1.Cord blood CD133+ cell transplantation improves cognitive function and survival of transgenic mice with dementia
Fengwu TANG ; Bo XIU ; Cungang FAN ; Zhiqiang CUI ; Kai XIAO ; Zhongchao HAN
Chinese Journal of Tissue Engineering Research 2008;12(47):9394-9400
BACKGROUND:Human umbilical cord blood (CB)-derived CD133+ cells are a minority population of primitive cells with extensive proliferation and differentiation potentials,which are considered to have ability of neural differentiation.OBJECTIVE:We hypothesized a possible application of CB CD133+ cells in the cognitive and survival function of mice with dementia,the present study observed the changes of the cognitive function and survival of amyloid precursor protein(APP)transgenic mice after CB CD 133+ cells transplantation to verify the above assumption.DESIGN,TIME AND SETTING:A completely randomized block design of animal experiments was performed in the Hematology Institute of Tianjin Hematology Hospital from September 2005 to December 2007.MATERIALS:Forty-eight eight-month-old male APP 695 transgenic C57BL/6 (BDF1/KM) mice were selected in this experiments All mice were divided randomly into three groups:control group (n=8),CD133+ transplantation group (n=20) and CD133 transplantation group (n=20).METHODS:Mice in control groups received an intraventricular injection of 10 μL phosphate buffered saline (PBS).The transgenic mice that received an intraventricular injection of 10 μL CD133+ (5×104/μL) and CD133 CB cells (5×104/μL) respectively.MAIN OUTCOME MEASURES:Radial ann water maze (RAWM) was used to evaluate cognitive function of the mice and the survival days of mice in different groups were recorded,lmmunohistochemical assessments and Dil Fluorescence labeled way was used to detect the differentiation phenotype of transplanted cells.RESULTS:The cognitive function of the mice in CD133+ transplantation group was significantly improved compared with the mice in CD 133- transplantation and control groups both 30 and 180 days after transplantation (P<0.05).The mean survival time of the mice in CD133+ transplantation group was significantly increased compared with CD133 transplantin group and control group (P<0.05).It was observed that the transplantation CB CD133+ cells labeled with Dil migrated into several brain regions at day 30 post-transplantation.These cells were stained for human βⅢ-tubulin,neuralfilement(NF),neuron specific enolase (NSE),and glial fibriliary acidic protein(GFAP).However,in the brain of mice that received CD133 cells transplantation,CB cells were distributed mainly in and around the lateral ventricle at day 30 and 180 post-transplantation and GFAP-,βⅢ-tubulin- and NSE-positive cells were rarely detected.After intraventricular transplantation of CB CD133+ cells,the percentage of transplanted Dil-labeled CB cells expressing βⅢ-tubulin was significant higher at day 30 than at day 180,and the percentage of CB cells expressing NSE was significant lower at day 30 than that at day 180 (both P<0.01).The percentage of CB cells expressing GFAP was relatively constant between the days 30 and 180 after transplantation (P>0.05).CONCLUSION:The result of this experiment suggested that the cognitive and survival function improvement achieved by transplantation of CB CD133+ cells is mainly due to a replacement of dysfunctional cells or augmentation of neural circuit by CB CD133+ cells transplantation.
2.Human umbilical cord blood AC133+ cells expressed neural marker in different inductions
Fengwu TANG ; Bo XIU ; Cungang FAN ; Zhiqiang CUI ; Kai XIAO ; Zhongchao HAN
Chinese Journal of Tissue Engineering Research 2008;12(38):7566-7572
BACKGROUND: At present, hemopoietic stem cells have been proved to differentiate into nerves in rodents animals. As for the human, this topic is in debate.OBJECTIVE: To investigate the neural differentiation potential of human umbilical cord blood-derived AC133+ cells. DESIGN, TIME AND SETTING: Control experiments by grouping were performed in the Hematology Institute of Tianjin Hematology Hospital and Central Laboratory of Neurosurgery in Yuquan Hospital of Tsinghua University from August 2005 to December 2007.MATERIALS: Human umbilical cord blood was sampled from full-term newborn infant. Fetal brain-derived trophic support cells were harvested from aborted fetus of 22 weeks old.MAIN OUTCOME MEASURES: After the induction, human cord blood cells were collected at weeks 1, 2 and 4. RT-PCR was used to detect the expression of nestin, bone morphogenetic protein-2 and neural cell adhesion molecule. Immunocytochemistry method was applied to detect the cytotype-specific antigen. RESULTS: In the culture medium containing epidermal growth factor and basic fibroblast growth factor, human cord blood AC133+ cells could express nestin and bone morphogenetic protein-2, which were down-regulated even closed up in suboptimal condition. In the DMEM/F12 medium supplemented with epidermal growth factor, basic fibroblast growth factor and brain-derived neurotrophic factor, the gene expression of bone morphogenetic protein-2 and nestin continued in optimal condition at 2 weeks. Moreover neural cell adhesion molecule, another gene of neural cells, also expressed in this condition. AC133+ cells co-cultured with fetal brain-derived trophic support cells exhibited similar expressions. In the optimal non-cell-cell contact co-culture system, glial fibrillary acidic protein-positive cells were found by immuocytochemistry, while neuronal marker β-tubulin Ⅲwas expressed in the cell-cell direct contact system. These outcomes indicated that human cord blood isolated AC133+ cells may have an effect through gene rearrangement on inducing stem cells to express nerve cell development factors.CONCLUSION: The human umbilical cord blood-derived AC133+ cells contain some multipotential stem cells with differentiation potential, neural differentiation-related antigen when exposed to a suitable microenvironment.
3.Timing of evoked potentials forecasting the prognosis of patients with severe cerebrovascular disease
Jincong ZHANG ; Shizhong SUN ; Dong WANG ; Kaiyang CHI ; Fengwu TANG ; Yongqing ZHAO
Chinese Critical Care Medicine 2016;28(12):1135-1140
Objective To investigate the effectiveness and the best assessment time of the short-latency somatosensory evoked potential (SLSEP) and brainstem auditory-evoked potential (BAEP) in the prognosis prediction of patients with severe cerebrovascular disease. Methods A prospective trial was conducted. The patients with severe cerebrovascular disease and Glasgow coma scale (GCS) ≤ 8 and admitted to the neurological intensive care unit (NICU) of Armed Police Logistics College Affiliated Brain Hospital from December 2014 to May 2015 were enrolled. The patients received SLSEP and BAEP nerve electrophysiological examinations within 24 hours and on 3, 7, 15 days after admission respectively and were graded according to Cant method. GCS was evaluated within 24 hours and on 15 days after admission. The prognosis was evaluated by Glasgow outcome scale (GOS) at six months after the onset of the disease. At different time windows after the onset of the disease, the correlations between different predictive indexes (GCS, SLSEP and BAEP) and outcome (GOS) were analyzed using spearman rank correlation; in the mean time, the efficacy for predicting the prognosis by single index or combined indexes was compared by receiver operator characteristic (ROC) curve. Results Seventy-eight patients were enrolled [men 46, women 32, age range (60.79±12.50) years old]. There were 78, 64, 44 and 19 patients observed at 24 hours and on 3, 7, 15 days after admission because the short-term death of some patients. The graded abnormal rate of SLSEP was 75.64%, 82.81%, 79.55% and 73.98% respectively; and the graded abnormal rate of BAEP was 82.05%, 84.38%, 85.94% and 73.68% respectively. ① Correlation analysis: all the predictors were correlated with GOS within 24 hours and on 3, 7, 15 days after admission, and SLSEP and BAEP grading were moderately correlated with GOS (0.4≤|R| < 0.7). ② The accuracy of the predicting prognosis: the area under the curve (AUC) of GCS on 15 days after admission [AUC = 0.772, 95% confidence interval (95%CI) = 0.561-0.984, P = 0.045] was the maximum when predicting survival. AUC of SLSEP (AUC = 0.825, 95%CI = 0.695-0.955, P = 0.000) and BAEP (AUC = 0.786, 95%CI = 0.646-0.927, P = 0.002) were the maximum on 7 days after admission when predicting death. ③ The effectiveness of the prognosis prediction: the sensitivity of SLSEP grading and BAEP grading were 92.6% and 96.3% respectively, while the sensitivity, specificity and accuracy of SLSEP and BAEP combined prediction were 100% on 7 days after admission. The specificity of GCS was 100% on 15 days after admission. Conclusions SLSEP and BAEP have more close correlation with prognosis compared with the GCS; Continuous dynamic combined evaluation of SLSEP and BAEP has important clinical value for patients with severe cerebrovascular disease possess in the prognosis assessment, the accuracy and the effectiveness of SLSEP and BAEP combined prediction were higher on 7 days especially.
4.Optimization of culture conditions for oligodendrocytes of the rat cerebral cortex
Kai YANG ; Yipeng LI ; Yingfu LIU ; Yuanchi CHENG ; Fengwu TANG ; Bing LIANG ; Zhongwei XU ; Xuyi CHEN
Chinese Journal of Tissue Engineering Research 2016;20(29):4328-4333
BACKGROUND:Oligodendrocytes are mostly differentiated from oligodendrocyte precursor cel s. A suitable medium and cel seeding density have a significant impact on the process of the isolation of oligodendrocyte precursor cel s to obtain oligodendrocytes. OBJECTIVE:To explore the optimization of oligodendrocyte culture conditions. METHODS:Oligodendrocyte precursor cel s isolated from the newborn rats 48 hours after birth were cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cel s/cm2, 4×104 cel s/cm2, 8×104 cel s/cm2, 16×104 cel s/cm2, 32×104 cel s/cm2, and 64×104 cel s/cm2, respectively. Oligodendrocyte precursor cel s were induced to differentiate into oligodendrocytes at 72 hours after cel adhesion. Morphology of differentiated oligodendrocyte precursor cel s were observed under a light microscope, and the differentiation results were identified by immunofluorescence staining after 7-day induced differentiation. RESULTS AND CONCLUSION:Morphology of oligodendrocyte precursor cel s were recognized when cultured in DMEM/high glucose medium or DMEM/F12 medium using seeding densities of 2×104 cel s/cm2, 4×104 cel s/cm2, and 8×104 cel s/cm2, respectively. Immunofluorescence staining showed that myelin basic protein-positive cel s were found after 7-day induced differentiation, and the positive cel number were 16.40±3.30, 49.95±2.33, and 76.95±4.86 in DMEM/F12 medium, and 12.65±2.53, 32.10±1.17, and 54.05±1.56 in DMEM/high glucose medium (P<0.05). These findings indicate that DMEM/F12 medium is more suitable for culturing oligodendrocyte precursor cel s compared with DMEM/high glucose medium to some extent. The number of differentiated oligodendrocytes was gradual y increased with the enhanced seeding density of oligodendrocyte precursor cel s, and the seeding densities from 4×104 to 8×104 cel s/cm2 were appropriate for the observation of cel morphology.
5.Analysis on therapeutic effect of radiofrequency ablation on discogenic lumbar spinal nerve posterior branch neuralgia after vertebral endoscope surgery
Chen YUN ; Guang HAN ; Xianfeng JIANG ; Haibin TIAN ; Cuiyun MIAO ; Jiamin LIANG ; Feng FU ; Jie LI ; Fengwu TANG ; Shuang TAI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2018;25(6):646-649
Objective To observe the clinical efficacy of radiofrequency ablation for treatment of discogenic lumbar spinal nerve posterior branch neuralgia after vertebral column endoscope operation. Methods Thirty-six patients with discogenic lumbar spinal nerve posterior branch neuralgia after vertebral column endoscope surgery admitted to the Affiliated Hospital of Logistics University of People's Armed Police from December 2011 to December 2017 were enrolled. According to difference in therapeutic methods, they were randomly divided into two groups, 18 cases in each group. The radiofrequency ablation group was treated with X-ray imaging guided lumbar spinal nerve posterior branch radiofrequency thermo-coagulation; the drug group received oral diclofenac sodium conservative treatment, 75 mg twice daily for 3 weeks. Both groups were followed up for 6 months, visual analogue scores (VAS) were used to evaluate the pain before and after treatment, the Oswestry dysfunction index was used to assess the degree of lumbar function recovery, and the surgical complications and adverse drug reactions were observed. Results The VAS scores in the two groups were similar before treatment; after treatment for 1 month, the VAS scores in both groups were significantly lower than those before treatment (radiofrequency ablation group: 1.83±0.71 vs. 5.67±0.77; drug group: 2.22±0.43 vs. 5.28±0.67, both P < 0.05); after treatment for 3 months and 6 months, the VAS scores were increased gradually, however, the scores of radiofrequency ablation group were significantly lower than those in the drug group (3 months was 2.00±0.59 vs. 3.39±0.70, 6 months was 2.17±0.51 vs. 3.61±0.50, both P < 0.05), moreover, the excellent and good rates of postoperative pain efficacy and of Oswestry dysfunction index improvement in the radiofrequency ablation group were significantly higher than those in the drug group [excellent and good rates of postoperative pain efficacy: 94.44% (17/18) vs. 22.22% (4/18), excellent and good rates of Oswestry dysfunction index improvement: 77.78% (14/18) vs. 44.44% (8/18), both P < 0.05]. There were no complications of infection and spinal nerve anterior branch injury in the radiofrequency ablation group, and 6 patients in the drug group presented mild gastric discomfort, which was relieved after symptomatic treatment. Conclusion The radiofrequency ablation is an effective method for treatment of discogenic lumbar neuralgia after vertebral column operation, compared with the conservative therapy, the ablation is more effective to relieve pain for a long time, promote the recovery of neural function, and the operation is safe with very few adverse reactions.
6.Regulatory mechanism underlying mycelium aggregation during filamentous fungi submerged fermentation.
Ruisang LIU ; Yajie TANG ; Fengwu BAI
Chinese Journal of Biotechnology 2019;35(5):749-758
Filamentous fungi are one of the platforms for producing fermented products. The specific characteristic of their submerged fermentation is the aggregation of mycelia that is affected by environmental conditions, leading to significantly different rheology for fermentation broth. Such a rheological change not only affects the transfer of mass, heat and momentum, but also the biosynthesis of target products and the efficiency of their production. In this article, strategies for morphological regulation of filamentous fungi are reviewed, and the impact of calcium signal transduction and chitin biosynthesis on apical growth of hyphae and branching of mycelia for their aggregation are further commented.
Fermentation
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Fungi
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physiology
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Hot Temperature
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Mycelium
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metabolism
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Rheology