Objective To establish and optimize ISSR-PCR systems of Chrysanthemum morifolium and lay the foundation for its genetic diversity research. Methods Based on the analysis of variance, an orthogonal design was used to optimize the ISSR-PCR amplification system on C. morifolium by four factors (Taq polymerase, Mg2+, dNTP, and primer) at three concentration levels, respectively. Results A suitable ISSR reaction system was constructed with the 20 ?L reaction system containing 1.00 U Taq polymerase, 2.00 mmol/L Mg2+, 0.20 mmol/L dNTP, and 0.50 ?mol/L primer. Conclusion ISSR-PCR is significantly influenced by the concentration of Taq polymerase, Mg2+, and dNTP. This ISSR-PCR system could provide clear bands, reliable reaction system, and abundant polymorphisms . It is proved to be suitable for the study of the genetic diversity of C. morifolium

AIM: To explore the properties of the acetylcholine(ACh)-sensitive potassium channel in type Ⅱ vestibular hair cells(VHCs Ⅱ) in mice saccular macula and the modulation effect of calcium ions.METHODS: Under the whole-cell patch mode,the pharmacology properties of ACh-sensitive potassium channel and the modulation of calcium ions on ACh-sensitive potassium channel were investigated.RESULTS: Following extracellular perfusion of ACh,VHCs Ⅱ displayed a slow and sustained outward current,which was sensitive to tetraethylammonium(TEA,5 mmol/L) and charybdotoxin(CTX,100 nmol/L),but not sensitive to 4-aminopyride(4-AP,15 ?mol/L).ACh-sensitive potassium current was inhibited by intracellular application of ethylene glycol-bis(B-aminoethylether)-N,N,N',N'-tetraacetic-acid(EGTA,5 mmol/L) and extracellular perfusion of Cd2+ and Ni2+,respectively.Intracellular application of heparin(8 g/L) failed to inhibit ACh-sensitive potassium current.CONCLUSION: Extracellular application of ACh activates the big conductance,calcium-dependent potassium current(BK) in VHCs Ⅱ of mice,which is potently modulated by extracellular Ca2+ ions.However,intracellular IP3-dependent Ca2+ ions release mechanism is not involved in the activation of the ACh-sensitive BK channel.

Objective To evaluate the dosimetric differences of one RapidPlan Model on different Radiotherapy devices.Methods A RapidPlan Model was built based on 30 reoptimization IMRT plans of cervical cancer patients on typeA LA.Dosimetric differences of automatic optimized IMRT plans using this model on 4 different type LAs,named respectivelyA,B,C andD,were compared with 12 test cervical cancer cases.These four LAs were well commissioned in the treatment planning system (TPS).Student t test was applied for statistical analysis on dosimetric differences.Results Dosimetric differences between A vs.B,C and D were observed on Dmean,HI,CI of PTV50 and PTV45,as well as on V50,V40,V30 of rectum and bladder.Significant dosimetric differences were observed between A and D (P<0.05).Conclusions Automatic planning with RapidPlan model may result in dosimetric differences on different Radiotherapy devices.These differences should be aware of with caution in its clinical application.

Objective To investigate the 4-year follow up of cognitive function outcomes and characteristics in patients after stroke.Methods Sixty three cases according with the diagnostic standard of acute unifocal subcortical stroke were consecutively collected in neurological ward from December 2009 to November 2010.They were followed up for average four years.Forty one out of them completed the neuropsychology tests identical to the baseline,which covered the general cognition function,attention,execution,memory,language,spatial,etc.According to the standard of clinical diagnosis,cognition function is divided into five degrees,including normal,VCI-ND,mild VaD,moderate VaD,and severe VaD.The improved group had 13 cases whose cognition function was improved by one or more ranks.The progressive group had 12 cases whose cognition function progressed by one or more ranks.The stable group had 16 cases whose cognition function remained the same as the baseline.Results According to qualitative analysis on the baseline versus 4-year follow-up outcome,in 13 improved cases,8 were VCI-ND and 5 were mild VaD.In 16 stable cases,11 were normal,4 were VCI-ND and 1 was mild VaD.In 12 progressive cases,3 were normal (change to mild VaD after follow-up),5 were VCI-ND (change to mild and moderate VaD after follow-up) and 4 were mild VaD (change to moderate VaD after 4-year follow-up).In the comparison of baseline cognition function among the improved,progressive and stable group,there was only one significantly different score (the right number of SCWT-A) in the improved and progressive group.The cognition function of improved group had significant differences in CFT-copy,right number of SCWT-C and the time of TMT-B before versus after follow-up.The cognition function of progressive group had significant differences in AVLT-Delay Recall and CFT-Recall.Conclusion Long-term cognitive function outcome after stroke is heterogenetic.The location of cognitive impairment or progression is not the same model for different cognitive outcome group.

Objective To compare and analyze clinical effects of Bipolar transurethral plasma kinetic enucleation of prostate (PKEP) and transurethral resection of the prostate(TURP) on the treatment huge benign prostatic hyperplasia.Methods Nine-six cases of huge benign prostatic hyperplasia were selected in this hospital from March 2012 to March 2015.All the patients were divided into two groups according to different operation method,namely PKEP group and TURP group.Then the operative time,bleeding amount,bladder washing time,hospital stay,complications between two groups were compared,and the international prostate symptom score (IPSS),quality of life score (QOL),maximal urinary flow rate (Qmax),residual urine volume 6 months before and after operation were compared between the two groups.Results The operative time [(100.0 ± 3.5)min],bleeding amount [(161.0 ± 9.2) mL],bladder washing time[(15.2 ± 1.2) h],hospital stay[(10.8 ± 2.6) d],complications (6 cases) in PKEP group were less than that in the TURP group,which were(132.0±4.2)min,(198.0±12.1)mL,(36.8±1.3)h,(13.6±2.9)d,complications (18 cases)respectively(P＜0.05).The IPSS,QOL,Qmax,residual urine volume in both group were significantly improved compared with surgery before(P＜0.05),and there were no significant differences between the two groups(P＞0.05).Conclusion PKEP and TURP both are effective surgeries for the treatment of huge BPH,while PKEP has short operation time,less intraoperarive bleeding and low incidence of complications,it is worthy of further clinical promotion.

OBJECTIVETo explore the character of inorganic elements in Flos Chrysanthemi indici and look for relationship between the element concentration and regions.

METHODThe contents of elements, including borum (B), sodium (Na), magnesium (Mg), phosphorus (P), potassium (K), calcium(Ca), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), strontium (Sr), cesium (Se), barium(Ba) and lead(Pb) in Chinese traditional herb Flos Chrysanthemi indici from different regions were determined by ICP-AES. The element distrubution diagram were plotted. The principal component analysis and correlation analysis of SPSS were applied for the study of characteristic elements.

RESULTSimilar curves of element concentration have been acquired. It is observed that the content of elements in the samples shows regional diversity. There are 15 correlative element pairs in correlation analysis. Four principal components which accounted for over 84.437% of the total variance were extracted from the original data. The first and second factors accounted for 60.090% of the total variance, which means that P, K, Ca, Mn, Fe, Cu, Sr, B, Na and Se may be the characteristic elements.

CONCLUSIONThe showed that element content in Flos Chrysanthemi Indici display special distributing diagram. Remarkable correlation is presented in some element pairs. The elements contents of Flos Chrysanthemi indici gained from Yunan, Hunan are higher than those from other regions.

China ; Chrysanthemum ; chemistry ; Drugs, Chinese Herbal ; analysis ; Flowers ; chemistry ; Principal Component Analysis ; Trace Elements ; analysis

Objective Methylation of anti-oncogene can be demethylated by related drugs which can help the inactivated gene to express again .This study aims to study the effects of the demethylating agent 5-Aza-2′-deoxycytidine on the growth of human thyroid papillary cancer cell line TPC-1 and mRNA and protein expres-sion of KLF4.Methods TPC-1 cells were treated with different concentration of 5-Aza-CdR.MTT was used to detect the influence of 5-Aza-CdR on cell proliferation .RT-PCR was used to detect mRNA and protein expression levels of KLF4.Results After being treated with 5-Aza-CdR for 24 hours, 48 hours, and 72 hours, the growth of TPC-1 cells was inhibited and the inhibition was in time and concentration depended manner .After treatment with 5-Aza-CdR, mRNA and protein expression levels of KLF 4 were increased, and the difference had statistical significance(P<0.05).Conclusion 5-Aza-CdR can inhibit the cell viability of TPC-1 cells through upregulat-ing KLF4 expression , which may provide experimental basis for 5-Aza-CdR in treating thyroid cancer .

Objective To research the clinical effect of transurethral plasmakinetic enucleation of prostate (PKEP) in the treatment of high-risk huge benign prostate hyperplasia(BPH).Methods Fifty-two cases of high-risk huge(＞120 g) BPH in this hospita1 from May 2010 to May 2015 were selected and performed PKEP.International prostate symptoms score(IPSS),quality of life(QOL) score,residual urine(RUV) and biggest urine flow rate(Qmax) were observed after operation.Results The mean operation time was (130.12 ± 12.14) min,the mean intraroperation bleeding amount was (120.24±9.81) mL,the mean hospital stay was (14.52 ± 1.82)d,the mean weight of resected prostate tissues was (113.42 ± 12.53)g.Follow-up lasted for 6 months without serious complications.IPSS、QOL,RUV and Qmax after operation were improved obviously,the difference was statistically significant compared with before operation(P＜0.05).Conclusion PKEP is safe and effective in the treatment of high-risk huge BPH.

BACKGROUND:Present studies mainly focused on in vitro culture of bone marrow mesenchymal stem cells(BMSCs)and cell transplantation for treating intracalvarium diseases.However,the understanding of survival,differentiation,migration and structure of transplanted cells in the damaged spinal cord is limited.OBJECTIVE:To explore effects of local BMSC transplantation in repair of spinal cord damage and feasibility of replacement therapy of BMSCs.METHODS:Adult healthy female Sprague-Dawley rats were randomly assigned to cell transplantation and control groups.Rat models of spinal cord transection damage were established.Rat BMSC suspension or calcium and magnesium phosphate buffer were transplanted immediately after injury to the damage zone.At 1 day,1,2,3,4 and 8 weeks before and after transplantation,BBB score motor function was observed in rats,and at 1 week after transplantation,immunohistochemical staining was utilized to observe BrdU-labeled BMSC survival in the spinal cord damaged site.At 4 weeks after transplantation,the general observation and histological detection were observed.RESULTS AND CONCLUSION:At 1-8 weeks after transplantation,BBB scores were higher in the cell transplantation group than in the control group.At 1 week following surgery,immunohistochemical staining showed that BrdU-positive cells were detected in the distal end of rat spinal cord in the cell transplantation group.At 4 weeks following surgery,nerve fibers were found in the damaged spinal cord.These verified that BMSCs were transplanted into rat damaged spinal cord immediately following damage,and the transplanted cells could survive.Living BMSCs can differentiate into neurons,and formed neuron pathway in the local region of damage,which will promote the recovery of conduction function of spinal nerve fibers,and contribute to the recovery of rat hindlimb motor function following high-level spinal cord injury.

BACKGROUND: Acidic fibrobiast growth factor can regulate cell proliferation, migration, differentiation and survival, also can down-regulate the known inhibitor of axon regeneration, such as proteoglycan, help axons overcome these inhibitory factors, and have significant role on the regeneration of nerve fibers.OBJECTIVE: To study the feasibility and effect of the acidic fibroblast growth factor combined with peripheral nerve transplantation in the treatment of high-level spinal cord injury in rats.METHODS: A total of adult 108 female SD rats were randomly divided into autologous nerve group, autologous nerve combined .with acidic fibroblast growth factor group, and high-level spinal cord injury group. The rat T_(8-10) spinous process and lamina were bite, revealing dural sac, high-level spinal cord was resected at a horizon level, cutting 3 mm, no nerve fibers were confirmed to be attached under the microscope. In the autogenous nerve group and autologous nerve combined with acidic fibroblast growth factor group, bilateral the 8~(th) to 10~(th) pairs of intercostal nerves were harvested 2 cm, then cross-transplanted into high-level spinal cord defect (proximal white matter and distal gray matter, distal white matter and proximal gray matter), fibrin gel and fibrin gel containing acidic fibroblast growth factor were used respectively to fix the implanted intercostal nerve, followed by dural suture.High-level spinal cord transection group was subjected to exclusion between stumps. At 90 days postoperation, somatosensory evoked potential and motor evoked potential were used to test nerve electrophysiological recovery. At 76 days postoperation,biotinylated dextran amine anterograde tracing was applied to observe the motor conduction bundle recovery. At 60 days postoperation, hindlimb motor function recovery was assessed by BBB score.RESULTS AND CONCLUSION: The somatosensory and motor evoked potential waveforms were not elicited in rats of high-level spinal cord transaction group, but did elicit in autogenous nerve group and autologous nerve combined acidic fibroblast growth factor group. The average latency and amplitude of somatosensory and motor evoked potentials, as well as BBB scores in autologous nerve combined acidic fibrobiast growth factor group were significantly superior to autologous nerve group (P ＜ 0.01).In the autogenous nerve group and autologous nerve combined acidic fibroblast growth factor group, many more biotinylated daxtran amine-positive nerve fibers passed in the damage zone, compared with high-level spinal cord transection group (P ＜0,01), the autologous nerve combined acidic fibrobiast growth factor group was more than autogenous nerve group (P ＜ 0.01). It is indicated that autologous peripheral nerve graft acidic flbroblast growth factor can better restore the limb motor functions of rats after high-level spinal cord injury.