Identifying and validating MMP family members(MMP2,MMP9,MMP12,and MMP16)as therapeutic targets and biomarkers in kidney renal clear cell carcinoma(KIRC)

Kidney Renal Clear Cell Carcinoma(KIRC)is a malignant tumor that carries a substantial risk of morbidity and mortality.The MMP family assumes a crucial role in tumor invasion and metastasis.This study aimed to uncover the mechanistic relevance of the MMP gene family as a therapeutic target and diagnostic biomarker in Kidney Renal Clear Cell Carcinoma(KIRC)through a comprehensive approach encompassing both computational and molecular analyses.STRING,Cytoscape,UALCAN,GEPIA,OncoDB,HPA,cBioPortal,GSEA,TIMER,ENCORI,DrugBank,targeted bisulfite sequencing(bisulfite-seq),conventional PCR,Sanger sequencing,and RT-qPCR based analyses were used in the present study to analyze MMP gene family members to accurately determine a few hub genes that can be utilized as both therapeutic targets and diagnostic biomarkers for KIRC.By performing STRING and Cytohubba analyses of the 24 MMP gene family members,MMP2(matrix metallopeptidase 2),MMP9(matrix metallopeptidase 9),MMP12(matrix metallopeptidase 12),and MMP16(matrix metallopeptidase 16)genes were denoted as hub genes having highest degree scores.After analyzing MMP2,MMP9,MMP12,and MMP16 via various TCGA databases and RT-qPCR technique across clinical samples and KIRC cell lines,interestingly,all these hub genes were found significantly overexpressed at mRNA and protein levels in KIRC samples relative to controls.The notable effect of the up-regulated MMP2,MMP9,MMP12,and MMP16 was also documented on the overall survival(OS)of the KIRC patients.Moreover,targeted bisulfite-sequencing(bisulfite-seq)analysis revealed that promoter hypomethylation pattern was associated with up-regulation of hub genes(MMP2,MMP9,MMP12,and MMP16).In addition to this,hub genes were involved in various diverse oncogenic pathways.The MMP gene family members(MMP2,MMP9,MMP12,and MMP16)may serve as therapeutic targets and prognostic biomarkers in KIRC.

Jasmonic acid regulates the biosynthesis of medicinal metabolites via the JAZ9-MYB76 complex in Salvia miltiorrhiza

Jasmonic acid(JA)signaling pathway plays an important role in tanshinone and phenolic acid biosynthesis in Salvia miltiorrhiza.However,the specific regulatory mechanism remains largely unclear.Previous work showed that a JASMONATE ZIM-domain(JAZ)protein,SmJAZ9,acted as a repressor of tanshinone production in S.miltiorrhiza.In this study,we revealed that SmJAZ9 reduced both phenolic acid accumulation and related biosynthetic gene expression,confirming that SmJAZ9 also negatively affected phenolic acid biosynthesis.Then,we identified a novel MYB transcription factor,SmMYB76,which interacted with SmJAZ9.SmMYB76 repressed phenolic acid biosynthesis by directly downregulating SmPAL1,Sm4CL2,and SmRAS1.Further investigation demonstrated that JA mediated phenolic acids biosynthesis via SmJAZ9-SmMYB76 complex.Taken together,these findings state the molecular mechanism that SmJAZ9-SmMYB76 regulated phenolic acid biosynthesis at the transcriptional and protein levels,which provided new insights into JA signaling pathway regulating plant metabolism.

Effects of Biological Fertilizer and Paclobutrazol on Total Flavonoids in Ophiopogon japonicus

[Objectives] The research aimed to study the effects of applying biofertilizer instead of paclobutrazol on the content of flavonoids in Ophiopogon japonicas and determine its quality characteristics,in order to provide the scientific basis for the quality control of GAP and the comprehensive utilization of resources. [Methods] O. japonicas collected in Lingxing Town and Huayuan Town of Sichuan Province was divided into three groups: blank control group,paclobutrazol group and biofertilizer group. Root tuber of O. japonicas was obtained,and the enzyme was killed by the microwave,and then it was dried in 60℃. The content of flavonoids in root tuber of O. japonicus was determined by ultraviolet spectrophotometry. Then,SPSS19. 0 was used to analyze the results. [Results]The average contents of flavonoids in different treatments were as follows: 0. 184%-0. 198% in control group; 0. 283%-0. 330% in the paclobutrazol group; 0. 404%-0. 428% in the biofertilizer group. Biological bacterial fertilizer had a significant role in promoting the synthetic accumulation of homoisoflavonoids in O. japonicus( P <0. 001**). [Conclusions]The application of biological fertilizer can not only significantly improve the content of homoisoflavonoids in root ruber of O. japonicus,but also significantly optimize the micro ecological environment of soil. Therefore,biofertilizer should be widely used to gradually replace paclobutrazol in the main production areas of O. japonicas.

A Preliminary Study on the Infrared Spectra Characteristics of Radix Chuanmingshinis Violacei

[Objectives] To carry out infrared spectroscopic study of different growth years and different parts of the traditional Chinese medicine Radix Chuanmingshinis Violacei. [Methods]Using potassium bromide squash method and Fourier transform infrared spectroscopy on the different growth years and different parts of Radix Chuanmingshinis Violacei for infrared spectrum and second derivative spectrum. [Results]With the increasing growth time,Radix Chuanmingshinis Violacei accumulated in the secondary metabolism substances gradually increased,the characteristic absorption peak appeared in the position such as 3 366,2 927,1 634,1 416,1 516,1 080,1 020,and 928 cm^(-1),the main chemicals as polysaccharide,coumarin,saponins and steroids. The difference of chemical position between root tuber and cauline leaf is obvious. [Conclusions]Infrared spectroscopy was applied in studying different growing periods and different parts of Radix Chuanmingshinis Violacei,of obvious characteristics,identification and quality evaluation method,is simple and suitable for Radix Chuanmingshinis Violacei.

Application of Multivariate Analysis Method in Evaluation of Quality and Traits of Sichuan Ophiopogon japonicus

[Objectives] To study the relationship between the yield and total saponin content of Sichuan Ophiopogon japonicus. [Methods]The multi-indicator function model of Sichuan O. japonicus was established by using the multivariate analysis method and taking the yield of and total saponin content of Sichuan O. japonicus as indicators. [Results] In the multivariate non-linear fitting,the Pearson correlation test was used to reduce the dimension of the model,and the significant correlated variables were rejected,leaving three independent variables: the total fresh weight of the plant( x_1),the fresh weight of the aboveground part(x_2) and the fresh weight of fibrous roots(x_3),established the total saponin( y) function model for Sichuan O. japonicus: y = a_1+a_2x_2+ a_3x_3+ a_4x_1x_2+ a_5x_1x_3+ a_6x_2x_3+ a_7x_1~2+ a_8x_2~2+ a_9x_3~2.[Conclusions]When the total fresh weight of plant,fresh weight of aboveground part and fresh weight of fibrous roots were known in Sichuan O. japonicus root tuber,the total saponin content could be estimated by polynomial of these three variables. The establishment of this functional model system is expected to provide a scientific basis for the scientific prediction of the yield and total saponin content of Sichuan O. japonicus.

Quality Characteristics and FTIR Research of Amorphophallus konjac K. Koch Cultivated under Forest

[Objectives]This study aimed to evaluate the quality characteristics of Amorphophallus konjac K.Koch and study the tubers of A.konjac K.Koch by infrared spectroscopy and second derivative method.[Methods]Referring to industrial standards of China for agriculture and related documents,the quality characteristics of A.konjac K.Koch were determined;and using potassium bromide tableting method and Fourier transform infrared spectroscopy,the infrared spectrum and second derivative spectrum of A.konjac K.Koch were studied.[Results]The drying rate,total ash content,acid-insoluble ash content,water extract content,ethanol extract content and crude polysaccharide content of A.konjac K.Koch were above 14.00%,4.57%-11.78%,0.04%-4.87%,26.31%-36.98%,2.20%-7.95%and 40.25%-61.18%,respectively.The infrared spectra of A.konjac K.Koch of different origins were relatively close,and the differences were mainly in peak intensity.The peaks at 1642,1322,1241,1154,1022,863,770 and 576 cm-1 were the main specific peaks.The main component was konjac glucomannan.[Conclusions]FTIR technology is simple and quick,and is suitable for quality control and sample identification of A.konjac K.Koch.This provides a scientific basis for the quality control and comprehensive utilization of A.konjac K.Koch cultivated under forest.

Experimental Study on Content of Aqueous Extract and Total Flavonoids in Different Parts of Sichuan Ophiopogon japonicus 1

[Objectives]To study the differences in the content of aqueous extract and total flavonoids in different parts of Sichuan Ophiopogon japonicus 1,and provide a scientific basis for the comprehensive utilization of O. japonicus resources. [Methods] According to Volume I of 2015 Chinese Pharmacopoeia,and with reference to literature methods,taking water as extraction solvent,hot dip method was used to determine the content of aqueous extract; taking ethanol as extraction solvent,the content of total flavonoids was determined using hot dip method,Ultraviolet-Visible Spectrophotometry. [Results]The content of aqueous extract and total flavonoids in the root tubers of Sichuan O. japonicus1 was higher than that of traditional O. japonicas; the content of aqueous extract in the root tubers of Sichuan O. japonicus 1 was higher than that in the leaves of Sichuan O. japonicus 1; the content of total flavonoids in leaves of Sichuan O. japonicus 1 was higher than that in the root tubers; the content of aqueous extract and total flavonoids in Sichuan O. japonicus 1 in the second year was higher than that in the first year.[Conclusions]The quality of new cultivar of Sichuan O. japonicas 1 was significantly higher than the traditional O. japonicus. Nowadays,the main part of drug use in O. japonicas is root,but as to the content of total flavonoids,fibrous roots and leaves are more dominant,it is feasible to develop and utilize the resources of flavonoids in the fibrous roots and leaves. The content of aqueous extract and total flavonoids from the second year O. japonicus was higher than that in the first year O. japonicus,but its increased amount did not meet the requirements of economic benefits.

Arabidopsis DET1 Represses Photomorphogenesis in Part by Negatively Regulating DELLA Protein Abundance in Darkness

Arabidopsis De-etiolated 1 （DET1） is one of the key repressors that maintain the etiolated state of seedlings in darkness. The plant hormone gibberellic acid （GA） also participates in this process, and plants deficient in GA synthesis or signaling show a partially de.etiolated phenotype in darkness. However, how DET1 and the GA pathway work in concert in repressing photomorphogenesis remains largely unknown. In this study, we found that the abundance of DELLA proteins in detl-1 was increased in comparison with that in the wildtype plants. Mutation in DET1 changed the sensitivity of hypocotyl elongation of mutant seedlings to GA and paclobutrazol （PAC）, an inhibitor of GA synthesis. However, we did not find obvious differences between detl-1 and wild-type plants with regard to the bioactive GA content or the GA signaling upstream of DELLAs. Genetic data showed that removal of several DELLA proteins suppressed the detl-1 mutant phenotype more obviously than GA treatment, indicating that DET1 can regulate DELLA proteins via some other mechanisms. In addition, a large-scale transcriptomic analysis revealed that DET1 and DELLAs play antagonistic roles in regulating expression of photosynthetic and cell elongation-related genes in etiolated seedlings. Taken together, our results show that DET1 represses photomorphogenesis in darkness in part by reducing the abundance of DELLA proteins.

Dynamic full-field refractive index distribution measurements using total internal reflection terahertz digital holography

Massive usage scenarios prompt the prosperity of terahertz refractive index(THz RI) measurement methods.However, they are very difficult in measuring the full-field dynamical RI distributions of either solid samples without a priori thickness or liquid samples. In this study, we propose total internal reflection THz digital holography and apply it for measuring RI distributions for both solid and liquid samples dynamically. An RI measurement model is established based on an attenuated total reflection prism with a pitching angle. The pitching angle and the field of view can be numerically calculated from the spectrogram of the off-axis Fresnel hologram,which solves the adjustment of the visually opaque prism irradiated by the invisible THz beam. Full-field RI distributions of the droplets of solid-state soy wax and distilled water are obtained and compared with THz time-domain spectroscopy. The evaporation of an ethanol solution droplet is recorded, and the variation of the RI distribution at the sample–prism interface is quantitatively visualized with a temporal resolution of 10 Hz. The proposed method greatly expands the sample range for THz RI measurements and provides unprecedented insight into investigating spontaneous and dynamic THz phenomena.