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.

AIM: To observe the immunoregulation of all ogeneic cornea on the human peripheral blood T lymphocytes in vitro. METHODS: After Co-culture of human peripheral blood lymphocytes and allogeneic cornea in vitro, T lymphocytes were labeled by monoclonal ant ibody, and analyzed by fluorescent activated cell sorter (FACS). RESULTS: CD25 expression on T lymphocytes in control was 25.2%, after stimulated by the allogeneic cornea or PDB, CD25 expression on T lymphocy tes was 56.8% and 80.9%, respectively. After stimulated by the allogeneic corn ea, CD25 expression on CD 4+ or CD 8+ T lymphocytes were 67.3% and 52.3% , respectively. CONCLUSION: Allageneic cornea stimulates CD25 expression on huma n peripheral blood T lymphocytes, and the CD25 expression on CD 4+ T lymphocy tes is more prominent than CD 8+ T lymphocytes.

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.

Objective To investigate the application of the full-appointment and time-phased appointment with doctor in nursery triage management.Methods From November 2012 to December 2013, Hangzhou Xixi hospital had carried out the full-appointment and time-phased appointment medical service.The coincidence rate of seeing a doctor in every half hour was considered to be the indicator of the results of management mechanism.Obtained the per-month outpatient attendances and the coincidence rate by HIS software and then calculated the per-month increase rate of outpatient attendances created the line chart.Results During the assessment, the coincidence rate of seeing a doctor at the outpatient mainly was above 90%every month and accumulated to 93.75%.The overall coincidence rate of seeing a doctor at the main outpatient departments reached 90.75%, and also it improved as the outpatient attendances.Conclusions The coincidence rate can be an evaluation indicator in time-phased appointment management system, which helps shorten the waiting time and relieve the patients′difficulty in seeing doctors.

Objective To explore the inhibiting effect of superoxide dismutase (SOD) on the growth of hepatocellular cancer cell line HepG2. Methods By gene transfer technique, hepatocellular cancer cells (HepG2) were transfected with a retroviral vector containing human CuZnSOD cDNA. The elevated SOD gene expression of the transfected cells was compared with the parental and neo control cells. Results Compared to the control cells, cancer cells transfected with SOD gene showed an inhibited cell growth, a reduced number of cells in S phase and decreased clone forming ability in soft agar, as well as a smaller tumor size formed in nude mice.Conclusion The overexpression of CuZnSOD gene could, to certain extent, suppress the cell growth of hepatocellular cancer cell line HepG2.

Objective To study the effects of the p16 gene on tumor cell growth inhibition and cell cycle arrest. Methods The recombinant retroviral vector pLp16SN was constructed by cloning the human p16 cDNA into the retroviral vector pLXSN. Retroviruses with or without the p16 gene were obtained by transfecting pLp16SN and pLXSN vectors into PA317 cells. Bcap-37 human breast cancer cells were infected with these retroviruses followed by selection with G418. The expression of p16 was detected by Northern and Westem blots. Cell biological characteristics, including cell growth rate, cell cycle and tumorigenesis in nude mice were assessed.Results Both mRNA and protein expression of p16 in Bcap-37 cells transfected with retroviral vector containing the p16 gene were much higher than that in Bcap-37 cells transfected with empty vector or parental Bcap-37 cells. Cell overexpressing the p16 gene exhibited a slower rate of growth, a higher percentage of cells in the G1 phase, and smaller tumors in nude mice, compared with parental Bcap-37 cells and cells transfected with empty vector.Conclusion Overexpression of the p16 gene could suppress the growth of Bcap-37 breast cancer cells by arresting the cell cvcle at the G1 to S-phase.

The development of synthetic biology has greatly promoted the construction of microbial cell factories, providing an important strategy for green and efficient chemical production. However, the bottleneck of poor tolerance to harsh industrial environments has become the key factor hampering the productivity of microbial cells. Adaptive evolution is an important method to domesticate microorganisms for a certain period by applying targeted selection pressure to obtain desired phenotypic or physiological properties that are adapted to a specific environment. Recently, with the development of technologies such as microfluidics, biosensors, and omics analysis, adaptive evolution has laid the foundation for efficient productivity of microbial cell factories. Herein, we discuss the key technologies of adaptive evolution and their important applications in improvement of environmental tolerance and production efficiency of microbial cell factories. Moreover, we looked forward to the prospects of adaptive evolution to realize industrial production by microbial cell factories.
Metabolic Engineering ; Industrial Microbiology/methods* ; Synthetic Biology ; Environment ; Industry