We examined eutopic,ectopic endometria and peritoneal fluid macrophages from 22 patients with endometriosis(EMS) and 14 women without EMS.To obtain evidence for the induction of programmed cell death,apoptotic cells were identified using a modified terminal deixynucleotidyltransferasebiotin nick end-labeling method(TUNEL).To evaluate cell death repressor activity,bcl-2,bax and fas genes expression was examined using immunohistochemical staining. The results showed that bcl-2 expression in eutopic,ectopic endometrium and peritoneal fluid macrophages with EMS was significantly increased compared with no EMS(P＜0.01).Bax expression in eutopic,ectopic and peritoneal fluid macrophages with EMS was significantly decreased compared with no EMS(P＜0.05). Fas expression in eutopic,ectopic endometrium was decreased compared with no EMS(P＜0.05). The expression of apoptotic genes were different in eutopic,ectopic endometrium and peritoneal fluid macrophages from EMS and no EMS.Apoptotic rate in EMS was lower than no EMS,and its acceptance was decreased,which could bear implications for the growth and survival of ectopic endometrial tissue.

We examined eutopic,ectopic endometria and peritoneal fluid macrophages from 22 patients with endometriosis(EMS) and 14 women without EMS.To obtain evidence for the induction of programmed cell death,apoptotic cells were identified using a modified terminal deixynucleotidyltransferasebiotin nick end labeling method(TUNEL).To evaluate cell death repressor activity,bcl 2,bax and fas genes expression was examined using immunohistochemical staining. The results showed that bcl 2 expression in eutopic,ectopic endometrium and peritoneal fluid macrophages with EMS was significantly increased compared with no EMS( P

0.05), whereas the value of new bone grafted with pDBM was significantly lower than that of the normal group (P0.05); but the CUS of the pDBM grafted group was significantly lower than that of normal radius (P0.05). Histological analysis exhibited that most of the DBM was absorbed and substituted by matured new cortical bone in the treated defects of both groups 6 weeks postoperatively, whereas in the untreated group, the defects were only filled with fibrous connective tissue in their mid-portion. Conclusion The sDBM and pDBM are both effective in repairing segmental bone defects. The properties of new bone induced by grafts with sDBM are superior to that of pDBM in biomechanics. These materials can be used in clinical practice as bone graft extenders or enhancers.

[Objective]To observe the histological characteristics of autologous chondrocytes/human articular cartilage derived scaffold (HACDS) composites for repairing chondral defects in rabbit models. [Methods]A full-thickness articular-cartilage defect (diameter 4 mm,thickness 2 mm) was created in the femoral condyle of rabbits. The rabbits were divided into two groups: control group,HACDS scaffold only,and experimental group,chondrocytes/HACDS scaffold. Fifteen months after implantation,the specimens were observed by inverted phase contrast microscopy,and assessed by staining with haematoxylin-eosin,alcian blue,toluidine blue,safranin O,as well as by the immunohistochemistry of collagen type Ⅱ.[Results]Histologically,the generated neo-cartilage integrated well with its surrounding normal cartilage and subchondral bone in the defects of experimental group at 15 months post-implantation,whereas only fibrous tissue was filled in the defects of control group. Histochemical and immunohistochemical analysis revealed that alcian blue,toluidine blue,safranin,and were obviously positive in the experimental groups. However,alcian blue,toluidine blue,and safranino O were negative in the control group.[Conclusion]The current histological examination demonstrated that an engineered cartilage composed of autologous chondrocytes on HACDS scaffold could be successfully obtained and further applied to repair an articular cartilage defect in a rabbit model.

Objective To find out a way for culturing the myocardial cells of neonatal rats in vitro, and to study their biological characters. Methods Cardiomyocytes from the heart of 1-3 days old Sprague-Dawley rats were prepared by a modified protocol. Hearts were harvested and cut into pieces of about 1mm3 in size, and placed into cell culture bottles with nylon without pre-treatment of digestive enzyme. DMEM supplemented with 10% (v/v) FCS was used for culture, with thymidine (6mg/ml) added to inhibit fibroblast growth. Cells in monolayer were formed on the gauze and the bottom of the culture flask, and then cells were isolated by 0.25% trypsin digestion for 30 seconds. Cell suspension was transferred to 100cm2 cell culture flasks at a density of 104 cells /cm2 for identification, viability test and morphologic observation. The cells obtained were stained with monoclone anti-a-sarcomeric actinin and FITC to evaluate the purity of the myocardial cell preparation by flow cytometry, AO-PI fluorescein stain was used to evaluate the viability and Giemsa staining for examining the morphology of the myocardial cells. Results 24-48 hours after culture, the myocardial cells became adherent to bottle wall to form a sheet, and began to pulsate. The purity of myocardial cells in culture cell population was 94.13%, the ratio of viable myocardial cells was 95.3%, and 95%CI was 91.6%-99%. The output of myocytes from each neonatal heart was 3.3?106 and 95%CI is 2.1? 106- 4.5?106. Conclusion Neonatal myocytes culture in this way can generate large amount of viable single myocardial cells suitable for myocardial research in a short period.

BACKGROUND:Titanium implants as a safe biological material have been used to produce the artificial Russian titanium cornea, but complications stil exist, including artificial cornea shift, leakage, corneal tissue melting and artificial cornea discharge.
OBJECTIVE:To evaluate in vivo biocompatibility of hydroxyapatite modified titanium skirt for keratoprosthesis in alkali burn cornea.
METHODS:A total of 30 alkali burned New Zealand white rabbit corneas were divided into three group groups. Hydroxyapatite modified titanium skirt (experimental group) and titanium skirt (control group) were respectively inserted into the corneal stroma of rabbits. In the blank control group, only a lamel ar corneal incision was made.
RESULTS AND CONCLUSION:Al skirts were stable without necrosis, melting and exclusion during the observation period. The number of inflammatory cells in the experimental and control groups was significantly higher than that in the blank control group at 2 and 8 weeks postoperatively (P<0.05), but there was no difference in inflammatory cellinfiltration among different groups by the 16th week. The number of corneal fibroblasts increased significantly in the experimental group compared with the control and blank control group after 2, 8, 16 weeks (P<0.05). The extracellular matrix deposited on the surface of hydroxyapatite modified titanium skirt was denser and tighter than that on the surface of titanium skirt. It indicates that hydroxyapatite modified titanium skirt for keratoprosthesis can promote the interfacial biointegration of skirt and host cornea.

Objective To detect the spreading scope of rectal cancer to mesorectum by RT PCR using carcinoembryonic antigen (CEA) mRNA as a marker and to investigate the excision scope of mesorectum in resection of rectal cancer. Methods Forty specimens from 40 rectal cancer patients who underwent curative operation was employed to detect the metastatic deposits scattered in the mesorectum by RT PCR using CEA as a marker. Results Nine of 40 (22.5%) specimens contained metastatic deposits scattered in the mesorectum. The metastasis was just within the range of 4cm mesorectum under the verge of tumor. The tumor spreading to mesorectum is correlated with Dukes stages,the infiltrated depth of bowel wall, tumor differentiation and tumor type( P 0.05). Conclusion The excision of mesorectum should be within the range of 5cm under the verge of tumor in surgical management of rectal cancer.

BACKGROUND:Because human cells for culturing alveolar bone cell line are from alveolar bone, which is in oral cavity,and easily polluted, so laboratory study is often unsuccessful. Because the samples are from adults, so cell division index and the successful rate of culture are low.OBJECTIVE: To compare the biological characteristics of survived cell line established through passage,cryopreservation and revitalization following in vitro culturing the alveolar bone tissue obtained from normal persons and patients with chronic periodontitis accompanied with osteoporosis in aseptic operation; To compare the biological characteristics of two kinds of cells so as to provide theoretical and related experimental evidence for defect, repair and treatment of alveolar bone.DESIGN: Controlled observation.SETTING: Department of Stomatology, Beijing Chaoyang Hospital, Capital Medical University; Institute of Orthopaedics,General Hospital of Chinese PLA.MATERIALS: Alveolar bone tissue obtained from normal persons and patients with chronic periodontitis confirmed in clinic was used in aseptic operation.METHODS: Alveolar bone tissue from normal persons and chronic periodontitis accompanied with osteoporosis were cultured in vitro. In the four cell lines (H-171, H-258, 261, 262) cultured primarily, cell lines H-171 and H-258 were chosen from periodonitis patients group and normal group respectively, and stained with histochemical and immunohistochemical methods. Cell morphology was observed. Doubling time and division index of two kinds of cells were calculated with cytometry. After several circles of passage, cryopreservation and revitalization, growth and aging rule of cells were compared.MAIN OUTCOME MEASURES: Passage and biological characteristics of two groups of cell lines.RESULTS: ①In the abnormal alveolar bone group, there was one successful primary culture and cells presented short-spindle shape. There were 3 times of cryopreservation and 3 times of revitalization. Its doubling time was 53.4 hours. The average division index was about 4‰. Cells well grew after 20 times of passages. ②In the normal alveolar bone group, there were 26 cases of cell lines cultured primarily, but passage was found in only 3 cases of cell lines due to various causes. There were 10 passages and the cells presented long-spindle shape. After two circles of cryopreservation and revitalization, the survival and growth rate of cells were inferior as compared with cell line H-171.Doubling time was 65.9 hours and the average division index was 3.5‰. ③Both two kinds of cells adhered the wall, with the characteristics of osteoblasts: AKP, toluidine blue, PAS, tetracycline-labeled mineralized nodus, type Ⅰ collagen and BMP-2 immunohistochemical staining all presented positive.CONCLUSION: Both two kinds of cultured cells have the characteristics of osteoblasts. The growth speed of cell line H-171 is faster than that of cell line H-258. No obvious mutation is found in 20 passages. In the 8th generation of H-258,aging appears and growth speed becomes slow.

BACKGROUND: Elimination of antigenic substances from natural extracellular matrix with the integrity of the tissue structure retained renders the matrix to possess better biocompatibility and provides a cell culture environment close to conditions of the internal environment. Such materials are the primary choice for cell culture scaffold in tissue engineering.OBJECTIVE: To prepare human articular cartilage acellular matrix so as to provide a methodological basis for further study of articular cartilage acellular matrix as cell scaffold materials.DESIGN: A single sample study of bone tissues.SETTING: The experiment was performed in Institute of Orthopedics, General Hospital of PLA, between January and May in 2004. The specimens were obtained from patients requiring joint replacement for femoral neck fracture.MATERIAIS: The experiment was conducted in the Department of Orthopedics, General Hospital of PLA from January to May in 2004. Human articular cartilage specimens were obtained from the femoral head of patients with total hip arthroplasty for femoral neck fracture.METHODS: Totally 10 specimens of fresh articular cartilage(3.5 mm × 4. 5 mm × 2.0 mm) were obtained and freeze-dried for 12 hours. Cartilage acellular matrix was prepared using Triton X-100, Dnase and Rnase and identified by means of hematoxylin-eosin(HE) and safranine O staining and immunohistochemical staining for cartilage proteoglycan.MAIN OUTCOME MEASURES: Histological observation of the articular cartilage acellular matrix and immunohistochemical staining of cartilage proteoglycan.RESULTS: HE and safranine O staining both showed no cellular structure in the matrix with only recesses left by the removed cells. Immunohistochemical staining for cartilage proteoglycan yielded positive results, suggesting the presence of cartilage proteoglycan in the acellular matrix.CONCLUSION: Human articular cartilage acellular matrix can be prepared using the modified four-step procedures with detergent and enzymatic extraction with lyophilization, and the preserved cartilage proteoglycan in the material may retain good pressure resistance.

Objective To study the cooperative method of goat's cruciate ligament reconstruction by bone-tendon autograft with interference fixation. Methods A customized reamer was used to make the bottleneck-like femoral tunnel, and the patellar tendon-tibial tuberosity autograft was harvested. Make sufficient preparation for this operation. Results The operation was successful, the laboratory animals all survived without any postoperative infection. Conclusion Scientific and careful operative-cooperation can guarantee the successful operation.