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Mycena, a symbiont of Gastrodia elata, promotes seed germination of G. elata and plays a crucial role in the sexual reproduction of G. elata. However, the lack of genetic transformation system of Mycena blocks the research on the interaction mechanism of the two. In order to establish the protoplast transformation system of Mycena, this study analyzed the protoplast enzymatic hydrolysis system, screened the resistance markers and regeneration medium, and explored the transient transformation. After hydrolysis of Mycena hyphae with complexes enzymes for 8 h and centrifugation at 4 000 r·min~(-1), high-concentration and quality protoplast was obtained. The optimum regeneration medium for Mycena was RMV, and the optimum resistance marker was 50 mg·mL~(-1) hygromycin. The pLH-HygB-HuSHXG-GFP-HdSHXG was transformed into the protoplast of Mycena which then expressed GFP. The established protoplast transformation system of Mycena laid a foundation for analyzing the functional genes of Mycena and the molecular mechanism of the symbiosis of Mycena and G. elata.
Subject(s)
Agaricales , Gastrodia/genetics , Protoplasts , Symbiosis/genetics , Transformation, GeneticABSTRACT
Objective: To investigate the chemical constituents from the whole plant of Incarvillea sinensis var. przewalskii, and a systematically phytochemical investigation was carried out. Methods: The chemical constituents were isolated by column chromatographies, such as silica gel and Sephadex LH-20 columns as well as semi-preparative HPLC, and their structures were determined on the basis of MS, NMR spectroscopic analyses, and/or comparison with literature data. Results: Thirteen compounds, including alkaloids and phenylethanoid glycosides, were isolated from the 80% ethanol extract of the whole plants. Their structures were elucidated as p-butoxyl-salidroside (1), incasine C (2), incarvillateine (3), 7-hydroxyskytanthine (4), incarvilline (5), phenethylalcohol glucoside (6), martynoside (7), acteoside (8), deacylisomartynoside (9), decaffeoy1acteoside (10), carceorioside B (11), salidroside (12), and deacylacteoside dimethyl ether (13). Conclusion: Compound 1 is a new one, and the other compounds are isolated from this species for the first time.
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Objective: To investigate the chemical constituents from the seeds of Illicium wardii. Methods: The chemical constituents were separated and purified by silica gel, Sephadex LH-20, MCI, etc. Results: Eleven sesquiterpenoids were isolated from the 95% ethanol extract in the seeds of I. wardii. Their structures were elucidated as wardinol C (1), 7, 8-epoxy-1(12)-caryophyllene-9β-ol (2), merrilliortholactone (3), 2α-hydroxyneoanisatin (4), 2, 6-dihydroxyhumlan-9(E)-3(12), 7(13), 9-trien (5), anisatin (6), pseudomajucin (7), 8α-hydroxy-10-deoxycyclomerrillianolide (8), 2α-hydroxyanisatin (9), rel-5-(3S, 8S-dihydroxy-1R, 5S-dimethyl-7-oxa-6-oxobicyclo-[3, 2, 1]-oct-8-yl)-3-methyl-2Z, 4E-pentadienoic acid (10), and phaseic acid (11). Conclusion: Compound 1 is a new one, and the other compounds are isolated from this species for the first time.
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<p><b>OBJECTIVE</b>To compare the clinical efficacy of trapezoid pad coordinate with small splint fixation treatment with plaster slab fixation treatment for comminuted Colles fracture.</p><p><b>METHODS</b>From October 2010 to March 2012,80 outpatients with comminuted Colles fracture were divided into two groups. There were 40 patients in trapezoid pad splint fixation treatment group and 40 patients in pipe type plaster control group. There were 13 males and 27 females in trapezoid pad splint fixation treatment group,with an average age of (62.1 +/- 19.2) years old. And there were 15 males and 25 females in pipe type plaster control group,with an average age of (60.2 +/- 19.5) years old. After treated with manipulative reduction,the fractures were fixed with pipe plaster in the control group, and were fixed with trapezoid pad joint by small splint in the treatment group. The patients in both groups were regularly taken X-ray recheck and changed dressings to obtain the clinical healing. Patients were guided to do functional exercise after splints were taken off. The therapeutic effects were evaluated according to Gartland-Werlley wrist score when splints taken off and after 2 months.</p><p><b>RESULTS</b>According to wrist score, the average scores in the treatment group were (8.1 +/- 2.8) when splints taken off and (5.3 +/- 4.5) after 2 months. And the average scores in the control group were (10.3 +/- 2.6) when splints taken off and (8.4 +/- 5.8) after 2 months. The score in the treatment group was higher than that of control group. The average shortened lengths of radius were (4.8 + 1.7) mm in the treatment group and (8.2 +/- 2.1) mm in the control group. The shortened length of control group was larger than that of treatment group.</p><p><b>CONCLUSION</b>Trapezoid pad coordinated with small splint fixation is better than traditional pipe type plaster for treating comminuted Colles fracture.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Case-Control Studies , Casts, Surgical , Colles' Fracture , General Surgery , Fractures, Comminuted , General Surgery , SplintsABSTRACT
<p><b>OBJECTIVE</b>To probe the mechanism of electro-scalp acupuncture in treatment of Parkinson's disease (PD) by single photon emission computer tomography (SPECT).</p><p><b>METHODS</b>Five cases of PD received electro-scalp acupuncture at Dingnie Qianxiexian (MS 6), Epangxian III (MS 4), Dingpangxian I (MS 8), Dingpangxian II (MS 9) and Zhenxia Pangxian (MS 14). Contralateral points were selected for pathologic change on one side and bilateral points were selected for pathologic lesion on both sides. All the patients received 99mTc-TRODAT-1 SPECT examination before and after acupuncture treatment of 6 weeks. And activities of dopamine transporter (DAT) were analyzed by the ratio of striatum/occipital lobe (ST/OC), which was evaluated by means of technique of regional of interesting (ROI).</p><p><b>RESULTS</b>The ratio of ST/OC on the same side of the affected extremity before and after treatment were 1.19 +/- 0.15 and 1.24 +/- 0.31 respectively. And on the other side were 0.90 +/- 0.12 and 0.95 +/- 0.25 respectively. They were increased after treatment (P > 0.05).</p><p><b>CONCLUSION</b>Electro-scalp acupuncture can decrease the loss of DAT and improve the activities of DAT in the striatum of the patient of PD.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Corpus Striatum , Chemistry , Dopamine Plasma Membrane Transport Proteins , Electroacupuncture , Parkinson Disease , Metabolism , Therapeutics , Scalp , Tomography, Emission-Computed, Single-PhotonABSTRACT
<p><b>OBJECTIVE</b>To observe the therapeutic effect of scalp electroacupuncture on Parkinson disease (PD) of grade 1.5-3.0 on Hoehn-Yahr scale.</p><p><b>METHODS</b>Thirty patients with Parkinson disease were randomized equally into the treatment group and control group. Patients in the treatment group were treated by scalp eletroacupuncture at the contralateral points of MS6, MS4, MS8, MS9 and MS14 in cases of unilateral lesions or at the bilateral points for bilateral lesions, with also medication with Madopar. The patients in the control group were given Madopar only. The scores of Webster scale and the motor function of United Parkinson's Disease Rating Scale (UPDRS) were used to for assessment before and after the therapeutic course (6 weeks).</p><p><b>RESULTS</b>Tremor, rigidity and bradykinesia were obviously improved in both the groups (P<0.05). The difference in the score of Webster scale was not significant between the two groups (P>0.05), but the treatment group showed greater improvement in motor functions than the control group (P<0.05).</p><p><b>CONCLUSION</b>Scalp electroacupuncture is effective and safe for therapy of Parkinson disease.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Electroacupuncture , Methods , Parkinson Disease , Therapeutics , ScalpABSTRACT
<p><b>OBJECTIVE</b>To study on the anatomical layers and depth of Yifeng (TE 17) so as to provide anatomical basis for clinical treatment.</p><p><b>METHODS</b>Fifteen fresh adult corpse samples, 10 males and 5 females, were randomly taken for investigating the anatomical structures and nerve-blood vessel relationship at the acupoint area of Yifeng (TE 17) with layer anatomical method layer by layer.</p><p><b>RESULTS</b>Anterior of Yifeng (TE 17) is posterior border of condylion, the part backward is the aponeurosis of sternocleidomastoid muscle and anterior border of papillary. Structures of this point are skin, subcutaneous fascia, the posterior border of parotid, venous plexus of infratemporal fossa in turn. And the superficial layer of the point are the branches of great auricular nerve and external jugular vein. The structures of the deep layer are occipital artery, branches of upper jaw's artery and vein, pterygoid venous plexus and facial nerve, mandibular nerve. The average dangerous depth is (35.52 +/- 6.31) mm.</p><p><b>CONCLUSION</b>There are important nerves and blood vessels around Yifeng (TE 17), which should be noted in acupuncture.</p>
Subject(s)
Humans , Facial Nerve , Neck MusclesABSTRACT
Objective To evaluate the expression of RAR-? gene in cervical carcinoma cell lines SiHa,HeLa,C33A and Caski and to analyze the relation between their gene expression and the promoter methylation of RAR-? DNA.Methods The expression of mRNA and protein of RAR-? gene in the four cell lines were analyzed by RT-PCR,western blot and immunofluoreseence,respectively.Methylation specific PCR(MSP)was used to check whether there was methylation in the promoter of RAR-? gene.The demethylating agent 5-aza-2'-deoxycytidine(5-Aza-cdR)was used to treat methylated cell lines and the change of RAR-? gene methylation and RAR-? gene expression defects were observed.The cell proliferation was assayed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.Results The mRNA and protein expression levels of RAR-? in cell lines SiHa,HeLa,Caski and C33A were 0.25 ?0.08,0,0.60?0.19,3.12?0.92 and 0.23?0.07,0,0.14?0.05,0.68?0.21,respectively.The mRNA and protein expression of RAR-? in SiHa,HeLa and Caski cell lines were decreased or silenced, whereas its expression increased in C33A cell line.MSP method showed that there were RAR-? gene methylation in SiHa,HeLa and Caski cell lines,while there was no RAR-? gene methylation in C33A cell line.After treated with 5-Aza-cdR,the mRNA and protein expression levels of RAR-? in SiHa,HeLa, Caski and C33A cell lines were 1.82?0.59,2.13?0.62,1.67?0.43,2.95?0.89 and 0.69?0.21, 0.83?0.29,0.56?0.16,0.64?0.20 respectively.The mRNA and protein levels of RAR-? had a significant difference between before and after interference with 5-Aza-cdR in SiHa,Helm,and Caski cell lines(P0.05).The 5-Aza-cdR treatment could suppress cell proliferation.Conclusions The RAR-? gene expression defects play an important role in the carcinogenesis of cervical cancer.Aberrant methylation in promotor region of RAR-? gene may be an important mechanism for the loss of expression of RAR-? gene.