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1. The expression and function of P-glycoprotein (P-gp) is associated with the phenotype of multidrug resistance (MDR). Saikosaponin A (SSA) is a triterpenoid saponin isolated fromRadix Bupleuri. This study was mainly designed to understand effects of SSA on MDR in MCF-7/ADR and HepG2/ADM cells. 2. MDR reversal was examined as the alteration of cytotoxic drugs IC50 in resistant cells in the presence of SSA by MTT assay, and was compared with the non-resistant cells. Apoptosis and uptake of P-gp substrates in the tumor cells were detected by flow cytometry. Western blot was performed to assay the expression of P-gp. 3. Our results demonstrate SSA could increase the chemosensitivity of P-gp overexpressing HepG2/ADM and MCF-7/ADR cells to doxorubicin (DOX), vincristine (VCR) and paclitaxel. SSA promoted apoptosis of MCF-7/ADR cells in the presence of DOX. Moreover, it could also increase the retention of P-gp substrates DOX and rhodamine 123 in MCF-7/ADR cells, and decrease digoxin efflux ratio in Caco-2 cell monolayer. Finally, a mechanistic study showed that SSA reduced P-gp expression without affecting hydrolytic activity of P-gp. 4. In conclusion, our findings suggest that SSA could be further developed for sensitizing resistant cancer cells and used as an adjuvant therapy together with anticancer drugs to improve their therapeutic efficacies. [ABSTRACT FROM AUTHOR]

[Objective]The effects of exogenous ABA and GA on protein expression of developing seed were studied to discovery the physiological mechanism of exogenous ABA and GA on regulating protein accumulation in Momordica charantia seeds. [Method]A small-warty fruit type genotype Momordica charantia variety MC26 was used as tested material, exogenous ABA (100 mg/L), GA(5 mg/L) and water (CK) were sprayed on fruits at 5 days after anthesis (DAA) at flowering stage, and the proteome changes of developmental Momordica charantia seeds harvested at 16, 22 and 28 DAA were analyzed by two-dimensional gel electrophoresis. [Result]The results showed that, the protein expression and bands of seeds harvested at 22 DAA treated with ABA were found to be the hightest, while those of seeds harvested at 28 DAA treated with GA were found to be the lowest. As compared to the control, the protein expression presented lower in seeds of GA treatment, while low molecular weight (18-35 kD) protein expression showed higher in seeds of ABA treatment. In total, 23 protein spots were found to be differentially expressed between exogenous ABA and water treatments, which 13 proteins were up-regulated, and 10 proteins were down-regulated; 26 protein spots were found to be differentially expressed between exogenous GA and water treatments, which 15 proteins were up-regulated, and 11 proteins were down-regulated. The 9 differentially expressed protein spots were identified out from 19 protein spots by using MALDI-TOF-TOF MS, which were grouped into four functional categories, viz., stress response, glycolysis, storage and unknown protein. [Conclusion]Significant differences in storage protein expression are found among seeds of exogenous ABA, GA and water treatments, which reveal that exogenous ABA and GA have regulation functions on storage protein accumulation of Momordica charantia seeds. [ABSTRACT FROM AUTHOR]

Bitter gourd (Momordica charantia L.) cultivated in China is regarded as an important vegetable crop and is of considerable economic importance. However, it is susceptible to fusarium wilt, which causes heavy economic losses. Forty-eight isolates were isolated from diseased bitter gourd plants that displayed typical fusarium wilt symptoms. Based on the morphological features, the rDNA internal transcribed space (ITS) sequences, pathogenicity and host biotypes, all of the isolates tested were pathogenic to the susceptible bitter gourd plants species (cv. 'Guinongke No. 2') and were identified as Fusarium oxysporum f. sp. momordicae (FOM). Our results classified different isolates as slightly, moderately or highly virulent. Among the isolates tested, 43 isolates slightly infected bottle gourd (Lagenaria siceraria var. clavata), whereas they did not infect other species from the family Cucurbitaceae. Genetic diversity among 48 isolates was characterized using amplified fragment length polymorphism (AFLP) analysis. The number of bands amplified by each primer pairs ranged from 41 to 66, with sizes ranging from 200 to 500 bp. A total of 366 bands were observed, out of which 363 were polymorphic (99.14%). The Nei's genetic identity of the six geographical populations varied from 0.7362 to 0.9707. The mean Nei's gene diversity index (H = 0.2644) and the mean Shannon's information index (I = 0.4071) at species level were higher than ones at populations level, indicated that the variation within populations was greater than that among populations. The Nei's GST (0.5103) and gene flow (Nm = 0.4923) revealed that genetic differentiation was mainly among populations and few gene exchanges. The dendrogram obtained from AFLP marker showed that there was a good correlation between isolates from different geographical locations and their pathogenicity. The AFLP marker effectively distinguished the high virulent isolates from the less virulent isolates. The highly virulent isolates were distinctly separated in different clusters, which indicated a significantly high correlation with the geographical origin in the AFLP dendrogram. The pathogenicity and molecular marker analysis confirmed the presence of variation in virulence as well as genetic diversity among the FOM isolates studied. [ABSTRACT FROM AUTHOR]

[Objective]The present study was conducted to establish an efficient method for germplasm innovation and genetic improvement of Momordica charantia via 60Co-γ irradiation on seeds of different species and molecular marker selection of mutants, in order to provide references for its breeding. [Method]The dry seeds of M. charantia species viz., MCI, MC42 and MC52 were treated with different doses (0, 600, 700, 800, 900, 1200 and 1500 Gy) of 60Co-γ rays, the germination rate, seedling emergency rate, and normal seedling rate were investigated to determine the optimal dosage. The mutants were detected in the M2 generation by SRAP marker, and were subsequently selected based on female flower rate phenotypic traits and disease resistances to breed gynociuos inbreed lines via directional selection of successive generations in the field. [Result]As compared to the control (0 Gy), the germination rate, emergency rate and normal seedling rate of different materials presented decreased change trend by using 600-1500 Gy 60Co-γ rays radiation. The VID50 of MCI, MC42 and MC52 seeds were 900, 700 and 600 Gy, respectively. The optimal irradiation dosage for M. charantia ranged from 600 to 700 Gy, and the normal seedling rate was the sensitive and reliable parameter of irradiation dosage indicator. The 18571 individuals of M2 generation were screened by SRAP marker, and a total of 508 mutant individuals were obtained, which comprised of 116 gynoecious individuals and 162 sub-gynoecious individuals (with more than 50% female flower rate). Through successive generations of directional selection, 8 gynoecious inbred lines with desirable traits were bred, which included 2 individuals of MC1-M5, 3 of MC42-M5 and 3 of MC52-M5. [Conclusion]Selecting optimal dosage of 60Co-γ rays radiation, and using "Plant Direct PCR Kit + SRAP marker" as a high throughput method for mutant screening in the M2 generation combining with directional selection of successive generations is an efficient and convenient method to innovate bitter gounl germplasm. [ABSTRACT FROM AUTHOR]

[Objective] In order to provide anti-wilt source to the breeding program, bitter gourd germplasms were subjected to evaluation of the resistant level to the pathogen of Fusarium oxysporum f. sp. Momordicae. [Method] 143 inbred lines of bitter gourd were inoculated with the pathogen of Fusarium oxysporum f. sp. Momordicae, and the reistance to Fusaiium wilt was identified acconling to the seedling-inoculation method. [ Result ] The resistant levels to bitter gourd Fusaiium wilt of all the germplasm materials were evaluated as 1 high resistance (IIR), 2 resistance (R), 5 moderate resistance (M R ), 17 susceptibility (S) and 118 high susceptibility (HS), accounting for 0.70%. 1.40%, 3.50%. 11.89% and 82.52%, respectively. The resistant germplasm mainly originated from mainland (China, and their characteristics of fruit were short club in shape, light green in skin color, stripe or grain-stripe in fruit ribbing. [(Conclusion] Most inbred lines of bitter gourd evaluated in this study were susceptible to Fusaiium wilt. Therefore, more efforts need to be added to promote germplasm collection, identification and genetic improvement to excavate anti-wilt source to bitter gourd I'usarium wilt. [ABSTRACT FROM AUTHOR]

We performed a meta-analysis of cohort studies to determine whether promoter methylation of the death-associated protein kinase ( DAPK) gene contributes to the pathogenesis of nonsmall cell lung cancer (NSCLC). A range of electronic databases were searched: MEDLINE (1966 ∼ 2013), the Cochrane Library Database (Issue 12, 2013), EMBASE (1980 ∼ 2013), CINAHL (1982 ∼ 2013), Web of Science (1945 ∼ 2013), and the Chinese Biomedical Database (CBM; 1982 ∼ 2013) without any language restrictions. Meta-analysis was conducted using the STATA 12.0 software. Crude odds ratio (OR) with 95 % confidence interval (95 % CI) was calculated. Our meta-analysis integrated results from 12 clinical cohort studies that met all inclusion criteria with a total of 1,027 NSCLC patients. We observed that the frequency of DAPK gene methylation in cancer tissues were significantly higher than that in the adjacent normal and benign tissues (cancer tissues vs. benign tissues: OR = 8.50, 95 % CI = 5.88 ∼ 12.28, P < 0.001; cancer tissues vs. adjacent tissues: OR = 5.95, 95 % CI = 4.11 ∼ 8.60, P < 0.001; cancer tissues vs. normal tissues: OR = 4.75, 95 % CI = 3.28 ∼ 6.87, P < 0.001; respectively). Subgroup analysis by ethnicity demonstrated that DAPK gene methylation was closely associated with the development and progression of NSCLC among both Asians and Caucasians (all P < 0.05). Furthermore, we conducted a subgroup analysis based on sample source and discovered that DAPK gene methylation was implicated in the pathogenesis of NSCLC in both blood and tissue subgroups (all P < 0.05). Our results suggest that DAPK promoter methylation may be involved in NSCLC carcinogenesis. Thus, the detection of aberrant DAPK methylation may be helpful in the diagnosis and prognosis of NSCLC. [ABSTRACT FROM AUTHOR]

We performed a meta-analysis of cohort studies to determine whether promoter methylation of the death-associated protein kinase (DAPK) gene contributes to the pathogenesis of nonsmall cell lung cancer (NSCLC). A range of electronic databases were searched: MEDLINE (1966 ∼ 2013), the Cochrane Library Database (Issue 12, 2013), EMBASE (1980 ∼ 2013), CINAHL (1982 ∼ 2013), Web of Science (1945 ∼ 2013), and the Chinese Biomedical Database (CBM; 1982 ∼ 2013) without any language restrictions. Meta-analysis was conducted using the STATA 12.0 software. Crude odds ratio (OR) with 95 % confidence interval (95 % CI) was calculated. Our meta-analysis integrated results from 12 clinical cohort studies that met all inclusion criteria with a total of 1,027 NSCLC patients. We observed that the frequency of DAPK gene methylation in cancer tissues were significantly higher than that in the adjacent normal and benign tissues (cancer tissues vs. benign tissues: OR=8.50, 95 % CI=5.88 ∼ 12.28, P<0.001; cancer tissues vs. adjacent tissues: OR=5.95, 95 % CI=4.11 ∼ 8.60, P<0.001; cancer tissues vs. normal tissues: OR=4.75, 95 % CI=3.28 ∼ 6.87, P<0.001; respectively). Subgroup analysis by ethnicity demonstrated that DAPK gene methylation was closely associated with the development and progression of NSCLC among both Asians and Caucasians (all P<0.05). Furthermore, we conducted a subgroup analysis based on sample source and discovered that DAPK gene methylation was implicated in the pathogenesis of NSCLC in both blood and tissue subgroups (all P<0.05). Our results suggest that DAPK promoter methylation may be involved in NSCLC carcinogenesis. Thus, the detection of aberrant DAPK methylation may be helpful in the diagnosis and prognosis of NSCLC.