The nutritional components and utilization value of such 6 wild vegetables as Gynura divaricata, Kalimeris indica, Artemisia argyi H. Lev. & Vaniot, Artemisia selengensis, Lysimachia clethroides and Taraxacum mongoli...The nutritional components and utilization value of such 6 wild vegetables as Gynura divaricata, Kalimeris indica, Artemisia argyi H. Lev. & Vaniot, Artemisia selengensis, Lysimachia clethroides and Taraxacum mongolicum were introduced, so as to promote the development and utilization of wild vegetable resources. The culture management of these 6 wild vegetables was also introduced, so as to provide reference for culture of wild vegetables.展开更多
This study aimed to explore simple culture techniques for symbiotic germination-promoting fungi and Armillariella mellea in artificial planting of Gastrodia elata, i.e., how to use a corner of ordinary house as inocul...This study aimed to explore simple culture techniques for symbiotic germination-promoting fungi and Armillariella mellea in artificial planting of Gastrodia elata, i.e., how to use a corner of ordinary house as inoculation room and cultivating site, how to use a pressure cooker for cooking food as sterilization tool, and how to use ordinary household heating as heating measures.展开更多
The new high-yielding cultural technique of rice was constituted by the high percentage of ear-bearing til-ler,high filled-grain number,high economic index and stable high-yielding panicle number,abbreviatedas"Th...The new high-yielding cultural technique of rice was constituted by the high percentage of ear-bearing til-ler,high filled-grain number,high economic index and stable high-yielding panicle number,abbreviatedas"Three Highs and One Stable"(THOS).It has been deliberated and summarized by Zhejiang Academy展开更多
After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resist...After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.展开更多
AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth pote...AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous immortalization.METHODS: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift assay.Activation of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme immunoassay.RESULTS: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely immortalized.SIPS cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP kinases.Interleukin-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP kinases.CONCLUSION: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.展开更多
AIM: To investigate FasL expression in hilar cholangiocarcinoma tissues and cultured cholangiocarcinoma cells, and to assess its ability to induce apoptosis. METHODS: We studied the expression of FasL by human hilar c...AIM: To investigate FasL expression in hilar cholangiocarcinoma tissues and cultured cholangiocarcinoma cells, and to assess its ability to induce apoptosis. METHODS: We studied the expression of FasL by human hilar cholangiocaroinomas tissues by immunohistochemistry, and the QBC939 cholangiocarcinoma cell line by RT-PCR, immunohistochemistry, and Western Blot. TUNEL and flow cytometry were used to detect apoptotic cells. RESULTS: Prevalent expression of FasL was detected in 39 resected hilar cholangiocarcinoma tissues. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumor. FasL mRNA and protein expressions in cholangiocarcinoma cells could induce Jurkat cells. CONCLUSION: Hilar cholangiocarcinomas may elude immunological surveillance by inducing, via Fas/FasL system, the apoptosis of activated lymphocytes.展开更多
Repetitive transcranial magnetic stimulation (rTMS) has been utilized as a therapeutic tool for neurodegenerative disorders including Alzheimer's disease. However, the precise mechanisms of its clinical effects rem...Repetitive transcranial magnetic stimulation (rTMS) has been utilized as a therapeutic tool for neurodegenerative disorders including Alzheimer's disease. However, the precise mechanisms of its clinical effects remain unknown. β-amyloid (Aβ) exhibits direct neurotoxic effects and is closely related to neuronal degeneration in Alzheimer's disease. Therefore, it has been hypothesized that the neuroprotective effects of rTMS are related to the mechanisms of protection against Aβ neurotoxicity. Organotypic hippocampal slices were prepared from 8-day old, Sprague Dawley rats. The tissue slices were exposed to 100 μmol/L Al3142 since day 12 in vitro with and without high-frequency (20 Hz) magnetic stimulation. Magnetic stimulation efficacy was evaluated by measuring neuronal nuclei (NeuN) protein expression and by observing cultures following propidium iodide fluorescence staining and bromodeoxyuridine (BrdU) immunohistochemistry. Lactate dehydrogenase activity was detected in the culture media to evaluate hippocampal neuronal damage. Our results demonstrated that high-frequency magnetic stimulation significantly reversed the reduction of NeuN protein expression because of Aβ1-42 exposure (P 〈 0.05) and significantly reduced the number of damaged cells in the hippocampal slices (P 〈 0.05). However, lactate dehydrogenase levels and anti-BrdU staining results did not reveal any statistical differences These findings indicate that high-frequency magnetic stimulation might have protective effect on hippocampal neurons from Aβ1-42 neurotoxicity.展开更多
INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisola...INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisolated and cultured crypt cells from embryonalintestines and aseptic animal intestine, but failed.We introduced normal rat epithelial cell line-IEC-6from the USA and its living condition for stablepassage was successfully established after trials. Thecell line was testified to be the small intestinalepithelial cell by electron microscopy,immunihistochemistry and enzymatic histoch-emistry. It has been applied to some relatedresearch work[17-21]. It was found that manyfactors were involved in the culture system. Ourpresent study focuses on the culture method and theinfluencing factors on IEC-6.展开更多
Objective\ To test the effect of hypothermia on Na+/H+ exchange, activated by shrinkage and cytoplasmic acidosis. Method\ Amiloride-sensitive Na+ influx in guinea pig red cells was traced with isotope 22Na an...Objective\ To test the effect of hypothermia on Na+/H+ exchange, activated by shrinkage and cytoplasmic acidosis. Method\ Amiloride-sensitive Na+ influx in guinea pig red cells was traced with isotope 22Na and intracellular Na+ concentration was measured by emission flame photometry. Result\ Amiloride-sensitive Na+ influx decreased linearly as a function of temperatures (about 37℃) in shrunken cells, but increased in acidified cells. The up-regulation of acid-induced Na+/H+ exchange by elevated temperature was enhanced by hypo-osmolarity. Less sensitivity of intracellular H+ site at 41℃ may be the mechanism for the inhibition of shrinkage-induced Na+/H+ exchange by elevated temperature. Heating-mediated explosive increase in the activity of acid-induced Na+/H+ exchange may be due to enhanced extracellular Na+ sensitivity and lower intracellular pH caused by acidic metabolites. Acid-induced Na+/H+ ewxchange contributes to cytoplasmic Na+ accumulation. Conclusion\ These two modes of Na+/H+ exchange with different response to elevated temperature may play different roles in the cellular pathogenesis of heatstroke.展开更多
OBJECTIVE: To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burn...OBJECTIVE: To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burns. METHODS: The proliferation and differentiation of corneal stem cells in vitro had been examined using colony-forming efficiency and immunohistochemistry. The stem cells had been cultured on amniotic membranes and transplanted to the limbal area for treating corneal burns. RESULTS: Corneal stem cells had a high proliferation capacity in primary and first passage, cytokeratin 3 was not expressed in primary culture but partly in first passage. The stem cells could proliferate to form cell layer on an amniotic membrane. When transplanted, stem cells could survive on limbus. After transplantation, ocular inflammation resolved, the cornea re-epithelialized, the stromal opacity reduced, the superficial neovascularity was lessened and the conjunctival fornix re-established. CONCLUSIONS: Ocular surface conditions could be improved by allograft of corneal stem cells cultured on amniotic membranes.展开更多
OBJECTIVE: To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts. METHODS: Osteoblas...OBJECTIVE: To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts. METHODS: Osteoblasts were isolated from the calvaria of a fetal rabbit through sequential collagenase digestion. In the attachment study, the osteoblasts labeled with 3H-leucine were incubated with the bio-derived bone materials in sterile microcentrifugable tubes for 15, 90 and 180 minutes, and 24 hours, respectively. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. In the proliferation study, the osteoblasts were cultured with the bio-derived bone materials for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. Osteoblasts were seeded on the bone graft materials for 60 or 120 minutes, 24 or 48 hours, and 3 or 7 days, then the co-culture was processed for scanning electron microscopy to observe the interaction of osteoblasts and the bio-derived bone materials. RESULTS: Osteoblasts attached to the bio-derived bone materials in a time-dependent manner. There were significantly (P展开更多
Background Control of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a pro...Background Control of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a promising method for the treatment of pituitary adenomas. In the present study, transfection efficiency of small interfering RNA (siRNA) was optimized in human prolactinoma cells. Methods First, a method was optimized to extract highly purified human prolactinoma cells in vitro. The extracted cells were verified to retain the physiological features of prolactin (PRL) secretion. Second, three conditions for siRNA transfection were tested by the evaluation of transfectfon efficiency and cell viability. The proper transfection condition was verified for human prolactinoma cells. Third, the siRNA for prolactin was transfected into the human prolactinoma cells, and the suppression of PRL mRNA was evaluated by quantitative real-time reverse transcription-PCR. Results The siRNA of 100 pmol with Lipofectamine 2000 of 5 μl for 1×10^6 cells was proved preferable, with transfection efficiency being 53.3% and cell viability being 69.7%. In the preliminary experiment the siRNA against PRL decreased the mRNA of PRL by 34.0%. Conclusion It is possible to inhibit hormone hypersecretion by RNA interference, that may eventually enable therapeutic siRNA drugs developed.展开更多
Objective To investigate the culture method of cytokines induced killer cells using CD3 monoclonal antibody,IL-2,IFN-γ and IL-1α in vitro.Methods Peripheral blood mononuclear cells (PBMC) were collected by a blood c...Objective To investigate the culture method of cytokines induced killer cells using CD3 monoclonal antibody,IL-2,IFN-γ and IL-1α in vitro.Methods Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 8 patients with refractory lymphoma,then expanded by priming them with recombinant IFN-γ,monoclonal antibody (mAb) to CD3 and human recombinant IL-1α,followed by adding recombinant IL-2 the next day.The CIK cells were counted on d1,4,7,10,13 of incubation,respectively.The phenotypic patterns were characterized before culture and on d13 of culture.Results CIK cells colony formed on d3.On d13,the CIK cells counting reached up to (7-18)×109(mean 12.7×109),multiplying 44-140 times(mean 98 times).Cell survival rate was over 90%.The average percentage of cells expressing CD3+,CD4+,CD8+ and CD3+ CD56+ were also increased from (50.9±3.5)%,(29.9±1.7)%,(41.3±3.2)%,(1.6±0.2)% to (90.2±1.6)%,(40.6±5.5)%,(52.8±4.9)% and (33.1±4.0)%,respectively.Conclusions CIK cells developed by the culture method have high in vitro proliferate rate and tumor-killing capacity.The simplicity of the operation and autogenic source of the cells indicate the method could be applied clinically treating patients with refractory lymphoma.展开更多
Several studies demonstrated the favorable effects of platelet rich plasma(PRP)on the skin and promoted its wide use in clinical practice.The growth factors stored in platelet alfa-granules allow for the tissue regene...Several studies demonstrated the favorable effects of platelet rich plasma(PRP)on the skin and promoted its wide use in clinical practice.The growth factors stored in platelet alfa-granules allow for the tissue regeneration and the main fields of application of PRP in current clinical practice are the cartilage and musculoskeletal defects,osteoarthritis and other bone disorders,chronic and difficult to heal wounds,and aesthetic procedures.The relevant number of different PRP preparation protocols may explain the inconsistency of the different clinical outcomes reported in the literature.Despite the technological advances in PRP preparation,the objective assessment of the clinical efficacy of PRP from the literature reports still is difficult due to the low homogeneity of the samples in terms of both inclusion criteria and size.Therefore,it might be useful to establish standardized and reproducible experimental models to confirm and objectively measure the effectiveness of the available clinical results.Many experimental investigations have been carried out to objectively assess the effectiveness of PRP and platelet gel on several tissues.As far as the skin is concerned,the studies carried out to date are limited to fibroblasts in in-vitro culture models or to collagen,vascular supply,epithelium,and hair follicle in in-vivo models.The skin,however,is a very complex organ,where different cell lines coexist and feature complex mutual interaction.A model that combines the advantages of both in-vitro and in-vivo cultures is the ex-vivo model.The demonstration of the platelet derived growth factors effects through the ex-vivo human full-thickness skin culture model is a keystone to support the evidence of the PRP effectiveness,as it represents an objective,fast,reproducible,and ethical investigational method.展开更多
The application of arbuscular mycorrhizal (AM) inocula in severely degraded wetlands could ensure success in restoration.Mycorrhizal fungi play an important role in plant individual's survival and development in a...The application of arbuscular mycorrhizal (AM) inocula in severely degraded wetlands could ensure success in restoration.Mycorrhizal fungi play an important role in plant individual's survival and development in a low nutrient condition.Based on the importance that mycorrhizal fungi have to their host plants,mycorrhizal inocula have been produced and applied in terrestrial ecosystems in order to let the plants become mycorrhizal.However,mycorrhizal inocula have not been used in wetland restorations,despite increasing evidence that mycorrhizal fungi are commonly found in various wetland systems and have the ability to survive under anoxic conditions.Evidence also shows that mycorrhizal fungal inocula in the soil could have been destroyed in the degraded wetland or could be destroyed during traditional wetland restoration process.Therefore,AM inocula production is strongly recommended for wetland restoration.In this paper,I will argue that AM inocula production is required when introduced recovery is necessary,and aeroponic culture technique is a preferable method to produce AM inocula.Last,a renewed wetland restoration flow chart is summarized.展开更多
基金Supported by Fund for Independent Innovation of Agricultural Science in Jiangsu Province[CX(15)1050]~~
文摘The nutritional components and utilization value of such 6 wild vegetables as Gynura divaricata, Kalimeris indica, Artemisia argyi H. Lev. & Vaniot, Artemisia selengensis, Lysimachia clethroides and Taraxacum mongolicum were introduced, so as to promote the development and utilization of wild vegetable resources. The culture management of these 6 wild vegetables was also introduced, so as to provide reference for culture of wild vegetables.
基金Supported by Key Natural Science Research Project of Sichuan Provincial Departmen of Education(12ZA102)~~
文摘This study aimed to explore simple culture techniques for symbiotic germination-promoting fungi and Armillariella mellea in artificial planting of Gastrodia elata, i.e., how to use a corner of ordinary house as inoculation room and cultivating site, how to use a pressure cooker for cooking food as sterilization tool, and how to use ordinary household heating as heating measures.
文摘The new high-yielding cultural technique of rice was constituted by the high percentage of ear-bearing til-ler,high filled-grain number,high economic index and stable high-yielding panicle number,abbreviatedas"Three Highs and One Stable"(THOS).It has been deliberated and summarized by Zhejiang Academy
基金the Natinnal Biotechnology Reseaxch Project of 863 High Technology, contract No. 101-01-01-02.
文摘After pre-culture and treatment of osmosis, cotyledons of immature peanut (Arachis hypogaea L.) zygotic embryos were transformed via particle bombardment with a plasmid containing a chimeric hph gene conferring resistance to hygromycin and a chimeric intron-gus gene. Selection for hygromycin resistant calluses and somatic embryos was initiated at 10th d post-bombardment on medium containing 10-25 mg/L hygromycin. Under continuous selection, hygromycin resistant plantlets were regenerated from somatic embryos and were recovered from nearly 1.6% of the bombarded cotyledons. The presence and integration of foreign DNA in regenerated hygromycin resistant plants was confirmed by PCR (polymerase chain reaction) for the intron-gus gene and by Southern hybridization of the hph gene. GUS enzyme activity was detected in leaflets from transgenic plants but not from control, non-transformed plants. The production of transgenic plants are mainly based on a newly improved somatic embryogenesis regeneration system developed by us.
基金Grant-in-Aid for Encouragement of Young Scientists from Japan Society for the Promotion of Science(to A.M.)Pancreas Research Foundation of Japan(to A.M.)
文摘AIM: Activated pancreatic stellate cells (PSCs) have been implicated in the pathogenesis of pancreatic fibrosis and inflammation. Primary PSCs can be subcultured only several times because of their limited growth potential. A continuous cell line may therefore be valuable in studying molecular mechanisms of these pancreatic disorders. The aim of this study was to establish a cell line of rat PSCs by spontaneous immortalization.METHODS: PSCs were isolated from the pancreas of male Wistar rats, and conventional subcultivation was performed repeatedly. Telomerase activity was measured using the telomere repeat amplification protocol. Activation of transcription factors was assessed by electrophoretic mobility shift assay.Activation of mitogen-activated protein (MAP) kinases was examined by Western blotting using anti-phosphospecific antibodies. Expression of cytokine-induced neutrophil chemoattractant-1 was determined by enzyme immunoassay.RESULTS: Conventional subcultivation yielded actively growing cells. One clone was obtained after limiting dilution,and designated as SIPS. This cell line has been passaged repeatedly more than 2 years, and is thus likely immortalized.SIPS cells retained morphological characteristics of primary,culture-activated PSCs. SIPS expressed α-smooth muscle actin, glial acidic fibrillary protein, vimentin, desmin, type Ⅰ collagen, fibronectin, and prolyl hydroxylases. Telomerase activity and p53 expression were negative. Proliferation of SIPS cells was serum-dependent, and stimulated with platelet-derived growth factor-BB through the activation of extracellular signal-regulated kinase. Interleukin-1β activated nuclear factor-κB, activator protein-1, and MAP kinases.Interleukin-1β induced cytokine-induced neutrophil chemoattractant-1 expression through the activation of nuclear factor-κB and MAP kinases.CONCLUSION: SIPS cells can be useful for in vitro studies of cell biology and signal transduction of PSCs.
文摘AIM: To investigate FasL expression in hilar cholangiocarcinoma tissues and cultured cholangiocarcinoma cells, and to assess its ability to induce apoptosis. METHODS: We studied the expression of FasL by human hilar cholangiocaroinomas tissues by immunohistochemistry, and the QBC939 cholangiocarcinoma cell line by RT-PCR, immunohistochemistry, and Western Blot. TUNEL and flow cytometry were used to detect apoptotic cells. RESULTS: Prevalent expression of FasL was detected in 39 resected hilar cholangiocarcinoma tissues. TUNEL staining disclosed a high level of cell death among lymphocytes infiltrating FasL positive areas of tumor. FasL mRNA and protein expressions in cholangiocarcinoma cells could induce Jurkat cells. CONCLUSION: Hilar cholangiocarcinomas may elude immunological surveillance by inducing, via Fas/FasL system, the apoptosis of activated lymphocytes.
文摘Repetitive transcranial magnetic stimulation (rTMS) has been utilized as a therapeutic tool for neurodegenerative disorders including Alzheimer's disease. However, the precise mechanisms of its clinical effects remain unknown. β-amyloid (Aβ) exhibits direct neurotoxic effects and is closely related to neuronal degeneration in Alzheimer's disease. Therefore, it has been hypothesized that the neuroprotective effects of rTMS are related to the mechanisms of protection against Aβ neurotoxicity. Organotypic hippocampal slices were prepared from 8-day old, Sprague Dawley rats. The tissue slices were exposed to 100 μmol/L Al3142 since day 12 in vitro with and without high-frequency (20 Hz) magnetic stimulation. Magnetic stimulation efficacy was evaluated by measuring neuronal nuclei (NeuN) protein expression and by observing cultures following propidium iodide fluorescence staining and bromodeoxyuridine (BrdU) immunohistochemistry. Lactate dehydrogenase activity was detected in the culture media to evaluate hippocampal neuronal damage. Our results demonstrated that high-frequency magnetic stimulation significantly reversed the reduction of NeuN protein expression because of Aβ1-42 exposure (P 〈 0.05) and significantly reduced the number of damaged cells in the hippocampal slices (P 〈 0.05). However, lactate dehydrogenase levels and anti-BrdU staining results did not reveal any statistical differences These findings indicate that high-frequency magnetic stimulation might have protective effect on hippocampal neurons from Aβ1-42 neurotoxicity.
基金Supported by the National Natural Science Foundation of China, No.39100119
文摘INTRODUCTIONCrypt epithelial cells in normal small intestineproliferate at a high speed. But they are verydifficult to culture in vitro and passage stably. A lotof studies have been done[1-16]. Some domestic labsisolated and cultured crypt cells from embryonalintestines and aseptic animal intestine, but failed.We introduced normal rat epithelial cell line-IEC-6from the USA and its living condition for stablepassage was successfully established after trials. Thecell line was testified to be the small intestinalepithelial cell by electron microscopy,immunihistochemistry and enzymatic histoch-emistry. It has been applied to some relatedresearch work[17-21]. It was found that manyfactors were involved in the culture system. Ourpresent study focuses on the culture method and theinfluencing factors on IEC-6.
文摘Objective\ To test the effect of hypothermia on Na+/H+ exchange, activated by shrinkage and cytoplasmic acidosis. Method\ Amiloride-sensitive Na+ influx in guinea pig red cells was traced with isotope 22Na and intracellular Na+ concentration was measured by emission flame photometry. Result\ Amiloride-sensitive Na+ influx decreased linearly as a function of temperatures (about 37℃) in shrunken cells, but increased in acidified cells. The up-regulation of acid-induced Na+/H+ exchange by elevated temperature was enhanced by hypo-osmolarity. Less sensitivity of intracellular H+ site at 41℃ may be the mechanism for the inhibition of shrinkage-induced Na+/H+ exchange by elevated temperature. Heating-mediated explosive increase in the activity of acid-induced Na+/H+ exchange may be due to enhanced extracellular Na+ sensitivity and lower intracellular pH caused by acidic metabolites. Acid-induced Na+/H+ ewxchange contributes to cytoplasmic Na+ accumulation. Conclusion\ These two modes of Na+/H+ exchange with different response to elevated temperature may play different roles in the cellular pathogenesis of heatstroke.
文摘OBJECTIVE: To investigate the proliferation and differentiation of cultured corneal stem cells and determine the effect of corneal stem cells cultured on amniotic membranes on the limbal area for treating corneal burns. METHODS: The proliferation and differentiation of corneal stem cells in vitro had been examined using colony-forming efficiency and immunohistochemistry. The stem cells had been cultured on amniotic membranes and transplanted to the limbal area for treating corneal burns. RESULTS: Corneal stem cells had a high proliferation capacity in primary and first passage, cytokeratin 3 was not expressed in primary culture but partly in first passage. The stem cells could proliferate to form cell layer on an amniotic membrane. When transplanted, stem cells could survive on limbus. After transplantation, ocular inflammation resolved, the cornea re-epithelialized, the stromal opacity reduced, the superficial neovascularity was lessened and the conjunctival fornix re-established. CONCLUSIONS: Ocular surface conditions could be improved by allograft of corneal stem cells cultured on amniotic membranes.
文摘OBJECTIVE: To study the effect of bio-derived bones, as substitutes of autogenous bone grafts and demineralized cadaver bones, on the attachment, spreading and proliferation of isolated osteoblasts. METHODS: Osteoblasts were isolated from the calvaria of a fetal rabbit through sequential collagenase digestion. In the attachment study, the osteoblasts labeled with 3H-leucine were incubated with the bio-derived bone materials in sterile microcentrifugable tubes for 15, 90 and 180 minutes, and 24 hours, respectively. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. In the proliferation study, the osteoblasts were cultured with the bio-derived bone materials for 24 hours and 3H-thymidine was added during the last 2 hours of the incubation. The attached cells were collected and the radioactivity was measured with liquid scintillation spectrometry. Osteoblasts were seeded on the bone graft materials for 60 or 120 minutes, 24 or 48 hours, and 3 or 7 days, then the co-culture was processed for scanning electron microscopy to observe the interaction of osteoblasts and the bio-derived bone materials. RESULTS: Osteoblasts attached to the bio-derived bone materials in a time-dependent manner. There were significantly (P
文摘Background Control of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a promising method for the treatment of pituitary adenomas. In the present study, transfection efficiency of small interfering RNA (siRNA) was optimized in human prolactinoma cells. Methods First, a method was optimized to extract highly purified human prolactinoma cells in vitro. The extracted cells were verified to retain the physiological features of prolactin (PRL) secretion. Second, three conditions for siRNA transfection were tested by the evaluation of transfectfon efficiency and cell viability. The proper transfection condition was verified for human prolactinoma cells. Third, the siRNA for prolactin was transfected into the human prolactinoma cells, and the suppression of PRL mRNA was evaluated by quantitative real-time reverse transcription-PCR. Results The siRNA of 100 pmol with Lipofectamine 2000 of 5 μl for 1×10^6 cells was proved preferable, with transfection efficiency being 53.3% and cell viability being 69.7%. In the preliminary experiment the siRNA against PRL decreased the mRNA of PRL by 34.0%. Conclusion It is possible to inhibit hormone hypersecretion by RNA interference, that may eventually enable therapeutic siRNA drugs developed.
文摘Objective To investigate the culture method of cytokines induced killer cells using CD3 monoclonal antibody,IL-2,IFN-γ and IL-1α in vitro.Methods Peripheral blood mononuclear cells (PBMC) were collected by a blood cell separator from 8 patients with refractory lymphoma,then expanded by priming them with recombinant IFN-γ,monoclonal antibody (mAb) to CD3 and human recombinant IL-1α,followed by adding recombinant IL-2 the next day.The CIK cells were counted on d1,4,7,10,13 of incubation,respectively.The phenotypic patterns were characterized before culture and on d13 of culture.Results CIK cells colony formed on d3.On d13,the CIK cells counting reached up to (7-18)×109(mean 12.7×109),multiplying 44-140 times(mean 98 times).Cell survival rate was over 90%.The average percentage of cells expressing CD3+,CD4+,CD8+ and CD3+ CD56+ were also increased from (50.9±3.5)%,(29.9±1.7)%,(41.3±3.2)%,(1.6±0.2)% to (90.2±1.6)%,(40.6±5.5)%,(52.8±4.9)% and (33.1±4.0)%,respectively.Conclusions CIK cells developed by the culture method have high in vitro proliferate rate and tumor-killing capacity.The simplicity of the operation and autogenic source of the cells indicate the method could be applied clinically treating patients with refractory lymphoma.
文摘Several studies demonstrated the favorable effects of platelet rich plasma(PRP)on the skin and promoted its wide use in clinical practice.The growth factors stored in platelet alfa-granules allow for the tissue regeneration and the main fields of application of PRP in current clinical practice are the cartilage and musculoskeletal defects,osteoarthritis and other bone disorders,chronic and difficult to heal wounds,and aesthetic procedures.The relevant number of different PRP preparation protocols may explain the inconsistency of the different clinical outcomes reported in the literature.Despite the technological advances in PRP preparation,the objective assessment of the clinical efficacy of PRP from the literature reports still is difficult due to the low homogeneity of the samples in terms of both inclusion criteria and size.Therefore,it might be useful to establish standardized and reproducible experimental models to confirm and objectively measure the effectiveness of the available clinical results.Many experimental investigations have been carried out to objectively assess the effectiveness of PRP and platelet gel on several tissues.As far as the skin is concerned,the studies carried out to date are limited to fibroblasts in in-vitro culture models or to collagen,vascular supply,epithelium,and hair follicle in in-vivo models.The skin,however,is a very complex organ,where different cell lines coexist and feature complex mutual interaction.A model that combines the advantages of both in-vitro and in-vivo cultures is the ex-vivo model.The demonstration of the platelet derived growth factors effects through the ex-vivo human full-thickness skin culture model is a keystone to support the evidence of the PRP effectiveness,as it represents an objective,fast,reproducible,and ethical investigational method.
文摘The application of arbuscular mycorrhizal (AM) inocula in severely degraded wetlands could ensure success in restoration.Mycorrhizal fungi play an important role in plant individual's survival and development in a low nutrient condition.Based on the importance that mycorrhizal fungi have to their host plants,mycorrhizal inocula have been produced and applied in terrestrial ecosystems in order to let the plants become mycorrhizal.However,mycorrhizal inocula have not been used in wetland restorations,despite increasing evidence that mycorrhizal fungi are commonly found in various wetland systems and have the ability to survive under anoxic conditions.Evidence also shows that mycorrhizal fungal inocula in the soil could have been destroyed in the degraded wetland or could be destroyed during traditional wetland restoration process.Therefore,AM inocula production is strongly recommended for wetland restoration.In this paper,I will argue that AM inocula production is required when introduced recovery is necessary,and aeroponic culture technique is a preferable method to produce AM inocula.Last,a renewed wetland restoration flow chart is summarized.