ABSTRACT
Direct acid hydrolysis of Dioscorea zingiberensis rhizomes for preparation of diosgenin is wildly used in the traditional industry， which uses a large amount of inorganic acid catalysts， with high wastewater discharge and serious environmental pollution. Therefore， exploring clean and efficient preparation methods and processes has become an inevitable choice to realize the sustainable development of industrial production of diosgenin. Herein， the author reviewed and analyzed the research progress and problems of enzymatic hydrolysis， microbial transformation and modified acid hydrolysis in the preparation of diosgenin from D. zingiberensis rhizomes during the last ten years， and their application prospects are analyzed. Enzymatic hydrolysis has mild reaction conditions， but the yield of diosgenin is low， the economic cost is high， and the purification process of active enzyme is complicated. Microorganism shows specific activity to the substrate and high efficiency for diosgenin production， and microbial transformation is clean and environmentally friendly， but microbial transformation is time-consuming and the metabolic intermediates are complicated. For the modified acid hydrolysis， two-phase acid hydrolysis can reduce the amount of acid catalyst， and sulfonic acid-functionalized ionic liquid displays good recyclable performance by replacing the traditional inorganic acid， however， the wastewater discharge should still be considered. Solid acid catalysts are non-corrosive and easy to be recycled， but the need to use ethanol as the reaction solvent has certain safety hazards， and the catalyst preparation process is cumbersome. In conclusion， exploring clean and efficient conversion methods is an important research trend for preparation of diosgenin from D. zingiberensis rhizomes. For the enzymatic hydrolysis， the key glycoside hydrolases in the bioconversion process should be explored in depth， the conversion pathway of enzymatic saponins and enzyme specificity should be fully elucidated， and efforts should be made to improve the efficiency of enzymatic hydrolysis. For the microbial transformation， we should accelerate its industrial application process based on selecting and breeding efficient transformation strains， and optimizing stable transformation systems and processes， and in-depth investigation of the mechanism of microbial transformation， fully elucidating the specific key hydrolases and its catalytic properties， and striving to improve the efficiency of microbial transformation. For the modified acid hydrolysis， novel acid catalytic system with simple structure， stable performance and good biodegradability should be explored and applied， which can effectively solve the problems of environmental pollution and production safety.
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Objective:To explore the inhibitory effects of gomisin A(GA)in combination with carboplatin (CBP)on the proliferation,invasion and metastasis of human ovarian cancer Skov3 cells and their mechanisms,and to illustrate the synergistic antitumor effect of GA.Methods:The ovarian cancer Skov3 cells were treated with different concentrations of GA combined with different concentrations of CBP.MTT assay was used to detect the inhibitory rates of proliferation.According to the results,the appropriate concentrations of GA and CBP were conformed.The Skov3 cells were treated with PBS,GA(0.04 μmol·L-1),CBP(16 mg·L-1)and GA (0.04 μmol·L-1)in combination with CBP(16 mg·L-1),respectively,and used as control group,GA group, CBP group and GA+CBP group.After 48 h of treatment,the cell morphology was observed by optical microscope, the cell reproductive ability was determined by clonogenic cell survival assay,and the cell migration ability was measured by cell wound healing test;Transwell experiment was used to detect the cell invasion ability.The expression levels of MMP-2 and MMP-9 mRNA were determined by RT-PCR and the protein expression levels of MMP-2,MMP-9,AKT1 and pAKT1 were determined by Western blotting method.Results:Compared with GA and CBP groups,the inhibitory rate of proliferation of Skov3 cells in GA + CBP group was significantly increased (P<0.01).The concentrations of GA and CBP for the best dose ratio were 0.04 μmol·L-1and 16 mg·L-1, respectively.The results of clone formation assay showed that the colony formation rates in CBP and GA+CBP groups were decreased compared with control group(P<0.05 or P<0.01);the colony formation rate of cells in GA + CBP group was significantly decreased compared with GA and CBP groups(P<0.01);the wound scratch assay results suggested that the number of metastic Skov3 cells in GA and CBP groups were decreased compared with control group;and the number of metastic Skov3 cells in GA+ CBP group was decreased compared with GA and CBP groups.The results of Transwell expriment showed that the capacities of cells passing the ECM in GA and CBP groups were decreased compared with control group;the capacity of cells passing the ECM was decreased compared with GA and CBP groups.The RT-PCR and Western blotting results showed that the expression levels of MMP-2 and MMP-9 mRNA in GA+CBP group were down-regulated compared with GA and CBP groups(P<0.01),and the expression levels of MMP-2,MMP-9,AKT1 and pAKT1 protein were down-regulated(P<0.01).Conclusion:GA can enhance the ability of CBP to inhibit the proliferation,invasion and metastasis of human ovarian cancer Skov3 cells,and thus play a role in decreasing the toxicity and increasing the efficacy of chemotherapy.
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Objective:To explore the effect of gomisin A (GA) in combination with carboplatin (CBP) on the apoptosis of ovarian cancer Skov3 cells,and to illustrate the synergistic antitumor effect of GA.Methods:The ovarian cancer Skov3 cells were treated with GA combined with CBP.MTT assay was used to detect the inhibitory rates of proliferation and the appropriate concentrations of GA and CBP were selected according to the results.The Skov3 cells were divided into control group,GA (0.04 μmol · L-1) group,CBP (16 mg · L 1) group and GA (0.04 μmol· L-1)+ CBP (16 mg · L-1) group.After 48 h treatment,the cell morphology was observed by microscope,the apoptotic rate was measured by flow eytometry,the apoptotic index (AI) was observed by TUNEL staining,the mitochondrial membrane potential was detected by JC-1 staining,and the mRNA and protein expression levels of apoptosis-related genes (Bax,caspase-3,Stat3) were determined by RT PCR and Western blotting method.Results:The inhibitory rate of proliferation of Skov3 cells in GA + CBP group (52.1％) was significantly higher than those in GA and CBP groups (P＜0.01).The concentrations of GA and CBP for the best dose ratio were 0.04 μmol· L-1 and 16 mg · L-1,respectively.Compared with control group,the cell refractivities were decreased,the cells retracted,and part of cells were suspended in GA group and CBP groups;there were more suspended cells and cell retraction phenomenon was more prominent in GA + CBP group.The results of TUNEL staining and flow cytometry showed that the AI and apoptotic rate of cells in GA + CBP group were significantly increased compared with control group,GA and CBP group (P＜0.05 or P＜0.01);the JC-1staining results suggested that the mitochondrial membrane potential of Skov3 cells in GA + CBP group was decreased (P＜0.05 or P＜0.01).The RT-PCR and Western blotting results showed that the expression levels of Bax and caspase-3 mRNA and protein were up-regulated (P＜0.05) and the expression levels of Bcl-2 and Stat3mRNA and protein in GA + CBP group were down-regulated compared with control,GA and CBP groups (P＜0.05).Conclusion:GA can enhance the ability of CBP to induce the apoptosis of human ovarian cancer Skov3 cells,and its mechanisms may be related to the up-regulation of the Bax and Caspase-3 expressions and the down-reguation of the Bcl-2 expression;GA can synergize the induction of CBP on apoptosis.
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Purpose To detect the expression of FOXO3 in lung cancer specimens and lung cancer stem cell, for diagnosis and treat-ment of lung cancer. Methods Immunohistochemistry, semi-quantitative PCR and Western blot were used to detect FOXO3 protein expression in clinical specimens of lung cancer. PCR and Western blot verified FOXO3 mRNA and protein expression of lung cancer cells. Results Immunohistochemistry showed that the FOXO3 was significantly lower expressed in lung cancer tissues and the corre-sponding lung cancer stem cells;Detection of FOXO3 mRNA and protein by RT-PCR and Western blot showed both had significantly lower expression;as progression of the disease, there was a clear decreasing trend, in which negative expression rate of FOXO3 protein accounted for 66. 7% in all patients;at the same times, the rate of the expression gradually decreased with loss of differentiation and metastasis of the tumor. Conclusions FOXO3 shows low expression in lung tissue and lung cancer stem cells, while it is closely relat-ed to the degree of malignancy and lung cancer metastasis, which may be a new target for the treatment.
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Objective To understand the dietetic habits,the knowledge level of nutrition and food hygiene,the understanding of the relationship between food and disease,and their sources of and the demands for nutrition and food hygiene in the residents of Tianjin.Method 1 206 subjects(605 male and 601 female) aged 18 to 69 yrs were selected by the stratified multi-stage random sampling and investigated with the questionnaire.Results of all the subjects,no one's diet was consistent with the Chinese balanced diet guideline.Most subjects had some knowledge about food and nutrition,but only 60.2% of them passed the test.The situation of the knowledge awareness varied with their living area,education background and occupation.87.5% of the subjects had a good awareness for the relationship between food and disease,and varied with their education and occupation.Television was the most important approach for their acquiring the knowledge about nutrition and food hygiene.81.5% subjects were interested in learning more about nutrition and food hygiene.Conclusions The dietetic habits in residents of Tianjin was still not satisfied.It is necessary to strengthen the propaganda and education of the nutrition and food hygiene in residents and explore an effective education model for them.
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2 score,20.0% vs 8.5%) between two groups (P0.05). After a mean post-biopsy follow-up of (34.6?33.3) months, the 3-year and the 5-year renal survival rates for elder group were 74.6% and 62.2%, respectively, which were lower than those of non-elder group (100% and 92.9%, P=0.002). Conclusion:Elder patients with IgA nephropathy were more likely to suffer from hypertension, hyperlipidemia, renal insufficiency and chronic pathologic lesions, which might be the risk factors for the patient's unfavorable prognosis.