MW-9, a chalcones derivative bearing heterocyclic moieties, ameliorates ulcerative colitis via regulating MAPK signaling pathway / 中国药理学通报

Aim To investigate the therapeutic effect of the MW-9 on ulcerative colitis(UC)and reveal the underlying mechanism, so as to provide a scientific guidance for the MW-9 treatment of UC. Methods The model of lipopolysaccharide(LPS)-stimulated RAW264.7 macrophage cells was established. The effect of MW-9 on RAW264.7 cells viability was detected by MTT assay. The levels of nitric oxide(NO)in RAW264.7 macrophages were measured by Griess assay. Cell supernatants and serum levels of inflammatory cytokines containing IL-6, TNF-α and IL-1β were determined by ELISA kits. Dextran sulfate sodium(DSS)-induced UC model in mice was established and body weight of mice in each group was measured. The histopathological damage degree of colonic tissue was assessed by HE staining. The protein expression of p-p38, p-ERK1/2 and p-JNK was detected by Western blot. Results MW-9 intervention significantly inhibited NO release in RAW264.7 macrophages with IC50 of 20.47 mg·L-1 and decreased the overproduction of inflammatory factors IL-6, IL-1β and TNF-α(P<0.05). MW-9 had no cytotoxicity at the concentrations below 6 mg·L-1. After MW-9 treatment, mouse body weight was gradually reduced, and the serum IL-6, IL-1β and TNF-α levels were significantly down-regulated. Compared with the model group, MW-9 significantly decreased the expression of p-p38 and p-ERK1/2 protein. Conclusions MW-9 has significant anti-inflammatory activities both in vitro and in vivo, and its underlying mechanism for the treatment of UC may be associated with the inhibition of MAPK signaling pathway.

Targeting cAMP in D1-MSNs in the nucleus accumbens, a new rapid antidepressant strategy

Studies have suggested that the nucleus accumbens (NAc) is implicated in the pathophysiology of major depression; however, the regulatory strategy that targets the NAc to achieve an exclusive and outstanding anti-depression benefit has not been elucidated. Here, we identified a specific reduction of cyclic adenosine monophosphate (cAMP) in the subset of dopamine D1 receptor medium spiny neurons (D1-MSNs) in the NAc that promoted stress susceptibility, while the stimulation of cAMP production in NAc D1-MSNs efficiently rescued depression-like behaviors. Ketamine treatment enhanced cAMP both in D1-MSNs and dopamine D2 receptor medium spiny neurons (D2-MSNs) of depressed mice, however, the rapid antidepressant effect of ketamine solely depended on elevating cAMP in NAc D1-MSNs. We discovered that a higher dose of crocin markedly increased cAMP in the NAc and consistently relieved depression 24 h after oral administration, but not a lower dose. The fast onset property of crocin was verified through multicenter studies. Moreover, crocin specifically targeted at D1-MSN cAMP signaling in the NAc to relieve depression and had no effect on D2-MSN. These findings characterize a new strategy to achieve an exclusive and outstanding anti-depression benefit by elevating cAMP in D1-MSNs in the NAc, and provide a potential rapid antidepressant drug candidate, crocin.

Histone methylation and diabetic cardiomyopathy / 生理学报

Histone methylation is one of the key post-translational modifications that plays a critical role in various heart diseases, including diabetic cardiomyopathy. A great deal of evidence has shown that histone methylation is closely related to hyperglycemia, insulin resistance, lipid and advanced glycation end products deposition, inflammatory and oxidative stress, endoplasmic reticulum stress and cell apoptosis, and these pathological factors play an important role in the pathogenesis of diabetic cardiomyopathy. In order to provide a novel theoretical basis and potential targets for the treatment of diabetic cardiomyopathy from the perspective of epigenetics, this review discussed and elucidated the association between histone methylation and the pathogenesis of diabetic cardiomyopathy in details.

Research progress on relationship between medial prefrontal cortex and neuropsychiatric diseases / 中国药理学通报

The medial prefrontal cortex (mPFC) received input from multiple cortical and subcortical areas, and integrated relevant information to other cortical and subcortical areas. mPFC play an important role in neuropsychiatric events such as depression, anxiety, schizophrenia, Alzheimer's and Parkinson's disease, as well as in neuropsychiatric processes such as cognitive, social, and reward behaviors. This article reviews the role and molecular mechanism of the neural circuitry in the medial prefrontal cortex in neuropsychiatric diseases, especially the recent research progress.

Effect of CysLT / 中国药理学通报

Aim To investigate the effect of CysLT

Effectiveness of in vitro fenestration versus bypass surgery for type B aortic dissection involving the left subclavian artery / 中国胸心血管外科临床杂志

@#Objective    To analyze the effectiveness of in vitro fenestration versus bypass surgery techniques in the treatment of type B aortic dissection involving the left subclavian artery by thoracic endovascular aortic repair (TEVAR). Methods    Among the 53 patients with type B aortic dissection involving the left subclavian artery admitted to our center from January 2017 to October 2020, 23 underwent in vitro fenestration + TEVAR (a fenestration group with 18 males and 5 females aged 53.6±5.3 years), and 30 patients underwent left common carotid artery-left subclavian artery bypass + TEVAR (a bypass group with 24 males and 6 females aged 51.8±3.8 years). The effectiveness and safety between the two groups were compared. Results    The surgical success rate was 100.0% in both groups. And there was no death within postoperative 30 days and during the follow-up. There was no endoleak immediately postoperatively and during 1-year follow-up in the two groups. The operation time and hospitalization expenses in the fenestration group was less or shorter than those in the bypass group (P<0.05). The reduction in blood pressure of the left upper limb in the fenestration group was greater than that in the bypass group (P<0.05). There was no symptom of left upper limb ischemia, dizziness or hoarseness in both groups. Conclusion    The two methods of reconstruction of the left subclavian artery are safe and effective. In vitro fenestration can reduce surgical trauma and costs, and bypass surgery can provide better forward blood flow for the left subclavian artery.

Epigenetic Landscape Changes Due to Acupuncture Treatment: From Clinical to Basic Research / 中国结合医学杂志

Acupuncture has been widely used for treating diseases since the ancient days in China, but the mechanism by which acupuncture exerts such powerful roles is unclear. Epigenetics, including DNA methylation, histone modification, and post-transcriptional regulation of miRNAs, is the study of heritable changes in gene expression that do not include DNA sequence alterations. Epigenetics has become a new strategy for the basic and clinical research of acupuncture in the last decade. Some investigators have been trying to illustrate the mechanism of acupuncture from an epigenetics perspective, which has shed new lights on the mechanisms and applications of acupuncture. Moreover, the introduction of epigenetics into the regulatory mechanism in acupuncture treatment has provided more objective and scientific support for acupuncture theories and brought new opportunities for the improvement of acupuncture studies. In this paper, we reviewed the literatures that has demonstrated that acupuncture could directly or indirectly affect epigenetics, in order to highlight the progress of acupuncture studies correlated to epigenetic regulations. We do have to disclose that the current evidence in this review is not enough to cover all the complex interactions between multiple epigenetic modifications and their regulations. However, the up-to-date results can help us to better understand acupuncture's clinical applications and laboratory research.

Problem-based learning method combined with the case-based learning in clinical teaching of orthopedic interns / 中华医学教育探索杂志

Objective:To explore the effect of the problem-based learning method (PBL) combined with the case-based learning (CBL) in clinical practice teaching of orthopedic interns.Methods:Totally 60 interns of clinical medicine who studied from July 2018 to December 2019 were randomly divided into two groups: experimental group ( n=30) receiving PBL combined with CBL teaching and control group ( n=30) receiving traditional teaching. The theory examination and satisfaction survey were conducted to assess the effects two teaching methods, and t-test was performed for data analysis using SPSS 15.0. Results:No significant difference was found in the average score of examination ( P>0.05), but the examination group showed a higher average score in the clinical case analysis than the control group ( P<0.05). The satisfaction survey showed that the students in the experimental group were more satisfied with the improvement in learning interests, self-study ability, and cooperation ability. Conclusions:PBL combined with CBL teaching method achieves good teaching effect and it is worth being recommended in clinical teaching.

Effects of electroacupuncture on inflammatory response of cardiac muscle tissue in mice with acute myocardial ischemia / 中国针灸

<p><b>OBJECTIVE</b>To observe the effects of electroacupuncture (EA) on inflammatory reaction of acute myocardial ischemia (MI) in mice, and to explore its action mechanism.</p><p><b>METHODS</b>Forty adult male C57BL/6 mice were randomly divided into a control group, a sham operation group, a model group and an EA group, 10 mice in each one. The model was established in the model group and EA group by ligating the left anterior descending branch of coronary artery. The mice in the EA group were treated with EA at "Neiguan" (PC 6) with 2 mA of intensity and 2 Hz /100 Hz of frequency; EA was given 30 min per treatment, once a day for totally 5 days. The mice in the control group and model group were treated with immobilization and no EA was given. The mice in the sham operation group were not treated with ligating at the left anterior descending branch of coronary artery, but the remaining procedure was identical to the model group. The electrocardiogram was recorded and △ST was calculated to evaluate the model. TTC and HE staining methods were applied to evaluate the infarct size and pathologic change of myocardial tissue, respectively. Western blot method was applied to test the protein expression levels of tumor necrosis factor-α (TNF-α), nuclear factor-κB p65 (NF-κB p65), interleukin-1β (IL-1β) and interleukin-8 (IL-8).</p><p><b>RESULTS</b>Compared with the sham operation group, the S-T segments in the model group and EA group were increased obviously after modeling (both <0.01), indicating the MI model was established successfully. The TTC and HE staining results indicated, compared with the sham operation group, the model group had larger infarction size (<0.01), more myocardial fibers injury and inflammatory infiltration; compared with the model group, the infarction size of the EA group was significantly reduced (<0.01), and the myocardial fibers injury and inflammatory infiltration were improved. Compared with the control group, the protein expression levels in the sham operation group were similar (all >0.05); compared with the sham operation group, the expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 were significantly increased in the model group (<0.01, <0.05); compared with the model group, the expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 were significantly reduced in the EA group (all <0.05).</p><p><b>CONCLUSION</b>EA might reduce the protein expression levels of TNF-α, NF-κB p65, IL-1β and IL-8 in cardiac muscle tissue to inhibit inflammatory reaction and achieve myocardial protective effect in mice with acute myocardial ischemia.</p>

Electroacupuncture for myocardial ischemia injury in rats via AMPK-HDAC5-HIF-1α signaling / 中国针灸

OBJECTIVE@#To explore the impact of electroacupuncture (EA) on the AMPKα-HDAC5-HIF-1α signaling in the heart of the rats with myocardial ischemia (MI) via detecting the expressions of AMP-activated protein kinase α (AMPKα), histone deacetylase 5 (HDAC5), hypoxia inducible factor (HIF-1α) and vascular endothelial growth factor (VEGF).@*METHODS@#Thirty-six healthy male SD rats were randomized into a sham operation group (6 rats), a sham + EA group (6 rats), a model group (12 rats) and an EA group (12 rats). We ligated the left anterior descending artery (LAD) for MI model, and exposed the heart of rats after opening the chest without ligation for the rats in the sham operation gorup and the sham + EA group. On the 2nd day after LAD ligation, EA was applied at "Neiguan" (PC 6) with 2 Hz/15 Hz and 1.5-2 mA for 30 min in the EA group and sham+EA group, once a day for 4 days. The same fixation was used in the sham operation group and the model group, without EA. Myocardial infarction area was observed by TTC staining and serum cardiac troponin T (cTnT) was detected by radioimmunoassay. The expression of VEGF mRNA was detected by real time PCR. The protein expressions of AMPKα, HDAC5, HIF-1α and VEGF were detected by western blot.@*RESULTS@#Compared with the sham operation group, 4 days after LAD ligation, the myocardial infarction was obvious and the expression of serum cTnT increased in the model group (0.05). After EA for 4 days, the myocardial infarction area and cTnT expression decreased in the EA group (both <0.01); the VEGF mRNA and protein expressions and AMPKα, HDAC5, HIF-1α protein expressions increased (<0.05, <0.01).@*CONCLUSION@#EA could regulate the AMPKα-HDAC5-HIF-1α signaling in myocardial tissue, which may activate VEGF expression for angiogenesis signaling, reduce myocardial infarction area so as to achieve cardioprotective effect.

Neuroprotective effects of Ginkgo biloba extract and Ginkgolide B against oxygen-glucose deprivation/reoxygenation and glucose injury in a new in vitro multicellular network model / 医学前沿

Acute ischemic stroke (AIS), as the third leading cause of death worldwide, is characterized by its high incidence, mortality rate, high incurred disability rate, and frequent reoccurrence. The neuroprotective effects of Ginkgo biloba extract (GBE) against several cerebral diseases have been reported in previous studies, but the underlying mechanisms of action are still unclear. Using a novel in vitro rat cortical capillary endothelial cell-astrocyte-neuron network model, we investigated the neuroprotective effects of GBE and one of its important constituents, Ginkgolide B (GB), against oxygen-glucose deprivation/reoxygenation and glucose (OGD/R) injury. In this model, rat cortical capillary endothelial cells, astrocytes, and neurons were cocultured so that they could be synchronously observed in the same system. Pretreatment with GBE or GB increased the neuron cell viability, ameliorated cell injury, and inhibited the cell apoptotic rate through Bax and Bcl-2 expression regulation after OGD/R injury. Furthermore, GBE or GB pretreatment enhanced the transendothelial electrical resistance of capillary endothelial monolayers, reduced the endothelial permeability coefficients for sodium fluorescein (Na-F), and increased the expression levels of tight junction proteins, namely, ZO-1 and occludin, in endothelial cells. Results demonstrated the preventive effects of GBE on neuronal cell death and enhancement of the function of brain capillary endothelial monolayers after OGD/R injury in vitro; thus, GBE could be used as an effective neuroprotective agent for AIS/reperfusion, with GB as one of its significant constituents.

GTS40, an active fraction of Gou Teng-San (GTS), protects PC12 from HO-induced cell injury through antioxidative properties / 中国天然药物

Oxidative stress, a predominant cause of apoptosis cascades triggered in neurodegenerative disorders, has been regarded as a critical inducement in the pathogenesis of Alzheimer's disease (AD). Gou Teng-San (GTS) is a traditional Chinese herbs preparation commonly utilized to alleviate cognitive dysfunction and psychological symptoms of patients with dementia. The present study aimed to investigate the protective effects of GTS40, an active fraction of GTS, on HO-induced oxidative damage and identify the potential active ingredients. Our results revealed that GTS40 exhibited radical scavenging activity, elevated cell viability, decreased the levels of intracellular reactive oxygen species (ROS), and stabilized mitochondrial transmembrane potential (MMP) in HO-treated PC12 cells. In addition, GTS40 blocked the apoptotic cascade by reversing the imbalance of Bcl-2/Bax and inhibiting the activity of caspase-3. Furthermore, an HPLC-QTOFMS method was developed to characterize major chemical constituents in GTS40. Our results revealed twenty-seven identified or tentatively characterized compounds through comparing their retention time (t) and MS spectra with reference standards. These results suggested that GTS40 was a promising active fraction that may be beneficial in the prevention and treatment of oxidative stress-mediated neurodegenerative disorders.

Recent progress of 89 Zr tracers for PET imaging / 中华核医学与分子影像杂志

89Zr is a positron-emitting metal with a long half-life (78.4 h) which is compatible to the labeling of slow-accumulating bioactive vectors. Many researchers have adopted 89 Zr for medical research. DFO is the most prevalent chelator used to link 89 Zr with bioactive vectors to synthesize stable PET agents, which gives high-resolution PET images with exceptional T/NT ratio. This review describes five methods based on DFO to synthesize 89 Zr-based PET agents, among which isothiocyanate ( Df-Bz-NCS) and N-succi-nyl-DFO ( N-suc-DFO) have been used in clinical studies. The targeting bioactive vectors labeled by 89 Zr for PET imaging include antibodies (monoclonal antibodies and their derivatives), polypeptides (RGD pep-tides), proteins (transferrin, albumin, etc.), bioengineering structures (nanoparticles, liposomes, micro-spheres, etc.), cells (stem cells, immune cell subsets) and so on. The application of 89Zr PET in drug de-velopment, cell tracking and tumor imaging has become increasingly prominent.

Role of glycogen synthase kinase-3βin pathogenesis and treatment of bipolar disorder:a review / 中国药理学与毒理学杂志

Bipolar disorder(BD)is a serious mood disorder with high prevalence,morbidity and mortality rates. Glycogen synthase kinase-3β(GSK-3β) is a multifunctional serine/threonine protein kinase that is generally located in eukaryotic cells with such functions as the adjustment of the synthe?sis of glycogen metabolism,cell proliferation and differentiation and gene expression. It is involved in multiple signaling pathways and regulates cell signaling proteins,structural proteins and transcription factors through phosphorylation,affecting the survival of the neurons and plasticity. According to gene poly?morphism and clinical studies,GSK-3β may be associated with BD. As a GSK-3β inhibitor,lithium is an effective BD therapeutic drug,and the small molecule inhibitor targeting GSK-3βis also a hotspot of BD treatment. GSK-3βmay be a potential target in the treatment of BD.

Effect of pranlukast on memory impairment and nerve injury in streptozocin-induced type 1 diabetic mice / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect of pranlukast(Pran) on learning and memory impairment and neuroinflammatory and apoptotic response in streptozocin(STZ)-induced type 1 diabetic mice. METHODS Male ICR mice were injected through the tail vein with STZ(150 mg·kg-1)to induce the type 1 diabetes model. Diabetic mice were administered orally with Pran. After 4 consecutive weeks of administration,the escape latency in hidden platform trials,number of platform crossings and time spent in the target quadrant of mice were assessed by the Morris water maze(MWM)test. Western blot was used to detect the proteins of cysteinyl-leukotrienes receptor-1(CysLT1R)and pro-inflammatory factors,nuclear factor-κB p65 subunit(NF-κB p65),interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α),and cleaved caspase 3,Bax and Bcl-2 in the hippocampus and prefrontal cortex of diabetic mice. We also determined fasting blood glucose,serum insulin and lipids such as triglyceride,total cholesterol,high density lipoprotein cholesterol,and low density lipoprotein cholesterol. RESULTS The data of the MWM test showed that untreated diabetic mice displayed a higher escape latency in hidden platform trials(P<0.05),and a smaller number of platform crossings(P<0.05)as well as shorter per?centage of time spent in the target quadrant(P<0.05). The data of Western blotting showed that treat?ment with Pran 0.6 and 1.2 mg·kg-1 significantly reduced the levels of CysLT1R,nuclear NF-κB p65, IL-1βand TNF-α,cleaved caspase 3,and the ratio of Bax and Bcl-2 in the hippocampus and prefrontal cortex of diabetic mice(P<0.05). However,Pran did not improve the fasting blood glucose,serum insulin or lipid metabolism disorder in diabetic mice. CONCLUSION Pran improves memory impairment and nerve injury in STZ-induced type 1 diabetic mice.

Advances in research of the role of microglia polarization in the mechanisms of pathogenesis of multiple sclerosis / 中国比较医学杂志

Multiple sclerosis ( MS) is an autoimmune disease of the central nervous system ( CNS) characterized by demyelination and inflammation lesions.MS predominantly affects young adults with a high incidence of disability. However, the exact pathogenesis of MS is still not clear.Studies found that microglia polarization tending to pro-inflammatory M1-like state during the onset of MS, causing the M1/M2 ratio imbalance, forming pro-inflammatory microenvironment state, and which further leading to nervous tissue damage ultimately.Microglia polarization may be considered as the initiator of pathologic alterations by releasing pro-inflammatory cytokines and secondarily trigger the initial microglia response.Given the pivotal role of imbalanced microglia polarization in MS initiation, a critical review of microglia polarization is presented here, in order to elucidate the pathogenesis of MS and highlight the noteworthy candidate therapeutic targets for clinic treatment.

Effect of 18-β Glycyrrhetinic Acid on the Endoplasmic Reticulum of Nasal Epithelial Cells in Allergic Rhinitis Model Rats / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect of 18-β glycyrrhetinic acid (GA) on the endoplasmic reticulum of nasal epithelial cells in allergic rhinitis (AR) model rats.</p><p><b>METHODS</b>Totally 96 Wistar rats were randomly divided into the blank group, the AR model group, the loratadine group, the GA group, 24 in each group. AR models were established by peritoneally injecting ovalbumin (OVA). Morphological scoring was performed. GA at 21. 6 mg/kg was intragastrically administered to rats in the GA group. Nasal mucosal tissues were taken for electron microscopic examinations at the second, fourth, sixth, and tenth week after drug intervention.</p><p><b>RESULTS</b>The overlapping score was 2.10 ± 0.45 in the blank group, 5.10 ± 0.56 in the loratadine group, 5.10 ± 0.56 in the AR model group, 5.20 ± 0.78 in the GA group, showing statistical difference when compared with the blank group (P < 0.01). Results under transmission electron microscope showed that the number of the endoplasmic reticulum increased in the AR model group, with obvious cystic dilatation, a lot of vacuole formation, and degranulation. A large number of free ribosomes could be seen in cytoplasm. With persistent allergen exposure, changes mentioned above was progressively aggravated in the endoplasmic reticulum of nasal mucosal epithelium in the AR model group. But the dilation of endoplasmic reticulum, vacuole formation, and degranulation were relieved in the GA group, and got close to those of the blank group.</p><p><b>CONCLUSION</b>18-β GA could improve the expansion, vacuolization, and degranulation of the endoplasmic reticulum of nasal epithelial cells in AR model rats.</p>

The effect of 18β-sodium glycyrrhetinic acid on the nasal mucosa epithelial cilia in rat models of allergic rhinitis / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate 18β-sodium glycyrrhetinic acid impact on nasal mucosa epithelial cilia in rat models of allergic rhinitis (AR).@*METHOD@#AR models were established by ovalbumin-induction. Wister rats were randomly divided into groups as normal group, model group, budesonide (0.2 mg/kg) group and sodium glycyrrhetinic acid (20 mg/kg and 40 mg/kg) group after the success of AR models. At 2 weeks and 4 weeks after treatment, the behavioral changes of rats were observed and recorded, and nasal septum mucosae were collected after 2 week and 4 week intervention, and the morphological changes of nasal mucosae were observed by electron microscope.@*RESULT@#Model group developed typical AR symptoms, the total score in all animals was > 5. With budesonide and sodium glycyrrhetinic acid treatment, the AR symptoms were relieved, and the total scores were reduced significantly (P < 0.01). Compared with the model group: after 2 weeks' intervention, thick mucous secretions on the top of columnar epithelium cilia in rat nasal mucosa was significantly reduced, and cilia adhesion, lodging, shedding were relieved in budesonide group and sodium glycyrrhetinic acid group, the relieve in budesonide group was slightly better than that in sodium glycyrrhetinic acid group; after 4 week intervention, Cilia adhesion, lodging, shedding were completely vanished, and the cilia were ranged in regular direction in budesonide group and sodium glycyrrhetinic acid group. Cilia in sodium glycyrrhetinic acid (20 mg/kg) group was more orderly, smooth than that in budesonide group and sodium glycyrrhetinic acid group (40 mg/kg), and the condition of cilia in sodium glycyrrhetinic acid group (20 mg/kg) was similar to the normal group.@*CONCLUSION@#18β-sodium glycyrrhetinic acid is effective to restrain the pathological changes of nasal mucosa cilia in rat models of AR.

Determination of markers from characteristic HPLC chromatogram of phenols in three official origins of Ephedrae Herba and quantitative analysis of four phenols / 中国中药杂志

This study is to establish the characteristic HPLC chromatogram of phenols in Ephedrae Herba, from which to pick out the marker peaks, followed by the analysis of the regularity of their distribution and content in the herbaceous stems of Ephedra sinica, E. intermedia and E. equisetina. The HPLC-DAD method for the characteristic chromatogram as well as quantitative analysis was established. The separation was carried out on a YMC-Pack ODS-A column (4.6 mm x 250 mm, 5 µm), eluted with the mobile phases as 0.01% formic acid aqueous solution (A) and acetonitrile (B) in a linear gradient (0-10 min, 17% B; 10-25 min, 17%-19% B; 25- 33 min, 19%-48% B; 33-35 min, 48%-51% B; 35-44 min, 51% B). The flow rate was kept at 1.0 mL · min⁻¹. The column tem- perature was 40 °C, and the detection wavelength was set at 350 nm (0-16 min) and 330 nm (16-44 min). Forty-six batches of collected samples from three official origins of Ephedrae Herba were detected, whose liquid chromatograms proven to be helpful to the differentiation of different origins. With principal component analysis and the analysis of distribution of peak area, twelve key peaks from the chromatogram were discussed in details on their contributions to the characteristics and differences of three official origins of the herb: peak area of peak 10, 11, 12 were found out to be significantly higher in E. equisetina than in other two origins, whose sum (higher than 146 mAU in E. equisetina) was useful for the discrimination between E. equisetina and the other two origins; peak area of 1 and 4 were respectively higher in E. sinica and E. intermedia than in other official origins, indicating their important effect on the differen- tiation of corresponding origins; peak 8 and 9 were picked out as two characteristic common peaks in three official origins of the herb, whose peak area showed little difference among different origins; further, peak area of other key peaks in the chromatogram also showed some difference among three origins, which make contributions to the differentiation of origins as well. Then, four phenols as 2"-O-α- L-rhamnosyl-isovitexin (1), vitexin (2), pollenitin B (5) and herbacetin-7-O-β-D-glucoside (6) were quantitative analyzed with the above-mentioned method, with good linear relationship and accuracy (recoveries in a range of 97.8%-102.5%). The content of the four phenols were firstly reported in Ephedrae Herba from official origins, which were respectively trace-1.55 (1), trace-0.160 (2), trace-0.284 (5) and trace-0.620 (6) mg · g⁻¹ in all of the tested samples. In addition, the content of these phenols showed differences in three official origins, especially 1, whose content in E. sinica [(0.670 ± 0.88) mg ± g⁻¹] were significantly higher than in other two origins (lower than 0.16 mg ± g⁻¹ besides sample Ei-060630-2-2), and 6, whose average content in E. equisetina [(0.260 ± 0.039 2) mg · g⁻¹] were twice as high as in E. sinica [(0.120 ± 0.270) mg · g⁻¹] and E. intermedia [(0.136 ± 0.485) mg g⁻¹], indicating the important effects of the two constituents on the differentiation among three official origins of the herb. The method established for the characteristic HPLC chromatogram and quantitative analysis of phenols was simple and accurate, and the marker constituents selected may provide new guides for the discrimination of official origins as well as the improvement of quality criteria of EphedraeHerba.

Icariin Promoted Osteogenic Differentiation of SD Rat Bone Marrow Mesenchymal Stem Cells: an Experimental Study / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe features of Icariin in promoting osteogenic differentiation of SD rat bone marrow mesenchymal stem cells (BMSCs) in vitro.</p><p><b>METHODS</b>(1) SD rats' BMSCs were isolated and purified by mechanically isolated and cultured by whole bone marrow adherent method. Effects of various concentrations Icariin on serum activities of alkaline phosphatase (ALP) were detected using amino antipyrine phenol determination method at day 3, 6, 9, 12, 15, 18, and 21. Calcium nodes of each groups were detected using alizarin red staining. Roles of various concentrations Icariin in promoting osteogenic differentiation of BMSCs were observed. (2) BMSCs were divided into the blank control group, the osteogenic induced group, and the Icariin group (0.5 microg/mL). ALP activities were detected at day 7, 14, and 21 of culture. Meanwhile, ALP positive staining rate and calcium nodes were detected at day 14 and 21 respectively. Additionally, mRNA expressions of Runt-related transcription factor-2 (Runx2) and Osteocalcin were detected at day 7, 14, and 21 by real-time fluorescent quantitative PCR.</p><p><b>RESULTS</b>(1) 0.05-5.0 microg/mL Icariin could significantly elevate serum ALP activities. Of them, 0.2-2.0 microg/mL Icariin significantly increased calcium nodes numbers (P < 0.01). (2) When Icariin promoted osteogenic differentiation of BMSCs, Runx2 mRNA expression levels and ALP activities increased earlier and then decreased, while osteocalcin mRNA expression levels continued to increase (P < 0.01). Compared with the osteogenic induced group, ALP activities and ALP positive staining rate were both elevated after 14 days of Icariin treatment in the Icariin group (P < 0.05, P < 0.01).</p><p><b>CONCLUSIONS</b>Icariin could promote the differentiation of BMSCs to osteoblasts by up-regulating Runx2 mRNA expression levels. It also could promote the mineralization by increasing ALP secretion and Osteocalcin mRNA expression levels, thereby promoting mature of newly generated osteoblasts.</p>