Fecal microbiota as a predictor of acupuncture responses in patients with postpartum depressive disorder.

Background: There are several clinical and molecular predictors of responses to antidepressant therapy. However, these markers are either too subjective or complex for clinical use. The gut microbiota could provide an easily accessible set of biomarkers to predict therapeutic efficacy, but its value in predicting therapy responses to acupuncture in patients with depression is unknown. Here we analyzed the predictive value of the gut microbiota in patients with postpartum depressive disorder (PPD) treated with acupuncture. Methods: Seventy-nine PPD patients were enrolled: 55 were treated with acupuncture and 24 did not received any treatment. The 17-item Hamilton depression rating scale (HAMD-17) was used to assess patients at baseline and after eight weeks. Patients receiving acupuncture treatment were divided into an acupuncture-responsive group or non-responsive group according to HAMD-17 scores changes. Baseline fecal samples were obtained from the patients receiving acupuncture and were analyzed by high-throughput 16S ribosomal RNA sequencing to characterize the gut microbiome. Results: 47.27% patients responded to acupuncture treatment and 12.5% patients with no treatment recovered after 8-week follow-up. There was no significant difference in α-diversity between responders and non-responders. The ß-diversity of non-responders was significantly higher than responders. Paraprevotella and Desulfovibrio spp. were significantly enriched in acupuncture responders, and these organisms had an area under the curve of 0.76 and 0.66 for predicting responder patients, respectively. Conclusions: Paraprevotella and Desulfovibrioare may be useful predictive biomarkers to predict PPD patients likely to respond to acupuncture. Larger studies and validation in independent cohorts are now needed to validate our findings.

Correlation between TCM Syndromes and Type 2 Diabetic Comorbidities Based on Fully Connected Neural Network Prediction Model.

OBJECTIVE: To predict the major comorbidities of type 2 diabetes based on the distribution characteristics of syndromes, and to explore the relationship between TCM syndromes and comorbidities of type 2 diabetes. METHODS: Based on the electronic medical record data of 3413 outpatient visits from 995 type 2 diabetes patients with comorbidities, descriptive statistical methods were used to analyze the basic characteristics of the population, the distribution characteristics of comorbidities, and TCM syndromes. A neural network model for the prediction of type 2 diabetic comorbidities based on TCM syndromes was constructed. RESULTS: Patients with TCM syndrome of blood amassment in the lower jiao were diagnosed with renal insufficiency with 95% test sensitivity. The patients with spleen deficiency combined with ascending counterflow of stomach qi and cold-damp patterns were diagnosed with gastrointestinal lesions with 92% sensitivity. The patients with TCM syndrome group of spleen heat and exuberance of heart fire were diagnosed as type 2 diabetes complicated with hypertension with a sensitivity of 91%. In addition, the prediction accuracy of combined neuropathy, heart disease, liver disease, and lipid metabolism disorder reached 70∼90% in TCM syndrome groups. CONCLUSION: The fully connected neural network model study showed that syndrome characteristics are highly correlated with type 2 diabetes comorbidities. Syndrome location is commonly in the heart, spleen, stomach, lower jiao, meridians, etc., while syndrome pattern manifests in states of deficiency, heat, phlegm, and blood stasis. The different combinations of disease location and disease pattern reflect the syndrome characteristics of different comorbidities forming the characteristic syndrome group of each comorbidity. Major comorbidities could be predicted with a high degree of accuracy through TCM syndromes. Findings from this study may have further implementations to assist with the diagnosis, treatment, and prevention of diabetic comorbidities at an early stage.

Study on the TCM Syndromes Evolution and Chinese Herbal Characteristics of Type 2 Diabetes Patients with Different Courses of Disease in TCM "Heat Stage": A Real-World Study.

OBJECTIVE: The purpose of this study is to analyze and summarize the syndrome distribution, syndrome evolution, and Chinese herb medicine characteristics of T2D in heat stage. METHOD: In this study, 228 heat-stage T2D patients were divided into three groups based on the course of disease. Group 1 (the course of disease ≤5 years) included 118 patients. Group 2 (5< the course of disease ≤10 years) had 73 patients. Group 3 (the course of disease >10 years) consisted of 37 patients. The main methods used in our study were complex network community partitioning algorithms and Sankey diagram visualization, based on the clinical electronic medical record data we collected. RESULT: In the three groups, the nodes with the highest node degree are all "heat syndrome." Edge weight between "heat" and "dampness," "qi stagnation," "phlegm," "liver," and "stomach" is the largest. During the whole course of treatment, 60.17%, 63.01%, and 62.16% of the patients' syndromes in groups 1, 2, and 3, respectively, were ascribed to the heat stage all the time. The patients' syndromes in groups 1 and 2 easily transformed to the syndrome of deficiency of both qi and yin of the spleen and stomach. In group 3, 27% of the patients' syndromes were easily transformed into kidney yin deficiency and qi deficiency and blood stasis syndrome. The largest Chinese herb communities of the patients whose syndromes did not change after treatment in the three groups were all heat-clearing drugs. The proportion of blood-activating drugs in patients with syndrome changes increased significantly after treatment. CONCLUSION: (1) The basic syndrome of T2D patients in the heat stage is liver-stomach heat syndrome. (2) T2D patients in the heat stage tend to deteriorate towards the direction of qi and yin deficiency syndrome. However, the longer the course of the disease is, the more likely it is to deteriorate to the direction of kidney yin deficiency syndrome and blood stasis syndrome. (3) Drugs that can help T2D patients in the heat stage to maintain their condition stably are heat-clearing drugs represented by Coptis chinensis, which usually need to be combined with warming interior drugs such as Zingiberis Rhizoma and Pinelliae Rhizoma.

Ligilactobacillus Salivarius LCK11 Prevents Obesity by Promoting PYY Secretion to Inhibit Appetite and Regulating Gut Microbiota in C57BL/6J Mice.

SCOPE: Obesity is a common disease worldwide and there is an urgent need for strategies to preventing obesity. METHODS AND RESULTS: The anti-obesity effect and mechanism of Ligilactobacillus salivarius LCK11 (LCK11) is studied using a C57BL/6J male mouse model in which obesity is induced by a high-fat diet (HFD). Results show that LCK11 can prevent HFD-induced obesity, reflected as inhibited body weight gain, abdominal and liver fat accumulation and dyslipidemia. Analysis of its mechanism shows that on the one hand, LCK11 can inhibit food intake through significantly improving the transcriptional and translational levels of peptide YY (PYY) in the rectum, in addition to the eventual serum PYY level; this is attributed to the activation of the toll-like receptor 2/nuclear factor-κB signaling pathway in enteroendocrine L cells by the peptidoglycan of LCK11. On the other hand, LCK11 supplementation effectively reduces the Firmicutes/Bacteroidetes ratio and shifts the overall structure of the HFD-disrupted gut microbiota toward that of mice fed on a low-fat diet; this also contributes to preventing obesity. CONCLUSION: LCK11 shows the potential to be used as a novel probiotic for preventing obesity by both promoting PYY secretion to inhibit food intake and regulating gut microbiota.

Computationally identified novel agonists for GPRC6A.

New insights into G protein coupled receptor regulation of glucose metabolism by ß-cells, skeletal muscle and liver hepatocytes identify GPRC6A as a potential therapeutic target for treating type 2 diabetes mellitus (T2D). Activating GPRC6A with a small molecule drug represents a potential paradigm-shifting opportunity to make significant strides in regulating glucose homeostasis by simultaneously correcting multiple metabolic derangements that underlie T2D, including abnormalities in ß-cell proliferation and insulin secretion and peripheral insulin resistance. Using a computational, structure-based high-throughput screening approach, we identified novel tri-phenyl compounds predicted to bind to the venus fly trap (VFT) and 7-transmembrane (7-TM) domains of GPRC6A. Experimental testing found that these compounds dose-dependently stimulated GPRC6A signaling in a heterologous cell expression system. Additional chemical modifications and functional analysis identified one tri-phenyl lead compound, DJ-V-159 that demonstrated the greatest potency in stimulating insulin secretion in ß-cells and lowering serum glucose in wild-type mice. Collectively, these studies show that GPRC6A is a "druggable" target for developing chemical probes to treat T2DM.

GPCR6A Is a Molecular Target for the Natural Products Gallate and EGCG in Green Tea.

SCOPE: The molecular mechanisms whereby gallates in green tea exert metabolic effects are poorly understood. METHODS AND RESULTS: We found that GPRC6A, a multi-ligand-sensing G-protein-coupled receptor that regulates energy metabolism, sex hormone production, and prostate cancer progression, is a target for gallates. Sodium gallate (SG), gallic acid (GA) > ethyl gallate (EG) > octyl gallate (OG) dose dependently activated ERK in HEK-293 cells transfected with GPRC6A but not in non-transfected controls. SG also stimulated insulin secretion in ß-cells isolated from wild-type mice similar to the endogenous GPRC6A ligands, osteocalcin (Ocn) and testosterone (T). Side-chain additions to create OG resulted in loss of GPRC6A agonist activity. Another component of green tea, epigallocatechin 3-gallate (EGCG), dose-dependently inhibited Ocn activation of GPRC6A in HEK-293 cells transfected with GPRC6A and blocked the effect of Ocn in stimulating glucose production in CH10T1/2 cells. Using structural models of the venus fly trap (VFT) and 7-transmembrane (7-TM) domains of GPRC6A, calculations suggest that l-amino acids and GA bind to the VFT, whereas EGCG is calculated to bind to sites in both the VFT and 7-TM. CONCLUSION: GA and EGCG have offsetting agonist and antagonist effects on GPRC6A that may account for the variable metabolic effect of green tea consumption.

[The "regulating conception-governor vessel" acupuncture method for infertility of polycystic ovarian syndrome].

OBJECTIVE: To compare clinical efficacy differences between"regulating conception-governor vessel" acupuncture method and clomiphene for infertility of polycystic ovarian syndrome (PCOS). METHODS: One hundred patients of PCOS were randomly assigned into an observation group and a control group, 50 cases in each one. The patients in the observation group were treated with"regulating Conception-Governor Vessel" acupuncture method at Zhongwan (CV 12), Guanyuan (CV 4), Qihai (CV 6), Zhongji (CV 3), Mingmen (GV 4), Yaoyangguan (GV 3) and Yaoshu (GV 2). The acupuncture treatment started at the end of menstruation, once every other day; after four treatments, the follicle was tested with B ultrasound; when follicle was longer than 18 mm or above, the acupuncture treatment was given once a day until follicle was discharged and acupuncture treatment finished. The patients in the control group were treated with oral administration of clomiphene from 5 days into menstruation, 50 mg per day for 5 consecutive days. Both groups were treated for three menstruation cycle. The menstrual cycles, endometrial thickness, endometrium types (A, B and C), cervical mucus scores, basal body temperature (BBT), cases of ovulation, cases of pregnancy were observed.The levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2), prolactin (PRL), progesterone (P) and testosterone (T) in serum were detected before and after the treatment. RESULTS: After the treatment, the menstrual cycles were shortened (both P<0.05),the endometrial thickness and cervical mucus scores were increased in the two groups (P<0.05, P<0.01); the improvement of endometrial thickness and cervical mucus score in the observation group were higher than those in the control group (both P<0.01). The percentages of type-A endometrium in both groups were higher than those before the treatment (both P<0.05); the number of type-A endometrium in the observation group was higher than that in the control group (P<0.05). The ovulation rate was 88.0% (44/50) in the observation group, which was superior to 70.0% (35/50) in the control group (P<0.05). After treatment, the levels of LH were reduced in the two groups (P<0.05, P<0.01), which was more significant in the observation group (P<0.05). CONCLUSIONS: The"regulating Conception-Governor Vessel" acupuncture method could improve menstrual cycles, increase endomet-rial thickness and promote the development of follicles in PCOS patient; in addition, it could decrease serum LH level, improve the ovarian functions and increase ovulation rate, which is superior to oral administration of clomiphene.

Electro-acupuncture at Conception and Governor vessels and transplantation of umbilical cord blood-derived mesenchymal stem cells for treating cerebral ischemia/reperfusion injury.

Mesenchymal stem cell transplantation is a novel means of treating cerebral ischemia/reperfusion, and can promote angiogenesis and neurological functional recovery. Acupuncture at Conception and Governor vessels also has positive effects as a treatment for cerebral ischemia/reperfusion. Therefore, we hypothesized that electro-acupuncture at Conception and Governor vessels plus mesenchymal stem cell transplantation may have better therapeutic effects on the promotion of angiogenesis and recovery of neurological function than either treatment alone. In the present study, human umbilical cord blood-derived mesenchymal stem cells were isolated, cultured, identified and intracranially transplanted into the striatum and subcortex of rats at 24 hours following cerebral ischemia/reperfusion. Subsequently, rats were electro-acupunctured at Conception and Governor vessels at 24 hours after transplantation. Modified neurological severity scores and immunohistochemistry findings revealed that the combined interventions of electro-acupuncture and mesenchymal stem cell transplantation clearly improved neurological impairment and up-regulated vascular endothelial growth factor expression around the ischemic focus. The combined intervention provided a better outcome than mesenchymal stem cell transplantation alone. These findings demonstrate that electro-acupuncture at Conception and Governor vessels and mesenchymal stem cell transplantation have synergetic effects on promoting neurological function recovery and angiogenesis in rats after cerebral ischemia/reperfusion.

[Efficacy and safety evaluation of fire needling for rats with rheumatoid arthritis].

OBJECTIVE: To explore the therapeutic effect and safety of fire needling on rat with rheumatoid arthritis. METHODS: Forty female Wistar rats were randomly divided into a normal group, a model group, a methotrexate (MTX) group and a fire needling group, 10 rats in each group. The 0.1 mL of normal saline was subcutaneously injected in rat's right back paw in the normal group, while the other groups received subcutaneous injection of 0.1 mL of complete Freund's adjuvant to establish the model. The pricking method was applied in the fire needling group at "Jiaji" (EX-B 2), "Zusanli" (ST 36) and Ashi points with a depth of 5 mm, once every three days, and totally eight times were required. In the MTX group, with intragastric administration of 2.0 mg/kg per rat, the treatment was given once every seven days, and totally four times were required. The weight, swelling rate of foot, joint pain score and polyarthritis index of rats in each group as well as the pathological change of liver tissue under light microscope were observed. RESULTS: The weight, swelling rate of foot, joint pain score and polyarthritis index in the model group were significantly higher than those in the normal group (P < 0.05, P < 0.01), after the treatment, the above-mentioned indexes in the two treatment groups were obviouly dereased as componed with those in the model group (all P < 0.01), and there were statistical differences between the fire needling group and MTX group (P < 0.05, P < 0.01). Compared with the normal group, inflammatory cells were appeared with a cluster distribution in the liver cells of rats in the model group. The inflammatory cells were reduced in the MTX group and fire needling group, and liver cells had degenerative edema and cell nucleus were largen and deformed in the MTX group. CONCLUSION: The fire needling has significant efficacy for rats with adjuvant arthritis (AA) without any damage to the liver, which coud have a better control of disease progression of rheumatoid arthritis .

Combinatorial effects of conception and governor vessel electroacupuncture and human umbilical cord blood-derived mesenchymal stem cells on pathomorphologic lesion and cellular apoptosis in rats with cerebral ischemia/reperfusion.

OBJECTIVE: To investigate the combinatorial effects of conception and governor vessel electroacupuncture (EA) and human umbilical cord blood-derived mesenchymal stem cells (HUCB-MSCs) on pathomorphologic lesion and cellular apoptosis in rats with cerebral ischemia/reperfusion. METHODS: With the HUCB-MSCs isolated, cultured and identified and the models of cerebral ischemia-reperfusion established, the HUCB-MSCs of passage three were intracranially transplanted and the EA at conception and governor vessels was applied. The pathomorphologic lesion by hematoxylin-eosin staining and the cellular apoptosis by terminal deoxynucleotidyl transferase-mediated nick-end labeling method around the ischemic focus were observed. RESULTS: The cultured adherent HUCB-MSCs exhibited a spindle shape and expressed MSC-specific markers, with the cell purity and proliferation rate significantly increasing after the primary passage. HE staining showed that there were no pathological changes observed in the sham surgery group. However, in the PBS transplantation group, degeneration and necrosis of a great number of nerve cells were seen. In both the HUCB-MSCs transplantation group and the HUCB-MSCs transplantation + EA group, reparative changes of the pathomorphism of the tissue were found. Both combination treatment and simple MSCs treatment were able to improve the pathomorphorlogic lesion following cerebral ischemia and reduce the abnormal TUNEL-positive numbers, with former better than latter. CONCLUSION: HUCB-MSCs improve pathological lesions and inhibit the cellular apoptosis around the cerebral ischemic area. EA at conception and governor vessels also improve pathological lesion and inhibit the cellular apoptosis in rats treated with HUCB-MSCs transplantation, which effects were superior to that of simple HUCB-MSCs transplantation.

Efficacy and safety evaluation of fire needling for rats with rheumatoid arthritis / 中国针灸

<p><b>OBJECTIVE</b>To explore the therapeutic effect and safety of fire needling on rat with rheumatoid arthritis.</p><p><b>METHODS</b>Forty female Wistar rats were randomly divided into a normal group, a model group, a methotrexate (MTX) group and a fire needling group, 10 rats in each group. The 0.1 mL of normal saline was subcutaneously injected in rat's right back paw in the normal group, while the other groups received subcutaneous injection of 0.1 mL of complete Freund's adjuvant to establish the model. The pricking method was applied in the fire needling group at "Jiaji" (EX-B 2), "Zusanli" (ST 36) and Ashi points with a depth of 5 mm, once every three days, and totally eight times were required. In the MTX group, with intragastric administration of 2.0 mg/kg per rat, the treatment was given once every seven days, and totally four times were required. The weight, swelling rate of foot, joint pain score and polyarthritis index of rats in each group as well as the pathological change of liver tissue under light microscope were observed.</p><p><b>RESULTS</b>The weight, swelling rate of foot, joint pain score and polyarthritis index in the model group were significantly higher than those in the normal group (P < 0.05, P < 0.01), after the treatment, the above-mentioned indexes in the two treatment groups were obviouly dereased as componed with those in the model group (all P < 0.01), and there were statistical differences between the fire needling group and MTX group (P < 0.05, P < 0.01). Compared with the normal group, inflammatory cells were appeared with a cluster distribution in the liver cells of rats in the model group. The inflammatory cells were reduced in the MTX group and fire needling group, and liver cells had degenerative edema and cell nucleus were largen and deformed in the MTX group.</p><p><b>CONCLUSION</b>The fire needling has significant efficacy for rats with adjuvant arthritis (AA) without any damage to the liver, which coud have a better control of disease progression of rheumatoid arthritis .</p>

Study strategies for acupuncture treatment of chronic nonbacterial prostatitis.

By retrospectively reviewing the current status of traditional Chinese medicine (TCM) and Western medicine treatments of chronic nonbacterial prostatitis (CNP), the TCM understanding of its etiologies and pathogenesis, the therapeutic principles and the mechanisms of acupuncture treatment of CNP, the clinical study strategies of acupuncture treatment for CNP were further proposed, which could provide more scientific basis and support for the definite longer-term therapeutic efficacy of acupuncture treatment of CNP. Breakthrough in the treatment of CNP will be achieved with the application of acupuncture therapy both in clinical practice and experimental research.

Multiligand specificity and wide tissue expression of GPRC6A reveals new endocrine networks.

Emerging evidence supports the hypothesis that the skeleton is an endocrine organ that regulates energy metabolism through the release of the osteoblast-derived hormone, osteocalcin (Ocn). This bone-pancreas endocrine network is controversial because important gaps remain to be filled in our knowledge of the physiological effects of Ocn in multiple organs and the complex alterations in other hormonal networks induced by Ocn administration. A key step toward understanding the integrative regulation of energy metabolism by bone is the identification of GPCR family C group 6 member A (GPRC6A) as the Ocn receptor. GPRC6A is an amino acid-sensing G protein-coupled receptor highly expressed in ß-cells and is activated by recombinant Ocn in vitro and in vivo but that is widely expressed in tissues other than the pancreas and is capable of sensing multiple structurally unrelated ligands, including l-amino acids, cations, and anabolic steroids in addition to Ocn. The broad expression and multiligand specificity of GPRC6A is identifying both systemic and paracrine regulation of seemingly disparate biological processes, ranging from energy metabolism, sexual reproduction, hypothalamic-pituitary function, bone formation, and prostate cancer. Consistent with the existence of more complex endocrine networks, ablation of GPRC6A in Gprc6a(-/-) mice results in complex metabolic abnormalities, including obesity, glucose intolerance, hepatic steatosis, insulin resistance, hyperphosphatemia, osteopenia, plus several hormonal abnormalities, including decreased circulating testosterone, IGF-I, and insulin and increased estradiol, LH, GH, and leptin. Recombinant Ocn also regulates testosterone production by the testes and male fertility through a GPRC6A-dependent mechanism, and testosterone regulation of LH secretion is abnormal in Gprc6a(-/-) mice. Thus, GPRC6A, as the biologically relevant receptor for Ocn, defines not only a molecular mechanism for linking bone metabolism with metabolic regulation of ß-cells and sexual reproduction but also as a receptor shared by testosterone and dietary factors, and it is also involved in multiple endocrine networks integrating the functions of pancreas, muscle, liver, fat, testes, bone, and the hypothalamic-pituitary axis with alterations in both environmental and endogenous ligands.

Research advances in treatment of cerebral ischemic injury by acupuncture of conception and governor vessels to promote nerve regeneration.

Cerebral ischemia is one of the most common diseases treated by acupuncture therapeutics. Recent studies indicated that acupuncture treatment by needling the conception and governor vessels had positive effects in promoting neural regeneration in patients after cerebral ischemia injury. Acupuncture intervention could continuously promote the proliferation and differentiation of the neural stem cells in the brain, obviously up-regulate expression of growth factors, accelerate angiogenesis and inhibit apoptosis. Hence, it is necessary to present an exhaustive review on the mechanisms. The present review gives a detailed description of pathological changes of cerebral ischemia and acupuncture intervention applied to the conception and governor vessels, and proposes research prospects in the future.

Effects of electroacupuncture at the conception vessel on proliferation and differentiation of nerve stem cells in the inferior zone of the lateral ventricle in cerebral ischemia rats.

OBJECTIVE: To observe the effects of electroacupuncture (EA) at the Conception Vessel on proliferation and differentiation of the nerve stem cells in the inferior zone of the lateral ventricle in cerebral ischemia rats. METHODS: The model rats were prepared by occlusion of the middle cerebral artery for 2 hours and then by reperfusion. They were randomly divided into two groups: a control group and an EA group. Changes in differentiation and proliferation of the nerve stem cells were observed 7, 14 and 28 days after successful modeling. RESULTS: As compared with the 7-day control group (C-7d group), there was no significant difference (P > 0.05) in the numbers of 5-bromodeoxyuridine (Brdu) positive cells, Brdu/GFAP, Brdu/Nestin and Brdu/Nse double-labeled cells in the inferior zone of the lateral ventricle in the EA group 7 days after modeling. However, in the 14-day EA group (R-14d group) and the 28-day EA group (R-28d group), the numbers of Brdu positive cells and Brdu/GFAP, Brdu/Nestin, Brdu/Nse double-labeled cells significantly increased as compared respectively with the 14-day control (C-14d group) and the 28-day control (C-28d) group (P < 0.05 or P < 0.01). CONCLUSIONS: EA at the Conception Vessel promotes differentiation and proliferation of the nerve stem cells in the inferior zone of the lateral ventricle in the cerebral ischemia rats, and may stimulate differentiation of the proliferous nerve stem cells towards the astrocytes.

Correlativity of the therapeutic effect of acupuncture on prolapse of lumbar intervertebral disc with the area location of image / 中国针灸

<p><b>OBJECTIVE</b>To probe into the correlativity of the therapeutic effect of acupuncture combined with traction on prolapse of lumbar intervertebral disc with the three-dimensional location.</p><p><b>METHODS</b>According to the three-dimensional location developed by HU You-gu, the position of the prolapse of lumbar intervertebral disc in the spinal canal was located from the sagittal plain, frontal plain and horizontal plain. One hundred and thirty patients with prolapse of lumbar intervertebral disc and completion of clinical data were treated with acupuncture, and the therapeutic effects were analyzed with three-dimensional location.</p><p><b>RESULTS</b>Among 98 cases in the effective group, 73 cases had the prolapse at the I plane in sagittal location, accounting for 74.49%, 82 cases had the prolapse at the 1, 2 zone in horizontal location, accounting for 83.68%, 52 cases had the prolapse at the a area in frontal location, accounting for 53.06%; among the 32 cases in the ineffective group, 10 cases had the prolapse at the I plane, accounting for 31.25%, 12 cases in the 1, 2 zone, accounting for 37.50%, 3 cases in the a area, accounting for 9.38%. There was a statistically significant difference between the two groups in the location.</p><p><b>CONCLUSION</b>The three-dimensional location can objectively show the size, direction, position, and the characteristics of the protruded discs, and correctly infer the pathological changes of the affected nerve root. Acupuncture combined with tract has a better effect on I plane, 1-2 zone, and a area.</p>

Calcyclin mediates serum response element (SRE) activation by an osteoblastic extracellular cation-sensing mechanism.

UNLABELLED: The molecular mechanism of sensing extracellular cations in osteoblasts is controversial. Using an expression-cloning strategy, the calcium-binding protein calcyclin was found to mediate the response of MC3T3-E1 osteoblasts to extracellular cations, but not the calcimimetic NPS-568, indicating the presence of another cation-sensing mechanism. Further understanding of calcyclin function in osteoblasts may identify novel targets for regulating bone formation. INTRODUCTION: Extracellular calcium and other cations seem to regulate the function of osteoblasts through a distinct calcium-sensing mechanism that is coupled to activation of c-fos gene transcription. The identity of this calcium-sensing mechanism is unknown. METHODS: To identify molecules that participate in this extracellular cation-sensing pathway, we developed an expression cloning strategy in COS-7 cells using cation stimulation of a serum response element (SRE) luciferase reporter derived from the c-fos promoter to screen a mouse MC3T3-E1 osteoblast cDNA library. RESULTS AND CONCLUSIONS: We identified calcyclin (S100A6), a calcium-binding protein of the EF-hand type belonging to the S100 family, as being responsible for transferring a cation-sensing response from osteoblasts to COS-7 cells. Transfection of the calcyclin cDNA into COS-7 and HEK-293 cells confirmed that the overexpression of calcylin caused these cells to gain the ability to sense extracellular cations, including aluminum, gadolinium, calcium, and magnesium. Conversely, we found that an antisense calcyclin construct reduced calcyclin expression and partially inhibited the cation-sensing response in MC3T3-E1 osteoblasts. These results implicate calcyclin in the activation of SRE and establish a role for calcyclin as an accessory protein involved in the cation-sensing pathway in osteoblasts.