In order to study the effect of phytohormone on growth and isoflavones contents of Pueraria phaseoloides hairy roots, we cultured the hairy roots with different concentrations of 6-benzylaminopurine (6-BA) alone or in combination with α-naphthaleneacetic acid (NAA). Then we determined the effects of 6-BA alone or in combination with NAA on the growth and the contents of isoflavones compounds and levels of antioxidase activities of hairy roots by spectrophotometry. The results show that 6-BA inhibited the growth, and decreased biomass and total isoflavones compounds of P. phaseoloides hairy roots. Furthermore, the inhibition was increased with the concentrations of 6-BA. Compared with the controls, different concentrations of 6-BA in combination with NAA 2.0 mg/L could inhibit the growth of hairy roots and decrease the content of total isoflavone compounds, and also significantly enhanced the contents of soluble protein and levels of peroxidase (POD) activities, but decreased the activities of superoxide dismutase (SOD). DNA ladders detected by agarose gel electrophoresis can be observed after hairy roots of P. phaseoloides were cultured with 6-BA alone for 30 days, but can appear on the 20th day after culture with 6-BA in combination with NAA 2.0 mg/L. This result indicates that 6-BA or 6-BA in combination with NAA can both stimulate appearance of programmed cell death (PCD), and NAA may play a synergistic role on PCD.
Benzyl Compounds ; Isoflavones ; chemistry ; Kinetin ; pharmacology ; Naphthaleneacetic Acids ; pharmacology ; Plant Growth Regulators ; pharmacology ; Plant Roots ; chemistry ; drug effects ; growth & development ; Pueraria ; drug effects ; growth & development ; Purines

In this article the prophylactic effects of the compound injection of tanshinone to panereatic tissues in the experimental white rats with acute pancreatitis was reported. The compound injection of tanshinone could retard the degrec of pancreatic hemorrhage and necrosis of the experimental white rats. It could also reduce the fatality rate of the animal, The value of serum superoxide dismutase in tanshinone protected group were significantly low or than that of nonprotected group The result suggested that the compound injection tanshinone might minimize the pathological change degrees of the panereatie tissue by means of improving microcirculation of the pancreatic tissues, enhencing the tolerence of the pancrease to anoxia, reducing generation of the free redical.

To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.
Agrobacterium ; Chromatography, Thin Layer ; Culture Media ; Dianthus ; growth & development ; Plant Growth Regulators ; Plant Leaves ; Plant Roots ; growth & development ; Plants, Genetically Modified ; Rhizobium ; Tissue Culture Techniques ; Transformation, Genetic

Objective To investigate the antishock effect of oxygenate solution and its possible mechanism. Methods The protective effects of oxygenates hypertonic hypercolloid solution on lipid peroxidation injures in hemorrhagic shock in rabbits at 4 700 m high altitude spot was observed. Results The oxygenated solution treatment can obviously reduce the malondialdehyde(MDA) and glutathione(GSH) level in plasma and tissue ; and increase the superoxide diamutase(SOD) and glutathione peroxidase(GSH-PX) level in plasma and tissue of the hemorrhagic shock animals. Conclusions Oxygenatea solution treatment can reduce the lipid peroxidation injure, recover the equilibrium of oxidation and antioxidation with shock body in time, and promote the resuscitation of shock.

Objective To explore the effect of hemeoxygenease/carbon monoxide system on hypoxic pulumonary vascular structural remodeling. Methods Twenty six Wister rats were randomly divided into four groups:control groups( n =6),hypoxia group( n =7),hypxia+ZnPP group( n =6) and hypoxia+CO group( n =7).Pulmonary artery mean pressure(mPAP) of each rat was elaluated by using right cardiac catheterization,the ultrastructaral changes in intra acinar pulmonary muscularized arteries were observed. Results mPAP was significantly increased in hypoxic rats(22?2 31?mmHg)as compared with that of normal controls(16 57?2 51mmHg)( P

[ ABSTRACT] AIM:To cultivate stem-like spheres from SW620 cell line in the specific serum-free medium and evaluate the features of the cancer stem cells, and to investigate the effects of docosahexaenoic acid ( DHA) and eicosapen-taenoic acid ( EPA) on the growth of SW620 stem cell-like cells.METHODS: Human colon cancer stem cell-like cells ( CSCLC) were obtained from SW620 spheres cultured in serum-free medium.These cells were tested for the expression of SSEA-1 and TRA-1-81 by immunofluorescence staining.The mRNA expression of Sox-2 and Oct-4 was detected by real-time PCR.The efficiency of colony formation on a soft agar gel and tumor formation in the nude mice was compared between SW620 adherent cells and CSCLC.The inhibitory effects of 5-fluorouracil (5-FU) and mitomycin C on both types of cells were measured by MTS assay.MTS assay, Annexin V/PI staining and trypan blue staining were used to determine the effects of DHA and EPA on both types of cells.MTS assay was also used to analyze the combined effect of DHA or EPA with chemotherapeutic drugs on SW620 CSCLC.RESULTS:SW620 cells formed spheres in serum-free culture.The cells from spheres highly expressed SSEA-1 and TRA-1-81, transiently expressed Sox-2 and Oct-4 genes and were more resistant to 5-FU and mitomycin C treatments.These cells exhibited a greater ability in clone formation and tumorigenicity, indica-ting that these cells carried stem cell-like features, hence were considered SW620-derived CSCLC.DHA and/or EPA sup-pressed SW620 CSCLC by inhibiting cell growth, inducing cell apoptosis and sensitizing them to chemotherapeutic drugs. CONCLUSION:The cells with stem cell-like features, such as high efficiency in clonogenicity, tumorigenicity and resist-ance to chemotherapeutic drugs, can be obtained from SW620 spheres cultured in serum-free condition.DHA and EPA in-duce apoptosis in SW620-derived CSCLC and sensitize them to chemotherapeutic drugs.

Abstract: In order to enhance the content of secondary metabolites patchouli alcohol in Pogostemon cablin, we induced polyploid hairy roots and their plant regeneration, and determined the content of patchouli alcohol through artificial chromosome doubling with colchicine. The highest rate of polyploidy induction was more than 40% when hairy roots were treated with 0.05% colchicine for 36 h. The obtained polyploid hairy roots formed adventitious shoots when cultured in an MS medium with 6-BA 0.2 mg/L and NAA 0.1 mg/L for 60 d. Compared with the control diploid plants, the polyploid hairy root-regenerated plants of P. cablin had more developed root systems, thicker stems, shorter internodes and longer, wider and thicker leaves. Observation of the chromosome number in their root tip cells reveals that the obtained polyploid regenerated plants were tetraploidy, with 128 (4n = 128) chromosomes. The leaves contained around twice as many stomatal guard cells and chloroplasts as the controls, but the stomatal density declined with increasing ploidy. The stomatal density in diploid plants was around 1.67 times of that in polyploid plants. GC-MS analysis shows that the content of patchouli alcholol in the hairy root-derived polyploid plants was about 4.25 mg/g dry weight, which was 2.3 times of that in diploid plants. The present study demonstrates that polyploidization of hairy roots can stimulate the content of patchouli alcholol in medicinal plant of P. cablin.
Colchicine ; Diploidy ; Lamiaceae ; genetics ; growth & development ; Plant Roots ; growth & development ; Plants, Medicinal ; genetics ; growth & development ; Polyploidy ; Regeneration ; Sesquiterpenes ; chemistry

By genetic transformation with Agrobacterum rhizogenes and artificial chromosome doubling techniques, we studied the induction of hairy roots and their polyploidization, and subsequent plant regeneration and nicotine determination to enhance the content of nicotine in Nicotiana tabacum. The results show that hairy roots could be induced from the basal surface of leaf explants of N. tabacum 8 days after inoculation with Agrobacterium rhizogenes ATCC15834. The percentage of the rooting leaf explants was 100% 15 days after inoculation. The hairy roots could grow rapidly and autonomously on solid or liquid phytohormones-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and paper electrophoresis of opines from N. tabacum hairy roots. The highest rate of polyploidy induction, more than 64.71%, was obtained after treatment of hairy roots with 0.1% colchicine for 36 h. The optimum medium for plant regeneration from polyploid hairy roots was MS+2.0 mg/L 6-BA +0.2 mg/L NAA. Compared with the control diploid plants, the hairy roots-regenerated plants had weak apical dominance, more axillary buds and more narrow leaves; whereas the polyploid hairy root-regenerated plants had thicker stems, shorter internodes and the colour, width and thickness of leaves were significantly higher than that of the control. Observation of the number of chromosomes in their root tip cells reveals that the obtained polyploid regenerated plants were tetraploidy, with 96 (4n = 96) chromosomes. Pot-grown experiments showed compared to the control, the flowering was delayed by 21 days in diploid hairy roots-regenerated plants and polyploid hairy root-regenerated plants. GC-MS detection shows that the content of nicotine in polyploid plants was about 6.90 and 4.57 times the control and the diploid hairy roots-regenerated plants, respectively.
Agrobacterium ; Plant Roots ; growth & development ; Plants, Genetically Modified ; growth & development ; Polyploidy ; Regeneration ; Tobacco ; genetics ; growth & development ; Transformation, Genetic

Objective To observe the expression of cytokeratin 14,15 (CK14,CK15) expression level in normal esophageal tissues and esophageal squamous cell carcinoma tissues of different differentiation degree and to analyze the relationship between occurrence,development of esophageal squamous cell carcinoma and CK14,CK15 expression level.Methods Esophageal squamous epithelial tissue from 55 cases of carcinoma tissues and 55 cases of adjacent tissues were collected.Immunohistochemical method was used to compare CK14,CK15 and PCNA expression levels in esophageal squamous carcinoma.Results Expression positive rates of CK14,CK15 and PCNA in esophageal squamous carcinoma were 72.7 ％ (40/55),63.6 ％ (35/55) and 65.5 ％ (36/55),respectively,and PCNA expression was correlated with CK14 or CK15 expression (C =0.585,P ＜ 0.001; C =0.405,P ＜ 0.001).CK14 and CK15 levels were higher in high differentiation carcinoma tissue than those in low differentiation carcinoma tissue,and PCNA expression level was increased in low differentiated carcinoma tissue.CK14,CK15 and PCNA were expressed located in base layer of esophageal squamous epithelial adjacent to carcinoma tissue,and their expression positive rates were 56.4 ％ (31/55),52.7 ％ (29/55) and 56.4 ％ (31/55).CK14 and CK15 levels were higher in esophageal squamous epithelial tissues of far-cut edge than those in tissues of near-cut edge (intraepithelial neoplasia).There were no associations between CK14,CK15 expression and the clinical parameters (P ＞ 0.05).Postoperative survival time in patients with CK14 or CK15 positive expression was shorter than that of patients with negative expression (P ＜ 0.05).Conclusions CK14 or CK15 positive expressions localized to base layer of esophageal squamous epithelial adjacent to carcinoma tissue may play some roles in generation and differentiation of esophageal squamous cell cancer.CK14 or CK15 positive expression in esophageal squamous carcinoma involves in differentiation process.Joint detection of CK14 and CK15 expression has clinical application value for early diagnosis,the degree of differentiation and prognostic judgment in esophageal squamous carcinoma.

Objective To evaluate the value of urinary normal and modified nucleosides in the diagnosis of primary gallbladder carcinoma.Methods Between March 2011 to September 2012,28 patients with primary carcinoma of the gallbladder (PCG) were included in this study.Spontaneous urine samples were collected and 10 kinds of urinary nucleosides in the sample were determined by reversed-phase high performance liquid chromatography method.Another 62 patients with benign gallbladder disease and 70 healthy volunteers were enrolled as controls.Routine clinical tumor markers,including serum CA19-9 and CA125 levels of the PCG patients were also evaluated.Results 10 kinds of nucleoside had a good linear correlation (r＞0.99),and the recovery rate was between 87.4％ ～121.5％ range,and the accuracy rate was between 87.7％ ～121.3％,and the RSD of precision was between 1.5％～10.5％ range.In addition to adenosine and cytidine,the mean levels of the rest of the urinary nucleosides in the PCG group were much higher than those of the controls (P＜0.01).Based on principal component analysis,89.3％ of the PCG patients were correctly identified,which was much higher than those detected by CA19-9 (60.7％) and CA125 (67.9％) (P＜0.01).Conclusion Urinary nucleosides may become additional tumor markers which when combined with other clinical methods may help in the screening and early diagnosis of primary gallbladder cancer.