Objective To lay a foundation for the cultivation of disease-free plantlet of Citrus medica L.var.sarcodactylis Swingle(CMS) by discussion of the factors affecting survival rate for micro-grafted shoot-tip of CMS.Methods With citrus limon seedlings cultured in the dark as the rootstock,shoot-tip of CMS was grafted to the inverse T-shaped cut in the cotyledon of rootstocks.Then an investigation was carried out on the factors which affected the survival rate for micrografted shoot-tip of CMS.The factors included age of rootstocks,scion size,culture medium with different concentrations of phytohormones,and rootstocks with or without cotyledon.Results The survival rate for micro-grafted shoot-tip of CMS was closely related to the factors mentioned above.A higher survival rate was obtained when CMS shoot-tip with 4 pieces of leaf primordia was grafted to the inverse "T" shaped cut in the cotyledon of parental stock(citrus limon seedlings) which was cultured in the dark for 14 days,and when the citrus limon seedlings were cultivated in MT-based medium with ?-naphthylacetic acid 0.1 mg?L-1 added.Conclusion The optimized micro-grafting conditions for CMS shoot-tips improve the propagation of micro-grafted plantlet and the survival rate of micro-grafted shoot-tip of CMS.

Objective To provide a new approach of preservation for germplasm resources of Citrus medica var.sarcodactylis(CMS).Methods A micrografting test was made on CMS shoot-tips after vitrified cryopreservation,resulting in living shoot-tip.Results First the CMS shoot-tip is inoculated in medium consisting of MS,5%DMSO,and 5%sucrose for 48 h preincubation.Then the shoot-tips were cut off 3—4 mm long and fore-treated by 60%PVS_2 at room temperature(25℃) for 30 min.After that,they were treated by PVS_2 at 0℃for another 80 min and conserved in liquid nitrogen for 24 h.Next the shoot-tips were defrosted by aqueous bath at 40℃and cleaned twice at room temperature(25℃) by MT minimal medium with additive 1.2 mol/L sucrose,10 min once.Finally data collected recorded a higher survival rate of 81.25%by TTC inspectation and a better regeneration rate of 52.94%if the CMS shoot-tips had been grafted on darkly-cultured parental stock,growing,and differentiating normally afterwards.The plantlets detected by polymerase chain reaction(PCR) showed that the pathogens of Huanglong disease had been removed.Conclusion Therefore a conclusion is made that the approach of vitrified cryopreservation may be applied to the preservation of CMS germplasm resources.

The sterile stem tips, nodular and non-nodular stem of Morinda officinalis How were cultured in vitro. The results showed that the most suitable procedure for sterilization of explants was soaking into 0.1% HgCl 2 solution for 10 to 15 minutes after pretreating with 70% ethanol for 30 seconds. MT culture medium with BA was effective to induce direct shooting of stem tips and nodular stem, the shooting rate of nodular stem being 97.5% . The optimal BA concentration was 1 mg?L -1 , the shooting rate would decrease when the concentration of BA increased. As for the induction of rooting, MT culture medium adding with 0.2 to 0.5 mg?L -1 NAA was optimal, the rooting rate being over 80.0% .

Objective To study the effects of the recipe for activating blood circulation and nourishing qi on expression of mitochondrial ribosomal protein L51 (MRPL51) in CVB3 infection model in rat cardiac myocytes, to reveal the pathogenesis of CVB3 myocarditis in the genetic level, to explore the therapeutic mechanism of the recipe for activating blood circulation and nourishing qi on CVB3 myocarditis, and to confirm the validity of the recipe for activating blood circulation and nourishing qi on CVB3 myocarditis. Methods After establishing CVB3 infection model and treatment model with recipe for activating blood circulation and nourishing qi by culturing neonatal rat myocardial cells, a modified suppression subtractive hybridization (SSH) was used to isolate differentially expressed genes between two model groups. These results were further verified by fluorescence RT-PCR. Results The results of SSH showed that gene expression of the treatment group was lower than that of the CVB3 infection group. The results of fluorescent RT-PCR which agreed with that of SSH displayed the threshold cycle number (Ct) in the treatment group was higher than the virus group. Conclusion Up-regulation of MRPL51 might be one of the pathogenesis of CVB3 myocarditis. The recipe for activating blood circulation and nourishing qi could treat viral myocarditis by regulating the expression of MRPL51.

[ Objective ] To supply a technological evidence for the rapid reproduction of Andrographis paniculata (Burm. f.) Nees (APN). [Methods] Different explants from sterile seedling of Andrographis paniculata (Burm. f.) Nees were cultured in vitro to induce the production of fascicular buds. [ Results ] Cotyledon with nodes and half cotyledon with nodes were the most suitable explants for culture, and the budding rate was 100% and over 90% respectively. 6-Benzylaminopurine (BA) in the concentrations of 0.5 ~ 1.0mg/L were beneficial for the reproduction and growth of buds. The age of explants had no correlation with budding. The optimal rooting medium was MT (Murashige and Tucher) medium added with Img/L NAA (naphthaleneacetic acid) or IBA (indolebutyric acid). A highest rooting rate as much as 61.9% was achieved after 10 days of culture. [Conclusion] An effective plant regeneration system has been established for explants of Andrographis paniculata (Burm. f.) Nees.

To investigate the quality of different parts of Radix Fici Hirtae. The content and components of volatile oils and ethanol-extracts from different parts of Radix Fici Hirtae were analyzed by the combination of gas chromatography and mass spectrometry (GC-MS). The content of volatile oils in the cortex was higher than that in the xylem and the chemical components either in the cortex or in the xylem are similar. The main components in volatile oils are hexadecanoic acid, oleic acid+linoleic acid, linoleic acid amide, palmitic acid amide,stearic acid amide and dibutyl phthalate. The main components in ethanol-extracts are psoralen, hexadecanoic acid, hexadecanoic acid ethyl ester, bergapten, oleic acid and linoleic acid. The content of psoralen in the cortex was higher than that in the xylem.[Conclusion]The chemical components in different parts of Radix Fici Hirtae are similar but the contents of volatile components and psoralen are higher in the cortex than those in the xylem.

Objective To investigate the expressions of CDX2 and β-catenin in acute lymphoblastic leukemia (ALL) and their correlation. Methods Real-time PCR was used to determine the expressions of CDX2 and β-catenin mRNA in 43 de novo ALL and 30 non-malignant patients (used as control). Results The positivity rate of CDX2 mRNA expression in ALL group was 93%, but CDX2 mRNA expression couldn′t be detected in the control group (P<0.01). The mRNA expression of β-catenin could be detected in patients in both two groups, butβ-catenin mRNA expression in the ALL group was significantly higher than in the control group (P < 0.01). And mRNA expressions of CDX2 andβ-catenin were significantly correlated with WBC counts and LDH level (P<0.01). When the ALL patients acquired complete remission (CR), the mRNA expressions of CDX2 and β-catenin were significantly decreased compared with their newly diagnosed status , while disease-relapsed the mRNA expressions of CDX2 andβ-catenin were increased again. There was significantly positive correlation between CDX2 and β-catenin mRNA expressions (r = 0.835, P = 0.000). Conclusion Up-regulation of CDX2 and activation of Wnt/β-catenin pathway coexist in the ALL patients and the mRNA expressions of CDX2 and β-catenin are positively correlated.

AIM:To investigate the effects of atorvastatin reloading in pre-percutaneous coronary intervention ( PCI) period on endothelial progenitor cell ( EPC) count and inflammatory cytokine expression in the stable angina pectoris patients who had previously received long-term statin treatment.METHODS:The patients with stable angina pectoris that had received long-term statin therapy and planned to accept PCI were randomized into 3 groups:80 mg atorvastatin 12 h and 40 mg 2 h before coronary angioplasty (80 mg reloading), pre-operatively with 40 mg/d atorvastatin for 7 d (40 mg re-loading) , and without atorvastatin reloading ( no reloading ) .CD45 -/CD133+/CD34 +, CD45 -/CD34 +/KDR+ and CD45 -/CD144 +/KDR+EPCs in 100 μL peripheral blood were determined by flow cytometry 1 h prior to PCI and 1 h, 6 h and 24 h after PCI.The serum concentrations of soluble intercellular adhesion molecule 1 ( sICAM-1) , C-reactive protein ( CRP) and troponin I ( TnI) were analyzed immediately prior to and 24 h after PCI.RESULTS:(1) In 80 mg reloading group, the numbers of circulating CD45 -/CD133 +/CD34 +and CD45 -/CD34 +/KDR+early differentiation stage EPCs 1 h and 6 h after coronary angioplasty was significantly elevated compared with those before PCI (P<0.05).(2) In control group, the serum concentrations of sICAM-1 and CRP 24 h after PCI were significantly elevated ( P<0.05) compared with preoperative values.(3) The rise in serum TnI concentration from pre-to post-operation in 80 mg reloading group was lowerthan that in control group.CONCLUSION: The method of atorvastatin reload before PCI affects the number of EPCs inperi-operative period.High dose of atorvastatin application before PCI triggers early EPC circulation.The serum levels ofpost-operative inflammatory cytokine sICAM-1 as well as CRP are reduced by atorvastatin reloading before PCI.

Objective To investigate the chemical constituents of Citrus medica var. sareodactylis from Sichuan province, and to provide evidence for the development and utilization and quality evaluation of the medicinal material.Methods Various chromatographic techniques were used to purify the components of this herb. Compounds were identi-fied by their physical characteristics and spectral feature. Results Seven compounds were isolated from Citrus medica var. sarcodactylis, and they were identified as sibiricol (Ⅵ), 7-methylesculetin (Ⅶ), bergapten (Ⅷ), sigmasteryl acetate(Ⅸ), 5-methoxyfurfural(Ⅹ), limonin(Ⅺ), daucosterol (Ⅻ). Conclusion Compound Ⅵ, Ⅶ and Ⅸ are isolated from plants of Rutaceae for the first time.

Objective To find out technological evidence for the rapid propagation of Aloe vera L.var.chinensis (Haw.)Berber. Methods The technique of plant tissue culture was used to study the rapid propagation of Aloe vera L.Var.Chinesis(Haw.)Berger.The shoot growing from the underground stem of Aloe vera L.Var.chinensis(Haw.)Berger was taken as the explant to induce fascicular bud. Results The optimal culture medium was MT.MT culture medium added with 6-benzylaminopurine(BA) was effective to induce the shoot formation and increase the direct shooting rate.The optimal BA concentration for the induction and formation of shoot was 2 mg?L -1 ,the shooting rate being 100%.As for the induction of rooting,MT culture medium adding with 0.5 mg?L -1 naphthalene acid was optimal,the rooting rate being 100%. Conclusion The culture of Aloe shoot growing from the underground stem,proper culture medium and BA concentration are important for the rapid propagation of Aloe.