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    09 April 2013, Volume 17 Issue 15 Previous Issue    Next Issue
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    Dynamic observation of rat hippocampal neuronal death process by hopping probe ion conductance microscopy
    Yang Guo-wei, Zhu Hui, Zhou Zi-wei, Li Ying, Zhang Yan-jun, Zhang Jian-ning
    2013, 17 (15):  2661-2668.  doi: 10.3969/j.issn.2095-4344.2013.15.001
    Abstract ( 465 )   PDF (2559KB) ( 580 )   Save

    BACKGROUND: The molecular mechanism underlying neuronal cell death has been made some progress in the past years. However, the cell morphology dynamics after neuron death is still missing because of technical shortcomings. Even the time required for the execution of the death program is still unknown. However, neurons cultured in vitro can be observed continually in a non-contact manner by a hopping probe ion conductance microscopy, therefore the morphology changes of a death process can be obtained.
    OBJECTIVE: To dynamically observe morphological changes of rat hippocampus neurons during a cell necrotic process by the hopping probe ion conductance microscopy.
    METHODS: Primary Wistar rat hippocampal neurons were used for the experiment, and cell necrosis was induced by 1 mmol/L hydrogen peroxide solution for 30 minutes. Neurons without hydrogen peroxide-treatment were used as control. The morphology of neuronal soma and neurites were continually imaged by the hopping probe ion conductance microscopy after hydrogen peroxide treatment, and the height and volume of a neuron were simultaneously measured.
    RESULTS AND CONCLUSION: Continuous imaging for 9 hours after hydrogen peroxide treatment under the hopping probe ion conductance microscopy showed that: Hippocampal neurons gradually transformed from a normal triangular or fusiform shape to a more spherical shape due to cell swelling. Axon beading and dendritic beading were gradually formed after scanning, and finally the neurites were ruptured and dissolved. Measurement of the cell height and volume showed that: The height and volume of hydrogen peroxide-treated neurons elevated after 1 hour of scanning, and continued increasing up to 7 hours after scanning (P < 0.05). The final height and volume were approximately twice as beginning. The increase of height and volume manifested as linear correlation with time within 1 to 7 hours by linear-regression analysis (cell height b=0.15, P < 0.005; cell volume b=0.17, P < 0.05). Meanwhile, no significant morphological change was observed during the time of scanning a normal neuron. It is successful to observe necrotic process of hippocampus neurons by the hopping probe ion conductance microscopy. Higher-quality cell image and more accurate information about cell height and volume can be obtained.

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    Turtle deer glue, epimedium, and rhodiola pills enhance bone mineral density of ovariectomized rats
    Yang Hao-xia, Yang Yang, Xue Peng, Zhou Fu-yi, Wang Sheng-jun
    2013, 17 (15):  2669-2676.  doi: 10.3969/j.issn.2095-4344.2013.15.002
    Abstract ( 537 )   PDF (920KB) ( 717 )   Save

    BACKGROUND: At present, there are no systematic pharmacodynamic studies addressing osteoporosis
    treatment with Chinese medicine. Chinese prescription is not simple enough and research on the mechanism is not close to the guiding of Chinese medicine theory.
    OBJECTIVE: To observe the therapeutic effects of turtle deer glue, epimedium, and rhodiola pills on serum indices, bone mineral density and osteoclast regulation in ovariectomized rats.
    METHODS: Ovariectomized rats were prepared and subjected to intragastric administration of turtle deer glue, epimedium and rhodiola in comparison with sham-surgery and model groups. The body mass was detected once a week starting from week 4. Bone mineral density, serum calcium, phosphorus content, and blood alkaline phosphatase activity were detected at week 12. Osteoclast inhibiting rates were evaluated by in vitro osteoclast culture model.
    RESULTS AND CONCLUSION: The body mass increased for the model group, the turtle deer glue could significantly reduce the increase rate (P < 0.05). After 12 weeks, compared with the model group, the serum calcium content was significantly higher in the three Chinese medicine groups (P < 0.05), whereas the blood phosphorus content and alkaline phosphatase activity were obviously lower (P < 0.05). Bone mineral density in the turtle deer glue and epimedium groups was significantly increased as compared with that in the model group (P < 0.05). The number of resorption pits was decreased significantly in the three Chinese medicine groups, especially in the rhodiola group, as compared with the sham-surgery group (P < 0.05). Rhodiola pills could prohibit the absorptive function of osteoclasts, the inhibitory rate of which was 86.85% (P < 0.01). These findings indicate that turtle deer glue, epimedium, and rhodiola all have significant effects on ovariectomized osteoporosis. Turtle deer glue and epimedium mainly increase bone mineral density, while rhodiola pills prohibit the absorptive function of osteoclasts.

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    Molecular mechanism by which Jiangu granules promote osteoblast proliferation
    Lin Yu, Lu Tian-xiang, Wu Yin-sheng, Huang Yun-mei, Lin Yan-ping
    2013, 17 (15):  2677-2684.  doi: 10.3969/j.issn.2095-4344.2013.15.003
    Abstract ( 434 )   PDF (905KB) ( 512 )   Save

    BACKGROUND: Extracellular signal-regulated kinase is a protein kinase dependent on Ras pathway activation, and plays an important role in the proliferation and differentiation of osteoblasts. PD98059 is a specific MEK inhibitor, and inhibits phosphorylation of extracellular signal-regulated kinase through inhibition of MEK activity, thus blocking extracellular signal-regulated kinase signaling pathway. At present, the role of extracellular signal-regulated kinase in the process of proliferation and differentiation of rat osteoblasts is reported less. It is still unclear about the role of extracellular signal-regulated kinase in the regulation of osteoblast proliferation and differentiation promoted by Jiangu granules.
    OBJECTIVE: To observe the role of extracellular signal-regulated kinase signaling pathway in the promotion of osteoblast proliferation and differentiation after intervention with Jiangu granules. 
    METHODS: Passage 3 osteoblasts from the skull of Sprague-Dawley rats were harvested and divided into four groups. Control group was intervened with saline serum. Drug group was intervened with the best concentration of serum containing Jiangu granules. Inhibitor group was intervened by PD98059. Combination group was intervened with PD98059 plus serum containing Jiangu granules. 3-(4,5-Dimethylthiazol-2- yl)-2,5- diphenyltetrazolium bromide assay was used to determine proliferative capacity of osteoblasts. Alkaline phosphatase and hydroxyproline levels were measured by colorimetric method. Expression of core binding factor alpha 1, type Ⅰ collagen, Osterix mRNA was measured by PCR-SYBR GREEN. Expression of extracellular signal-regulated kinase in osteoblasts was detected using western blot method.
    RESULTS AND CONCLUSION: After blocking extracellular signal-regulated kinase signal pathway, the expression of extracellular signal-regulated kinase in the inhibitor and combination groups was significantly lower than that in the control and drug groups. Levels of alkaline phosphatase and hydroxyproline, as well as core binding factor alpha 1, type Ⅰ collagen, Osterix mRNA expression in osteoblasts, in the inhibitor group were significantly lower than those in the control and drug groups. It is indicated that extracellular signal-regulated kinase exerts an important role in promoting osteoblast proliferation and differentiation after intervention with Jiangu granules.

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    Early diagnosis value of combination detection of four serum biomarkers for osteoarthritis
    Yu Wei-guang, Zhang Xin-chao, Zhu Xing-fei, Liu Zhi-yong
    2013, 17 (15):  2685-2692.  doi: 10.3969/j.issn.2095-4344.2013.15.004
    Abstract ( 622 )   PDF (1272KB) ( 770 )   Save

    BACKGROUND: Studies have shown that biomarkers in the diagnosis of early osteoarthritis have their unique advantages, but so far, it is not clear on application value of biomarkers in the diagnosis of early 
     osteoarthritis in human being.
    OBJECTIVE: To investigate the value and significance as early diagnosis and condition assessment indicators of osteoarthritis by means of quantitative detection of the changes in C-telopeptide of collagen typeⅡ, matrix metalloproteinase protein-3, tumor necrosis factor-α, and interleukin-1. 
    METHODS: Thirty-five New Zealand white rabbits were divided into experimental group (n=28) and control group (n=7). Resection of the ligament and meniscus was performed in the experimental group for modeling. No treatment was done in the control group. C-telopeptide of collagen type Ⅱ, matrix metalloproteinase protein-3, tumor necrosis factor-α, and interleukin-1 levels were detected at 1, 2, 3, 4 weeks after modeling in the experimental group, and these biomarkers were also detected in the control group at the same time points. Knee MRI examination, joint sample general observation and histopathological observation were conducted.
    RESULTS AND CONCLUSION: Levels of four biomarkers in the experimental group were significantly higher than those in the control group at each time point after modeling (P < 0.05). Histopatological changes and knee MRI variation did not occur at 1 week after operation; however, these changes appeared at 2 weeks, and strengthened at 3-4 weeks. In addition, the levels of four biomarkers began to increase at 1 week after operation. The results suggest that combination detection of C-telopeptide of collagen type II, matrix metalloproteinase protein-3, tumor necrosis factor-α, and interleukin-1 in serum can become a reference indicator for early diagnosis and assessment of osteoarthritis.

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    A novel co-culture system for interaction of mouse osteoblasts and osteoclasts
    Li Jian-yu, Liu Lu, Wan Zong-ming, Hao Qing-xin, Li Rui-xin, Guo Yong,
    2013, 17 (15):  2693-2967.  doi: 10.3969/j.issn.2095-4344.2013.15.005
    Abstract ( 346 )   PDF (324KB) ( 3174 )   Save

    BACKGROUND: It is well known that bone remodeling is an important biological process in which osteoblasts and osteoclasts play the critical role. However, the detailed mechanisms of osteoblast-osteoclast communication in bone remodeling remain unclear.
    OBJECTIVE: To in vitro establish a novel co-culture system for interaction of mouse osteoblasts and osteoclasts using the Transwell technology.
    METHODS: Osteoblastic cell line MC3T3-E1 and preosteoclasts RAW264.7 were selected, and then the cells were induced to differentiate into osteoblasts and osteoclasts, respectively. The co-culture system for osteoblasts and osteoclasts interaction was established with Transwell co-culture plate (0.4 µm polyester film), and the osteoblasts and osteoclasts were co-cultured for 6 days. After that, the proliferation and differentiation activities of osteoblasts were analyzed through testing the cell activity and alkaline phosphatase activity, and the differentiation and bone resorption of osteoclasts were observed using tartrate-resistant acid phosphatase staining, toluidine blue staining, tartrate-resistant acid phosphatase activity detecting and scanning microscope technology.  
    RESULTS AND CONCLUSION: In the co-culture system, the unlimited proliferation ability of osteoblasts was decreased, while the differentiation activity was increased. Meanwhile, the preosteoclasts could differentiate into mature osteoclasts with bone resorption function. Thus, this co-culture system can be applied in the further study of osteoblast-osteoclast communication in bone remodeling.

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    Bone extrusion effects on implant primary stability
    Sun Song, Zhao Feng, Sun Yong
    2013, 17 (15):  2698-2702.  doi: 10.3969/j.issn.2095-4344.2013.15.006
    Abstract ( 635 )   PDF (1270KB) ( 1302 )   Save

    BACKGROUND: Primary stability of implants is the key factor to the success of implantation, and improving primary stability can raise the rate of successful implantation when bone quality is poor in implant region.
    OBJECTIVE: Through the establishment of the cancellous bone model to simulate the clinical dental implant surgery, to measure the primary stability of the implant so as to assess the bone extrusion operation effect as compared with the traditional hole prepared surgery.
    METHODS: Bone segments from the crest of scapular spine of fresh pigs were made into cuboid bone blocks (n=12) of 10 mm×10 mm×10 mm. After fixation, bone extrusion operation and traditional hole prepared surgery were performed to implant BEGO implants with the same specifications and the same type. Primary stability of implants was recorded, and t test was performed.
    RESULTS AND CONCLUSION: Compared with the traditional hole prepared surgery, a better primary stability for the implant was obtained following bone extrusion operation (P < 0.05), indicating that bone extrusion operation is better for primary stability of implants.

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    Effect of icariin and simvastatin with different dosage ratios on rabbits’ radial fracture healing
    Wang Hui-ping, He Ai-yong, Chang Xiao-fei
    2013, 17 (15):  2703-2713.  doi: 10.3969/j.issn.2095-4344.2013.15.007
    Abstract ( 471 )   PDF (3226KB) ( 510 )   Save

    BACKGROUND: Icariin and simvastatin alone or in combination can promote the radial fracture healing of rabbits. But the best dosage ratio is still lack of basis.
    OBJECTIVE: To investigate the effect of icariin and simvastatin with different dosage ratios on rabbits’ radial fracture healing.
    METHODS: Forty-five healthy rabbits were enrolled to prepare bilateral radial fracture models, and then randomly divided into five groups: model group; bone morphogenetic protein-2 control group; and three experimental groups (ratio of icariin and simvastatin: 3:1, 1:1, 1:3). Model and bone morphogenetic protein-2 control groups were given normal saline by gastric perfusion. The three experimental groups were given icariin and simvastatin solution with dosage ratios of 3:1, 1:1, 1:3 by gastric perfusion. Model group and three experimental groups were injected normal saline at fracture site, while bone morphogenetic protein-2 control group was injected bone morphogenetic protein-2 solution. Gastric perfusion was performed for 6 consecutive weeks and local injection at fracture site lasted for 2 weeks.
    RESULTS AND CONCLUSION: Bone morphogenetic protein-2 control group and 1:1 experimental group were the best followed by the 3:1 and 1:3 experimental groups, and the model group was the worst in the following aspects: growth and alteration of callus, recanalization of bone marrow cavity, bone mineral density and morphology of the fracture site. Icariin and simvastatin with different dosage ratios all can promote the radial fracture healing of rabbits. When the dosage ratio is 1:1, the combination of icariin and simvastatin can further promote fracture healing.

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    Labial and lingual alveolar bone thickness of adult tooth root
    Ding Ji-qun, Fang Jian-qiang, Yuan Chang-qing, Chen Jie
    2013, 17 (15):  2714-2722.  doi: 10.3969/j.issn.2095-4344.2013.15.008
    Abstract ( 592 )   PDF (472KB) ( 975 )   Save

    BACKGROUND: Oral treatment is affected by the root position and the surrounding bone plate thickness of the alveolar bone, and the improper teeth control can cause iatrogenic complications. The scholars have conducted the research about the jaw, such as anatomical observation, bone thickness or bone density. However, the research regarding the spatial position of the root within the alveolar bone and its relationship with the surrounding bone is less of a concern.
    OBJECTIVE: To establish a digital three-dimensional computer model of the jaw, and then to measure the thickness of labial and lingual alveolar bone around the tooth root, providing a reference for orthodontic tooth movement design and tooth extraction.
    METHODS: A total of 70 young adult orthodontic patients were selected with complete dentition and with no bone absorption shown on panoramic radiographs. The maxillofacial region was scanned using dental cone beam CT machine. Collected data were input into a computer workstation to implement coronal or sagittal multi-planar reconstruction with high-resolution three-dimensional images, and then raw data at DICOM format were outputted to the integrated three-dimensional design software, Invivo5 software, for measurement.
    RESULTS AND CONCLUSION: The digital and virtual reconstruction model of the jaw could be observed and measured from the multi-plane, and the mean alveolar thickness was measured with every root in 70 patients. The mean lingual alveolar bone thickness of anterior tooth was thicker than the labial (P < 0.05). Besides the dental cervix of upper premolars, the mean lingual alveolar bone thickness of premolars was thicker than the labial (P < 0.05). There were no differences when the bilateral measurements were compared for upper molars and mandibular first molar (P > 0.05), but a significant difference between labial alveolar bone and lingual alveolar bone was noted in the mandibular second molar (P < 0.01). The results confirmed that the significant difference exists between lingual and labial alveolar bone thickness of the young adults at different tooth positions.

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    Regular exercise improves age-related decline in arterial elasticity by enhancing number and activity of endothelial progenitor cells
    Yang Zhen1, Zhang Yuan-yuan1, Xia Wen-hao, Luo Chu-fan, Chen Long, Jin Ya-fei, Ou Zhi-jun, Liao Xin-xue, Tao Jun
    2013, 17 (15):  2723-2728.  doi: 10.3969/j.issn.2095-4344.2013.15.009
    Abstract ( 374 )   PDF (615KB) ( 630 )   Save

    BACKGROUND: Recent studies have shown that exercise can increase the number and ability of endothelial progenitor cells which is beneficial to the improvement of endothelial function. However, it is still unclear whether exercise can improve age-related decline in arterial elasticity by enhancing number and activity of circulating endothelial progenitor cells.
    OBJECTIVE: To investigate whether regular exercise-induced enhanced circulating endothelial progenitor cells improves age-related decline in arterial elasticity in healthy men.
    METHODS: Effects of regular exercise on circulating endothelial progenitor cells of 10 older and 10 young healthy men undergoing 3 months regular exercise were observed. Flow cytometry analysis was performed to evaluate the number of CD34 and KDR double-positive labeled circulating endothelial progenitor cells before and after exercise, and acetylated low-density lipoprotein; and lectin fluorescent staining method was used to evaluate the number of cultured endothelial progenitor cells. The migratory and proliferative activities of endothelial progenitor cells were also evaluated. In addition, brachial-ankle pulse wave velocities of older and young sedentary healthy men were measured.
    RESULTS AND CONCLUSION: Compared with young healthy men, the older healthy men exhibited decreased number and activity of circulating endothelial progenitor cells and increased brachial-ankle pulse wave velocities. Flow cytometry analysis showed that the circulating endothelial progenitor cells positive for CD34 and KDR in the peripheral blood increased after regular exercise. The acetylated-low-density lipoprotein and lectin fluorescent staining method indicated that the cultured endothelial progenitor cells also increased after exercise. The proliferative and migratory activities of cultured endothelial progenitor cells were significantly higher after exercise. However, the increased number and activity of circulating endothelial progenitor cells and decreased brachial-ankle pulse wave velocities of older sedentary healthy men were higher. There was a closely positive correlation between increased circulating endothelial progenitor cells and decreased brachial-ankle pulse wave velocities. Multivariate analysis identified increased proliferative activity of circulating endothelial progenitor cells as an independent predictor of decreased brachial-ankle pulse wave velocities. Regular exercise-induced enhanced number and activity of circulating endothelial progenitor cells attenuates age-related decline in arterial elasticity, indicating that the modulation of exercise on circulating endothelial progenitor cells may be the mechanism underlying exercise-exhibited protection against age-related vascular injury.

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    Mitochondria membrane potentials and reactive oxygen species of vascular endothelial cells during senescence process
    Zhao Hai-mei, Yang Bin, Cheng Bei
    2013, 17 (15):  2729-2734.  doi: 10.3969/j.issn.2095-4344.2013.15.010
    Abstract ( 607 )   PDF (759KB) ( 826 )   Save

    BACKGROUND: Balance among aging, apoptosis and regeneration of vascular endothelial cells exerts a critical role in maintenance of the function of normal vessels. Mitochondria are a key cell organelle in the body, which cannot only support energy for the cells via adenosine triphosphate synthesis, but also control the procedure death, aging of cells as well as many pathophysical metabolism processes.
    OBJECTIVE: To investigate the relationship between mitochondria membrane potentials and reactive oxygen species of endothelial cells during growing and senescence process in order to explore the dysfunction occurring during the senescence process.
    METHODS: Human umbilical vein endothelial cells were cultured in vitro and passaged. Passages 2, 4, 6 and 8 human umbilical vein endothelial cells were selected to detect those mitochondria membrane potentials and reactive oxygen species. Levels of reactive oxygen species and mitochondria membrane potentials were checked by flow cytometry. At the same time, passage 2 and 8 cells were selected for observation of normal and aging ultramicro-morphoses by electron microscope.
    RESULTS AND CONCLUSION: During the passage and senescence, mitochondria membrane potentials were decreased gradually with cell passage, while the levels of reactive oxygen species were increased firstly and then decreased. The number of mitochondria and reticulum was reduced along with cell passage. In replicate aging process, the mitochondrial membrane potential was decreased and the mitochondrion was damaged. At the first time the production of reactive oxygen species was increased, while decreased along with the cell degeneration.

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    Three-dimensional CT angiography for anatomic measurements of the carotid artery bifurcation
    Duan Shao-yin1, 2, Lin Chang-hua1, Jing Jing1, Yang Lie1, Lin Qing-chi1, 2
    2013, 17 (15):  2735-2742.  doi: 10.3969/j.issn.2095-4344.2013.15.011
    Abstract ( 1420 )   PDF (1202KB) ( 573 )   Save

    BACKGROUND: Anatomic measurement is the basis of clinical medicines. It can provide basis and reference for clinical diagnosis and surgical operation. The anatomical measurement with 64-spiral CT angiography has the obvious technical superiority and good application prospect.
    OBJECTIVE: To measure the anatomical structure of the carotid artery bifurcation with 64-spiral CT angiography, and to provide anatomic basis for relevant researches. 
    METHODS: A total of 92 subjects, who underwent 64-spiral CT angiography in head and neck without any pathology of carotid artery bifurcation from June 2008 to June 2010, were selected from Department of Radiology, Zhongshan Hospital Xiamen University. There were 40 male cases (≤40 years old) and 52 female cases (> 40 years old). The cross-sectional images were processed with three-dimensional imaging to obtain the three-dimensional image and to measure the anatomical structure of the carotid artery bifurcation.
    RESULTS AND CONCLUSION: The three-dimensional images could clearly display the structure of carotid artery bifurcation and could achieve the mono- or multi-structure and multi-directional observation of structure. The measurement results showed the carotid artery bifurcation angle was (43.5±12.3)°, the distal inner diameter of the common carotid artery was (6.83±0.65) mm, the proximal inner diameter of the internal carotid artery enlargement area was (7.25±1.04) mm, the width of the internal carotid artery enlargement area was    (8.15±1.35) mm, the distal inner diameter of the internal carotid artery enlargement area was (5.03±0.55) mm, and the inner diameter of the external carotid artery was (4.22±0.60) mm. Compared with the ≤ 40 years old group, the carotid artery bifurcation angle, proximal inner diameter of the internal carotid artery enlargement area, width of the internal carotid artery enlargement area and the distal inner diameter of the internal carotid artery enlargement area in the > 40 years old group were increased significantly, while the inner diameter of the external carotid artery was small (P < 0.05), and there was no significant difference in inner diameter of common carotid artery between two groups (P > 0.05). Compared with the male group, the measurement indexes of carotid artery bifurcation in the female group were significantly smaller (P < 0.05). The carotid artery bifurcation angle on the left side was bigger than that on the right side, and there were no significant differences in the other measurement indexes of carotid artery bifurcation between left and right group (P > 0.05). Three-dimensional CT image can objectively and accurately measure the anatomical values of carotid artery bifurcation. It has individual characteristics that can provide the objective basis for applied anatomy, disease diagnosis and surgery treatment.

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    Fuzi Lizhong Decoction and its disassembled prescriptions for rats with spleen-yang deficiency syndrome
    Tang Han-qing, Huang Cen-han, Huang Zan-song, Huang Xiu-feng
    2013, 17 (15):  2743-2748.  doi: 10.3969/j.issn.2095-4344.2013.15.012
    Abstract ( 340 )   PDF (1550KB) ( 463 )   Save

    BACKGROUND: Chinese herbal compounds mostly pose pharmacodynamic characteristics of “two-way adjustment” and “corrective bias” to restore the body’s balance. This experiment is proposed to verify the effect of Fuzi Lizhong Decoction and its disassembled prescription on deficiency of spleen-yang syndrome via warming yang and invigorating the spleen, further providing basic data for understanding the multi-node mechanism.
    OBJECTIVE: To observe and evaluate the curative effects of Fuzi Lizhong Decoction and its disassembled prescription in the treatment of rats with spleen-yang deficiency syndrome under the conception of translational medicine.
    METHODS: A model of deficiency of spleen-yang syndrome was made in Wistar rats through interscapular removal of brown fat and high fat diet under cold environment that changed every other day. After 70 days, the rat models were administered with Fuzi Lizhong Decoction, Lizhong Decoction and Fuzi Ganjiang Decoction, respectively. Another Wistar rats without modeling were used as controls that were fed with normal diet. Body mass, rectal temperature and area under rectal temperature curve were measured in all the groups.
    RESULTS AND CONCLUSION: Body mass was increased slowest in the model group and fastest in the Fuzi Lizhong Decoction group (P < 0.05). The model group had lower rectal temperature than the other four groups and significantly differed from the control group in the rectal temperature (P < 0.05). There was a significant difference in the rectal temperature between the Fuzi Lizhong Decoction and model groups (P < 0.05). Area under rectal temperature curve in the model group was lower than that in the control group (P < 0.05). As compared with the models group, Fuzi Lizhong Decoction, Lizhong Decoction and Fuzi Ganjiang Decoction groups had the larger area under the rectal temperature curve (P < 0.05 or P < 0.01). Fuzi Lizhong Decoction, Lizhong Decoction and Fuzi Ganjiang Decoction can all improve the pathological conditions of rats with spleen-yang deficiency syndrome presenting with body mass reduction or slow growth and low temperature. Fuzi Lizhong Decoction has the best biologic effects among these three decoctions.

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    Establishing a nude mouse model of green fluorescence endometriosis
    Yuan Hua, Gong Jian, Wang Jia-yuan, Huang Wei
    2013, 17 (15):  2749-2754.  doi: 10.3969/j.issn.2095-4344.2013.15.013
    Abstract ( 555 )   PDF (356KB) ( 463 )   Save

    BACKGROUND: It is simple to fabricate an animal model of endometriosis by subcutaneous injection of green fluorescence labeled endometrium of nude mice. Meanwhile, in vitro ectopic lesions can be observed dynamically within a certain period, which contributes to the depth study of endometriosis.
    OBJECTIVE: To explore the method to establish an animal model of subcutaneous endometriosis in nude mice.
    METHODS: Enhanced green fluorescent protein adenovirus transfected endometrium of nude mice was injected subcutaneously into six nude mice (fluorescent group) and non-transfected endometrium was injected into another six nude mice (control group). Green fluorescence intensity, duration of fluorescence labeled lesions, and histological changes of ectopic lesions in vivo were detected under stereological fluorescence microscope at 5, 10, 15, 20, 25, 30 days after injection.
    RESULTS AND CONCLUSION: Five living fluorescence mouse models of endometriosis were induced successfully in the fluorescent group and another five living non-fluorescent nude mouse models of endometriosis were also prepared in the control group. There were 2-3 lesions per nude mouse. Development of ectopic lesions in the mice could be monitored under the stereological fluorescence microscope. The fluorescent area and intensity were decreased with time, which lasted for about 4 weeks. Fluorescence labeled lesions were invisible in the control group. Hematoxylin-eosin staining showed the typical endometrial glands and stroma structure. These findings indicate that this modified nude mouse model can enable the dynamic and quantitative observation of lesion growth and development without injury and contribute to the development of new drugs for endometriosis and the underlying mechanism of endometriosis.

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    Anti-Staphylococcus aureus substance purification from Wu Gu Chong and its related antibacterial mechanisms
    Zhang Zhen, Hong Liang, Wang Shou-yu, Lü De-cheng
    2013, 17 (15):  2755-2760.  doi: 10.3969/j.issn.2095-4344.2013.15.014
    Abstract ( 353 )   PDF (1793KB) ( 562 )   Save

    BACKGROUND: The antibacterial substances purification from Wu Gu Chong is controversial, and the antibacterial mechanism is unclear.
    OBJECTIVE: To isolate and purify antibacterial substances from Wu Gu Chong and to identify its related antibacterial mechanism.
    METHODS: After ultra-filtration, the Turbidometric assay and Kirby-Bauer disk diffusion susceptibility test were used to examine the antibacterial activity of the fractions against Staphylococcus aureus. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to examine the effect of the extracts of maggot that was screened on the proliferation of Staphylococcus aureus and to determine the minimum inhibitory concentration. Electron microscopy was used to examine the ultra-structural changes of Staphylococcus aureus induced by antimicrobial substances of the extracts.
    RESULTS AND CONCLUSION: The filtrate (Mr < 10 000) had the best antibacterial activity against Staphylococcus aureus (P < 0.05), which significantly inhibited the proliferation of Staphylococcus aureus. The minimum inhibitory concentration was 1 g/L. The antibacterial substances of maggot increased the permeability and destroyed the cell membrane of Staphylococcus aureus. The anti-Staphylococcus aureus substances from Wu Gu Chong was less than 10 000, and its related mechanisms of anti-bacterial effect was a disruption of the outer and cytoplasmic membranes leading to lysis of the target cells.

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    Muscle satellite cells transfected with acidic fibroblast growth factor gene in vitro
    Yang Shao-an, Cai Jin-kui, Zhou Chu-song, Xiao Xiao-tao, Xiao Sha, Zou Xiao-ying
    2013, 17 (15):  2761-2768.  doi: 10.3969/j.issn.2095-4344.2013.15.015
    Abstract ( 304 )   PDF (2279KB) ( 425 )   Save

    BACKGROUND: It has been reported that acidic fibroblast growth factor (aFGF) not only can mediate cell division and differentiation, but also can prevent motor endplate degeneration and muscular atrophy.
    OBJECTIVE: To calculate gene transfection efficiency and detect the target protein expression of muscle satellite cells which were transfected with aFGF gene, in purpose to further study the method to set up cell bank for preventing motor endplate degeneration and muscular atrophy.
    METHODS: Muscle satellite cells were extracted from adult Wistar rat, purified by difference-speed adherence method and identified by immunohistochemical assay. The recombinant eukaryotic expression plasmid pEGFP-N1-aFGF was transfected into cells by LipofectamineTM2000 Reagent as experimental group. Muscle satellite cells transfected with pEGFP-N1 served as negative controls. Blank control group was set by adding transfection reagent. Inverted fluorescent microscope was applied to observe green fluorescent protein expression in the cells to calculate transfection efficiency at 24-72 hours after transpection and passaging. Western Blot of aFGF was performed to detect the target protein. Total RNA was extracted at 72 hours after transfection. Real-time fluorescent quantitative PCR was employed in order to find out the changes of cells after transfection on mRNA level.
    RESULTS AND CONCLUSION: The immunohistochemical results showed that cultivated cells were muscle satellite cells. The expression of green fluorescent protein appeared as early as 6 hours after transfection, and the amount and intensity peaked at 72 hours. Green fluorescent protein was still seen in the subculture cells. Real-time fluorescent quantitative PCR proved stronger aFGF mRNA expression in the transfected cells with aFGF gene, while a little in the control groups. aFGF protein was highly expressed in the cells transfected with target gene detected by Western Blot. All the results indicate that the aFGF gene can be transfected efficiently and safely into muscle satellite cells and expressed normally, which can serve as the new seed cells for tissue engineering to prevent motor endplate degeneration and muscular atrophy.

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    Culture methods for primary cervical carcinoma cells
    Cheng Jing-xin, Ma Xiu-ping, Zhou Ping, Yuan Min, Huang Kun, Zhang Yi
    2013, 17 (15):  2769-2775.  doi: 10.3969/j.issn.2095-4344.2013.15.016
    Abstract ( 806 )   PDF (746KB) ( 896 )   Save

    BACKGROUND: In vitro isolation and culture of cervical epithelial carcinoma cells is the foundation for studying tumor etiology and treatment from the cellular level, which is the most basic aspect of cervical cancer tissue engineering research.
    OBJECTIVE: To investigate the culture methods for primary cervical carcinoma cells, and passaged cell growth, proliferation and shape in vitro.
    METHODS: We isolated and cultured primary cervical carcinoma cell by using enzyme joint digestion technique (0.25% trypsin and 2% type I collagenase), observed the shape of the tumor cell and condition of the cell proliferation using the inverted fluorescent microscope, and detected CK17 and Ki67 expression by immunofluorescence.
    RESULTS AND CONCLUSION: Cervical epithelial carcinoma cells harvested using the enzyme joint digestion technique had a higher purity, and this technique was relatively simple to be repeated. The passaged cervical epithelial carcinoma cells also had a stable cell phenotype. CK17 positive expression indicated the cultured cells were cervical epithelial carcinoma cells and Ki67 positive expression showed that the cultured cells had the proliferative characteristics of malignant tumor cells.

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    Deguelin regulates the expression of Wnt signaling pathway related genes in Zebrafish embryo
    Liu Rui-jin, Wu Xin-rong
    2013, 17 (15):  2776-2779.  doi: 10.3969/j.issn.2095-4344.2013.15.017
    Abstract ( 688 )   PDF (550KB) ( 585 )   Save

    BACKGROUND: Previous studies found that in different periods of early development of zebrafish, embryos treated with certain concentration of deguelin show varying degrees of developmental delay and abnormal phenotype.
    OBJECTIVE: To observe the effect of deguelin on expression of Wnt signaling pathway related genes in Zebrafish embryo during early embryo development.
    METHODS: Zebrafish embryos that incubated to the shield period were collected and put into the 6-well plate, 0.6 μmol/L deguelin solution were added and treated in dark, and then cultured in the incubator under 28 ℃. Zebrafish embryos treated with 0.1% dimethylsulfoxide solution were as control group, and cultured in the same conditions. At 6, 8, 10, 12, 14 and 24 hours after fertilization, phenotype of Zebrafish embryos was observed with upright fluorescence microscope. The expression levels of target genes before and after treatment were tested by quantitative real time PCR.
    RESULTS AND CONCLUSION: The embryos treated by deguelin on shield stage stop growing and stay at bud stage; the expressions of wnt9a, wnt10b, axin2 as well as snail1b and Jnk2 related with Wnt signal were changed significantly, among them, the expression of wnt9a, wnt10b, axin2 and snail 1b were decreased, while the expression of Jnk2 was increased. Deguelin has a negative effect on Zebrafish in vivo Wnt signal.

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    Processing of the retinal cone image based on the MATLAB
    Zhao Chao-yang, Yao Jun-ping, Liu Yong, Dai Yun, Gu Xin, Yin Zheng-qin
    2013, 17 (15):  2780-2785.  doi: 10.3969/j.issn.2095-4344.2013.15.018
    Abstract ( 874 )   PDF (794KB) ( 957 )   Save

    BACKGROUND: The cone cell for the image retina adaptive optical imaging system is characterized by the relatively concentrated gray distribution, vague window edge, the existing pseudo outline, and the adhesion of edge photograph. Looking for a suitable processing algorithm of the cone image to get the clear outline is the key content for the future work.
    OBJECTIVE: To perform edge extraction of the retina cone cell image and get clear edge profile of the cone cell based on the MATLAB.  
    METHODS: According to the retinal image of the different parts of 30 normal subjects, based on Mat lab image processing toolbox, we could preprocess the cone cell image, edge extraction and process morphological, and get the clear edge profile of the cone cell image. The cell numbers of processed image was counted, the cone distribution characteristics were analyzed, and then the feasibility of this study which designed image processing algorithm applied to the study of the distribution characteristics of cone cells were verified.
    RESULTS AND CONCLUSION: A clear outline of the retinal cones image was obtained successfully. From the results of cone cell’s density, as far away from the center fovea of macula, the cell density presented a trend of reduction; from the center fovea of macula partial range 0.5° to 1°, the retinal cones density decreased rapidly. The findings indicated that the algorithm of the image processing designed in this study can be used to study the retinal characteristics of the cones distribution.

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    Vascularized strategy for tissue-engineered bone
    Zheng Xiao-hui, Chen Zhen-guang
    2013, 17 (15):  2786-2794.  doi: 10.3969/j.issn.2095-4344.2013.15.019
    Abstract ( 370 )   PDF (711KB) ( 553 )   Save

    BACKGROUND: How to rebuild the blood supply during construction of tissue-engineered bone is becoming a critical problem reversed from basic research to clinical application.
    OBJECTIVE: To review a series of vascularization methods during tissue-engineered bone construction.
    METHODS: A computer-based search of PubMed and Wanfang databases was performed by the first author for the literature about vascularization methods during tissue-engineered bone construction published from 1974 to 2008. The key words were “tissue-engineered bone, vascularized, bone defects, blood supply” in English and Chinese, respectively. Repetitive articles or those with unrelated purpose were excluded, and finally 61 articles were included in result analysis.
    RESULTS AND CONCLUSION: Vascularized methods for construction of tissue-engineered bone are mainly classified as in vivo and in vitro mehtods. Vascular endothelial cells and growth factors belonged to in vitro methods, while vascular bundle, vascular pedicled fascial flap, vascular pedicled muscle flap, vascular pedicled periosteal flap categorized as in vivo methods. Although a great development has been made in vascularized tissue-engineered bone, there are still many inadequacies. We believe that, with the development of mechanism of angiogenesis, vasculared tissue engineered bone will have a promising prospect because of promoting the ability of release-controlled growth factors and their synergies, optimizing the choose of seed cells, and improving the structure and properties of materials

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    Pathogenesis, prevention and therapy of disuse osteoporosis
    Yuan Wei, Zhao Jin-zhu
    2013, 17 (15):  2795-2802.  doi: 10.3969/j.issn.2095-4344.2013.15.020
    Abstract ( 940 )   PDF (808KB) ( 489 )   Save

    BACKGROUND: Disuse osteoporosis is common. Due to its special etiology, the prevention and therapy is difficult, and the burden on the social and family is very heavy.
    OBJECTIVE: To analyze the etiology, pathogenesis and strategy of prevention and therapy of disuse osteoporosis in order to offer a reference for prevention of disuse osteoporosis effectively.
    METHODS: CNKI database and PubMed database were searched for the relevant papers of disuse osteoporosis published from January 2002 to September 2012 with the key word of “disuse osteoporosis” in Chinese and English, respectively. Totally 178 documents were retrieved. All retrieved articles were checked, and those that were closely relevant with the subject, original and reliable, had clear viewpoint and comprehensive analysis were included. Duplicate documents and experiments without randomized controlled trials were excluded.
    RESULTS AND CONCLUSION: There are 50 articles enrolled in this paper. The etiology of disuse osteoporosis is classified into three kinds: lack of mechanical loading, microgravity status, spinal cord injury and peripheral nerve injury. The prevention and therapy for disuse osteoporosis are shown as follows: (1) drug treatment, including calcitonin, Chinese herbs, diphosphonate, vitamin, calcium, strontium salt and nerve growth factors; (2) physical treatment, including pulse electric-magnetic field, ultrasound, resonance wave, passive movement and electroacupunture treatment. Currently, there is no ideal prevention and treatment strategy for disuse osteoporosis. Scholars have tended to investigate the pathogenesis of disuse osteoporosis from the aspect of genetics recently, and have obtained some progress in signal pathway proteins and special gene loci.

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    Anabolic drug application in osteoporosis
    Liu Yin, Tian Jing
    2013, 17 (15):  2803-2810.  doi: 10.3969/j.issn.2095-4344.2013.15.021
    Abstract ( 423 )   PDF (636KB) ( 930 )   Save

    BACKGROUND: Bone fracture and its complications caused by osteoporosis trigger a serious impact on the life quality of the elderly and menopause women.
    OBJECTIVE: To explore the research progress of anabolic drug application in the treatment of osteoporosis in order to promote its clinical application.
    METHODS: A computer-based online search of PubMed database, CNKI database, VIP database and Wanfang database between January 1997 and August 2012 was performed by the first author to search related articles with the key words of “bone formation, osteoporosis, osteoblast, osteoclast, bone metabolism, parathyroid hormone, Wnt signaling, medicine, target, inhibitor” in English or Chinese. Literatures related to osteoporosis were selected; in the same field, the articles published lately in authoritative journals were preferred. A total of 398 literatures were primarily selected, 40 of which addressing anabolic drug application in osteoporosis and its side-effects were involved for summary according to inclusion criteria.
    RESULTS AND CONCLUSION: In contrast with antiresorptive drugs, anabolic drug can increase bone mass more greatly and it can also reduce the risk of fractures. Anabolic drug has an important clinical value in the treatment for osteoporosis. Parathormone and parathormone related peptide, Wnt signaling and related inhibitors of secreted glycoprotein such as selerostin antibody, Dkk1, sFRP, GSK3β are research hotspots. But their limitations cannot be ignored in clinic.

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    Pulsed electromagnetic fields and cartilage metabolism
    Ruan Jia-li, Tian Jing
    2013, 17 (15):  2811-2818.  doi: 10.3969/j.issn.2095-4344.2013.15.022
    Abstract ( 404 )   PDF (741KB) ( 452 )   Save

    BACKGROUND: Peripheral pulsed electromagnetic fields have an obvious effect on cartilage metabolism; however, its molecular mechanism is still unclear.
    OBJECTIVE: To investigate the physical characteristics of pulsed electromagnetic fields and its mechanism and effect on cartilage formation.
    METHODS: A computer-based online search of PubMed database, CNKI database, VIP database andWanfang database was performed to search related articles. “Pulsed electromagnetic fields, cartilage, cartilage cells, matrix, bone marrow mesenchymal stem cells” were used for word retrieval in the title, abstract, and keywords. Repetitive studies or old literatures were excluded.
    RESULTS AND CONCLUSION: A total of 145 articles were retrieved in the initial search, and finally 46 articles were included in result analysis. The results showed that pulsed electromagnetic fields can promote the differentiation of bone marrow mesenchymal stem cells into chondrocytes and cartilage specific matrix such as type II collagen and proteoglycan synthesis, thus playing a cartilage inducing effect. Pulsed electromagnetic fields can also promote the expression of transforming growth factor beta 2 and other factors to regulate the chondrocyte growth and differentiation, which can realize the permanent repair of defects in clinic.

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    Tissue engineering technique to repair articular cartilage injury: Environment, material, safety and controllability
    Kou Jian-qiang, Wang Qian-qian, Wang Chang-yao, Wang Ying-zhen
    2013, 17 (15):  2819-2826.  doi: 10.3969/j.issn.2095-4344.2013.15.023
    Abstract ( 319 )   PDF (692KB) ( 377 )   Save

    BACKGROUND: The traditional methods for cartilage defect repair have their limitations. The application of tissue engineering methods to repair articular cartilage defects has shown promising prospects.
    OBJECTIVE: To review the progress of tissue engineering technique to repair articular cartilage at home and abroad in recent years.
    METHODS: PubMed and CNKI databases were searched online by the first author for papers concerning applying tissue engineering technique to repair articular cartilage defects in 1990-2011. Totally 187 Chinese papers and 211 English papers were retrieved, and finally 49 papers were included in result analysis.
    RESULTS AND CONCLUSION: The main method of cartilage tissue engineering is to apply the principle of engineering and life sciences to isolate, culture and proliferate needed seed cells. Then, the cells were implanted onto the appropriate biological scaffold, and the cytoskeleton was implanted into the defect site. Gradually, new cartilage tissues form under some inducing conditions. In recent years, the cartilage tissue engineering research focuses on the following aspects: (1) Selection of seed cells, including autologous chondrocytes, allogenic chondrocytes, embryonic stem cells and bone marrow mesenchymal stem cells; (2) Cell induction and conditional culture, including cytokines, cell culture conditions, transgenic technology; (3) Selection and research of biological scaffold materials. The following aspects have become hotspots in the tissue engineering research: finding ideal seed cells, combination with cytokines, literally imitating microenvironment for cell survival, safety, high efficiency, and controlled transfection of gene engineering, and constructing ideal scaffold materials.

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    Transforming growth factor beta 1 effects on cartilage tissue metabolism
    Guo Tie-feng1, Zhou Ming-wang2, Li Sheng-hua2, Sun Feng-qi1, Mu Huan-xi1
    2013, 17 (15):  2827-2834.  doi: 10.3969/j.issn.2095-4344.2013.15.024
    Abstract ( 462 )   PDF (815KB) ( 617 )   Save

    BACKGROUND: Transforming growth factor beta 1 can mediate cartilage synthesis and inhibit decomposition of collagen and protein polysaccharide, which has a most important effect on induction of cartilage differentiation in vitro and maintenance of cartilage phenotype, realizing the functional repair of cartilage defects.
    OBJECTIVE: Based on the biological characteristics, applications in biotechnology, gene polymorphism, signaling pathways and microRNA, to introduce research progress of transforming growth factor beta 1 influence on cartilage tissue metabolism.
    METHODS: The first author searched CNKI and SPRINGERLINK databases (2007/2012) to retrieve articles related to transforming growth factor beta 1 influence on cartilage tissue metabolism using the key words of “transforming growth factor beta 1, cartilage differentiation, cartilage matrix” in Chinese and English, respectively. A total of 130 articles were retrieved, and finally 54 were included.
    RESULTS AND CONCLUSION: Transforming growth factor beta 1 can induce chondrogenic differentiation of mesenchymal stem cells, promote cartilage-specific matrix synthesis, protect the cartilage matrix against proteolysis, enhance self-renew ability of cartilage tissues, and realize repair of cartilage injury. In the field of cartilage repair, transforming growth factor beta 1 demonstrates a great potential value.

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    Protein synthesis and resistance in aging skeletal muscle
    Wang Shun-li, Shi Reng-fei
    2013, 17 (15):  2835-2842.  doi: 10.3969/j.issn.2095-4344.2013.15.025
    Abstract ( 532 )   PDF (661KB) ( 620 )   Save

    BACKGROUND: Sarcopenia will become more serious with aging, presenting with the loss of muscle mass and muscle strength. Further studies reveal that the rates of synthesis and breakdown of muscle protein fail to change with aging, and it is the protein synthesis resistance which appears in aging skeletal muscle that leads to sarcopenia. 
    OBJECTIVE: To review the concepts, mechanisms and interventions of protein synthesis resistance, which could be the references for nutrition and exercise interventions to prevent and slow skeletal muscle aging.
    METHODS: The databases of PubMed and CNKI (2000-01/2011-12) were retrieved by computer with key words of “skeletal muscle, aging, resistance training, protein” both in Chinese and English. The following literatures were included: (1) Experimental papers with reliable arguments; (2) Papers concerning skeletal muscle aging. The papers unrelated to this paper’s aim or repeatability studies were excluded.
    RESULTS AND CONCLUSION: Protein synthesis resistance appears in aging skeletal muscle that leads to
    sarcopenia. Protein and resistance training are involved in protein synthesis. Further research is still needed on the amount of protein supplement, resistance exercise intensity and when to add protein in old people.

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    Bariatric surgery for non-obese type 2 diabetes mellitus in Mainland China: A meta-analysis
    Xu Jin-hua1, Pan Wen2, Gong Jian, Lu Suo-xing, Guan Shu-hui, Wang Dong-xia, Piao Zhe, Li Ning, Li Jing-shu
    2013, 17 (15):  2843-2850.  doi: 10.3969/j.issn.2095-4344.2013.15.026
    Abstract ( 549 )   PDF (852KB) ( 590 )   Save

    BACKGROUND: Bariatric surgery for obese type 2 diabetes mellitus has obtained better effects in Mainland China, but there is no clear report about bariatric surgery for non-obese type 2 diabetes mellitus.
    OBJECTIVE: To assess the clinical effectiveness of bariatric surgery for non-obese type 2 diabetes mellitus in Mainland China. 
    METHODS: We searched the Cochrane Central Register of Controlled Trials (Issue 2, 2012), MEDLINE (1990 to February 2012), EMbase (1990 to February 2012), CBMdisc (1990 to February 2012) and CNKI (1990 to February 2012). Manual search of relevant journals and conference proceedings was also performed. Clinical trials in which bariatric surgery (gastric bypass) was used to treat patients with type 2 diabetes mellitus in Mainland China were collected. Then we screened the retrieved studies according to predefined inclusion and exclusion criteria, evaluated the quality of the included studies, and performed meta-analyses by RevMan 5.1.0 software. 
    RESULTS AND CONCLUSION: A total of 30 articles were found and seven articles involving 307 patients were finally included. All these articles were regarded as low quality (grade C). We chose the random-effect model to conduct meta-analysis because significant heterogeneities were found among these articles. The results of meta-analysis showed that after treatment with bariatric surgery, there were significant reductions in fasting plasma glucose [3 months: mean difference (MD)= -2.84, 95% confidence interval (CI) (-3.60, –2.08), P < 0.000 01; 6 months: MD= -3.79, 95%CI (–4.56, –3.02), P < 0.000 01], glycated hemoglobin [3 months: MD= -1.52, 95%CI (-2.14, -0.89), P < 0.000 01; 6 months: MD= -1.90, 95%CI (-2.46, -1.34), P < 0.000 01] and insulin resistance index [6 months: MD= –1.80, 95%CI (–2.68, –0.91), P < 0.000 1]. Sensitivity analysis indicates that results were credible and stable. Funnel-plots display non-symmetrical figures, indicating publication bias. Bariatric surgery is effective for non-obese type 2 diabetes mellitus in Mainland China. However, due to lack of quality in the included studies, more studies with high-quality large-size samples should be conducted.

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