Effect of acupuncture on HIF-1α/NLRP3 inflammatory signaling pathway in the rats with cerebral ischemia-reperfusion injury / 中国针灸

OBJECTIVE@#To observe the effects of Xingnao Kaiqiao (regaining consciousness and opening orifices) acupuncture therapy on the expression of hypoxia-inducible factor 1α (HIF-1α) and Nod-like receptor protein 3 (NLRP3) in cerebral ischemia-reperfusion rats, and to explore the mechanism of acupuncture against cerebral ischemia-reperfusion injury.@*METHODS@#Seventy-two male SD rats were randomly divided into a sham-operation group, a model group, an acupuncture group and a non-point acupuncture group, with 18 rats in each one. Using modified Longa thread embolization method, the rat model of acute focal cerebral ischemia was prepared; and after 2 h ischemia, the reperfusion was performed to prepared the model of cerebral ischemia-reperfusion. Immediately after reperfusion, Xingnao Kaiqiao acupuncture method was applied to bilateral "Neiguan" (PC 6) and "Shuigou" (GV 26) in the acupuncture group, while in the non-point acupuncture group, acupuncture was delivered at non-points and all of the needles were retained for 30 min in these two groups. The samples were collected 24 h after reperfusion in the rats of each group. Zea-Longa neurological deficit score was used to evaluate the degree of cerebral neurological impairment, TTC staining was adopted to observe the volume percentage of cerebral infarction, HE staining was provided to observe the morphological changes of brain, and Western blot was applied for detecting the expression of HIF-1α and NLRP3 proteins in the cerebral cortex on the right side.@*RESULTS@#Compared with the sham-operation group, neurological deficit score and volume percentage of cerebral infarction were increased in the model group (P<0.01), and HIF-1α and NLRP3 protein expression was elevated (P<0.01). Compared with the model group, neurological deficit score and volume percentage of cerebral infarction were decreased (P<0.01), and HIF-1α and NLRP3 protein expression was lower (P<0.01) in the acupuncture group. There was no significant difference in above indexes in the non-point acupuncture group compared with the model group (P>0.05). Compared with the sham-operation group, the brain tissue of the rats in the model group and the non-point acupuncture group was loose and edema, and the nuclei were shriveled. The brain tissue morphology in the acupuncture group was similar to that of the sham-operation group.@*CONCLUSION@#Acupuncture can alleviate cerebral ischemia-reperfusion injury, and its mechanism may be related to the regulation of HIF-1α/NLRP3 signaling pathway to attenuate inflammatory response.

Mechanisms of Electroacupuncture on Alzheimer's Disease: A Review of Animal Studies / 中国结合医学杂志

Alzheimer's disease (AD) is the main type of dementia affecting elderly people. The medicines used for treating AD have limits in their ability to alleviate cognitive and functional decline. Electroacupuncture (EA), an economical alternative and complementary therapeutic measure used in Chinese medicine, has strong clinical applicability, and its treatment effects are more reliable as its stimulation factors can be standardized. EA has been effectively used for ameliorating spatial learning and memory impairment in AD patients and rodent AD models. Here, we summarized the mechanisms of EA on rodent AD models, aiming to provide a theoretical basis for identifying new targets and research directions, and exploring appropriate EA stimulation parameters, then provide a detailed methodology to optimize the effects of EA, ultimately reducing the heavy burden of treating AD patients on families and countries. Our review showed that 8 acupoints were used when treating AD animals; Baihui (GV 20) was the most frequently used. Four mechanisms of EA were studied: (1) protection of cerebral neurons from oxidative stress, apoptosis, neuroinflammation, synaptic plasticity and hormone levels; (2) regulation of metabolism, including glucose metabolism and plasma metabolism; (3) regulation of brain-derived neurotrophic factor; and (4) regulation of β-amyloid peptide protein deposition.

The effect of periodic dynamic mechanical load with different intensity on microgravity-induced osteoporosis / 医用生物力学

Objective To investigate the role and influence of physiological loading and overloading on microgravity-induced osteoporosis, so as to find a reliable way to prevent or treat related-orthopedic disorders in astronauts induced by long-time space activity. Methods The microgravity environment in space was simulated by tail-suspension experiment, then the osteoporosis models of mice were built. A total of 32 C57BL/6J mice were randomly and evenly separated into four groups: normal group (normal), tail-suspension group (TS), physiological loading group (loading) and overloading group (overloading). Periodic dynamic mechanical load was applied on the left tibia in loading group and overloading group during tail-suspension test. After four weeks, tibial mechanical properties, micro-parameters of bone trabecular, biochemical indices and osteogenesis-related gene expression in each group were compared and analyzed. Results A great loss of tibial cancellous bone, significantly lower tibial biomechanical expression, serious damage of microstructure and weaker osteogenic activity were found in tail-suspended mice as compared with those of normal group. Physiological loading could clearly improve mechanical properties of bones, microstructure of bone trabecular, osteogenic activity and relative gene expression (P＜0.05). Overloading could also improve the condition of microgravity-induced osteoporosis, but the effect was not obvious (P＞0.05). Conclusions Tail-suspension can successfully simulate microgravity environment and duplicate osteoporosis model. Physiological loading can effectively prevent the emergence and development of microgravity-induced osteoporosis, while overloading can also counter microgravity-induced osteoporosis, but the results have no significant differences.

The effect of periodic dynamic mechanical load with different intensity on microgravity-induced osteoporosis / 医用生物力学

Objective To investigate the role and influence of physiological loading and overloading on microgravity-induced osteoporosis,so as to find a reliable way to prevent or treat related-orthopedic disorders in astronauts induced by long-time space activity.Metbods The microgravity environment in space was simulated by tail-suspension experiment,then the osteoporosis models of mice were built.A total of 32 C57BL/6J mice were randomly and evenly separated into four groups:normal group (normal),tail-suspension group (TS),physiological loading group (loading) and overloading group (overloading).Periodic dynamic mechanical load was applied on the left tibia in loading group and overloading group during tail-suspension test.After four weeks,tibial mechanical properties,micro-parameters of bone trabecular,biochemical indices and osteogenesis-related gene expression in each group were compared and analyzed.Results A great loss of tibial cancellous bone,significantly lower tibial biomechanical expression,serious damage of microstructure and weaker osteogenic activity were found in tail-suspended mice as compared with those of normal group.Physiological loading could clearly improve mechanical properties of bones,microstructure of bone trabecular,osteogenic activity and relative gene expression (P ＜ 0.05).Overloading could also improve the condition of microgravity-induced osteoporosis,but the effect was not obvious (P ＞ 0.05).Conclusions Tail-suspension can successfully simulate microgravity environment and duplicate osteoporosis model.Physiological loading can effectively prevent the emergence and development of microgravity-induced osteoporosis,while overloading can also counter microgravity-induced osteoporosis,but the results have no significant differences.

The effect of periodic dynamic mechanical load with different intensity on microgravity-induced osteoporosis / 医用生物力学

Objective To investigate the role and influence of physiological loading and overloading on microgravity-induced osteoporosis,so as to find a reliable way to prevent or treat related-orthopedic disorders in astronauts induced by long-time space activity.Metbods The microgravity environment in space was simulated by tail-suspension experiment,then the osteoporosis models of mice were built.A total of 32 C57BL/6J mice were randomly and evenly separated into four groups:normal group (normal),tail-suspension group (TS),physiological loading group (loading) and overloading group (overloading).Periodic dynamic mechanical load was applied on the left tibia in loading group and overloading group during tail-suspension test.After four weeks,tibial mechanical properties,micro-parameters of bone trabecular,biochemical indices and osteogenesis-related gene expression in each group were compared and analyzed.Results A great loss of tibial cancellous bone,significantly lower tibial biomechanical expression,serious damage of microstructure and weaker osteogenic activity were found in tail-suspended mice as compared with those of normal group.Physiological loading could clearly improve mechanical properties of bones,microstructure of bone trabecular,osteogenic activity and relative gene expression (P ＜ 0.05).Overloading could also improve the condition of microgravity-induced osteoporosis,but the effect was not obvious (P ＞ 0.05).Conclusions Tail-suspension can successfully simulate microgravity environment and duplicate osteoporosis model.Physiological loading can effectively prevent the emergence and development of microgravity-induced osteoporosis,while overloading can also counter microgravity-induced osteoporosis,but the results have no significant differences.

Effect of 1, 25-dihydroxy vitamin D3 combined with mechanical strain treatment on proliferation, differentiation, and expression of OPG and RANKL in osteoblastic MC3T3-E1 cells / 医用生物力学

Objective To investigate the effect of 1,25-(OH)2-vitamin D3 (VD3) or mechanical strain alone and their combined treatment on proliferation and differentiation of pre-osteoblast MC3T3-E1 cells in vitro, as well as gene and protein expression of osteoprotegerin (OPG) and receptor activator of nuclear factor-кB ligand (RANKL) in those cells. Methods MC3T3-E1 cells were treated with 10 nmol/L VD3, intermitted mechanical strain or with a combination of these two factors. Cell proliferation was assessed with flow cytometry, and alkaline phosphatase (ALP) activity was measured using a fluorometric detection kit. The mRNA expression of ALP, runt-related transcriptional factor 2 (Runx2), OPG, and RANKL genes was determined by real-time PCR. The proteins expression of Runx2, OPG, and RANKL was determined by Western blotting. ResultsVD3 inhibited the proliferation of MC3T3-E1 cells, but the mechanical strain had no effect on cell proliferation. Mechanical strain, VD3, and the combined treatment enhanced the ALP activity of MC3T3-E1 cells as well as the protein expression of Runx2. The effect of combined treatment was less pronounced than the effect of VD3 or mechanical strain alone. Mechanical strain promoted the gene and protein expression of osteoprotegerin (OPG) and increased the ratio of OPG/RANKL. However, the combination of VD3 and mechanical strain led to a decrease in ratio of OPG/RANKL. Conclusions Mechanical strain might be effective in inducing osteogenic differentiation and increasing bone formation. A joint stimulation with VD3 and strain can decrease proliferation and osteogenic differentiation and increase RANKL expression, which might affect bone remodeling. This study supplies some new data, which might be important in theoretical and clinical research of osteoporosis (OP) and other related bone diseases.

Bone formation in rabbit cancellous bone explant culture model is enhanced by mechanical load.

BACKGROUND: When studying and designing an artificial bone in vitro with similar features and functionality of natural bone by tissue engineering technology, the culturing environment, especially the mechanical environment is supposed to be an important factor, because a suitable mechanical environment in vitro may improve the adaptability of the planted-in tissue engineering bone in the body. Unfortunately, up to now, the relationship between mechanical stimuli and natural bone growth has not yet been precisely determined, and it is so imperative for a prior study on effect of mechanical loading on growth of the natural bone cultured in vitro. METHODS: Under sterile conditions, explant models of rabbit cancellous bone with 3 mm in thickness and 8 mm in diameter were prepared and cultured in a dynamic loading and circulating perfusion bioreactor system. By Micro-CT scanning, a 3D model for finite element (FEM) analysis was achieved. According to the results of FEM analysis and physiological load bearing capacity of the natural bone, these models were firstly subjected to mechanical load with 1Hz frequency causing average apparent strain of 1000 µÎµ, 2000 µÎµ, 3000 µÎµ and 4000 µÎµ respectively for 30 min every day, activities of alkaline phosphatase (AKP) were detected on the 5th and the 14th loading day and on the 14th and the 21st day, mechanical properties, tissue mineral density (TMD) of the bone explant models were investigated and Von-kossa staining and fluorescence double labeling assays were conducted to evaluate whether there were fresh osteoid in the bone explant models. In addition, Western blot, Elisa and Real-time PCR were employed to analyze expression of Collagen-I (COL-1), bone morphogenetic protein-2 (BMP-2) and osteoprotegerin (OPG) protein and RNA. RESULTS: The explant models of rabbit cancellous bone prepared under sterile conditions grew well in the bioreactor system. With the increasing culturing time and load levels, bone explant models in groups with 1000 µÎµ and 2000 µÎµ average apparent strain experienced improving mechanical properties and TMD (P<0.05), and results of Von-kossa staining and fluorescence double labeling also showed apparent fresh osteoid formation. Under the same loading conditions, a up-regulations in protein and RNA of COL-1, BMP-2 and OPG were detected, especially, relative genes notably expressed after 21 days. CONCLUSION: Our study demonstrated that mechanical load could improve function and activity of osteoblasts in explant models of cancellous bone. Through regulations of COL-1, OPG and BMP-2 secreted by osteoblasts, the mechanical load could improve the tissue structural density and stiffness due to formation of fresh osteoid.

Meta analysis of acupuncture-moxibustion in treatment of irritable bowel syndrome / 中国针灸

<p><b>OBJECTIVE</b>To evaluate the clinical efficacy and safety of acupuncture-moxibustion in treatment of irritable bowel syndrome systematically.</p><p><b>METHODS</b>Clinical randomized controlled trials on treatment of irritable bowel syndrome with acupuncture-moxibustion were collected. Through retrieval of CNKI (1979 - December of 2011) and VIP (1979- December of 2011), randomized and quasi-randomized controlled clinical trials on treatment of irritable bowel syndrome with control study between acupuncture and sham acupuncture or western medication were included. The test bias risk and quality assessment of each experiment were carried out by two researchers in accordance with the Cochrane Handbook 5.1.0 standard. And RevMan 5.1.6 software was adopted for the Meta analysis.</p><p><b>RESULTS</b>Eleven researches were included with totally 969 patients. Meta analysis shows that the effective rate of the combined methods of acupuncture and moxibustion [RR = 1. 27, 95% CI ( 1.09, 1.49)] is superior to conventional western medication treatment.</p><p><b>CONCLUSION</b>Acupuncture-moxibustion for irritable bowel syndrome is better than the conventional western medication treatment.</p>

Effects of neuroactive steroid allopregnanolone on the damage of cortical neurons / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the protective mechanism of neuroactive steroid allopregnanolone on N-methyl-D-aspartate (NMDA) induced toxicity in primary mouse cortical neurons.</p><p><b>METHODS</b>Primary cultured mouse cortical neurons were subjected to allopregnanolone, the expression of beta-aminobutyric acid receptor beta2 subunit (beta2-GABA-R) mRNAs was detected by RT-PCR and Akt phosphorylation was assayed by Western blot using Akt-phosphoserine 473-specific antibody. After the cultured mouse cortical neurons were pretreated with or without allopregnanolone prior to treatment with NMDA , DNA isolated was analyzed by agarose gel electrophoresis and proteins collected were analyzed by Western blot with anti-cleaved-PARP, anti-cleaved caspase-3, and anti-cleaved caspase-9 antibodies.</p><p><b>RESULTS</b>When cultured mouse cortical neurons were exposed to allopregnanolone both the expression of beta2-GABA-R mRNAs and Akt phosphorylation increased. Allopregnanolone inhibited the NMDA-induced apoptosis and decreased the level of active-PARP, active-caspase-3 and active-caspase-9 notably at a final concentration of 5 x 10(6) mol/L.</p><p><b>CONCLUSION</b>Pretreatment with allopregnanolone may be neuroprotective on NMDA-induced neuronal cells apoptosis by increasing beta2-GABA-R expression and Akt phosphorylation.</p>

Expression of Helicoverpa armigera Cathepsin B in Pichia pastoris / 中国生物工程杂志

Cathepsin B from Helicoverpa armigera (HCB) belongs to the group of cysteine proteinases. HCB is proved being involved in the degradation of yolk proteins during embryonic development,which is an acidic preferring enzyme and is resistant to SDS. The expression of the proenzyme may offer a model for investigating the activation of the enzyme. The HCB gene was constructed into pPIC9K and expressed in Pichia pastoris KM71 strain . After induction by methanol, HCB was expressed and secreted into the medium. The molecular weight of the recombinant procathepsin B was determined as about 38 kDa. The expressed product was confirmed to be HCB by immunoblotting assay using specific rabbit anti-HCB polyclonal antibody. The activity of the product was assayed by in situ hydrolysis (gelatin-SDS-PAGE). These results showed that HCB with proteolytic activity was expressed in P. pastoris KM71. This proenzyme can be used for further research on the activation of the proenzyme or industrial production.