Objective To analyze the differential expression of caveolin-2 in the psoriasis vulgaris and normal skin tissues, and investigate the relationship between caveolin-2 and the development of psoriasis vulgaris. Methods The expression of caveolin-2 mRNA and protein in psoriasis vulgaris patients and normal skin tissues were detected by quantitative PCR, immunohistochemistry and Western blot respectively. Results The quantitative PCR showed that the expression of caveolin-2 mRNA significantly decreased in the psoriasis vulgaris skin tissues when compared with the normal skin tissues (P < 0.01). The immunohistochemistry demonstrated that the caveolin-2 protein was mainly expressed in the cytoplasm of the basal layer cells in the normal skin tissues, but the caveolin-2 protein was not expressed in the lesions of psoriasis vulgaris. And the results of Western blot showed that the expression of caveolin-2 protein was significantly reduced in the psoriasis vulgaris skin tissues compared with the normal skin tissues. Conclusion The expression of caveolin-2 was reduced or lost in lesional epidermis of psoriasis vulgaris patients, which may serve as an aetiological factor in the development and or progression of psoriasis.

Objective To identify the reliable reference genes for gene expression analysis of the pericarp and seed of Amomum villosum Lour. by using real-time fluorescence quantitative polymerase chain reaction ( qRT-PCR). Methods Using the fruits ( separated into peels and seeds) of A. villosum at three different developmental periods as the experimental material, 5 candidate reference genes (β-actin, EF-1α, GAPDH, PGK, TUA) with steady expression were screened out by the high throughout sequencing of transcriptome and expression profile data. The qRT-PCR technique was applied to study the expression levels of 5 candidate reference genes in different samples. The stability of the candidate reference genes were evaluated by GeNorm and NormFinder software. Results The 5 reference genes had different stabilities in the pericarp and seed of A. villosum Lour. at different development periods . The order of the steadiness of reference genes showed by GeNorm was EF-1α = TUA>PGK>GAPDH>β-actin. The results of NormFinder revealed that EF-1α was the most stable, followed by TUA, and the order of the other three genes was as same as the results of GeNorm. Conclusion EF-1αand TUA could be used as double reference genes for the normalization of gene expression in A. villosum fruits at different developmental periods by using qRT-PCR.

Objective To establish a molecular identification method for three cultivars of Amomum villosum Lour.(AVL),thus to provide scientific evidence for the identification,selection and breeding of AVL.Methods The fragments of 26S rDNA D1-D3 region and matK gene of three cultivars of AVL and Amomum longiligulare T.L.Wu were amplified by polymerase chain reaction(PCR) and with corresponding primers,and then their sequences were analyzed,and phylogenetic tree was constructed based on the sequences.Results We obtained 739 bp in 26S rDNA D1-D3 sequence.Differences in 4 basic sites of 739 bp were shown between AVL and Amomum longiligulare T.L.Wu.The two cultivars of AVL,Changguo and Yuanguo,had the same sequence,but there was a difference in one basic site of Changguo and Yuanguo from Chunxuan.The phylogenetic tree based on 26S rDNA D1-D3 sequence revealed the difference between Chunxuan and the other two cultivars of AVL.We also obtained 824 bp in matK gene sequence.The three cultivars of AVL showed the consistent sequence,but there was a difference in one basic site of three cultivars of AVL from Amomum longiligulare T.L.Wu.Conclusion We can identify the three cultivars of AVL through the sequence differences at the molecular level,and Chunxuan has a closer genetic relationship with Amomum longiligulare T.L.Wu.

AIM Aim to study the effect of PLC on human platelet cytoplasmic free calcium concentration, and then to explore the mechanism of PLC anti aggregation to platelet. METHODS Washed platelet of human was marked with fluorescence indicator Furu 2/AM, and then was treated with PSS, ASA and different dose of PLC respectively. Cytoplasmic free [Ca 2+ ] i of differently treated platelet at static state and activated sate induced by ADP was determined by double beam fluorescence spectrophotometric method. RESULTS Healthy human platelet was treated with PSS, ASA 334 ?mol?L -1 , 2 5, 3 75, 5, 10 and 20U PLC?ml -1 . Cytoplasmic [Ca 2+ ] i determined at static state was 152 55?15 07, 131 63?15 58, 140 27?12 03, 139 48?1 73, 121 11?9 58, 116 62?15 96 and 107 20?17 07 respectively. [Ca 2+ ] i determined at activated sate induced by ADP was 902 62?94 74, 687 99?62 86, 810 99?72 37, 701 73?21 37, 429 67?71 59, 342 82?44 86 and 263 27?25 46 respectively. Compared with PSS, inhibition rates ( % ) of ASA 334 ?mol?L -1 , 2 5, 3 75, 5,10, 20 U PLC?ml -1 to the increment of activated platelet cytoplasmic [Ca 2+ ] i were 25 83, 10 58, 25 04, 58 86, 69 84, 79 19 respectively. At the same condition, inhibition rate ( % ) of 10 U PLC?ml -1 to the increment of five hypertensive activated platelet cytoplasmic [Ca 2+ ] i were 74 25, 73 48, 61 32, 82 87,70 60 respectively. CONCLUSION PLC has significant effects on static cytoplasmic[Ca 2+ ] i of healthy and hypertensive human platelet ( P

Biological chemistry of Chinese herbal germplasm resources (BCCHGR) is a new cross-discipline formed from rapid development of modern science and technology and its application in the area of Chinese herbal resources. BCCHGR was defined as probing and understanding biological processes like heredity, gene transcription, expression and metabolism of Chinese herbal germplasm, at the interface of biochemistry, molecular biology and chemistry, elu-cidating the nature of Chinese herbal germplasm using as TCM medicine as well as the forming mechanism thereof. In this paper, the scientific background, definition, significance and contents of BCCHGR were discussed to depict a preliminary picture of BCCHGR and arouse popular consideration and discussions.

This study was aimed to discuss the dynamic variation of soluble sugar contents, sucrose metabolizing en-zyme activities and gene expression quantities during the fruits development of A momum villosum, in order to pro-vide the basis of improvement of the fruit yield. Fresh fruits at three different development processes (30 DAF, 60 DAF, 90 DAF) were used to investigate changes of soluble sugar components and sucrose metabolizing enzyme activ-ities by HPLC and UV spectrophotometry. Combining with the high-throughput sequencing expression profile data of three fruit development period, the trends of three key enzymes gene expressed in sugar metabolism were analyzed. The results showed that the fruit sugar components were dominated by fructose, glucose and sucrose. The concentra-tion of hexose (fructose and glucose) gradually decreased in peel. But in seeds the concentration of hexose decreased at first and then increased. The content of sucrose and the net activities of sucrose synthase (synthesizing direction minus decomposing direction) in peel and seeds were gradually increased. The expression trends of key enzyme gene in sugar metabolism examined by RNA-seq quantification showed that sucrose phosphate synthase and sucrose syn-thase gene increased and then kept constant, but the invertase gene expression trend was gradually rising. Conse-quently, sucrose synthase was the key enzyme catalyzing sucrose synthesis and decomposition. The activity of sucrose synthase and sucrose contents in peel and seeds reached the highest peak in the end of fruit mature.

This study was aimed to reveal the effects and molecular regulation mechanism of methyl jasmonate (Me-JA) on volatile terpenoids from Amomum villosum Lour. After the leaves and fruits of A momum villosum Lour. were treated with different concentrations of MeJA, the volatile terpenoids of fresh fruits from A . villosum Lour. were ex-tracted with microwave method and analyzed by GC-MS. Then, leaves and fruits treated with MeJA were sequenced by Illumina. The transcriptome data was analyzed by bioinformatic methods. The results showed that there were 20 and 33 volatile terpenoids detected in peels and seed groups, respectively. Contents of volatile terpenoids in peels and seed groups were both improved after 600 μmol·L-1 MeJA treating fruits for 24 h, such as bornyl acetate, cam-phor, borneol, and etc. While 200 μmol·L-1 MeJA treating different parts for 24 h can regulate the biosynthesis of some volatile terpenoids in peels differently. And 200 μmol·L-1 MeJA treating fruits can improve the content of ma-jor volatile terpenoids in seed groups. A total of 68 168 unigenes were obtained with de novo assembly, and 48 627 unigenes were annotated after comparison with public protein databases. Analysis of functional annotation against KEGG database showed that there were 208 unigenes closely related with metabolism of volatile terpenoids and 22 u-nigenes related with MYC2 transcription factors. It was concluded that MeJA can effectively regulate the metabolism of volatile terpenoids from A . villosum Lour. There were a lot of candidate genes related with the biosynthesis of volatile terpenoids obtained by analyzing the transcriptome data which also provided a large amount of data for the discovery and regulation of functional genes related with the biosynthesis of volatile terpenoids from A . villosum Lour.

HMGR and DXR are key enzymes of terpenoids biosynthesis pathway. This study was aimed to discuss the effects of overexpression of HMGR and DXR from A momum villosum Lour. on the biosynthesis of terpenoids in transgenic tobacco. The real-time fluorescence quantitative PCR (RT-qPCR) was used to analyze the expression level of AvHMGR and AvDXR. Then, enzyme activities of HMGR and DXR were determined by spectrometer using the substrate-specific method. Different terpenoids were detected by GC-MS. The results showed that individual overex-pression of HMGR/DXR can inhibit the enzyme activities of HMGR and DXR but promote the biosynthesis of men-thene, neophytediene, cembrenene and sterol. The co-overexpression of HMGR and DXR had different enzyme activ-ities and can promote the biosynthesis of sterol and phytol, but inhibit the biosynthesis of neophytadiene. It was con-cluded that the overexpression of HMGR and DXR had diverse effects when regulating the biosynthesis of different terpenoids. This study provided the basis for using A vHMGR and A vDXR to regulate the metabolism of terpenoids.

Objective: To investigate the relationship of PD-L1 protein expression and gene amplification in gastric cancer and their correlation with clinicopathologic factors. Methods: The cohort included 247 gastric cancer specimens with follow-up data and clinicopathologic data obtained from Shanxi Cancer Hospital in 2011. PD-L1 expression was detected by immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH). Results: PD-L1 protein was expressed in 25.9% (64/247) of the tumor cells and 26.7% (66/247) of the tumor infiltrating immune cells (IC). There was a correlation between the two (P<0.01). The expression of PD-L1 in tumor cells correlated with the degree of differentiation and tumor diameter(P<0.05). The PD-L1 expression in IC correlated with vascular tumor thrombi(P<0.05). The amplification rate of PD-L1 gene detected by FISH was 19.0% (47/247), and was associated with age, large/small curvature of the stomach, tumor location, tumor diameter, and lymph node metastasis(P<0.05). The positive coincidence rate of the two methods was 25.0% (16/64), negative coincidence rate was 83.0% (152/183), and total coincidence rate was 68.0% (168/247), suggesting that the coincidence of IHC and FISH was poor (P=0.157). There was a negative correlation between PD-L1 protein expression on tumor cells and prognosis in gastric cancer. There was no significant correlation between PD-L1 protein expression on IC and PD-L1 gene amplification with prognosis. Vascular tumor thrombi, tumor diameter, depth of invasion, and lymph node metastasis were all poor prognostic factors of gastric cancer(P<0.05). Multivariate Cox regression analysis showed that PD-L1 protein expression, depth of invasion and lymph node metastasis were all independent prognostic risk factors for gastric cancer. Conclusions Concordance between PD-L1 protein expression and gene amplification is poor. PD-L1 protein expression may signify poor prognosis. There is no significant correlation between PD-L1 gene amplification and prognosis of patients with gastric cancer.

Objective To establish a DNA extraction method for DNA barcoding analysis of Chinese medicinal materials.Methods Seven different DNA extraction methods were used to extract DNA from 6 medicinal recalcitrant plants which are rich in secondary metabolites.Results CTAB method 3 was fast,simple,universal and effective,by which a high DNA concentration and qualified ratio were obtained as compared with the other methods.The DNA extracted by this method could provide good results for DNA barcoding analysis.The main improved steps of this methods were as follows:①adoption of 3 %CTAB rather than 2 %CTAB in the exaction;②adding 1 %polyvinylpyrrolidone(PVP) and 0.2 %?-mercaptoethnoal in extraction solution to remove secondary metabolites and to prevent DNA degradation;③centrifuge at 10000 r/min for 15 min to remove protein and impurity.Conclusion CTAB method 3 is a proper method of DNA extraction for DNA barcoding analysis of Chinese medicinal materials.