OBJECTIVE:To establish the method for determining the contents of protocatechuic acid,rutin,gallic acid and kaempferol in Uygur medicine Capparis spinosa. METHODS:HPLC was performed on the column of Agilent C18 with the mobile phase of acetonitrile-0.1% phosphoric acid solution(7∶93,V/V)at the flow rate of 1.0 ml/min,the detection wavelength was 327 nm,column temperature was 30 ℃ and volume was 10 μl. RESULTS:The linear range was 0.260 0-50.0 μg for protocatechuic ac-id(r=0.995 6),3.109 0-102.0 μg for rutin(r=0.999 9),1.018 0-40.0 μg for gallic acid (r=0.998 9) and 0.063 0-36.0 μg for kaempferol(r=0.998 8);RSDs of precision,stability and reproducibility tests were all no more than 1.20%;average recoveries were respectively 101.51%(RSD=1.85%,n=6),99.70%(RSD=1.23%,n=6),98.28%(RSD=1.86%,n=6) and 100.97%(RSD=1.74%,n=6). CONCLUSIONS:The method is specific,and can fast and accurately determine the contents of protocate-chuic acid,rutin,gallic acid and kaempferol in Uygur medicine C. spinosa.

Moderate drought stress has been found to promote the accumulation of active ingredients in Glycyrrhiza uralensis root and hence improve the medicinal quality. In this study, the transcriptomes of 6-month-old moderate drought stressed and control G. uralensis root (the relative water content in soil was 40%-45% and 70%-75%, respectively) were sequenced using Illumina HiSeq 2000. A total of 80,490 490 and 82 588 278 clean reads, 94,828 and 305,100 unigenes with N50 sequence of 1,007 and 1,125 nt were obtained in drought treated and control transcriptome, respectively. Differentially expressed genes analysis revealed that the genes of some cell wall enzymes such as β-xylosidase, legumain and GDP-L-fucose synthase were down-regulated indicating that moderate drought stress might inhibit the primary cell wall degradation and programmed cell death in root cells. The genes of some key enzymes involved in terpenoid and flavonoid biosynthesis were up-regulated by moderate drought stress might be the reason for the enhancement for the active ingredients accumulation in G. uralensis root. The promotion of the biosynthesis and signal transduction of auxin, ethylene and cytokinins by moderate drought stress might enhance the root formation and cell proliferation. The promotion of the biosynthesis and signal transduction of abscisic acid and jasmonic acid by moderate drought stress might enhance the drought stress tolerance in G. uralensis. The inhibition of the biosynthesis and signal transduction of gibberellin and brassinolide by moderate drought stress might retard the shoot growth in G. uralensis.
Droughts ; Gene Expression Regulation, Plant ; Glycyrrhiza uralensis ; genetics ; Plant Roots ; Sequence Analysis, DNA ; Stress, Physiological ; Transcriptome

The dynamic changes of germination percentage, germination potential, thousand-seed weight, antioxidase activity in Desmodium styracifolium seeds with different storage time were tested, and electrical conductivity, contents of soluble sugar, soluble protein, starch in seed leach liquor were also determined in order to reveal the mechanism of seed deterioration. The results as the following. (1) The germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds declined, while the seed coat color darkened with the extension of storage time. (2) The activities of superoxide dismutase (SOD) and peroxidase (POD) decreased with the prolongation of storage period. The SOD activity declined fastest in 1,095-1,185 d of storage, while the POD activity declined significantly in 365-395 d of storage. (3) The electrical conductivity and the contents of soluble sugar, starch in seed leach liquor increased, while the content of soluble protein declined with the extension of storage time. (4) Correlation analysis indicated that the germination percentage, germination potential and thousand-seed weight of D. styracifolium seeds have a significantly positive correlation with SOD and POD activity, while have a significantly negative correlation with the electrical conductivity, contents of soluble sugar and starch. It can be concluded that during the storage of D. styracifolium seeds, physiological and biochemical changes including decrease in antioxidase activity, rise in electrical conductivity, degradation effluent of soluble sugar and starch, degradation of soluble protein were the main factors leading to the seed deterioration.
Color ; Fabaceae ; chemistry ; enzymology ; growth & development ; metabolism ; Germination ; Peroxidases ; metabolism ; Plant Proteins ; metabolism ; Seeds ; chemistry ; enzymology ; growth & development ; metabolism ; Starch ; metabolism ; Superoxide Dismutase ; metabolism ; Time Factors

Objective To retrospectively evaluate the utility of apparent diffusion coefficient (ADC) values in predicting aggressiveness of prostate cancer.Comparison was made with transrectal ultrasound-guided biopsy Gleason scores (GS) and prostatectomy GS.Methods Diffusion weighted images of 51 patients with biopsy-proven prostate cancer were obtained using 1.5 T MR with a pelvic phased-array coil.Regions of interest (ROIs) were drawn on areas of the suspicious lesion and the ADC values were calculated.The correlations between the ADC values and prostatectomy GS were assessed with Pearson correlation.The relationship between biopsy GS and prostatectomy GS were also evaluated.Meanwhile,receiver operating characteristic (ROC) curves were used to determine the ability of ADC values and biopsy GS in differentiating low-grade prostate cancer from intermediate/high grade prostate cancer.Results The accuracy of transrectal ultrasound-guided biopsy in predicting prostatectomy GS was 41.2％(21/51).Compared with prostatectomy GS,up to 11.8％ of the patients (n =6) was overestimated by biopsy,while 47.0％ (n =24) were underestimated.These 51 patients had a mean ADC value of (0.974 ±0.194) × 10-3 mm2/s.The mean ADC value of intermediate/high-grade tumors (n =35) was (0.907 ±0.160) ×10-3 mm2/s while that of low-grade tumors was (1.121 ±0.185) × 10-3 mm2/s (n =16).A significant negative correlation was found between mean ADC values of suspicious lesions and their prostatectomy GS (r =-0.761,P ＜ 0.01).No significant correlation was found between biopsy GS and prostatectomy GS (r =0.187,P =0.189).The area under the ROC curves of ADC and biopsy GS was 0.827 and 0.689,respectively.Conclusion The ADC values of cancerous areas in prostate perform better than biopsy GS in predicting aggressiveness of prostate cancer.

OBJECTIVETo investigate the effect of phosphoinositide 4-phosphate (PI4P) on human glioma U87 cells and the mechanism of action of PI4P in the development of human glioma through the overexpression or silencing of PI4P in human glioma U87 cells, and to provide a new target for basic research and clinical treatment of glioma.

METHODSLV-Helper1, LV-Helper2, pWPXLd-PI4P, and pLL3.7-shPI4P were used to package pWPXLd-PI4P and pLL3.7-shPI4P lentiviruses. The U87-GFP (PI4P-overexpression control group), U87-GFP-PI4P (PI4P-overexpression experimental group), U87-Scramble (PI4P-silencing control group), and U87-shPI4P (PI4P-silencing experimental group) cell lines were established. Wound-healing assay and Transwell assay were used to evaluate cell migration and invasion, and Western blot was used to measure the expression of PI4P in each group.

RESULTSWestern blot detected the expression of exogenous PI4P in the U87-GFP-PI4P cell line, and the U87-shPI4P cell line showed reduced expression of PI4P compared with the U87-Scramble cell line in the control group. The U87-GFP-PI4P cell line with PI4P overexpression had a significantly stronger ability of migration than the U87-GFP cell line in the control group (P<0.01); the U87-shPI4P cell line with PI4P silencing had a reduced ability of migration than the U87-Scramble cell line in the control group (P<0.01). The U87 cell line with PI4P overexpression had a significantly stronger invasion ability than the control group (P<0.05); after PI4P silencing, the experimental group showed a significant reduction in invasion ability compared with the control group (P<0.05).

CONCLUSIONSIn human glioma U87 cells, PI4P can promote the invasion and migration of glioma cells and may become a new target in the basic research and clinical treatment of glioma.

Cell Line, Tumor ; Cell Movement ; drug effects ; Glioma ; pathology ; Humans ; Neoplasm Invasiveness ; Phosphatidylinositol Phosphates ; pharmacology

Adolescent ; Child ; Collagen Type I ; metabolism ; Extracellular Matrix ; metabolism ; Female ; Fibrillin-1 ; Fibrillins ; Heart Defects, Congenital ; metabolism ; pathology ; Humans ; Male ; Microfilament Proteins ; metabolism

BACKGROUND: Surgical guides designed based on a three-dimensional cone-beam CT (CBCT) model have been reported. However, CBCT cannot remodel fine soft tissue such as gums, and it can only be used to design a simple dental retainer with relatively poor stability. OBJECTIVE: To establish a high-precision three-dimensional (3D) integrated maxillodental model by matching CBCT model with 3D digital maxillodental model using 3D automatic registration method, based on which, we designed and manufactured individualized miniscrew surgical guides. METHODS: CBCT maxillodental models and laser-scanned dentition models obtained from six malocclusion cases were matched and overlapped using the 3D automatic registration method to fabricate the 3D integrated maxillodental model. Then, we accurately positioned and virtually implanted a micro-implant into the 3D integrated maxillodental model. Subsequently we prepared a high-precision individualized resin surgical guide by rapid prototyping technology. The inner diameter of the guide track was detected by a vernier caliper. Patients tried on the resin surgical guide, and then occlusion condition, guide seating and retention were detected. RESULTS AND CONCLUSION: Due to the high-precision registration of the model, all the resin surgical guide plates were suitable. The plate retention was enhanced after tooth clinching, and all the patients felt comfort when wearing the surgical guide plate, with no compression or other discomforts. The inner diameter of the guide track was (1.79±0.23) mm, and the measurement error was not statistically significant (P >0.05). These findings demonstrate that the high-precision surgical guide plate based on the high-precision 3D integrated model can provide the foundation for further investigations on the clinical application of surgical guides.

The rhizome of Alpinia officinarum is a widely used Chinese herbal medicine. The essential oil in A. officinarum rhizome is mainly composed of 1, 8-cineole and other monoterpenes, as the major bioactive ingredients. In plants, monoterpenes are synthesized through the methylerythritol phosphate (MEP) pathway in the plastids, and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) is an enzyme catalyzing a committed step of the MEP pathway. In the present study, the full-length cDNA encoding DXR was cloned from the rhizome of A. officinarum, using homology-based RT-PCR and rapid amplification of cDNA ends (RACE) techniques. The new cDNA was designated as AoDXR and submitted to GenBank to be assigned with an accession number HQ874658. The full-length cDNA of AoDXR was 1 670 bp containing a 1 419 bp open reading frame encoding a polypeptide of 472 amino acids with a calculated molecular mass of 51.48 kDa and an isoelectric point of 6.15. Bioinformatic analyses revealed that AoDXR showed extensive homology with DXRs from other plant species and contained a conserved plastids transit peptide, a Pro-rich region and two highly conserved NADPH-binding motifs in its N-terminal region characterized by all plant DXRs. The phylogenetic analysis revealed that AoDXR belonged to angiosperm DXRs. The structural modeling of AoDXR showed that AoDXR had the typical V-shaped structure of DXR proteins. The tissue expression pattern analysis indicated that AoDXR expressed strongly in leaves, weak in rhizomes of A. officinarum. Exogenous methyl jasmonate (MeJA) could enhance the expression of AoDXR and the production of 1, 8-cineole in A. officinarum rhizomes. The cloning and characterization of AoDXR will be helpful to reveal the molecular regulation mechanism of monoterpene biosynthesis in A. officinarum and provides a candidate gene for metabolic engineering in improving the medicinal quality of A. officinarum rhizome.
Aldose-Ketose Isomerases ; genetics ; Alpinia ; chemistry ; enzymology ; genetics ; Amino Acid Sequence ; DNA, Complementary ; genetics ; Gene Expression Regulation, Enzymologic ; Gene Expression Regulation, Plant ; Molecular Sequence Data ; Monoterpenes ; metabolism ; Phylogeny

OBJECTIVETo explore the quality variation and genetic diversity of Desmodium styracifolium from different provenances, and lay a foundation for rational exploitation on germplasm resources and fine variety breeding of D. styracifolium.

METHODAmplified fragment length polymorphism (AFLP) markers were developed to analyze genetic diversity in D. styracifolium from 18 resources. NTSYSpc-2. 11F software was used to analyze the similarity among the D. styracifolium germplasms and construct the genetic phylogenetic tree. The schaftoside content in D. styracifolium from different provenances was determined by HPLC.

RESULTA total of 844 fragments were amplified with 8 primers, in which 717 were polymorphic bands, accounting for 84. 27% of the total detected variation. All the specimens from 18 resources could be grouped into 3 clusters by cluster analysis. The schaftoside contents of D. styracifolium germplasms differed significantly, with the highest content in the germplasm from Sanya, Hainan.

CONCLUSIONSignificant quality variation and genetic diversity can be observed among D. styracifolium germplasms. The diverse germplasm resources should be explored and the fine variety should be selected to breed.

Amplified Fragment Length Polymorphism Analysis ; Fabaceae ; classification ; genetics ; Genetic Variation ; genetics

OBJECTIVETo investigate the relationship between the changes of the copy numbers of mtDNA in peripheral blood mono-nucle- ar cell(PBMC) and the disordered of antioxidant capacity of hepatocellular carcinoma (HCC) patients.

METHODSThe Ficoll Hypaque method was used to isolate the PBMC from blood specimens. The ND1 gene of the mitochondrial was amplified by real-time PCR; meantime β-actin was served as a quantitative standard marker; the difference of mtDNA copy number in PBMC was compared between HCC and healthy control group. The level of reactive oxygen species (ROS) in PBMC was determined by flow cytometry. The change of total antioxidant capacity (T- AOC) of plasma was detected by the biochemistry examination.

RESULTSThe copy numbers of ND1 gene in PBMC of HCC was 73% that of the healthy control group,which suggested a decrease of the copy numbers of mtDNA in HCC. The levels of ROS of PBMC in HCC was (417. 82 ± 110.62) and (301.82 ± 75.54) in control group, which showed that the levels of ROS of PBMC in HCC were significant higher than that in control group (P < 0.01).Plasma T-AOC in HCC was (1.30 ± 0.85), and (3.20 ± 1.62) in control. The T-AOC of plasma of HCC was significantly lower than in control group (P < 0.01).

CONCLUSIONThere was a certain relationship between the decrease of the copy numbers of mtDNA and the disordered antioxidant capacity in hepatocellular carcinoma, which may be associated with the development of hepatocellular carcinoma.

Actins ; Antioxidants ; metabolism ; Carcinoma, Hepatocellular ; blood ; genetics ; Case-Control Studies ; DNA Copy Number Variations ; DNA, Mitochondrial ; genetics ; Humans ; Leukocytes, Mononuclear ; metabolism ; Liver Neoplasms ; blood ; genetics ; Reactive Oxygen Species ; metabolism