and HEL/ hUCBDSCs co-culture group, respectively. The expression of C-mpl protein in HEL/ hUCBDSCs co-culture group was higher than that in the two other groups (P0.05). Conclusion TPO-pathway might be the important intermediate process for hUCBDSCs to promote megakaryocyte proliferation. The promotion as such may be realized by up-regulating the c-Mpl expression in translation or post-translational modification phase.

Objective To investigate the relation between serum homocysteine(HCY) and the development of atherosclerosis(AS).Methods Diet contained 1% methionine was fed to make models of hyperhomocysteinemia and atherosclerosis in 18 male New Zealand rabbits. Hyperlipemia group was as control group. Ultrasonographic image was performed in 0,4,8 weeks respectively with HP 8500 and recorded the stenosis and plaque of abdominal aorta compared with pathology.Results Stenosis and plaque of abdominal aorta were observed in 4 weeks in high fat diet group. Stenosis and plaque of abdominal aorta were observed in 8 weeks in high methionine diet group.This results were confirmed by pathology anatomy.Satistically,significant correlation was found between AS and hypercholesterolemia or hyperhomocysteinemia respectively.But there was no correlation between hypercholesterolemia and hyperhomocysteinemia. Conclusions Hyperhomocysteinemia is an independent important risk factor of atherosclerosis.

Objective To observe the effect of the human umbilical cord blood-derived stromal cells(hUCBDSCs)and multiple myeloma bone marrow stromal cells(MM BMSCs)on the biological characteristics of KM3 cells.Methods After KM3 cells were co-cultured with hUCBDSCs or MM BMSCs,the proliferation,cell cycle and the apoptotic rate were determined by growth curve plotting and flow cytometry respectively.Real-time Q-PCR was used to measure the mRNA expressions of Bcl-2 and Bax.Results As compared with MM BMSCs,hUCBDSCs markedly suppressed the proliferation of the KM3 cells.KM3 cells in the KM3/MM-BMSCs co-culture group were arrested in G0/G1 phrase,while KM3 cells in the KM3/hUCBDSCs co-culture group predominantly in S phrase.The apoptotic rate of KM3 cells in the KM3/hUCBDSC co-culture group were higher than that in the KM3/MM-BMSCs co-culture group.Conclusion In comparison with MM BMSCs,hUCBDSCs inhibit the proliferation of KM3 cells and promote the apoptosis.

Objective To observe the efficacy of intense pulsed light(IPL)incorporated with optimal pulse technology (OPT)in the treatment of rosacea.Methods Thiris-two patients with erythematotelangiectatic rosacea and 53 patients with papulopustular rosacea were treated with IPL-OPT for 4 sessions with an interval of 3 weeks.Patients were assessed clinically and photographically by physicians before each treatment.Skin melanin index,erythema index,sebum secretion level and water content in stratum corneum were tested at the baseline,3 weeks after each treatment,and 6 months after the last treatment.Results Three weeks after the last treatment,the effective(more than 60%improvement)rate WaS 81.18%in total,75%in erythematotelangiectatic rosacea,and 84.91%in papulopustular rosacea;there WaS no significant difference between erythematotelangiectatic rosacea and papulopustular rosacea (x2=1.28,P＞0.05).Six months after the last treatment,the total effective rate still remained at 78.82%with no significant difierence from that observed at 3 weeks after the treatment(x2=1.62,P＞0.05).The value of melanin index,erythema index and sebum secretion level decreased significantly after the final treatment,however,water content in stratum corneum remained at the salne level as that before treatment.After 6-month follow-up,no significant change was noticed in the above 4 parameters compared with those obtained at 3 weeks after the treatment.Neither hyperpigmentation nor hypopigmentation was observed during the treatment and follow-up.Conclusion This study demonstrates that IPL-OPT is an effective treatment for rosacea with relatively few side effects.

Objective To assess the changes in frequency of peripheral T lymphocytes expressing different cytokines in patients with atopic dermatitis (AD) before and after treatment with BCG-PSN and their relationship with disease severity. Methods A randomized, double blinded and placebo cross-over control study was conducted. A total of 8 patients with AD were recruited in this study. Intramuscular BCG-PSN or placebo was given to patients every other day for 36 days. Flow cytometry was performed to measure the frequency of IL4-, IL5-, IFN-γ- and TNFα-expressing peripheral CD4+ T cells and CD8+ T cells before and after the therapy. Disease severity was evaluated by atopic dermatitis area and severity index score (ADASIS). Results The difference value in IFN-γ+CD8+ T cell frequency before and after therapy was significantly higher in patients treated with BCG-PSN than in those with placebo (8.056 ± 13.962 vs -6.549 ± 10.491, U = 2.26, P< 0.05). There was no statistical difference in the frequency of IL4-, IL5-, TNFa-expressing CD8+ T cells between BCG-PSN- and placebo-treated patients (all P > 0.05). The decrease in ADASIS was 1.56 ± 1.49 in patients treated with BCG-PSN, which was statistically higher than that in placebo-treated patients (-0.05 ± 1.54, U = 2.00, P< 0.05). Conclusion As an immunomodulator, BCG-PSN may control AD by restoring the balance of T-cell subsets.