Two new dihydropyrones,rhytismatones C(1)and D(2),and a known compound,penicillenol A1(3),were isolated from the co-culture broth of the deep-sea-derived fungus Penicillium crustosum PRB-2 and Suaeda salsa-derived end...Two new dihydropyrones,rhytismatones C(1)and D(2),and a known compound,penicillenol A1(3),were isolated from the co-culture broth of the deep-sea-derived fungus Penicillium crustosum PRB-2 and Suaeda salsa-derived endophytic fungus Peni-cillium citrinum HDN11-186.Their structures were elucidated through comprehensive analysis of nuclear magnetic resonance(NMR)spectra and mass spectra.The absolute configurations of new compounds were determined by calculating the electronic circular di-chroism(ECD)spectrum.UPLC-MS data showed that compounds 1–3 could only be detected in the media of co-culture,suggesting new biosynthetic pathways were activated in the co-cultured fungi.Compound 1 showed obvious antibacterial activities against Pro-teus sp.MMBC-1002 and Bacillus subtilis MMBC-1004 with minimum inhibitory concentration(MIC)both at 25μmolL^(-1).展开更多
Rice-duck co-culture is an integrated farming technology that benefits rice production, grain quality, and ecological sustainability in paddy fields. However, little is known about the effects of rice-duck co-culture ...Rice-duck co-culture is an integrated farming technology that benefits rice production, grain quality, and ecological sustainability in paddy fields. However, little is known about the effects of rice-duck co-culture on enzyme activity involved in the biosynthesis of 2-acetyl-1-pyrroline (2-AP), the volatile that gives fragrant rice its' distinctive and sought-after aroma. The present study aimed to examine the influence of rice-duck co-culture on the photosynthesis, yield, grain quality, rice aroma, and the enzymes involved in 2-acetyl-1-pyrroline biosynthesis in the cultivar Meixiangzhan 2 during the early and late rice growing seasons of 2016 in Guangzhou, China. We compared the rice grown in paddy fields with and without ducks. We found that rice-duck co-culture not only improved the yield and quality of fragrant rice grain, but also promoted the precursors of 2-AP biosynthesis formation and 2-AP accumulation in the grain. Grain 2-AP content in rice-duck co-culture was noticeably increased with 9.60% and 20.81% in early and late seasons, respectively. Proline and pyrroline-5-carboxylic acid (P5C) (precursors of 2-AP biosynthesis) and the activity of enzymes such as proline dehydrogenase (ProDH), ornithine aminotransferase (OAT) and Δ1 pyrroline-5-carboxylic acid synthetase (P5CS) were all improved by 10.15%–12.99%, 32.91%–47.75%, 17.81%–26.71%, 6.25%–21.78%, and 10.58%–38.87% under rice-duck co-culture in both seasons, respectively. Overall, our results suggest that rice-duck co-culture is an environmentally-friendly and sustainable approach to improving rice aroma and grain quality of fragrant rice.展开更多
Exchange of nitrogen and phosphorus across sediment-water interface plays an important role in the management of nutrient recycling in the aquaculture pond. In this study, a plot experiment was conducted to study the ...Exchange of nitrogen and phosphorus across sediment-water interface plays an important role in the management of nutrient recycling in the aquaculture pond. In this study, a plot experiment was conducted to study the effect of rice-catfish/shrimp co-culture on the micro-profile of oxygen (O2), pH and nutrient exchange across sediment-water interface in the intensive culture ponds. The results showed that rice-catfish co-culture increased the concentration and penetrating depth of O2, but decreased the pH value across the sediment-water interface, compared with catfish monoculture. Additional rice cultivation significantly reduced the flux rates of ammonium (NH4+) and nitrate (NO3-) across sediment-water interface in the catfish and shrimp ponds. The flux rates of NO2 - and soluble phosphorus (PO43-) showed no significant difference between rice-catfish/shrimp co-culture ponds and catfish/shrimp monoculture ponds. Rice only affected the dissolved inorganic nitrogen and phosphorus fractions in the sediment. The concentrations of NH4 + were significantly lower in the sediment of co-culture ponds than in the monoculture ponds. Additional rice cultivation also significantly reduced the content and percentage of dissolved inorganic phosphorus in the sediment of catfish ponds.展开更多
Rice-fish co-culture has gained increasing attention to remediate the negative environmental impacts induced by intensive aquaculture. However, the effect of rice-fish co-culture on oxygen depletion has rarely been in...Rice-fish co-culture has gained increasing attention to remediate the negative environmental impacts induced by intensive aquaculture. However, the effect of rice-fish co-culture on oxygen depletion has rarely been investigated. We constructed a rice-fish co-culture system in yellow catfish(Pelteobagrus fulvidraco) and freshwater shrimp(Macrobrachium nipponense) ponds using a new high-stalk rice variety, and conducted a field experiment to investigate the effect of rice-fish co-culture on water parameters and oxygen consumption. The results showed that rice-fish co-culture reduced the nutrients(total nitrogen, ammonia-N, total phosphorous and potassium) and the dissolved oxygen content in fish and shrimp ponds. However, they showed similar seasonal change of dissolved oxygen in the water of fish and shrimp ponds. Rice-fish co-culture reduced the total amount of oxygen consumption and optimized the oxygen consumption structure in pond. The respiration rates in water and sediment were significantly reduced by 66.1% and 31.7% in the catfish pond, and 64.4% and 38.7% in the shrimp pond, respectively, by additional rice cultivation. Rice-fish co-culture decreased the proportions of respiration in sediment and water, and increased the proportion of fish respiration. These results suggest that rice-fish co-culture is an efficient way to reduce hypoxia in intensive culture pond.展开更多
By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein en-dothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of trans-planted hMSCs for the treatment of avascu...By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein en-dothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of trans-planted hMSCs for the treatment of avascular ni ANFH, the effect of hMSCs on survival, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition were investigated in vitro. hMSCs and HUVECs were cultured and identified in vitro. Three kinds of conditioned media, CdM-CdMNOR, CdM-CdMHYP and HUVEC-CdMHYP were pre-pared. HUVECs were cultured with these conditioned media under hypoxia. The survival rate, apop-tosis rate, migration and angiogenesis of HUVECs were respectively detected by CCK-8, flow cy-tometry, Transwell and tube formation assay. The content of SDF-1α, VEGF and IL-6 in CdM was determined by ELISA. Our results showed that hMSCs and HUVECs were cultured and identified successfully. Compared with MSC-CdMNOR and HUVEC-CdMHYP groups, the survival rate, migra-tion and angiogenesis of HUVECs in MSC-CdMHYP group were significantly increased while the apoptosis rate was declined (P<0.05). Moreover, the expression of SDF-1α, VEGF and IL-6 in MSC-CdMHYP group was up-regulated. Under hypoxia, the apoptosis of HUVECs was inhibited while survival, migration and angiogenesis were improved by co-culture of hMSCs and HUVECs. The underlying mechanism may be that hMSCs could secrete a number of cytokines and improve niche, which might be helpful in the treatment of femoral head necrosis.展开更多
Summary: An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cu...Summary: An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0. 4 % bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73 % developed to the morula stage and 67.21 % cavitated to blastocysts with 59. 74 % hatching, as compared with 61. 34 % to morula stage, 48. 47 % to blastocysts and none hatching in the controls, respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.展开更多
Monoculture of sea cucumber(pond S) and polyculture of shrimp with sea cucumber(pond SS) were established to evaluate the effect of shrimp on the environmental conditions of sea cucumber farming pond. Contributions of...Monoculture of sea cucumber(pond S) and polyculture of shrimp with sea cucumber(pond SS) were established to evaluate the effect of shrimp on the environmental conditions of sea cucumber farming pond. Contributions of sediment organic matter(SOM2) resuspended from benthic sediment and the suspended particulate organic matter(SPOM) deposited from the water column to the precipitated organic matter(SOM1) collected with sediment traps were estimated with carbon stable isotope analysis. The results showed that the levels of SPOM and SOM2 in pond SS significantly decreased in comparison with those in pond S at the end of experiment(P < 0.05), indicating that co-culturing shrimp in sea cucumber farming pond could purify the farming water. Carbon stable isotope analysis showed that the proportion of SOM2 in SOM1 in pond SS(84.97% ± 0.38%) was significantly lower than that in pond S(95.20% ± 0.30%)(P < 0.05), suggesting that the resuspension of organic matter from benthic sediment into overlying water was reduced in polyculture pond. In contrast, the proportion of SPOM in SOM1 in pond SS(15.03% ± 0.38%) was significantly higher than that in pond S(4.80% ± 0.30%)(P < 0.05), indicating that the sedimentation of SPOM from water column was enhanced in pond SS owing to the biodeposition effect of shrimp.展开更多
The study investigated the effects of pulsed electromagnetic fields (PEMFs) of different frequencies on the gene expression of receptor activator of nuclear factor kappa B (RANK) and Nuclear factor of activated T-cell...The study investigated the effects of pulsed electromagnetic fields (PEMFs) of different frequencies on the gene expression of receptor activator of nuclear factor kappa B (RANK) and Nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) in rat osteoblast and osteoclast co-cultured model. Osteoblast-like cells were isolated from calvariae of Newborn Sprague Dawley rats (SD rats), while osteoclast-like cells were obtained from femora and tibiae of five weeks old SD rats. After 1 days of co-culture, the cells were exposed to premarin (E2) and different frequencies of PEMFs (8 Hz and 16 Hz, respectively) for 3 days. The expression of RANK and NFATc1 mRNA was analysed with realtime quantitative polymerase chain reaction. The gene expression of RANK and NFATc1 in the E2, PEMF with 8 Hz and 16 Hz group was significantly lower than that in the control group respectively. The gene expression of NFATc1 in the PEMF with 8 Hz group was significantly lower than that in the control group and PEMF with 16 Hz group. The study indicates that PEMF with 8 Hz could regulate the gene expression of RANK and NFATc1 in co-cultured model.展开更多
BACKGROUND: Animal experiments and clinical studies about tissue engineering method applied to repair nerve injury mainly focus on seeking ideal artificial nerve grafts, nerve conduit and seed cells. Autologous nerve,...BACKGROUND: Animal experiments and clinical studies about tissue engineering method applied to repair nerve injury mainly focus on seeking ideal artificial nerve grafts, nerve conduit and seed cells. Autologous nerve, allogeneic nerve and xenogeneic nerve are used to bridge nerve defects, it is one of the methods to promote the repair of nerve injury by culturing and growing Schwann cells, which can secrete various neurotrophic factor activities, in the grafts. OBJECTIVE: To observe the effect of acellular nerve grafts co-cultured with Schwann cells in repairing defects of sciatic nerve. DESIGN: An observational comparative study. SETTING: Tissue Engineering Laboratory of China Medical University. MATERIALS: The experiment was carried out in the Tissue Engineering Laboratory of China Medical University between April 2004 and April 2005. Forty neonatal Sprague-Dawley rats of 5-8 days (either males or females) and 24 male Wistar rats of 180-220 g were provided by the experimental animal center of China Medical University. METHODS: ① Culture of Schwann cells: The bilateral sciatic nerves and branchial plexus were isolated from the 40 neonatal SD rats. The sciatic nerves were enzymatically digested with collagenase and dispase, isolatd, purified and cultured with the method of speed-difference adhersion, and identified with the SABC immunohistochemical method. ② Model establishment: In vitro Schwann cells were microinjected into 10-mm long acellular nerve grafts repairing a surgically created gap in the rat sciatic nerve. According to the different grafted methods, the animals were randomly divided into three groups: autografts (n=8), acellular nerve grafts (n=8), or acellular nerve grafts with Schwann cells (n=8). ③ The regenerated nerve fiber number and average diameter of myeline sheath after culture were statistically anlayzed. MAIN OUTCOME MEASURES: ① The regenerated nerve ultrastructure, total number and density of myelinated nerve fibers, and the thickness of myeline sheath were observed under electron microscope. ② The images were processed with the Mias-1000 imaging analytical system to calculate the number of myelinated nerve fibers, and the thickness of myeline sheath. RESULTS: All the 24 Wistar rats were involved in the analysis of results. ① Results observed under transmission electron microscope: The regenerated myelinated nerve fibers in the group of acellular nerve grafts with Schwann cells were more even than those in the group of acellular nerve grafts, the number of myelinated nerve fibers and thickness of myelin sheath were close to those in the allografts group (P > 0.05), but significantly different from those in the group of acellular nerve grafts (P < 0.05). ② Results observed under scanning electron microscope: A great amount of Schwann cells with two polars were observed in the group of grafts with Schwann cells, the feature of cultured Schwann cells showed shoulder by shoulder, head to head. ③ The number of myelinated nerve fibers and thickness of myelin sheath analyzed by Mias-1000 imaging system in the group of acellular nerve grafts with Schwann cells were close to those in the autografts group (P > 0.05), but significantly different from those in the group of acellular nerve grafts (P < 0.05). CONCLUSION: Host axonal regeneration is significantly increased after implant of acellular nerve grafts. Acellular nerve grafts with Schwann cells offers a novel approach for repairing the gap of nerve defect.展开更多
Schwann cells play an important role in the peripheral nervous system, especially in nerve repair following injury, so artificial nerve regeneration requires an effective technique for obtaining purified Schwann cells...Schwann cells play an important role in the peripheral nervous system, especially in nerve repair following injury, so artificial nerve regeneration requires an effective technique for obtaining purified Schwann cells. In vivo and in vitro pre-degeneration of peripheral nerves have been shown to obtain high-purity Schwann cells. We believed that in vitro pre-degeneration was simple and controllable, and available for the clinic. Thus, we co-cultured the crushed sciatic nerves with bone marrow-derived cells in vitro. Results demonstrated that, 3 hours after injury, a large number of mononuclear cells moved to the crushed nerves and a large number of bone marrow-derived cells infiltrated the nerve segments. These changes promoted the degradation of the nerve segments, and the dedifferentiation and proliferation of Schwann cells. Neural cell adhesion molecule and glial fibrillary acidic protein expression were detected in the crushed nerves. Schwann cell yield was 9.08 ± 2.01 × 104/mg. The purity of primary cultured Schwann cells was 88.4 ± 5.79%. These indicate a successful new method for obtaining Schwann cells of high purity and yield from adult crushed sciatic nerve using bone marrow-derived cells.展开更多
In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth...In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth supporting substrate. Various physio-chemical and nutritional variables were optimized using classical and completely randomized designs for induced production of exoglucanase, and β-glucosidase from the co-culture of Trichoderma viride and Ganoderma lucidum in solid state fermentation (SSF). Analysis profile showed that when the conditions of the SSF medium containing 15 g corn stover substrate (50% w/w moisture) inoculated with 6 mL of inoculum were optimal, the maximum productions of exoglucanase (485 ± 6.5 U/mL) and β-glucosidase (255 ± 3.3 U/mL) were recorded after 5 days of incubation at pH 6 and 35°C.展开更多
Purpose:To examine selected putative virulent prop-erties of mutans streptococci in two groups with dif-ferent caries activity and to examine co-culture hy-bridization of the strains in vitro. Methods: A set of strain...Purpose:To examine selected putative virulent prop-erties of mutans streptococci in two groups with dif-ferent caries activity and to examine co-culture hy-bridization of the strains in vitro. Methods: A set of strains from caries-free subjects (115) and another set from caries-active subjects (165) were isolated. Each strain was characterized for three virulence determi-nants. The clinical bacteria were then cocultured by three strains exhibiting the highest levels of virulence. Isolate colonies of last filial generation bacteria were enrichment-incubated and estimated for virulence again. RAPD-PCR and MLEE analyses were processed for parental bacteria and last filial genera-tion one. Results: No difference associated with caries activity of the subjects from whom the isolates origi-nated. Virulent properties of a filial generation strains was not different in the same generation, but was very different from their parent strains. Conclu-sion: The coexist properties of virulent polyclonal strain of MS may hold in a very general conditional sense in a dental plaque ecosystem in vivo, however, one of the co-culture strains may became dominant and displa- ced the others as the result of continuous ecological succession in vitro.展开更多
Objectives: Estrogens significantly contribute toward the growth and development of endometrial cancers. Two principal pathways have been implicated in the final steps of estrogen synthesis: the steroid sulfatase (STS...Objectives: Estrogens significantly contribute toward the growth and development of endometrial cancers. Two principal pathways have been implicated in the final steps of estrogen synthesis: the steroid sulfatase (STS) and aromatase pathways. In this study, we aimed to evaluate the possible effects of tumor-stromal interactions on local estrogen biosynthesis in endometrial cancer. We also assessed the biological effects of inhibitors of steroid sulfatase and aromatase in the co-culture system compared with usual monocultures. Methods/Materials: We isolated stromal cells from endometrial cancer patients to examine local biosynthesis of estrogens and tumor-stromal interactions. Next we examined the effects of steroid sulfatase inhibitor and aromatase inhibitor in monoculture of endometrial cancer cell line (Ishikawa) and in a co-culture system involving an Ishikawa cells and stromal cells. Results: Estrogen receptor and steroid sulfatase mRNA levels in cancer cells were significantly higher in the co-cultures compared with the monocultures of endometrial cancer cells. Estradiol and androstenediol concentrations were also significantly higher in the co-cultured cells. Proliferation of the cancer cells was significantly increased through the steroid sulfatase pathway, which metabolizes androgens, estrone sulfate, and estradiol sulfate as its substrates. However, its proliferation was significantly decreased by the treatment of steroid sulfatase or aromatase inhibitors. The significant growth inhibition by the steroid sulfatase and aromatase inhibitors were also observed in the co-culture system. Conclusions: We evaluated the effects of STS inhibitor and aromatase inhibitors on the proliferation of estrogen-dependent endometrial cancer cells. Considering that intratumoral estrogen metabolism plays an important role, our co-culture systems provide an environment similar to that of the tumor in living patients in terms of metabolism and synthesis of intratumoral estrogens. The results of this study may aid in achieving improved clinical responses from patients treated with STS inhibitors.展开更多
Many researchers have described that mesenchymal stem cells conditioned medium and immune cells conditioned medium have a clear whitening effect when they are used as cosmetic ingredients. In this study, we confirmed ...Many researchers have described that mesenchymal stem cells conditioned medium and immune cells conditioned medium have a clear whitening effect when they are used as cosmetic ingredients. In this study, we confirmed the whitening efficacy of various concentrations of immune cells and stem cell conditioned media. The author tried to study a conditioned medium that has a strong whitening effect even with a composition of less than 20% (the most used concentration in cosmetics). Because of the fact that the conditioned medium contains various cytokines and growth factors secreted by stem cells or immune cells, it is known to have effects such as wound healing, antioxidant, and whitening effect. Recently, stem cells have been used not only in the development of cosmetic raw materials but also in skincare procedures, and there are reports being released of cosmetics using immune cells conditioned medium. The concentration-dependent whitening effect equivalently increased as the concentration of the mono-cultured conditioned medium was obtained through the stem cells or immune cells culture. In the case of co-culture, whitening results are like the effect of positive control such as arbutin in the medium carrying only 10% of the co-cultured conditioned medium. It is possible that enhanced whitening efficiency in co-cultured conditioned medium leads to a major innovation in the global cosmetic market.展开更多
Bibenzyls,a kind of important plant polyphenols,have attracted growing attention for their broad and remarkable pharmacological activities.However,due to the low abundance in nature,uncontrollable and environmentally ...Bibenzyls,a kind of important plant polyphenols,have attracted growing attention for their broad and remarkable pharmacological activities.However,due to the low abundance in nature,uncontrollable and environmentally unfriendly chemical synthesis processes,these compounds are not readily accessible.Herein,one high-yield bibenzyl backbone-producing Escherichia coli strain was constructed by using a highly active and substrate-promiscuous bibenzyl synthase identified from Dendrobium officinale in combination with starter and extender biosynthetic enzymes.Three types of efficiently postmodifying modular strains were engineered by employing methyltransferases,prenyltransferase,and glycosyltransferase with high activity and substrate tolerance together with their corresponding donor biosynthetic modules.Structurally different bibenzyl derivatives were tandemly and/or divergently synthesized by co-culture engineering in various combination modes.Especially,a prenylated bibenzyl derivative(12)was found to be an antioxidant that exhibited potent neuroprotective activity in the cellular and rat models of ischemia stroke.RNA-seq,quantitative RT-PCR,and Western-blot analysis demonstrated that 12 could up-regulate the expression level of an apoptosis-inducing factor,mitochondria associated 3(Aifm3),suggesting that Aifm3 might be a new target in ischemic stroke therapy.This study provides a flexible plug-and-play strategy for the easy-to-implement synthesis of structurally diverse bibenzyls through a modular co-culture engineering pipeline for drug discovery.展开更多
Potassium 2-(1-hydroxypentyl)-benzoate(D,L-PHPB), a new drug candidate for ischemic stroke at the phase II clinic trial, has been shown to protect neurons by inhibiting oxidative injury and reducing neuron apoptosis i...Potassium 2-(1-hydroxypentyl)-benzoate(D,L-PHPB), a new drug candidate for ischemic stroke at the phase II clinic trial, has been shown to protect neurons by inhibiting oxidative injury and reducing neuron apoptosis in previous studies. But the mechanisms of D,L-PHPB remain to be studied.In this study, a neuron–astrocytes co-culture system was used to elucidate the roles of astrocytes in neuroprotection of D,L-PHPB under oxygen-glucose deprivation/reoxygenation(OGD/R) condition. Our data showed that D,L-PHPB reduced neuronal apoptosis in mono-culture system and this effect was enhanced in neuron–astrocyte co-culture system under the OGD/R condition. Meanwhile, D,L-PHPB obviously increased the levels of brain-derived neurotrophic factor(BDNF) and nerve growth factor(NGF), which were mainly secreted from astrocytes, in the co-culture system after OGD/R. The PI3K/AKT and ERK signaling pathways as well as the p-TRKA/B receptors were involved in the process. In addition, the levels of TNF-α and IL-1β secreted from astrocytes after OGD/R were markedly reduced after D,L-PHPB treatment, which was mainly due to the suppression of phosphorylated p38. In conclusion, the present study demonstrates that the neuroprotective effects of D,L-PHPB were improved by astrocytes, mainly mediated by increasing the release of BDNF/NGF and attenuating inflammatory cytokines.展开更多
In this work,a multi-functional analysis platform by coupling a microfluidic chip to a mass spectrometry(MS) detector was described.We constructed a three-dimensional tumor-endothelial co-culture model for simulating ...In this work,a multi-functional analysis platform by coupling a microfluidic chip to a mass spectrometry(MS) detector was described.We constructed a three-dimensional tumor-endothelial co-culture model for simulating drug resistance during tumor treatment.On this specially designed integrated platform,the first step was to prepare heterogeneous cell-encapsulated alginate microcapsules for threedimensional co-culture,and the second step was to achieve on-line perfusion culture and continuous drug stimulation on chip.It facilitates cell proliferation analysis and the collection of metabolism medium.After micro solid phase extraction column(SPE) pretreatment,subsequent mass spectrometry could detect drug metabolism.The high activity of two kinds of cells(A549 and HUVEC) shows the biocompatibility of the platform.Paclitaxel was used as a model drug,the distinctions of drug absorption between the mono-culture group and co-culture group were clearly observed by electrospray ionization quadrupole time-of-flight mass spectrometry(ESI-Q-TOF MS).Therefore,the integrated platform has shown promise as a high throughput,low cost for cell metabolism research and drug screening processes.展开更多
基金supported by the National Natural Science Foundation of China(No.41806167)the High-Level Talents Research Fund of Qingdao Agricultural University(No.665/1120034)+4 种基金the NSFC-Shandong Joint Fund(No.U1906212)the Major Project of the 14th Five-Year Plan(No.2022QNLM030003-1)the Natural Science Foundation of Shandong Province(No.ZR2021ZD28)the Hainan Provincial Joint Project of Sanya Yazhou Bay Science and Technology City(No.2021CXLH0012)the Youth Innovation Plan of Shandong Province(No.2019KJM004).
文摘Two new dihydropyrones,rhytismatones C(1)and D(2),and a known compound,penicillenol A1(3),were isolated from the co-culture broth of the deep-sea-derived fungus Penicillium crustosum PRB-2 and Suaeda salsa-derived endophytic fungus Peni-cillium citrinum HDN11-186.Their structures were elucidated through comprehensive analysis of nuclear magnetic resonance(NMR)spectra and mass spectra.The absolute configurations of new compounds were determined by calculating the electronic circular di-chroism(ECD)spectrum.UPLC-MS data showed that compounds 1–3 could only be detected in the media of co-culture,suggesting new biosynthetic pathways were activated in the co-cultured fungi.Compound 1 showed obvious antibacterial activities against Pro-teus sp.MMBC-1002 and Bacillus subtilis MMBC-1004 with minimum inhibitory concentration(MIC)both at 25μmolL^(-1).
基金supported by the Science and Technology Project of Guangdong Province (2015B090903077, 2016A020210094, 2017A090905030), Chinathe Science and Technology Project of Guangzhou (201604020062), China+1 种基金the Innovation Team Construction Project of Modern Agricultural Industry Technology System of Guangdong Province (2016LM1100), Chinathe Overseas Joint Doctoral Training Program of South China Agricultural University (2018LHPY010), China
文摘Rice-duck co-culture is an integrated farming technology that benefits rice production, grain quality, and ecological sustainability in paddy fields. However, little is known about the effects of rice-duck co-culture on enzyme activity involved in the biosynthesis of 2-acetyl-1-pyrroline (2-AP), the volatile that gives fragrant rice its' distinctive and sought-after aroma. The present study aimed to examine the influence of rice-duck co-culture on the photosynthesis, yield, grain quality, rice aroma, and the enzymes involved in 2-acetyl-1-pyrroline biosynthesis in the cultivar Meixiangzhan 2 during the early and late rice growing seasons of 2016 in Guangzhou, China. We compared the rice grown in paddy fields with and without ducks. We found that rice-duck co-culture not only improved the yield and quality of fragrant rice grain, but also promoted the precursors of 2-AP biosynthesis formation and 2-AP accumulation in the grain. Grain 2-AP content in rice-duck co-culture was noticeably increased with 9.60% and 20.81% in early and late seasons, respectively. Proline and pyrroline-5-carboxylic acid (P5C) (precursors of 2-AP biosynthesis) and the activity of enzymes such as proline dehydrogenase (ProDH), ornithine aminotransferase (OAT) and Δ1 pyrroline-5-carboxylic acid synthetase (P5CS) were all improved by 10.15%–12.99%, 32.91%–47.75%, 17.81%–26.71%, 6.25%–21.78%, and 10.58%–38.87% under rice-duck co-culture in both seasons, respectively. Overall, our results suggest that rice-duck co-culture is an environmentally-friendly and sustainable approach to improving rice aroma and grain quality of fragrant rice.
基金supported by the Natural Science Foundation of China(Grant Nos.41877548 and 31400379)Natural Science Foundation of Zhejiang Province of China(Grant No.LY15C030002)Innovation Program of Chinese Academy of Agricultural Sciences
文摘Exchange of nitrogen and phosphorus across sediment-water interface plays an important role in the management of nutrient recycling in the aquaculture pond. In this study, a plot experiment was conducted to study the effect of rice-catfish/shrimp co-culture on the micro-profile of oxygen (O2), pH and nutrient exchange across sediment-water interface in the intensive culture ponds. The results showed that rice-catfish co-culture increased the concentration and penetrating depth of O2, but decreased the pH value across the sediment-water interface, compared with catfish monoculture. Additional rice cultivation significantly reduced the flux rates of ammonium (NH4+) and nitrate (NO3-) across sediment-water interface in the catfish and shrimp ponds. The flux rates of NO2 - and soluble phosphorus (PO43-) showed no significant difference between rice-catfish/shrimp co-culture ponds and catfish/shrimp monoculture ponds. Rice only affected the dissolved inorganic nitrogen and phosphorus fractions in the sediment. The concentrations of NH4 + were significantly lower in the sediment of co-culture ponds than in the monoculture ponds. Additional rice cultivation also significantly reduced the content and percentage of dissolved inorganic phosphorus in the sediment of catfish ponds.
基金supported by the Natural Science Foundation of China(Grant No.31400379)Natural Science Foundation of Zhejiang Province of China(Grant No.LY15C030002)Innovation Program of Chinese Academy of Agricultural Sciences
文摘Rice-fish co-culture has gained increasing attention to remediate the negative environmental impacts induced by intensive aquaculture. However, the effect of rice-fish co-culture on oxygen depletion has rarely been investigated. We constructed a rice-fish co-culture system in yellow catfish(Pelteobagrus fulvidraco) and freshwater shrimp(Macrobrachium nipponense) ponds using a new high-stalk rice variety, and conducted a field experiment to investigate the effect of rice-fish co-culture on water parameters and oxygen consumption. The results showed that rice-fish co-culture reduced the nutrients(total nitrogen, ammonia-N, total phosphorous and potassium) and the dissolved oxygen content in fish and shrimp ponds. However, they showed similar seasonal change of dissolved oxygen in the water of fish and shrimp ponds. Rice-fish co-culture reduced the total amount of oxygen consumption and optimized the oxygen consumption structure in pond. The respiration rates in water and sediment were significantly reduced by 66.1% and 31.7% in the catfish pond, and 64.4% and 38.7% in the shrimp pond, respectively, by additional rice cultivation. Rice-fish co-culture decreased the proportions of respiration in sediment and water, and increased the proportion of fish respiration. These results suggest that rice-fish co-culture is an efficient way to reduce hypoxia in intensive culture pond.
基金supported by agrant from the National Natural Sciences Foundation of China(No.30750010)
文摘By co-culturing humm mesenchymal stem cells (hMSCs) and human umbilical rein en-dothelial cells (HUVECs) under hypoxia and creating a microenvironment similar to that of trans-planted hMSCs for the treatment of avascular ni ANFH, the effect of hMSCs on survival, apoptosis, migration and angiogenesis of human umbilical vein endothelial cells (HUVECs) under the hypoxic condition were investigated in vitro. hMSCs and HUVECs were cultured and identified in vitro. Three kinds of conditioned media, CdM-CdMNOR, CdM-CdMHYP and HUVEC-CdMHYP were pre-pared. HUVECs were cultured with these conditioned media under hypoxia. The survival rate, apop-tosis rate, migration and angiogenesis of HUVECs were respectively detected by CCK-8, flow cy-tometry, Transwell and tube formation assay. The content of SDF-1α, VEGF and IL-6 in CdM was determined by ELISA. Our results showed that hMSCs and HUVECs were cultured and identified successfully. Compared with MSC-CdMNOR and HUVEC-CdMHYP groups, the survival rate, migra-tion and angiogenesis of HUVECs in MSC-CdMHYP group were significantly increased while the apoptosis rate was declined (P<0.05). Moreover, the expression of SDF-1α, VEGF and IL-6 in MSC-CdMHYP group was up-regulated. Under hypoxia, the apoptosis of HUVECs was inhibited while survival, migration and angiogenesis were improved by co-culture of hMSCs and HUVECs. The underlying mechanism may be that hMSCs could secrete a number of cytokines and improve niche, which might be helpful in the treatment of femoral head necrosis.
文摘Summary: An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0. 4 % bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73 % developed to the morula stage and 67.21 % cavitated to blastocysts with 59. 74 % hatching, as compared with 61. 34 % to morula stage, 48. 47 % to blastocysts and none hatching in the controls, respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.
基金funded by the National Natural Science Foundation of China (Nos. 31172426 and 31372549)Ministry of Science and Technology of China (Nos. 2011BAD13B03 and 2012GA740001)supported by the Program for New Century Excellent Talents in University (NCET-11-0466)
文摘Monoculture of sea cucumber(pond S) and polyculture of shrimp with sea cucumber(pond SS) were established to evaluate the effect of shrimp on the environmental conditions of sea cucumber farming pond. Contributions of sediment organic matter(SOM2) resuspended from benthic sediment and the suspended particulate organic matter(SPOM) deposited from the water column to the precipitated organic matter(SOM1) collected with sediment traps were estimated with carbon stable isotope analysis. The results showed that the levels of SPOM and SOM2 in pond SS significantly decreased in comparison with those in pond S at the end of experiment(P < 0.05), indicating that co-culturing shrimp in sea cucumber farming pond could purify the farming water. Carbon stable isotope analysis showed that the proportion of SOM2 in SOM1 in pond SS(84.97% ± 0.38%) was significantly lower than that in pond S(95.20% ± 0.30%)(P < 0.05), suggesting that the resuspension of organic matter from benthic sediment into overlying water was reduced in polyculture pond. In contrast, the proportion of SPOM in SOM1 in pond SS(15.03% ± 0.38%) was significantly higher than that in pond S(4.80% ± 0.30%)(P < 0.05), indicating that the sedimentation of SPOM from water column was enhanced in pond SS owing to the biodeposition effect of shrimp.
文摘The study investigated the effects of pulsed electromagnetic fields (PEMFs) of different frequencies on the gene expression of receptor activator of nuclear factor kappa B (RANK) and Nuclear factor of activated T-cells cytoplasmic 1 (NFATc1) in rat osteoblast and osteoclast co-cultured model. Osteoblast-like cells were isolated from calvariae of Newborn Sprague Dawley rats (SD rats), while osteoclast-like cells were obtained from femora and tibiae of five weeks old SD rats. After 1 days of co-culture, the cells were exposed to premarin (E2) and different frequencies of PEMFs (8 Hz and 16 Hz, respectively) for 3 days. The expression of RANK and NFATc1 mRNA was analysed with realtime quantitative polymerase chain reaction. The gene expression of RANK and NFATc1 in the E2, PEMF with 8 Hz and 16 Hz group was significantly lower than that in the control group respectively. The gene expression of NFATc1 in the PEMF with 8 Hz group was significantly lower than that in the control group and PEMF with 16 Hz group. The study indicates that PEMF with 8 Hz could regulate the gene expression of RANK and NFATc1 in co-cultured model.
基金the National Natural Science Foundation of China, No. 30070775 a grant from the Scientific Research Foundation of Liaoning Department of Education, No. 2005L5371
文摘BACKGROUND: Animal experiments and clinical studies about tissue engineering method applied to repair nerve injury mainly focus on seeking ideal artificial nerve grafts, nerve conduit and seed cells. Autologous nerve, allogeneic nerve and xenogeneic nerve are used to bridge nerve defects, it is one of the methods to promote the repair of nerve injury by culturing and growing Schwann cells, which can secrete various neurotrophic factor activities, in the grafts. OBJECTIVE: To observe the effect of acellular nerve grafts co-cultured with Schwann cells in repairing defects of sciatic nerve. DESIGN: An observational comparative study. SETTING: Tissue Engineering Laboratory of China Medical University. MATERIALS: The experiment was carried out in the Tissue Engineering Laboratory of China Medical University between April 2004 and April 2005. Forty neonatal Sprague-Dawley rats of 5-8 days (either males or females) and 24 male Wistar rats of 180-220 g were provided by the experimental animal center of China Medical University. METHODS: ① Culture of Schwann cells: The bilateral sciatic nerves and branchial plexus were isolated from the 40 neonatal SD rats. The sciatic nerves were enzymatically digested with collagenase and dispase, isolatd, purified and cultured with the method of speed-difference adhersion, and identified with the SABC immunohistochemical method. ② Model establishment: In vitro Schwann cells were microinjected into 10-mm long acellular nerve grafts repairing a surgically created gap in the rat sciatic nerve. According to the different grafted methods, the animals were randomly divided into three groups: autografts (n=8), acellular nerve grafts (n=8), or acellular nerve grafts with Schwann cells (n=8). ③ The regenerated nerve fiber number and average diameter of myeline sheath after culture were statistically anlayzed. MAIN OUTCOME MEASURES: ① The regenerated nerve ultrastructure, total number and density of myelinated nerve fibers, and the thickness of myeline sheath were observed under electron microscope. ② The images were processed with the Mias-1000 imaging analytical system to calculate the number of myelinated nerve fibers, and the thickness of myeline sheath. RESULTS: All the 24 Wistar rats were involved in the analysis of results. ① Results observed under transmission electron microscope: The regenerated myelinated nerve fibers in the group of acellular nerve grafts with Schwann cells were more even than those in the group of acellular nerve grafts, the number of myelinated nerve fibers and thickness of myelin sheath were close to those in the allografts group (P > 0.05), but significantly different from those in the group of acellular nerve grafts (P < 0.05). ② Results observed under scanning electron microscope: A great amount of Schwann cells with two polars were observed in the group of grafts with Schwann cells, the feature of cultured Schwann cells showed shoulder by shoulder, head to head. ③ The number of myelinated nerve fibers and thickness of myelin sheath analyzed by Mias-1000 imaging system in the group of acellular nerve grafts with Schwann cells were close to those in the autografts group (P > 0.05), but significantly different from those in the group of acellular nerve grafts (P < 0.05). CONCLUSION: Host axonal regeneration is significantly increased after implant of acellular nerve grafts. Acellular nerve grafts with Schwann cells offers a novel approach for repairing the gap of nerve defect.
基金supported by the Key University Natural Science Research Project of Anhui Province of China,No.KJ2016A870
文摘Schwann cells play an important role in the peripheral nervous system, especially in nerve repair following injury, so artificial nerve regeneration requires an effective technique for obtaining purified Schwann cells. In vivo and in vitro pre-degeneration of peripheral nerves have been shown to obtain high-purity Schwann cells. We believed that in vitro pre-degeneration was simple and controllable, and available for the clinic. Thus, we co-cultured the crushed sciatic nerves with bone marrow-derived cells in vitro. Results demonstrated that, 3 hours after injury, a large number of mononuclear cells moved to the crushed nerves and a large number of bone marrow-derived cells infiltrated the nerve segments. These changes promoted the degradation of the nerve segments, and the dedifferentiation and proliferation of Schwann cells. Neural cell adhesion molecule and glial fibrillary acidic protein expression were detected in the crushed nerves. Schwann cell yield was 9.08 ± 2.01 × 104/mg. The purity of primary cultured Schwann cells was 88.4 ± 5.79%. These indicate a successful new method for obtaining Schwann cells of high purity and yield from adult crushed sciatic nerve using bone marrow-derived cells.
文摘In the present study, a co-culture technique was adopted with an aim to investigate a hyper production of exoglucanase, and β-glucosidase using cheap and easily available agro-industrial residue corn stover as growth supporting substrate. Various physio-chemical and nutritional variables were optimized using classical and completely randomized designs for induced production of exoglucanase, and β-glucosidase from the co-culture of Trichoderma viride and Ganoderma lucidum in solid state fermentation (SSF). Analysis profile showed that when the conditions of the SSF medium containing 15 g corn stover substrate (50% w/w moisture) inoculated with 6 mL of inoculum were optimal, the maximum productions of exoglucanase (485 ± 6.5 U/mL) and β-glucosidase (255 ± 3.3 U/mL) were recorded after 5 days of incubation at pH 6 and 35°C.
文摘Purpose:To examine selected putative virulent prop-erties of mutans streptococci in two groups with dif-ferent caries activity and to examine co-culture hy-bridization of the strains in vitro. Methods: A set of strains from caries-free subjects (115) and another set from caries-active subjects (165) were isolated. Each strain was characterized for three virulence determi-nants. The clinical bacteria were then cocultured by three strains exhibiting the highest levels of virulence. Isolate colonies of last filial generation bacteria were enrichment-incubated and estimated for virulence again. RAPD-PCR and MLEE analyses were processed for parental bacteria and last filial genera-tion one. Results: No difference associated with caries activity of the subjects from whom the isolates origi-nated. Virulent properties of a filial generation strains was not different in the same generation, but was very different from their parent strains. Conclu-sion: The coexist properties of virulent polyclonal strain of MS may hold in a very general conditional sense in a dental plaque ecosystem in vivo, however, one of the co-culture strains may became dominant and displa- ced the others as the result of continuous ecological succession in vitro.
文摘Objectives: Estrogens significantly contribute toward the growth and development of endometrial cancers. Two principal pathways have been implicated in the final steps of estrogen synthesis: the steroid sulfatase (STS) and aromatase pathways. In this study, we aimed to evaluate the possible effects of tumor-stromal interactions on local estrogen biosynthesis in endometrial cancer. We also assessed the biological effects of inhibitors of steroid sulfatase and aromatase in the co-culture system compared with usual monocultures. Methods/Materials: We isolated stromal cells from endometrial cancer patients to examine local biosynthesis of estrogens and tumor-stromal interactions. Next we examined the effects of steroid sulfatase inhibitor and aromatase inhibitor in monoculture of endometrial cancer cell line (Ishikawa) and in a co-culture system involving an Ishikawa cells and stromal cells. Results: Estrogen receptor and steroid sulfatase mRNA levels in cancer cells were significantly higher in the co-cultures compared with the monocultures of endometrial cancer cells. Estradiol and androstenediol concentrations were also significantly higher in the co-cultured cells. Proliferation of the cancer cells was significantly increased through the steroid sulfatase pathway, which metabolizes androgens, estrone sulfate, and estradiol sulfate as its substrates. However, its proliferation was significantly decreased by the treatment of steroid sulfatase or aromatase inhibitors. The significant growth inhibition by the steroid sulfatase and aromatase inhibitors were also observed in the co-culture system. Conclusions: We evaluated the effects of STS inhibitor and aromatase inhibitors on the proliferation of estrogen-dependent endometrial cancer cells. Considering that intratumoral estrogen metabolism plays an important role, our co-culture systems provide an environment similar to that of the tumor in living patients in terms of metabolism and synthesis of intratumoral estrogens. The results of this study may aid in achieving improved clinical responses from patients treated with STS inhibitors.
文摘Many researchers have described that mesenchymal stem cells conditioned medium and immune cells conditioned medium have a clear whitening effect when they are used as cosmetic ingredients. In this study, we confirmed the whitening efficacy of various concentrations of immune cells and stem cell conditioned media. The author tried to study a conditioned medium that has a strong whitening effect even with a composition of less than 20% (the most used concentration in cosmetics). Because of the fact that the conditioned medium contains various cytokines and growth factors secreted by stem cells or immune cells, it is known to have effects such as wound healing, antioxidant, and whitening effect. Recently, stem cells have been used not only in the development of cosmetic raw materials but also in skincare procedures, and there are reports being released of cosmetics using immune cells conditioned medium. The concentration-dependent whitening effect equivalently increased as the concentration of the mono-cultured conditioned medium was obtained through the stem cells or immune cells culture. In the case of co-culture, whitening results are like the effect of positive control such as arbutin in the medium carrying only 10% of the co-cultured conditioned medium. It is possible that enhanced whitening efficiency in co-cultured conditioned medium leads to a major innovation in the global cosmetic market.
基金supported by the National Key Research and Development Program of China(2020YFA0908000)CAMS Innovation fund for Medical Sciences(CIFMS-2021-I2M1-028 and CIFMS-2021-I2M-1-029,China)Beijing Key Laboratory of non-Clinical Drug Metabolism and PK/PD Study(Z141102004414062,China)。
文摘Bibenzyls,a kind of important plant polyphenols,have attracted growing attention for their broad and remarkable pharmacological activities.However,due to the low abundance in nature,uncontrollable and environmentally unfriendly chemical synthesis processes,these compounds are not readily accessible.Herein,one high-yield bibenzyl backbone-producing Escherichia coli strain was constructed by using a highly active and substrate-promiscuous bibenzyl synthase identified from Dendrobium officinale in combination with starter and extender biosynthetic enzymes.Three types of efficiently postmodifying modular strains were engineered by employing methyltransferases,prenyltransferase,and glycosyltransferase with high activity and substrate tolerance together with their corresponding donor biosynthetic modules.Structurally different bibenzyl derivatives were tandemly and/or divergently synthesized by co-culture engineering in various combination modes.Especially,a prenylated bibenzyl derivative(12)was found to be an antioxidant that exhibited potent neuroprotective activity in the cellular and rat models of ischemia stroke.RNA-seq,quantitative RT-PCR,and Western-blot analysis demonstrated that 12 could up-regulate the expression level of an apoptosis-inducing factor,mitochondria associated 3(Aifm3),suggesting that Aifm3 might be a new target in ischemic stroke therapy.This study provides a flexible plug-and-play strategy for the easy-to-implement synthesis of structurally diverse bibenzyls through a modular co-culture engineering pipeline for drug discovery.
文摘Potassium 2-(1-hydroxypentyl)-benzoate(D,L-PHPB), a new drug candidate for ischemic stroke at the phase II clinic trial, has been shown to protect neurons by inhibiting oxidative injury and reducing neuron apoptosis in previous studies. But the mechanisms of D,L-PHPB remain to be studied.In this study, a neuron–astrocytes co-culture system was used to elucidate the roles of astrocytes in neuroprotection of D,L-PHPB under oxygen-glucose deprivation/reoxygenation(OGD/R) condition. Our data showed that D,L-PHPB reduced neuronal apoptosis in mono-culture system and this effect was enhanced in neuron–astrocyte co-culture system under the OGD/R condition. Meanwhile, D,L-PHPB obviously increased the levels of brain-derived neurotrophic factor(BDNF) and nerve growth factor(NGF), which were mainly secreted from astrocytes, in the co-culture system after OGD/R. The PI3K/AKT and ERK signaling pathways as well as the p-TRKA/B receptors were involved in the process. In addition, the levels of TNF-α and IL-1β secreted from astrocytes after OGD/R were markedly reduced after D,L-PHPB treatment, which was mainly due to the suppression of phosphorylated p38. In conclusion, the present study demonstrates that the neuroprotective effects of D,L-PHPB were improved by astrocytes, mainly mediated by increasing the release of BDNF/NGF and attenuating inflammatory cytokines.
基金supported by the National Natural Science Foundation of China(Nos.21727814,21435002 and 21621003)。
文摘In this work,a multi-functional analysis platform by coupling a microfluidic chip to a mass spectrometry(MS) detector was described.We constructed a three-dimensional tumor-endothelial co-culture model for simulating drug resistance during tumor treatment.On this specially designed integrated platform,the first step was to prepare heterogeneous cell-encapsulated alginate microcapsules for threedimensional co-culture,and the second step was to achieve on-line perfusion culture and continuous drug stimulation on chip.It facilitates cell proliferation analysis and the collection of metabolism medium.After micro solid phase extraction column(SPE) pretreatment,subsequent mass spectrometry could detect drug metabolism.The high activity of two kinds of cells(A549 and HUVEC) shows the biocompatibility of the platform.Paclitaxel was used as a model drug,the distinctions of drug absorption between the mono-culture group and co-culture group were clearly observed by electrospray ionization quadrupole time-of-flight mass spectrometry(ESI-Q-TOF MS).Therefore,the integrated platform has shown promise as a high throughput,low cost for cell metabolism research and drug screening processes.
