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Objective To investigate the causes of recurrent hemoptysis one week after interventional treatment.Methods 56 patients with massive hemoptysis were included in this study.All patients underwent emergent interventional therapy, including angiography and embolization therapy of bronchial artery, intercostal artery, internal thoracic artery, external thoracic artery and phrenic artery via femoral artery puncture.Results 6 cases had rebleeding within one week after interventional therapy,including 2 cases with primary lung cancer,1 case with bronchiectasis,1 case with pulmonary tuberculosis,1 case with esophageal cancer after surgery,1 case with esophageal cancer after radiotherapy.Then, these patients once again underwent angiography and embolization therapy of bronchial artery,intercostal artery,internal thoracic artery,external thoracic artery and phrenic artery.Conclusion The use of vasoconstrictive drugs before intervention, diversification of pulmonary feeding artery, wide range of lesions, inappropriate embolic material and poor image quality can lead to recurrent hemoptysis after interventional treatment.
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BACKGROUND:Cartilage extracelular matrix with a large number of signaling molecule proteins and factors is likely to be an ideal material for tissue engineering cartilage. OBJECTIVE: To investigate the possibility of calcium alginate and cartilage extracelular matrix combined with microencapsulated stem cels derived from human umbilical cord Wharton’s jely to construct ectopic tissue-engineered cartilage in nude mice. METHODS: Microfilament suspension of the cartilage extracelular matrix was prepared. Human stem cels derived from Wharton’s jely of the umbilical cord were inoculated in to calcium alginate and cartilage extracelular matrix gel microspheres as experimental group. Stem cels derived from human umbilical cord Wharton’s jely were incubated in simple alginate gel microspheres as control group. After in vitro culture, the microspheres wereimplanted into the dorsal subcutaneous tissue of nude mice. Samples were taken after 4 weeks, respectively, for gross and histological observation. RESULTS AND CONCLUSION:The stem cels exhibited paralel-chondrocyte morphology in microspheres, which grew and proliferated quite wel during in vitro culture. A new paralel-cartilaginous tissue was found in the subcutaneous tissue 4 weeks after surgery in the experimental group, and the tissue was positive for hematoxylin-eosin, safranine O, toluidine blue and colagen II. A large number of paralel-chondrocytes and cartilage lacuna-like structures were observed under a microscope with no obvious inflammatory reaction around the microspheres. The control group showed the partial degradation of microspheres, surrounded by only a smal number of inflammatory cels and lymphocytes. Calcium alginate and cartilage extracelular matrix microspheres have a rather good histocompatibility which can be used to construct paralel-cartilaginous tissues by implanting stem cel-microspheric compound into the subcutaneous tissue of nude mice.
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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.
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Objective To probe the blood supply of liver metastasis by celiac artery,proper hepatic artery DSA.portal vein perfusion CT during superior mesenteric arterial poaography(PCTAP).Methods One hundred patients with liver metastases were examined prospectively by plain CT scan,multiphase enhanced CT scan,celiac arteriography and proper hepatic artefiography.Of them,56 patients were examined by PCTAP.All primary lesions wero confirmed by operation and(or)pathology examination.In order to investigate the blood supply of metastasis lesions.the software of Photoshop Was used to obtain the time-attenuation cugves(TDC)of tumor center,tumor edge,portal vein and normal liver parenchyma adjacent to the tumor to talculate liver perfusion for DSA image analysis,while a deconvolution model from CT perfusion software Was designed for the dual blood supply.Results DSA findings:TDC of proper hepatic arteriography showed:the mean peak concentration(K value)in tumor centers was(67±12)%,and it was(76±15)%for peritumor tissue,(51±10)%in normal liver parenchyma.TDC of celiac arteriogaphy showed that the contrast concentration of tumor centers and tumor edge increased fast in early stage.then maintained a slight upward plateau,in the meanwhile,the contrast concentration of normal liver parenchyma kept increasing slowly.PCTAP findings:tumors exhibited no enhancement during 30 s continued scans.Conciusion The blood supply of liver metastasis mainly comes from hepatic artery,but barely from portal vein.
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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.
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BACKGROUND:Choose an ideal vector for amplified chondrocytes arouses more and more attention during the construction of epiphyseal cartilage tissue engineering. OBJECTIVE:To explore the feasibility and performance of epiphyseal cartilage tissue engineering scaffold constructed by chitosan and extracellular matrix of cartilage. DESIGN,TIME AND SETTING:An in vitro study was performed at Department of Orthopedics,General Hospital of Chinese PLA from December 2007 to March 2003. MATERIALS:Chitosan(deacetylation:90%;Mr10?105) was provided by Haihui Bioengineering Company,Qingdao;articular cartilage of swine was collected from market. METHODS:Fresh porcine articular cartilages were obtained and shattered in the iso-osmia liquid. After pulverization and gradient centrifugation,3% artilage microfilament suspension was equally mixed with 2% chitosan. Three-dimensional porous scaffolds were fabricated using a simple freeze-drying method. MAIN OUTCOME MEASURES:After the second gradient ethanol treatment,the scaffolds were investigated by histological staining and scanning electron microscopy to measure aperture,porosity,and water absorption rate. MTT test was also done to assess cytotoxicity of the scaffolds. After induced by transforming growth factor-?1(TGF-?1) ,bone marrow mesenchymal stem cells(BMSCs) of rabbits were incubated onto the scaffolds. Cell proliferation and differentiation were analyzed using inverted microscopy and scanning electron microscopy. RESULTS:The three-dimensional porous scaffold had good pore interconnectivity with pore diameter(161?31) ?m,(90.1?1.6) % porosity and(2 361?132) % water absorption rate. The histological staining showed that toluidine blue,safranin O and anti-collagen II immunohistochemistry staining were positive. The intrinsic cytotoxicity assessment of the scaffolds using MTT test showed that the scaffolds had no cytotoxic effect on BMSCs. Most of the BMSCs attached and covered the surface of the scaffolds with matrix secretion. CONCLUSION:The three-dimensional porous scaffold constructed by extracellular matrix of cartilage and chitosan has good pore diameter and porosity,non-toxicity and good biocompatibility,so it is a suitable scaffold for epiphyseal cartilage tissue engineering.
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[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.
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OBJECTIVE To observe the effect of electrolyzed oxidizing water(EOW) on the surface sterilization of indoor environment.METHODS Sterilized the surface of ground,wall,table,mob and thermostat-controlled water-bath in cell culture room and good laboratory practice(GLP) with EOW,then to draw the materials and culture with culture dish for 48 h in a 37℃ incubator,and count the colony number.Sterilization method and grouping: group A treated as a control,group B sterilized with EOW,group C sterilized with ultraviolate ray for 30 min and group D first treated with ultraviolate ray for 30 min,then sterilized with EOW.RESULTS In group A,the bacteria were overgrew and formed flakiness in 10/10 culture dishes;1 colony was formed in group B,the sterilization effective rate was 90%.The bacteria culture of group C found no bacteria growing after sterilization with ultraviolate ray,however,sample from surface and culture after sterilization were seen bacteria,though the number of bacteria was less than group A.The bacteria culture outcome of group D was negative.CONCLUSIONS The EOW has a good sterilization effect,it is safe and untoxic,costly cheap and convenient to use,and fit to claim of environmental protection.
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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.
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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.
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<p><b>OBJECTIVE</b>To investigate the biological characteristics of cell lines of healthy and diseased human dental alveoli.</p><p><b>METHODS</b>Primary cell lines from either healthy or diseased human dental alveoli were obtained. Two cell lines, H-258 and H-171 derived from healthy and diseased human tissues respectively, were selected for morphological study and research on their growth and aging, using cell counting, and histochemical and immunohistochemical staining.</p><p><b>RESULTS</b>Primary cell lines were successfully established from innormal dental alveoli. After freezing and thawing for three times, cell growth was continued and no morphological alterations were observed. The doubling time was 53.4 hours and mean division index (MDI) was 4 per thousand. Cells were kept normal after twenty generations with no obvious reduction of doubling time and MDI. Of twenty-six primary cell lines derived from healthy human dental alveoli, only three cell lines achieved generation. After freezing and thawing for twice, cultured cells were still alive at a decreased growth speed, with doubling time of 85.9 hours and MDI of 3 per thousand. Both cell lines, H-171 and H-258, shared the characteristics of osteoblast.</p><p><b>CONCLUSIONS</b>Primary cell lines of diseased human dental alveoli show greater growth potential. All cell lines of dental alveoli share characteristics of osteoblast. The technique we developed may be put into practice for the treatment of abnormal dental alveoli.</p>
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Humans , Cell Division , Physiology , Cell Line , Tooth Socket , Cell BiologyABSTRACT
e: Objective:To establish a useable animal model for the purification and immunotherapy of HSP with the observation of the growth rule of S180 sarcome tumor tissue and the immunohistochemical expression of different HSP in mice. Methods:The tumor incidence?the role of growth?the survival time and immunohistochemical expression of different HSP were observed after the S180 sarcome cells were inoculated at Balb/C mice back subcutaneous in different dose. Results:Sarcoma were formed in 100%.There are significant difference between vary dose in survival time, The expression was positive in HSP60?70?90? and grp94, and HSP70 expression was enhanced whereas HSP90? expression was decreased after heat shock treated. Conclusion:The use of S180 sarcome in mice is a good model to observe the expression of different HSP and the tumor tissue is suitable to purify. The model can be also used in the library study of HSP tumor vaccine.
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Objectives:To investigate the effects of verapamil on serum induced proliferation of rabbit pigment epithelial(RPE) cells so as to search for simple and effective medicine on PVR. Methods:The rabbit RPE cells(passage 2 to 5) were cultured with various concentrations of verapamil in DMEM.The effects of verapamil on the cell cycle of RPE were analyzed with flow cytometry. Results:Verapamil significantly inhibited the serum induced proliferation of RPE cells, prevented RPE from G 1 phase transiting to S phase. Conclusions:Verapamil significantly inhibits RPE cell proliferation, and it may become a promising drug on PVR.
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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.
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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.
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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.
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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