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    08 March 2019, Volume 23 Issue 7 Previous Issue    Next Issue
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    Exosomes in serum of ovariectomized rats promote primary osteoblast proliferation
    Gao Kun1, Zhu Wenxiu2, Li Heng1, Liu Weidong1, Li Quan1, Yu Weiji1, Wang Lixin1, Cao Yafei1
    2019, 23 (7):  958-989.  doi: 10.3969/j.issn.2095-4344.1071
    Abstract ( 334 )   PDF (1103KB) ( 147 )   Save

    BACKGROUND: During bone metabolism, changes in the number and function of osteoblasts influence the biological characteristics of bone tissues. Exosomes can mediate cell-to-cell transmission, and have the potential to promote cell proliferation and differentiation.
    OBJECTIVE: To study the properties of exosomes in serum of rats with osteoporosis and to investigate their effects on osteoblasts.
    METHODS: Several newborn Sprague-Dawley rats at 24 hours of birth were used for primary osteoblast isolation and culture. A rat model of ovariectomized osteoporosis was established in 32 healthy female adult Sprague-Dawley rats. Rats undergoing sham operation were used as control group. All the experimental animals were provided by Guangdong Medical Animal Experimental Center, China. Exosomes were extracted from the serum of control rats and ovariectomized rats using ExoQuick and identified by electron microscopy, nanosight and western blot. Primary osteoblasts in logarithmic phase were seeded into 96-well culture plates with an inoculation concentration of 2×104/well. Then, the cells were divided into blank control group, exosome group, and osteoporosis+exosome group, and DPBS, normal serum-derived exosomes, and osteoporosis serum-derived exosomes, 100 μL/well, were added, respectively. The cells in each group were cultured in a CO2 incubator for 1 day. The proliferation of osteoblasts was detected by MTT colorimetric assay. Enzyme-linked immunosorbent assay was used to detect the concentration of vascular endothelial growth factor in exosomes.
    RESULTS AND CONCLUSION: Successfully extracted exosomes particles were below 100 nm in size. The serum exosome protein concentration in osteoporotic rats was highest at 12 weeks after ovariectomized surgery. The number of exosome particles was also highest at 12 weeks after ovariectomized surgery, but there was no significant difference in the osteoporotic rats at different time points. Compared with the blank control and exosome group, exosomes from the serum of osteoporotic rats significantly promoted the proliferation of osteoblasts, and the level of vascular endothelial growth factor in exosomes increased. These findings indicate that exosomes from the serum of osteoporotic rats truly promote the proliferation of osteoblast.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Application of 25-question Geriatric Locomotive Function Scale combined with bone mineral density test in screening locomotive syndrome and risk of falls in the elderly: a survey of 1 458 people from 6 urban communities in Tianjin
    Song Chao1, Tian Xin1, Liu Lin1, Xu Yawen1, Meng Yuan1, Li Xiaoli1, Peng Meng2, Li Huadong2, Wang Zhitao3, Ge Sha1
    2019, 23 (7):  990-995.  doi: 10.3969/j.issn.2095-4344.1063
    Abstract ( 504 )   PDF (1281KB) ( 112 )   Save

    BACKGROUND: Subjective assessments, such as 25-question Geriatric Locomotive Function Scale (GLFS-25), balance function assessment, and gait test, are commonly used to screen for motor dysfunction and predict the risk of falls in the elderly. Therefore, the combination of impersonal assessment indicators and these subjective assessments can fully reflect the motor and balance functions of the elderly.
    OBJECTIVE: To explore the application of GLFS-25 combined with bone mineral density test in screening for locomotive syndrome and risk of falls in the older adults.
    METHODS: A total of 1 458 elderly people who resided in six communities of Heping District, Tianjin, China were enrolled by the method of zoning cluster random sampling. According to the GLFS-25 scores, the subjects were divided into locomotive syndrome group (198 cases, GLFS-25 ≥ 16) and non-locomotive syndrome group (1 260 cases, GLFS-25 < 16). The height, weight, body mass index, body fat percentage, and visceral fat area were compared between the two groups. The bone mineral density, fall frequency, knee flexibility and lower limb strength, balance function and gait parameters were also tested and compared between the two groups. The Pearson method was used to analyze the correlation between GLFS-25 score, bone mineral density and fall frequency, the frequency of sitting posture conversion, gait parameters and balance function score in 30 seconds. Receiver operating characteristic curve was used to analyze the efficacy of GLFS-25 combined with bone mineral density test in the diagnosis of locomotive syndrome and fall risk in the elderly.
    RESULTS AND CONCLUSION: (1) Compared with the non-locomotive syndrome group, height, body mass index, and fat percentage were significantly higher in the locomotive syndrome group (P < 0.05), while the bone mineral density was significantly lower (t=7.467, P=0.000). (2) Compared with the locomotive syndrome group, the non-locomotive syndrome group showed significantly less fall frequency within 1 year and lower risk of falls, lower 30-second sitting/standing conversion frequency, worse self-balance feeling score (P=0.000), but longer total distance, faster step speed, and larger step size, and lower variation coefficient of the left and right strides (P < 0.05). (3) Pearson correlation analysis showed that there was no significant correlation between GLFS-25 score, bone mineral density and height, body mass index, and body fat percentage (P > 0.05). GLFS-25 score was negatively correlated with bone mineral density, self-balance feeling, total distance, average step speed, and average step length, but positively correlated with fall frequency, fall risk index, and variation coefficient of the left and right strides (P < 0.05). (4) Receiver operating characteristic curve analysis showed that the area under the curve of GLFS-25 combined with bone mineral density for prediction of locomotive syndrome and risk of falls was 0.895 and 0.903, respectively, which was significantly higher than that of GLFS-25 or bone mineral density test alone. And their sensitivity increased to 90.73% and 90.15%, respectively. Overall, the proportion of older adults with locomotive syndrome in Tianjin urban communities is relatively high. The patients with locomotive syndrome have higher GLFS-25 score, lower bone mineral density, lower knee flexibility, limb strength and balance, and a higher fall risk. GLFS-25 combined with bone mineral density test is therefore more effective in the screening of locomotive syndrome and fall risk assessment in the elderly.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Isolation, culture and identification of osteoblasts from neonatal rabbit calvarium
    Ailimaierdan•Ainiwaer1, Li Peng1, Diao Zhaofeng1, Muhetaer•Huojia2
    2019, 23 (7):  996-1000.  doi: 10.3969/j.issn.2095-4344.0576
    Abstract ( 459 )   PDF (4010KB) ( 114 )   Save

    BACKGROUND: A publicly accepted way to isolate osteoblasts with high purity and efficiency has not been found.
    OBJECTIVE: To explore the efficacy of advanced enzymatic digestion method to isolate and culture osteoblasts from calvarial bone of neonatal New Zealand rabbit in vitro.
    METHODS: Improved enzymatic digestion method was used to isolate osteoblasts from calvarium of 3-day-old New Zealand rabbits and cells were purified via differential attachment methods. The morphology and growth of osteoblasts were observed by inverted microscope daily.  Cell proliferation was detected by MTT assay and the growth curve was drawn. Alkaline phosphatase activity was measured. The expression levels of collagen type I, osteocalcin and Runt related transcription factor 2 were detected by immunohistochemical staining. Alizarin red staining was carried out to testify mineralized matrix productivity.
    RESULTS AND CONCLUSION: Osteoblasts were successfully isolated, cultured and purified by enzyme digestion. The isolated osteoblasts adhered on the wall possessing typical characteristics of osteoblasts with good proliferation ability. Alkaline phosphatase staining and immunohistochemical staining on collagen type I, osteocalcin and Runt related transcription factor 2 presented positive results. Alizarin red staining showed mineralized nodules with different sizes after 21 days of culture. These findings suggest that the osteoblasts cultured by improved enzymatic digestion hold the typical characteristics of osteoblasts, with high purity and survival rate.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Micro-CT changes and vascular endothelial growth factor expression in reconstructed mandibular condylar cartilage under continuous mandibular advancement in growing and adult rats
    Wu Min1, Li Xue1, Gao Jie1, Yang Shuai1, Cai Yizhi2, Wang Mingguo1
    2019, 23 (7):  1001-1006.  doi: 10.3969/j.issn.2095-4344.1067
    Abstract ( 444 )   PDF (5066KB) ( 112 )   Save

    BACKGROUND: Previous experimental results have shown that mandibular advancement can remodel the condylar process in growing rats, which is shown to promote the growth of condylar tissue. However, the influence of mandibular advancement on condylar remodeling and ultrastructure of condylar cartilage in adult rats different from those in growing rats are still a key issue.
    OBJECTIVE: To observe the condylar remodeling and ultrastructural variations in the mandibular condylar cartilage in adult and growing rats undergoing mandibular advancement.
    METHODS: Thirty 9-week-old male Sprague-Dawley rats and thirty 4-week-old male Sprague-Dawley rats were randomly divided into experimental and control groups with 15 adult rats and 15 growing rats in each group. Rats in the experimental group were subjected to wearing mandibular advancement splints. Rats were sacrificed on days 3, 7, 14, 21, and 30. Sections from the condyles were cut, and vascular endothelial growth factor expression in the condylar cartilage was examined through immunohistochemical analysis. These samples were also observed under Micro-CT. 
    RESULTS AND CONCLUSION: Compared with the control group, the number of vascular endothelial growth factor positive cells in the condylar cartilage and the gray value gradually increased in the experimental group on day 7 of the intervention: for the adult rats, the positive cell number and gray value were at the maximum level on day 30 (P < 0.01), while for the growing rats, the highest number of positive cells and gray value appeared on day 21 (P < 0.01), and the gray value on day 30 was lower than that on day 21 but still higher than that in the control group. Micro-CT revealed that the trabecula and the newly formed bone in the experimental group gradually thickened, and the number and separation of trabeculae increased with time. To conclude, both adult and growing rats display hyperplastic remodeling in the condyle with the presence of high expression of vascular endothelial growth factor under continuous mandibular advancement. However, the adaptive remodeling is earlier in the growing rats than in the adult rats.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Comparison of the effects of two kinds of platelet-rich fibrins on the proliferation activity of human gingival fibroblasts
    Wang Canli1, 2, Jiao Zhili2, Sun Yong1, 2, Sun Song3
    2019, 23 (7):  1007-1012.  doi: 10.3969/j.issn.2095-4344.0589
    Abstract ( 357 )   PDF (4373KB) ( 84 )   Save

    BACKGROUND: Platelet-rich fibrin (PRF) prepared by autologous blood has become the hotspot in regeneration and repair of soft tissues.
    OBJECTIVE: To compare the effect of two kinds of PRFs on the proliferation activity of human gingival fibroblasts (HGFs), so as to provide reference for the selection of oral soft tissue regeneration and repair materials in the clinic.
    METHODS: Healthy gingival flaps from eight patients were collected when mandibular impacted teeth were removed. HGFs were obtained using tissue explant method, and then identified. The venous blood of the corresponding patients was drawn and made into advanced PRF and PRF, respectively, and passage 5 HGFs were divided into advanced PRF, PRF and blank groups for culture. The cell proliferation activity was detected by cell counting-kit 8 assay at 1, 3, 5, and 7 days.
    RESULTS AND CONCLUSION: The HGFs were positive for Vimentin staining and negative for Cytokeratin staining. Two kinds of PRFs were prepared successfully. The advanced PRF exhibited the deeper color, more thick membrane and better flexibility compared with the general PRF. Cell counting-kit 8 assay results revealed that at 1 day after culture, there was no significant difference in the proliferation activity among groups (P > 0.05). At 3 days, the cell proliferation activity in the advanced PRF and PRF groups was significantly higher than that in the blank group (P < 0.05). At 5 and 7 days, the order of cell proliferation activity was as follows: advanced PRF group > PRF group > blank group (P < 0.01, P < 0.05). To conclude, both advanced PRF and PRF can increase the proliferation activity of HGFs, especially the former. 

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Post-stroke dysarthria treated by acupuncture combined with speech-language training: bibliometric analysis and verification of clinical efficacy
    Han Jian
    2019, 23 (7):  1013-1017.  doi: 10.3969/j.issn.2095-4344.1119
    Abstract ( 436 )   PDF (1101KB) ( 183 )   Save

    BACKGROUND: There is a lack of effective drug for post-stroke dysarthria. Effect of single rehabilitation training is insignificant, and traditional acupuncture and moxibustion therapy obtains good treatment outcomes.
    OBJECTIVE: To analyze the research focus and prospects of post-stroke dysarthria by bibliometrics, and to explore the treatment efficacy of acupuncture and moxibustion combined with speech-language training for post-stroke dysarthria.
    METHODS: (1) A computer-based online research of CNKI database for the articles addressing post-stroke dysarthria from January 2004 to December 2018. The distribution of publication year, author, affiliation and funding in the literature was analyzed by group-level browsing and visualization tool. After reading the title and abstract, the review and the articles about nursing and radiography were excluded, and the articles concerning acupuncture and moxibustion were included to summarize acupuncture and moxibustion in the treatment of post-stroke dysarthria. (2) Ninety-six patients with post-stroke dysarthria admitted at the First People’s Hospital of Shenyang between June 2013 and August 2015 were enrolled, and then divided into control and treatment groups (n=48 per group). The control group underwent speech-language training, and the treatment group received speech-language training and acupuncture and moxibustion (acupuncture at tongue and neck), for two courses and 40 days in total. The clinical efficacy was evaluated by advanced Frenchay dysarthria assessment, and the adverse reactions were recorded.
    RESULTS AND CONCLUSION: (1) One hundred and twenty-one Chinese articles were retrieved. There was 1 article published in 2004, total 14 articles by the end of 2018. The articles (n=5) are mainly form Chengdu University of Traditional Chinese Medicine by Hu Kaming team. Sixty percent of the first ten affiliations and the first eight authors were form the Chinese Medicine Universities or their affiliated hospitals. Of them, 72 articles addressed acupuncture and moxibustion, including 7 in acupuncture at tongue, 8 in acupuncture at tongue and neck, 7 in acupuncture at tongue and head, 12 in acupuncture and electrostimulation, 3 in acupuncture and hyperbaric oxygen therapy, and 35 in acupuncture and rehabilitation training. (2) The total effctiveness rate in the treatment group (92%) was significantly higher than that in the control group (54%) (P < 0.05). In summary, in China, studies on the treatment of post-stroke dysarthria are mainly from Chinese Medicine Universities and their affiliated hospitals. Chinese medicine, especially acupuncture and moxibustion (acupuncture at tongue and neck mostly used) is an important means in the treatment of post-stroke dysarthria. Clinical trial results suggest that post-stroke dysarthria treated by acupuncture at tongue and neck combined with speech-language training can obtain satisfactory outcomes.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Inhibitory effect of capsaicin on fibroblast proliferation and its molecular mechanism
    Zhou Yanxing1, Peng Xinsheng2, Hou Gan1, Li Jiangbin1, Zhang Hua1, Zhou Zhikun2, Zhou Yanfang3
    2019, 23 (7):  1018-1022.  doi: 10.3969/j.issn.2095-4344.0574
    Abstract ( 381 )   PDF (1480KB) ( 116 )   Save

    Abstract
    BACKGROUND
    : Capsaicin has been reported to inhibit fibrosis and the proliferation and collagen synthesis of hypertrophic fibroblasts. However, the underlying mechanism is unclear.
    OBJECTIVE: To investigate the effect of capsaicin on inhibiting the proliferation of fibroblasts and the corresponding molecular mechanism.
    METHODS: Neonatal Sprague-Dawley rat epidermal fibroblasts were cultured in vitro, and were treated with capsaicin at different concentrations (0.5, 1, 2, 4, 8 mg/L) for 24 hours. The cells without treatment of capsaicin were included in the negative control group. Morphological changes of the epidermal fibroblasts were observed under phase-contrast microscope. The cell viability and IC50 value were detected by MTT assay. The cell apoptosis was detected by Hoechst 33258 staining and AnnexinV-FITC/PI staining. The cell cycle distribution was determined by flow cytometry. The protein expression levels of FoxO1, p-FoxO1, Akt and p-Akt were detected by western blot assay.
    RESULTS AND CONCLUSION: With capsaicin concentration increasing, cell survival rate decreased gradually in a concentration-dependent manner, and IC50 value was 3.55 mg/L. Compared with the negative control group, after 24 hours, capsaicin treated fibroblasts stopped in G1 phase and the apoptosis rate increased. Meanwhile, capsaicin could decrease the expression of P-Akt and P-FoxO1. Therefore, capsaicin can inhibit the proliferation of fibroblasts, promote cell apoptosis, and present a concentration-dependent effect. The Akt-FoxO1 signaling pathway may be involved in the inhibition of capsaicin on fibroblast proliferation.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Spinal cord stimulation alters the serum levels of inflammatory factors in canine atrial fibrillation models
    Shi Hui1, Zhao Lulu1, Hua Baotong1, Du Yunhui1, Guo Tao1, 2
    2019, 23 (7):  1023-1029.  doi: 10.3969/j.issn.2095-4344.1026
    Abstract ( 351 )   PDF (1234KB) ( 134 )   Save

    Abstract
    BACKGROUND
    : Some studies have shown that spinal cord stimulation can inhibit atrial fibrillation by regulating the balance of cardiac autonomic nerves. However, it is less reported on the effect of spinal cord stimulation on the inflammatory response to atrial fibrillation.
    OBJECTIVE: To investigate the effect of spinal cord stimulation on the serum levels of inflammatory factors in canine atrial fibrillation models.
    METHODS: Nine adult beagles from the Experimental Animal Center of Kunming Medical University were selected as laboratory canines and divided into control group (n=3), atrial fibrillation group (n=3) and atrial fibrillation+spinal cord stimulation group (n=3) at random. The canine atrial fibrillation models were made by rapid right atrial pacing in the latter two groups. The spinal cord stimulators were implanted into the canine atrial fibrillation models in the atrial fibrillation+spinal cord stimulation group and had released stimulation for 12 weeks. The miniaturized insertable cardiac monitors were implanted into the canine atrial fibrillation models for monitoring AT/AF burden. Venous blood samples of the experimental beagles were extracted, in which the serum levels of inflammatory factors (C-reactive protein, hypersensitive C-reactive protein, interleukin-1, interleukin-6 and complement-3) were tested by ELISA. In the atrial fibrillation group, the blood samples were taken before and after modeling (before pacemaker implantation and when atrial fibrillation had lasted for 15 minutes), while in the atrial fibrillation+spinal cord stimulation group, the blood samples were extracted before and after spinal cord stimulation therapy (before spinal cord stimulation therapy and when spinal cord stimulation had implemented for 12 weeks). Meanwhile, venous blood samples were taken from each beagle in the control group.
    RESULTS AND CONCLUSION: The serum levels of inflammatory factors in the atrial fibrillation group were significantly higher than those in the control group (P < 0.05), while the levels were significantly decreased in the atrial fibrillation+spinal cord stimulation group compared to the atrial fibrillation group (P < 0.05). For the canine models of atrial fibrillation, the serum concentration of inflammatory factors in the beagles which had received spinal cord stimulation therapy for 12 weeks were all lower than those without spinal cord stimulation (P < 0.05). The AT/AF burden of the atrial fibrillation+spinal cord stimulation group declined significantly compared with the atrial fibrillation (P < 0.05). To conclude, atrial fibrillation can induce inflammatory response, and spinal cord stimulation can suppress atrial fibrillation and further inhibit the inflammatory response caused by atrial fibrillation.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    A chemical radiculitis model in the rat: establishment and evaluation
    Zhao Caiping, Zhu Meiling, Wang Tingting, Liu Xuelei, Liu Cuiling
    2019, 23 (7):  1030-1034.  doi: 10.3969/j.issn.2095-4344.1065
    Abstract ( 456 )   PDF (4505KB) ( 123 )   Save

    BACKGROUND: Chemical nerve root modeling experiments provide a stable animal model for studying the intervertebral discs. 
    OBJECTIVE: To explore the establishment of lumbar disc herniation and chemical nerve root inflammation in rats by formaldehyde soaked filter paper, and to find the best operation plan and the solution concentration.
    METHODS: Ten Sprague-Dawley rats provided by Beijing Huafukang Bioscience Co., Inc., in China were randomly divided into two groups and subjected to biting off the spinous process or not. Then, 4% formaldehyde soaked filter papers were placed beside the dorsal root nerve to make the model. Based on behavioral performance, the optimal surgical plan was determined. Thereafter, the effect of different concentrations of formaldehyde soaked filter papers on the modeling. Another 20 Sprague-Dawley rats were randomized into 4 groups, 15 of which were subjected to the above-determined surgical plan of biting off the spinous process, and 1%, 2% and 4% formaldehyde soaked filter papers were placed beside the dorsal root nerve. Siegal neurological score, hematoxylin-eosin staining, and measurement of related inflammatory factors were used to assess modeling effects.
    RESULTS AND CONCLUSION: The Siegal neurological score and grading were significantly increased in the rats with biting off the L4, 5 than those without biting off the spinous process. Compared with the control group, the Siegal neurological score and grading were markedly increased in 1%, 2%, 4% formaldehyde groups at 2-8 days after modeling, but this increasing tendency remained only in the 2% and 4% formaldehyde groups within 2 weeks after modeling. Moreover, significant inflammatory cell infiltrations occurred in the rat’s nerve, muscle and intervertebral disc, and the levels of serum related inflammatory factors increased remarkably. These findings indicate that the improved modeling method has high successful rate and low mortality, which can provide ideal animal models for subsequent diseases and drug researches.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Ultrastructural evaluation of a rabbit model of steroid-induced osteonecrosis of the femoral head after treatment with panax notoginseng saponins 
    Han Jie1, Chen Yueping1, Mo Jian1, Wang Dawei1, Su Bo1, Li Shuzhen1, Xia Tian2, Wang Shixin2
    2019, 23 (7):  1035-1039.  doi: 10.3969/j.issn.2095-4344.1072
    Abstract ( 328 )   PDF (3780KB) ( 125 )   Save

    BACKGROUND: Preliminary studies have shown that panax notoginseng saponins can promote the repair of osteocyte ultrastructure in a rabbit with alcohol-induced osteonecrosis of the femoral head. 
    OBJECTIVE: To observe the structural changes of osteocytes in a rabbit with steroid-induced osteonecrosis of the femoral head treated with panax notoginseng saponins by transmission electron microscope.
    METHODS: Ninety 9-10 weeks old healthy male New Zealand rabbits (provided by Laboratory Center of Guangxi University of Chinese Medicine), weighting 3.5-4.0 kg were selected. Rabbit models of steroid-induced osteonecrosis of the femoral head were established by injecting modified horse serum combined with methylprednisolone sodium succinate. Fifteen rabbits died, so the remaining 75 rabbits were divided into three groups (n=25 per group), and then treated with normal saline, cervus and cucumis polypeptide or panax notoginseng saponins, respectively for 5 weeks. The structural changes of osteocytes were observed under transmission electron microscope.
    RESULTS AND CONCLUSION: In the normal saline group, the number of mature osteocytes was reduced, vascular endothelial cells were damaged, and mitochondria swelling, fuzzy cristae structure, nuclear fragmentation, and lipid droplets were observed. In the cervus and cucumis polypeptide and panax notoginseng saponins groups, there were more mature osteocytes and less vascular endothelial cell damage, swollen mitochondria alleviated with clear structure, and the numbers of mast adocytes and lipid droplets were decreased. The structural changes of osteocytes in the cervus and cucumis polypeptide and panax notoginseng saponins groups were significantly more encouraging than those in the normal saline group (P < 0.05). Our results indicate that show that, panax notoginseng saponins can repair the osteocyte structure of rabbits with steroid-induced osteonecrosis of the femoral head.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Proliferation and differentiation of MG63 cells transfected with recombinant adenovirus vector overexpressing estrogen-related receptor alpha after silencing Bak1 and Bcl2
    Huang Hong1, Huang Jiachun2, Huang Hongxing3, Wang Jili2, Liu Shaojin2, Wang Yuedong2
    2019, 23 (7):  1041-1045.  doi: 10.3969/j.issn.2095-4344.0570
    Abstract ( 184 )   PDF (4852KB) ( 167 )   Save

    BACKGROUND: Estrogen deficiency is the main pathogenesis of osteoporosis in postmenopausal women, but there are few researches on the correlation between estrogen-related receptor α (ERRα) and osteoporosis. Little is known about their mechanisms of action.
    OBJECTIVE: To investigate the effect of adenoviral overexpression of ERRα and silencing of Bak1/Bcl2 in MG63 cells.
    METHODS: Adenovirus vector overexpressing ERRα and silencing of Bak1/Bcl2 was constructed. MG63 cells were divided into blank control, Ad-shBak1, Ad-shBcl2, and Ad-shBak1+shBcl2 groups. MG63 cells of different groups were infected by ERRα, Bak1, and Bcl2 overexpressing recombinant adenovirus. The cell proliferation was detected by MTT assay. The alkaline phosphatase activity was measured by Coomassie brilliant blue method. The Ca2+ concentration was detected by flow cytometry. The expression levels of related bone-regulating proteins (bone morphologic protein 4, connective tissue growth factor, osteopontin, Runt 2 and tumor necrosis factor α) were tested by western blot assay.   .
    RESULTS AND CONCLUSION: Compared with the blank control group, the cell viability and alkaline phosphatase activity were significantly increased and Ca2+ concentration was significantly decreased, bone morphologic protein 4, connective tissue growth factor, osteopontin and Runt 2 were increased, and tumor necrosis factor α was decreased in the Ad-shBak1 group. The cell viability, alkaline phosphatase activity and connective tissue growth factor level were decreased, but not significant in the Ad-shBcl2 group (P > 0.05). In the Ad-shBcl2 group, the Ca2+ concentration, bone morphologic protein 4, and Runt 2 were significantly decreased, osteopontin and tumor necrosis factor α were significantly increased (P < 0.01). In the Ad-shBak1+shBcl2 group, the cell viability and alkaline phosphatase activity were increased, Ca2+ concentration was decreased, and level of each protein was increased (P > 0.05). Compared with the Ad-shBcl2 group, in the Ad-shBak1 and Ad-shBak1+shBcl2 groups, the cell viability and alkaline phosphatase activity were significantly increased, Ca2+ concentration was significantly decreased, bone morphologic protein 4, connective tissue growth factor, osteopontin and Runt 2 were increased, and tumor necrosis factor α was significantly decreased (P < 0.01 or P < 0.05). Our findings suggest that the overexpression of ERRα can increase the MG63 cell proliferation and alkaline phosphatase activity after transfection by Bak1 recombinant adenovirus, decrease Ca2+ concentration, and also has certain effects on related bone regulating proteins.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Human amniotic membrane repairs acute sciatic nerve injury in rat models
    Cao Peng, Wang Haonan, Tian Weifeng, Sun Naichao, Bai Jiangbo, Yu Kunlun, Tian Dehu
    2019, 23 (7):  1046-1051.  doi: 10.3969/j.issn.2095-4344.1073
    Abstract ( 416 )   PDF (5894KB) ( 137 )   Save

    BACKGROUND: Human amniotic membrane with low immunogenicity can reduce scar formation, inhibit inflammation and prevent angiogenesis.
    OBJECTIVE: To observe the effect of human amniotic membrane on repair and regeneration process after acute nerve injury.
    METHODS: Ninety adult male Sprague-Dawley rats provided by Laboratory Animal Center of Hebei Medical University were used for establishing the models of acute sciatic nerve injury and then randomly divided into amniotic membrane (nerve entrapment with amniotic membrane), chitosan (nerve entrapment with chitosan gel) and blank control (without any treatment) groups (n=30 per group). Gross, light microscope and transmission electric microscope observations, axon image analysis and electrophysiological examination were performed.
    RESULTS AND CONCLUSION: Gross, light microscope and transmission electric microscope observations revealed: compared with the blank control group, the mild limb swelling, neurohyperemia, and edema were observed in the amniotic membrane group. In addition, the adhesion with the surrounding tissue was mild in the amniotic membrane group, the inflammatory reaction was mild and there were few fibroblasts and newborn capillaries. In the amniotic membrane group, function of organelles was active, there were few fibroblasts and less collagen, and the repair time was short, which were similar with the chitosan group. The axon image analysis showed that thickness of the myelin sheath and the diameter of myelinated fibers in the amniotic membrane and chitosan groups were superior to those in the blank control group (P < 0.05). Neurophysiological stimulation showed that there was no significant difference in the nerve conduction velocity among groups. The amplitude was significantly different among groups (P < 0.05). The latent period in the amniotic membrane and chitosan groups was significantly shorter than that in the blank control group (P < 0.05). To conclude, amniotic membrane can provide a relatively stable microenvironment for nerve repair, reduce adhesion, inhibit inflammatory reaction, promote the repair and regeneration of nerve, and show good therapeutic effect. Amniotic membrane has a long retention time in vivo, which is beneficial to nerve regeneration and microenvironment maintenance, and contributes to the recovery of nerve function. Therefore, amniotic membrane is better than the chitosan.

     

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    Anesthesia management for aortic valve plus mitral valve replacement in Small-Tailed Han sheep
    Peng Xianjiang1, Liu Bozhi2, Duan Haifeng1, Yang Qianzi1, Hu Sheng1
    2019, 23 (7):  1052-1056.  doi: 10.3969/j.issn.2095-4344.1064
    Abstract ( 397 )   PDF (3980KB) ( 89 )   Save

    BACKGROUND: At present, there are few studies on the anesthetic methods for cardiac surgeries in Small-Tailed Han sheep, and further experimental verifications on the effectiveness and safety are still required.
    OBJECTIVE: To evaluate the safety and effectiveness of anesthetic methods for cardiac surgeries in Small-Tailed Han sheep.
    METHODS: Forty Small-Tailed Han sheep, aged (10.95±0.89) months, weighing (71.3±7.7) kg, underwent aortic valve plus mitral valve replacement after basic anesthesia with intramuscular ketamine (8-12 mg/kg)+scopolamine (10-20 μg/kg) or with intravenous ketamine (2.5-3.5 mg/kg)+scopolamine (10-15 μg/kg) and general anesthesia with intravenous propofol and fentanyl. The bucking response in endotracheal intubation, vital signs, hemodynamic changes, recovery condition and complications were observed during the perioperation (5 minutes before incision, 5 minutes after incision, 5 minutes before in vitro transfer, 5 minutes after in vitro shutdown, immediately after surgery, 1 day after surgery).
    RESULTS AND CONCLUSION: None of the animals failed in intubation. Perioperative sedation and analgesia were satisfactory, and vital signs were stable; perioperative blood gas analysis shows stable internal environment; no body movement occurred, and all cases well tolerated valve replacement; no regurgitation and aspiration occurred during operation; and there were no deaths during the procedure. Anesthesia with propofol and isoflurane plus low-dose fentanyl has the advantages of rapid recovery, stable hemodynamics and good anesthetic effect, which is a safe and effective anesthesia scheme for cardiac surgery in Small-Tailed Han sheep.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Expression of PLCgamma2 in a wild-type mouse model of chronic periapical periodontitis
    Yang Chunhe, Zhang Hong, Dong Ming, Wang Lina, Niu Weidong
    2019, 23 (7):  1057-1062.  doi: 10.3969/j.issn.2095-4344.1023
    Abstract ( 408 )   PDF (4601KB) ( 89 )   Save

    BACKGROUND: Inhibiting BTK-PLCγ2-Ca2+ signaling pathway has been shown to inhibit formation and function of osteoclasts, suggesting that PLCγ2 participates in cell formation and differentiation.
    OBJECTIVE: To study the expression of PLCγ2 in periapical tissues by establishing a model of periapical periodontitis in wild-type mice.
    METHODS: Twenty male wild-type C57BL/6L mice were selected. Bilateral mandibular first molar pulp cavities of 16 mice were exposed to the oral environment and the formation of apical periodontitis was induced (experimental group). The mice were randomly sacrificed at 1, 2, 3 and 4 weeks and the lower jaws were isolated. The remaining four mice were used as blank control group, and were sacrificed at 0 week to isolate the lower jaws. After being histologically processed, they were hematoxylin-eosin stained to observe the inflammation conditions in the apical area. Expression and distribution of PLCγ2 were analyzed by immunohistochemical staining. Enzyme-histochemical staining was used to detect the expression of osteoclasts in apical inflammatory tissue.
    RESULTS AND CONCLUSION: According to the histological examination, there was almost no inflammatory cell infiltration in the periodontal tissue of the mandibular first molar in the blank control group. In the experimental group, at 1 to 4 weeks after operation, the infiltration of inflammatory cells in the periodontal tissues and the destruction of alveolar bone of mandibular first molar increased gradually, indicating that the mouse periapical periodontitis model was established successfully. Immunohistochemical analysis found that the expression of PLCγ2 was detected at 0 to 4 weeks after periapical periodontitis in mice. A small amount of PLCγ2 expressed in normal dental apical tissues. PLCγ2 in the experimental group was mainly expressed in the nucleus of lymphocytes, plasma cells and other cells, and there was a small amount of expression in the cell membrane and cytoplasm. At the1st, 2nd, 3rd, 4th weeks, the positive expression of PLCγ2 also increased due to the gradual expansion of inflammatory cell infiltration. The positive cell counts were all significantly different (P < 0.05). The results of TRAP staining showed that almost no osteoclasts were observed in periapical periodontal tissues of blank control group. In the experimental group, at 1 week after operation, a small amount of multinucleated osteoclasts were observed. The number of osteoclasts continued to increase at 2 weeks after operation. At 3 weeks, the number of osteoclasts reached the peak. At 4 weeks postoperatively, a decrease in the number of osteoclasts was observed. The difference was significant between experimental and control groups (P < 0.05). Correlation analysis between TRAP positive cell count and PLCγ2 positive cell count showed a significant correlation (r=0.627, P < 0.001). These results indicate that PLCγ2 is expressed in periapical tissues of mice, implying that it may be related to periapical inflammatory reaction and bone resorption.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Making chronic ischemia rat models by high-fat diet and thread embolism technique: evaluation and comparison of modeling methods
    Chen Lei1, Ding Meng2, Zhang Qingyun1, Gao Jianguo1, Guo Fengling3, Yang Zhi1, Zhang Meng4
    2019, 23 (7):  1063-1067.  doi: 10.3969/j.issn.2095-4344.1048
    Abstract ( 352 )   PDF (3860KB) ( 108 )   Save

    BACKGROUND: Some studies have prepared a chronic ischemic animal model using the thread embolism technique, but the observation period is longer, and the survival time of the experimental animals may be shortened because of surgical trauma and disease. Herein, the authors further established a reliable animal model of chronic ischemia using the thread embolism technique by addition of high-fat diet.
    OBJECTIVE: To investigate the feasibility of establishing a reliable animal model of chronic ischemia in high-fat diet rats using the thread embolism method. 
    METHODS: Forty Sprague-Dawley rats, 2 weeks of age, weighing 50-70 g, were provided by the Animal Experimental Center of Chengde Medical University in China. The rats were randomly divided into two groups, and fed with high-fat diet (experimental group) and normal diet (control group), respectively. Chronic ischemia animal models were then established in the hind limbs of all the rats using the thread embolism technique. Muscle strength of the rats was detected at 1, 2, 4, and 6 weeks postoperatively. Meanwhile, blood flow changes in the hind limbs and histological changes of the gastrocnemius and vascular tissue were observed using laser Doppler and hematoxylin-eosin staining, respectively.
    RESULTS AND CONCLUSION: (1) Most of the rats in the experimental group had abnormal muscle performance at 1 week postoperatively. Most of the rats in the control group had nothing-unusual (P < 0.05). Most of the rats in both groups appeared with abnormal muscle performance at 2 and 4 weeks postoperatively, and there was no significant difference between the two groups. At 6 weeks postoperatively, the number of rats with abnormal muscle performance was lower in the control group than the experimental group. (2) Laser Doppler flow examination and histological examination both showed that the experimental group had ischemic symptoms earlier than the control group, and worse recovery from ischemia was found in the experimental group compared to the control group. These findings indicate that the thread embolism method can effectively make a rat model of chronic ischemia, and the addition of high-fat diet can cause an ischemic appearance earlier than the addition of normal diet, indicating the high-fat diet can prolong the state of chronic ischemia.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Expression of myeloid differentiation factor 88 in a rat model of experimental periapical periodontitis  
    Zuo Meina1, Wang Lina1, Han Shujuan2, Dong Ming3, Niu Weidong1
    2019, 23 (7):  1068-1072.  doi: 10.3969/j.issn.2095-4344.1049
    Abstract ( 303 )   PDF (4156KB) ( 124 )   Save

    BACKGROUND: Studies have found that myeloid differentiation factor 88 is expressed in rat dental follicles and affects the eruption of teeth and the number of osteoclasts.
    OBJECTIVE: To further verify the expression of myeloid differentiation factor 88 in an experimental rat model of periapical periodontitis.
    METHODS: Twenty-five male Sprague-Dawley rats, 6 weeks of age, were provided by the SPF Animal Experimental Center of Dalian Medical University in China. The pulp of the first mandibular molar was exposed to establish the rat model of experimental periapical periodontitis in each rat. Endodontic pulp exposure 0 week was set as control group, while endodontic pulp exposure 1, 2, 3, 4 weeks as experimental group. Five rats were randomly selected at each time point after exposure of the endodontic pulp. Then, paraformaldehyde fixation of the mandible, decalcification, and pathological frozen sections were mode. Hematoxylin-eosin staining and immunohistochemical staining were used to observe the pathological changes of the periapical tissues. RT-PCR was used to detect the expression of myeloid differentiation factor 88 in the rat model of experimental periapical periodontitis.
    RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining results showed inflammatory infiltration in the apical area of the experimental group, and spread around, indicating that we successfully established the rat model of experimental periapical periodontitis. (2) Immunohistochemical staining results showed the positive expression of myeloid differentiation factor 88 in the periapical inflammation area at 1, 2, 3, 4 weeks after modeling. The positive expression of myeloid differentiation factor 88 significantly increased within 1-3 weeks, peaked at 3 weeks, but decreased at 4 weeks. There were only a small number of myeloid differentiation factor 88 positive cells in the control group. (3) RT-PCR results showed that the mRNA expression of myeloid differentiation factor 88 increased gradually from 1 week to 3 weeks, and then decreased at 4 weeks. To conclude, myeloid differentiation factor 88 is expressed in the animal model of experimental periapical periodontitis and shows a tendency of expression over time. Myeloid differentiation factor 88 may be involved in the occurrence of periapical periodontitis and the resorption of alveolar bone in periapical periodontitis.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Evaluation of lower limb function in rat models of spinal cord injury at different segments
    Wei Weibing1, Zhou Xiangxing1, Zhou Binbin2, Li Yijun1, Zou Zhilong1, Yang Hui1
    2019, 23 (7):  1073-1077.  doi: 10.3969/j.issn.2095-4344.1058
    Abstract ( 412 )   PDF (4100KB) ( 172 )   Save

    BACKGROUND: An ideal animal model is essential for studies on the pathophysiologic and anatomical structures. Therefore, selecting injury method and a safe and effective segment of spinal cord is critical.
    OBJECTIVE: To explore the effects of different spinal cord injury segments on the recovery of lower extremity function.
    METHODS: Rat models of spinal cord injury at different segments were established. Seventy-two Sprague-Dawley rats were randomly divided into six groups: T8, T9, T10, T11, T12 and sham operation (laminectomy and spinal meninge exposure) groups. The rats in the T8, T9, T10, T11, T12 groups were used for establishing rat models of spinal cord injury by clipping the corresponding spinal cord segments using a microvessel clamp.
    RESULTS AND CONCLUSION: In the climbing grid experiment, the number of rats stepping on the air in each group decreased with time. Compared with the sham operation group, the Basso, Beattie, and Bresnahan scores in each group were significantly decreased. At 2-4 weeks after injury, the Basso, Beattie, and Bresnahan scores in the T9, T10 and T11 groups were higher than those in the T8 and T12 groups, and the scores in the T10 group were higher than those in the T9 and T11 groups. In summary, T10 group showed the best recovery of lower limb function after spinal cord injury.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Dominant T cell and B cell epitopes in EgA31 protein of Echinococcus granulosus by bioinformatics
    Zhao Xiao1, Zhang Fengbo2, Wang Hongying1, Yan Fang3, An Mengting1, Li Yujiao1, Pang Nannan4, Ding Jianbing1
    2019, 23 (7):  1078-1083.  doi: 10.3969/j.issn.2095-4344.1001
    Abstract ( 478 )   PDF (1232KB) ( 86 )   Save

    BACKGROUND: EgA31 protein participates in acetabulum muscle contraction, and is an important candidate vaccine in the protective immunity of Echinococcus granulosus.
    OBJECTIVE: To analyze the EgA31 protein of the Echinococcus granulosus using bioinformatics and to predict the potential dominant T cell and B cell epitopes.
    METHODS: The amino acid sequence of EgA31 from the NCBI GenBank database was obtained. The molecular mass, theory isoelectric point, amino acid composition, atomic composition, extinction coefficient, instability coefficient, and total average hydrophobicity were analyzed using ProtParam online program. The secondary structure of EgA31 protein was analyzed using protein module of DNAstar software and SOPMA online services. The tertiary structure model of EgA31 protein was established Phyre server. Finally, the dominant T cell and B cell epitopes were predicted using ABCpred, BepiPred, SYFPEITHI, and IDBE software.
    RESULTS AND CONCLUSION: EgA31 protein consisted of 601 amino acids, including 112 negatively charged residues and 121 positively charged residues, which were classified as the unstable and hydrophilic protein. Online service analysis revealed that the second structure of EgA31 protein comprised 82.36% of α-helixs, 4.16% of β-sheets, 3.16% of β-turns and 10.32% of random coils. According to ABCpred, BepiPred, SYFPEITHI and IDBE databases, four dominant T cell epitopes, six dominant T cell epitopes, and one T-B combined epitopes were predicted successfully. Bioinformatics can comprehensively predict the dominant T cell and B cell epitopes in EgA31 protein of the Echinococcus granulosus, which lays a foundation for developing vaccine and reagent in the future.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Protection of reduced glutathione and ulinastatin on frozen-thawed ovarian tissues after xenotransplantation
    Long Huidong1, 2, Long Lingli1, Mai Qingyun1, Zhao Wen1, Li Yubin1
    2019, 23 (7):  1084-1089.  doi: 10.3969/j.issn.2095-4344.1074
    Abstract ( 321 )   PDF (1383KB) ( 105 )   Save

    BACKGROUND: Reduced glutathione, as an effective antioxidant can inhibit inflammatory cytokines. Ulinastatin can directly remove oxygen free radicals and inhibit production of oxygen free radicals and various inflammatory mediators, sustain the stability of cell membrane and lysosomal membrane, and inhibit the activity of active enzymes, thus protecting the transplanted organ.
    OBJECTIVE: To evaluate the effect of reduced glutathione and ulinastatin on improving the xenotransplantation efficiency of frozen-thawed human ovarian tissue.
    METHODS: Ovarian tissue samples from 6 women aged 31.8±5.7 years old in the First Affiliated Hospital, Sun Yat-sen University, were collected and each tissue sample was randomly divided into four groups after slow freezing: reduced glutathione, ulinastatin, glutathione + ulinastatin and blank control (normal saline) groups. Ovarian tissues were xenotransplantated under the back skin of male NOD-SCID mice after 15-minute cultivation. The mice were administered with corresponding drugs for 5 consecutive days. The grafts were collected at 1, 3, 7, 14, 28, 56, and 85 days after transplantation. Histomorphology, growth situation and viability of the follicle were observed through hematoxylin-eosin staining and enzymatic isolation method.
    RESULTS AND CONCLUSION: The majority of the frozen-thawed ovarian tissues were primordial follicles (79.5±14.3%). The viability of the follicle in fresh ovarian tissue was 97.6%, and (84.4±5.7)% in thawed ovarian tissue. The reduced glutathione, ulinastatin, and glutathione + ulinastatin groups showed significantl improvement in the viability of the follicle compared with the control group (P=0.022). There was no significant difference between single and combined drug groups (P > 0.05). The follicle viability reached a lowest value at 3 days after transplantation (P < 0.05), and increased after 1 week later. The mean survival rate of the follicle in the isolated follicle at different time points showed no significant difference among groups (P=0.114). Compared with the histological sections, the isolated follicle showed highest survival rate. Our results suggest that reduced glutathione and ulinastatin can increase the early viability of the follicles after xenotransplantation, and combined administration shows no extra benefit.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Ginsenoside Rg1 protects against oxygen-glucose deprivation/reperfusion injury in PC12 cells
    Ye Jintao1, Li Fengtao1, Song Huanjin1, Xue Jianli1, Lin Lei2, Cheng Bin1
    2019, 23 (7):  1090-1096.  doi: 10.3969/j.issn.2095-4344.1066
    Abstract ( 307 )   PDF (5328KB) ( 150 )   Save

    BACKGROUND: Ginsenoside has anti-tumor, anti-oxidative, anti-fatigue, anti-aging, hypolipidemic, memory enhancement, immunity enhancement effects, and Rb1 and Rg1 are the most important active components of ginsenosides. However, it is unclear whether ginsenoside Rg1 also protects against apoptosis in PC12 cells induced by oxygen-glucose deprivation (OGD).
    OBJECTIVE: To investigate the effects of pretreatment with ginsenoside Rg1 on PC12 cell viability, survivin and caspase-3 expression and apoptosis in PC12 cells induced by OGD/reperfusion.
    METHODS: In the OGD/reperfusion model, the PC12 cells were pretreated with 5, 10, 20, 40 μmol/L ginsenoside Rg1. Normal cells were used as controls. Cell viability was determined by MTT assay. Expression of survivin and caspase-3 proteins was measured by immunocytochemistry. Apoptosis in PC12 cells was detected by TUNEL assay.
    RESULTS AND CONCLUSION: (1) The cell viability was significantly decreased after OGD/reperfusion (P < 0.05). Pretreatment with ginsenoside Rg1 could elevate the cell viability, but it was still lower than the normal level (P < 0.05). (2) Pretreatment with ginsenoside Rg1 increased survivin positive cells, but decreased caspase-3 positive cells and TUNEL positive cells. (3) Neuronal apoptosis was negatively correlated with the ginsenoside Rg1-induced survivin, but positively correlated with the caspase-3. All these findings indicate that ginsenoside Rg1 pretreatment can promote the expression of survivin in PC12 cells after OGD/reperfusion, and moreover, this promotion effect on survivin can further inhibit the expression of caspase-3, thereby suppression apoptosis in PC12 cells, in a dose-dependent manner.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Mechanism underlying adiponectin intervention on the apoptosis of human periodontal ligament cells mediated by high glucose
    Lin Yangdong, Yin Kai
    2019, 23 (7):  1097-1102.  doi: 10.3969/j.issn.2095-4344.1022
    Abstract ( 407 )   PDF (4228KB) ( 103 )   Save

    BACKGROUND: Adiponectin is a recently discovered specific protein secreted by adipocytes, which has been shown to regulate glucose and lipid metabolism and protect against oxidative stress. Thereafter, adiponectin is a promising therapy for diabetes mellitus and its complications. 
    OBJECTIVE: To investigate the effect of the classic Wnt/β-catenin signaling pathway on the apoptosis of human periodontal ligament cells mediated by high glucose after the intervention of adiponectin.
    METHODS: Human periodontal ligament cells were revived and cultured in vitro. The cells were randomly divided into control group (normal RPMI1640 culture medium containing 11 mmol/L glucose), high glucose group (40 mmol/L glucose in the RPMI1640 culture medium), mannitol group (19.5 mmol/L mannitol in RPMI1640 culture medium) and adiponectin group (40 mmol/L glucose in the RPMI1640 culture medium+20 mg/L adiponectin). In different time points (24-96 hours), the cell survival rate in each group was detected by MTT assay. The cell cycle distribution and the cell apoptosis was detected by flow cytometry. The Wnt1, Wnt10b and β-catenin gene and protein expression levels were determined by RT-PCR and western blot assay, respectively.
    RESULTS AND CONCLUSION: The high level glucose could inhibit the cell proliferation, improve the cell apoptosis and had no obvious effect on the cell cycle. The effect to decrease the cell proliferation was in a time-dependent manner. But the adiponectin intervention significantly increased the cell proliferation and reduced the apoptosis, compared with the high glucose group (P < 0.05). Compared with the control and mannitol groups, the mRNA and protein expression levels of Wnt1, Wnt10b and β-catenin were significantly decreased in the high glucose group, while the levels in the adiponectin group were obviously increased (P < 0.05). To conclude, high glucose can improve the apoptosis of human periodontal ligament cells and inhibit the proliferation, but have no obvious effect on the cell cycle. Adiponetin can remit the effect induced by high glucose by up-regulating Wnt1 and Wnt10b expression and then activate the Wnt/β-catenin signaling pathway.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Genotype identification and model application of apolipoprotein E knockout mice
    Zhang Wenjiang1, 2, Yi Jian3, Jia Ping1, Chen Bowei1, Wang Yong2, Liu Baiyan1
    2019, 23 (7):  1103-1108.  doi: 10.3969/j.issn.2095-4344.1059
    Abstract ( 888 )   PDF (6384KB) ( 202 )   Save

    BACKGROUND: Apolipoprotein E knockout mice are an ideal model of atherosclerosis, which has been reported in the other diseases.
    OBJECTIVE: To systematically summarize the application of apolipoprotein E knockout mice in disease models.
    METHODS: Six male and six female APOE-/- mice introduced from the Biomedical Research Center of Nanjing University were bred in each cage at 1:1 in the SPF laboratory. The genomic DNA was extracted from mouse tail tissue by the method of boiling lysis, and the genotypes were detected by PCR. The APOE-/- mice were fed with normal diet for 8 weeks, and then fed with a Western diet (provided by Beijing Huafukang Bioscience Co., Ltd., No. H10141, 21% of fat, 0.15% of cholesterol) to 25 week old. At the same time, 10 8-week-old C57/6J mice were fed with a normal diet to 25 weeks old.
    RESULTS AND CONCLUSION: Genetic identification: agarose gel electrophoresis results indicated that the size of molecular weight of the PCR products from APOE-/- gene knockout mice was consistent with the expected target gene fragment and of different genotypes successfully. Changes of related indicators: compared with the wild type mice, the body mass, liver weight, spleen weight, abdominal fat weight, liver index, and serum levels of total cholesterol, triacylglycerol, low density lipoprotein cholesterol, high density lipoprotein cholesterol, alanine transaminase, and aspartate transaminase in the APOE-/- mice were significantly increased (P < 0.05). The histological changes: APOE-/- mice showed obvious plaques in the aortic arch, aortic valve and common carotid artery by hematoxylin-eosin staining, but no plaques in the wild type mice. The liver of APOE-/- mice had vacuoles of varying sizes, and the nucleus was pushed aside. The nucleus of wild mice liver was in the middle, the cytoplasm was red, and there was no vacuole phenomenon. These results suggest that PCR can fast and reliably identify the genotypes of apolipoprotein E knockout mice using genomic DNA extracted by boiling lysis. APOE-/- mice are ideal models for studying atherosclerosis, fatty liver, hyperlipidemia, hypercholesterolemia, obesity and many other diseases.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Risk factors for multi-drug resistant organisms infection after liver transplantation
    Liu Mengyuan1, Fang Fang2
    2019, 23 (7):  1109-1114.  doi: 10.3969/j.issn.2095-4344.0572
    Abstract ( 364 )   PDF (1018KB) ( 78 )   Save

    BACKGROUND: Multi-drug resistant organisms (MDRO) infection after liver transplantation seriously affects the survival and mortality of patients after liver transplantation. High failure in MDRO infection treatment catches much attention.
    OBJECTIVE: To analyze the risk factors for MDRO infection after liver transplantation. 
    METHODS: A prospective cohort study was used to collect and analyze the clinical data of 114 patients after liver transplantation. There were 28 cases of MDRO infection, and 86 cases of non-MDRO infection. The MDRO infection rate was calculated. Univariate analysis and multivariate logistic regression analysis were used to screen independent risk factors.
    RESULTS AND CONCLUSION: The MDRO infection rate after liver transplantation was 24.6%. Multivariate logistic regression analysis results showed the following four factors: prolonged prothrombin time (OR: 3.102, 95% CI (0.066-9.024), P=0.038), ICU retention time ≥ 10 days (OR: 3.745, 95%CI (1.313-10.687), P=0.014), tracheal intubation time ≥ 72 hours (OR: 8.972, 95%CI (1.983-40.582), P=0.004), thoracentesis (OR: 3.564, 95%CI (1.223-10.387), P=0.020). These results imply that prolonged prothrombin time, ICU indwelling time ≥ 10 days, tracheal intubation time ≥ 72 hours and thoracentesis can increase the risk of MDRO infection in patients after liver transplantation.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Research progress and medical application of modeling and simulation of cardiovascular system
    Zhang Chaohui, Zhao Feng, Feng Yunpeng, Wang Wenbin, Kuang Baoping, Huang He
    2019, 23 (7):  1115-1121.  doi: 10.3969/j.issn.2095-4344.1075
    Abstract ( 630 )   PDF (1208KB) ( 127 )   Save

    BACKGROUND: Modeling and simulation technology has gained more and more attention and gradually is applied in fields of military drills, industrial tests and medical tests. Of them, there are various professionals have emerged in studies on medical cardiovascular system.
    OBJECTIVE: To systematically review the simulation method of cardiovascular system.
    METHODS: Databases of PubMed, VIP and CNKI were searched for the articles concerning the cardiovascular modeling and simulation published from 1974 to 2018 with the keywords of “cardiovascular system, modeling simulation, the heart model” in English and Chinese, respectively. The modeling methods and simulation types of cardiovascular system, and application of cardiovascular simulation in coronary heart disease, valvular disease, heart failure, hypertension and myocardial infarction were summarized, and then made prospects.
    RESULTS AND CONCLUSION: According to the clinical, medical teaching and research needs, the summary includes: (1) The hemodynamic model of coronary circulatory system simulates the coronary circulatory system in the case of coronary heart disease, which solves the problem of invasion and expensive const. Moreover, it is of great significance for the early diagnosis and treatment of coronary heart disease. (2) The mitral valve three-dimensional simulation model analyzes the stress distribution of the mitral valve. The simulation is more elaborate in the geometric structure, which can improve the treatment scheme and indirectly reduce the risk of surgery. (3) The cardiovascular system focuses on the many electrical network models, which helps further understand the clinical analysis and diagnosis of cardiovascular diseases. (4) The three-dimensional finite element mechanical model of human left ventricle simulates the electrical excitation and myocardial infarction in myocardial infarction, and can guide the clinical practice. (5) We comprehensively summarize the modeling and simulation of cardiovascular system, which contributes to further understanding the pathogenesis of cardiovascular disease, and is of great significance for the prevention, auxiliary diagnose and clinical treatment of cardiovascular disease. 

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Epoxyeicosatrienoic acids in tissue and organ regeneration: research status and prospects
    Dang Haixia1, Wang Fu2
    2019, 23 (7):  1122-1128.  doi: 10.3969/j.issn.2095-4344.1076
    Abstract ( 392 )   PDF (1006KB) ( 90 )   Save

    BACKGROUND: Epoxyeicosatrienoic acids are lipid mediators produced by arachidonic acid metabolism. Early clinical studies on epoxyeicosatrienoic acids mainly focus on inflammation, pain, cardiovascular and cancer diseases. In recent years, epoxyeicosatrienoic acids have been found to be closely related to regeneration of tissues and organs in vivo and in vitro.
    OBJECTIVE: To review the origin and metabolism of epoxyeicosatrienoic acids, the role of epoxyeicosatrienoic acids in the tissue and organ regeneration as well as related mechanisms associated with neovascularization and inflammation, so as to provide the basis for future research.
    METHODS: CNKI and PubMed databases were searched using the keywords of “epoxyeicosatrienoic acids, tissue regeneration, organ regeneration, angiogenesis” in Chinese and English, respectively, to retrieve relevant articles published from January 2000 to February 2018. Sixty-nine representative articles were selected for overview.
    RESULTS AND CONCLUSION: Epoxyeicosatrienoic acids, as important lipid molecules produced by arachidonic acid, play an important role in the growth and regeneration of tissues and organs. The related mechanisms mainly focus on the neovascularization. Tissue regeneration and repair involve the regulation of proinflammatory and anti-inflammatory cytokines. Epoxyeicosatrienoic acids have been shown to be the important signals in regulating inflammation, pain and neovascularization. Consequently, the further exploration of the relevant mechanisms of epoxyeicosatrienoic acids in tissue and organ regeneration, can provide novel treatment idea for soft tissue defects, and other defects caused by periodontitis, trauma, inflammation and tumor.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Research progress of ginsenosides Rb1 in flap ischemia/reperfusion injury
    Cao Chang, Ren Xiaoyan, Meng Fanjun, Cheng Fengrui, Cen Ying
    2019, 23 (7):  1129-1135.  doi: 10.3969/j.issn.2095-4344.1077
    Abstract ( 319 )   PDF (1105KB) ( 118 )   Save

    BACKGROUND: Ischemia/reperfusion injury is a common pathophysiological process in plastic surgery, which happens in random flap, free flap and composite tissue flap transplantation, even will lead to partial or whole flap necrosis. Ginsenosides Rb1 has been shown to exert protection in ischemia/reperfusion injuries of various tissues and organs.
    OBJECTIVE: To review the mechanism of ischemia/reperfusion injury of flap and action mechanism of ginsenosides Rb1.
    METHODS: CNKI and PubMed databases were searched for the articles published between January 1996 and June 2018 with the keywords of “ginsenosides, reperfusion injury, flaps” in English and Chinese, respectively. The articles addressing the pharmacological effects of ginsenosides Rb1, ginsenosides Rb1 alleviating tissue ischemia/reperfusion injury and flap ischemia/reperfusion injury were included.
    RESULTS AND CONCLUSION: Flap ischemia/reperfusion injury is complex non-specific cascade damage after ischemic tissues restoring blood supply, which Involves in excessive production of oxygen free radicals, inflammation infiltration and its interaction with endothelial cells, energy failure, intracellular calcium overload, and nitric oxide production. Ginsenosides Rb1 has the effects of anti-oxidation, anti-inflammation and antiapoptosis. Ginsenosides Rb1 exerts good protection in ischemia/reperfusion injuries of various tissues and organs through anti-oxidation, anti-inflammation, increasing nitric oxide production, antiapoptosis, inhibiting calcium overload.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Application and prospect of organoid technique in cancer research 
    Gao Jianjun, Qin Wei, Wang Hao, Zhong Xiangyu
    2019, 23 (7):  1136-1141.  doi: 10.3969/j.issn.2095-4344.1577
    Abstract ( 485 )   PDF (1048KB) ( 150 )   Save

    BACKGROUND: As a new kind of model in vitro, the organoid allows the in vitro long-term culture of patient-derived cancer cells that faithfully recapitulate the in vivo phenotype. This technology facilitates antineoplastic drug screening and personalized therapies.
    OBJECTIVE: To describe the development of organoid technology briefly and review the applications in cancer research.
    METHODS: Using the keywords of “organoid, three-dimensional culture, cancer research, stem cell” in English and Chinese, respectively, the first author retrieved relevant articles published from May 2013 to May 2018 in PubMed and CNKI databases. The literature types included clinical research, basic research and review articles. After removal of the articles that were not related to the purpose of the study or repetitive, 47 articles were finally analyzed.
    RESULTS AND CONCLUSION: Patient-derived organoids provide a reliable model for cancer research and treatment, and in particular, open up a new perspective for personalized cancer treatment. Patient-derived organoids are three-dimensional tissue cell clusters derived from tissues or tumor specific stem cells, which can recapitulate characteristics of tumors and heterogeneity of tumor cells in vivo. In cancer research, optimizing various culture conditions has resulted in the development of numerous patient-derived tumor organoids, including colon, prostate, gastric, breast, and pancreatic cancers, in addition to endometrial/ovary carcinomas, uterine carcinosarcoma, urothelial carcinoma, and renal carcinoma. However, there are obvious limitations in organoids culture, one of which is the lack of nerve, blood vessels and immune cells in this model. This cannot show all features of organs, and partially simulate the process of disease.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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    Application value of surface electromyography in the repair of peripheral nerve injury
    Xia Ling, Wang Pan, Wu Chunfang, Zhang Zhaobo
    2019, 23 (7):  1142-1148.  doi: 10.3969/j.issn.2095-4344.1078
    Abstract ( 383 )   PDF (1090KB) ( 96 )   Save

    BACKGROUND: Surface electromyography is an objective quantitative index that has many advantages, such as safety, non-invasion, painless, reliability, and objective quantification and plays an irreplaceable role in rehabilitation evaluation.
    OBJECTIVE: To summarize and analyze the application of surface electromechanical in rehabilitation medicine, thereby providing clues and directions for further clinical application.
    METHODS: Medline, PubMed, WanFang, VIP and CNKI databases were retrieved for the literature published between 2006 and 2018 with the keywords of “surface electromyography assessment, rehabilitation assessment, clinical” in English and Chinese, respectively. After screening, 59 eligible articles were reviewed.
    RESULTS AND CONCLUSION: Extensive application of surface electromyography in rehabilitation and sports medicine can objectively and quantificationally detect the physical activity of the limb of stroke patients, especially in motion analysis, and muscle function evaluation. Surface electromyography plays an important role in the evaluation of rehabilitation and the establishment of rehabilitation goals, which can directly reflect the changes of muscle function.

    中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

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