Production of bioactive ginsenoside Rg3(S) and compound K using recombinant Lactococcus lactis

Lingli Jiang,Soo Jin Lee,Qiu Ping Yuan,임완택,김선창,NAM SOO HAN 고려인삼학회 2018 Journal of Ginseng Research Vol.42 No.4

Background: Ginsenoside Rg3(S) and compound K (C-K) are pharmacologically active components of ginseng that promote human health and improve quality of life. The aim of this study was to produce Rg3(S) and C-K from ginseng extract using recombinant Lactococcus lactis. Methods: L. lactis subsp. cremoris NZ9000 (L. lactis NZ9000), which harbors b-glucosidase genes (BglPm and BglBX10) from Paenibacillus mucilaginosus and Flavobacterium johnsoniae, respectively, was reacted with ginseng extract (protopanaxadiol-type ginsenoside mixture). Results: Crude enzyme activity of BglBX10 values comprised 0.001 unit/mL and 0.003 unit/mL in uninduced and induced preparations, respectively. When whole cells of L. lactis harboring pNZBglBX10 were treated with ginseng extract, after permeabilization of cells by xylene, Rb1 and Rd were converted into Rg3(S) with a conversion yield of 61%. C-K was also produced by sequential reactions of the permeabilized cells harboring each pNZBgl and pNZBglBX10, resulting in a 70% maximum conversion yield. Conclusion: This study demonstrates that the lactic acid bacteria having specific b-glucosidase activity can be used to enhance the health benefits of Panax ginseng in either fermented foods or bioconversion processes.

Overexpression and Clinicopathological Contribution of DcR3 in Bladder Urothelial Carcinoma Tissues

Jiang, Yi-Qiang,Zhong, Teng-Fei,Dang, Yi-Wu,Zou, Ling-Song,Yang, Liu,Yang, Xia,Chen, Gang Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Background: To explore the expression of DcR3 protein and its clinicopathological significance in bladder urothelial carcinomas (BUC). Materials and Methods: Immunohistochemistry was performed to detect the expression of DcR3, caspase-3, Bcl-2, VEGF, Ki-67, PCNA and P53 in 166 BUC and 56 normal bladder tissues. Western blotting was used to detect the expression of DcR3 in the supernatants of cultured BUC cells. Results: Overexpression of DcR3 was found in BUC tissues and cell lines, with significant elevation as compared to normal bladder tissues (p<0.0001). Higher DcR3 expression was related to the status of invasion, lymph node metastasis and recurrence. Furthermore, DcR3 expression was negatively correlated with caspase-3 and positively associated with Bcl-2, VEGF, Ki-67 labeling index (LI), PCNA LI and P53 (all p<0.0001), respectively. Conclusions: DcR3 may play a crucial role as an oncogene in tumorigenesis, deterioration and progress of BUC via influencing related pathways of apoptosis, proliferation and angiogenesis. The detection of DcR3 protein in the formalinfixed and paraffin-embedded samples could assist to predict in prognosis of BUC patients.

A Novel Crawler Based on Loginning Simulation for Weibo Social Network

Ling Xing,Ling Jiang,Bao Peng 보안공학연구지원센터 2016 International Journal of Grid and Distributed Comp Vol.9 No.1

With the rapid development of Weibo, which is the most popular microblog in china, more and more attention was paid to relative studies about it. With the objective of gathering precise information data from Weibo, which is the groundwork of these researches, a novel high efficient Weibo crawler (WCrawler) based on loginning simulation is designed. The priority evaluation is described to ensure the correlation between entires. MD5 is introduced to check for duplicates of URL crawled. Experiments demonstrate that the novel crawler has an efficiency and integrity of information collecting compared with API crawler. In addition, we present a summary of the data that collected from Weibo social network by WCrawler.

Construction and Differential Screening of a cDNA Library Specific to Osmotic Stress of Haloxylon ammodendron Seedlings

Jiang, Xiao-Cheng,Guo, Xin-Hong,Pan, Xiao-Ling,Song, Song-Quan Korean Society for Biochemistry and Molecular Biol 2004 Journal of biochemistry and molecular biology Vol.37 No.5

A subtracted cDNA library specific to osmotic stress of Haloxylon ammodendron (Mey.) Bge seedlings was constructed by suppression subtractive hybridization (SSH) and T/A cloning. SSH was performed between two groups of H. ammodendron seedlings, one was cultivated in Hoagland (H) solution as a driver and the other group was treated with osmotic stress of the Hoagland solution by the addition of 400 mM mannitol (M), as a tester. The library consisted of about 400 recombinant clones, with the average size being of 500 bp, ranging from 300 bp to 1500 bp. Using a PCR-select differential screening kit, 100 recombinant clones were randomly chosen from the subtracted cDNA library and hybridized with forward,reverse subtracted and unsubtracted probes for two rounds. As a result, 21 positive clones specific to osmotic stress were obtained and some of them were verified by Northern blot analysis. The sequencing analysis of 6 positive clones and the following homology comparison to GenBank [blastx] non-redundant databases characterized that two sequences obtained in this experiment may contribute to novel drought-related genes.

High-sensitivity detection for cantharidin by ratiometric fluorescent sensor based on molecularly imprinted nanoparticles of quantum dots

Jiang Ling,Wenqi Zhang,Zijia Cheng,Yanjun Ding 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.112 No.-

Cantharidin (CTD), an effective component of Chinese herbal medicine with unique efficacy, caused poisoningor death constantly for incorrect use. Therefore, there is an intense need for a reliable, selective,sensitive and low-costing analytical method for monitoring the concentration of CTD in biological samples. In this work, a ratiometric fluorescent sensor (CdTe@MIPs/CDs@NIPs) was successfully constructedfor selective determination of CTD by simply integrated the non-imprinted polymers coated CDs(CDs@NIPs) with cadmium telluride quantum dots coated with molecular imprinted polymers(CdTe@MIPs). CDs@NIPs were directly used as reference signal without addition of CTD templates andthe as-prepared CdTe@MIPs were response signal. With the CTD addition, the fluorescence intensity ofCdTe@MIPs decreased, while the fluorescence of CDs@NIPs remained approximately unchanged, resultinga detection limit as low as 0.15 nM. By spiked human blood samples, the ratiometric sensor was successfullyapplied to CTD detection with good recoveries of 96.12–107.40% and relative standard deviation(RSD) of 2.87–3.96%. Moreover, after centrifuging and re-eluting, the developed sensor could be effectivelyrecycled and reused five times. Significantly, the proposed platform with low-cost and highsensitivityopens a door towards the practical applications of ratiometric sensor and shows immensepotential in pursuing the sensitive, reliable and low-cost determination of poisons in forensic toxicologyanalysis.

Pokemon Inhibits Transforming Growth Factor β-Smad4-Related Cell Proliferation Arrest in Breast Cancer through Specificity Protein 1

Ling Chen,Jing Zhong,Jiang-Hua Liu,Duan-Fang Liao,Ying-Ying Shen,Xiao-Lin Zhong,Xiao Xiao,Wen-Jun Ding,Xiu-Da Peng,Wei Xiong,Xu-Yu Zu 한국유방암학회 2019 Journal of breast cancer Vol.22 No.1

Purpose: Pokemon, also known as ZBTB7A, belongs to the POZ and Krüppel (POK) family of transcription repressors and is implicated in tumor progression as a key proto-oncogene. This present study aimed at determining the mechanism by which Pokemon inhibits transforming growth factor β (TGFβ)-Smad4 pathway-dependent proliferation arrest of breast cancer cells via specificity protein 1 (SP1). Methods: Over-expressing plasmid or small interfering RNA (siRNA) transfection was used to regulate Pokemon levels. The EdU incorporation assay, MTS assay, and clone formation were used to identify the inhibitory effect of Pokemon siRNA on cell proliferation. Quantitative real-time polymerase chain reaction assay confirmed that Pokemon deletion inhibited the expression of proliferation-associated genes. The dual-luciferase reporter assay, electrophoretic mobility shift assay, and co-immunoprecipitation assay were used to analyze binding between Pokemon, Smad4, and SP1. Results: Pokemon deletion induced proliferation arrest of breast cancer cells and inhibited the expression of proliferation-associated genes, especially Smad4. Pokemon bound with SP1 to interdict Smad4 promoter activity. Information on clinical samples was obtained from The Cancer Genome Atlas data, in which the Pokemon mRNA levels showed a negative correlation with Smad4 levels in different subtypes of breast cancer in two independent datasets. Conclusion: We demonstrated that Pokemon binds to SP1 to down-regulate Smad4 expression, thereby promoting proliferation of breast cancer cells. This suggests that Pokemon is a potential TGFβ-signaling participant in breast cancer progression.

Tentative identification of 20(S)-protopanaxadiol metabolites in human plasma and urine using ultra-performance liquid chromatography coupled with triple quadrupole time-of-flight mass spectrometry

Ling, Jin,Yu, Yingjia,Long, Jiakun,Li, Yan,Jiang, Jiebing,Wang, Liping,Xu, Changjiang,Duan, Gengli The Korean Society of Ginseng 2019 Journal of Ginseng Research Vol.43 No.4

Background: 20(S)-Protopanaxadiol (PPD), the aglycone part of 20(S)-protopanaxadiol ginsenosides, possesses antidepressant activity among many other pharmacological activities. It is currently undergoing clinical trial in China as an antidepressant. Methods: In this study, an ultra-performance liquid chromatography coupled with triple quadrupole time-of-flight mass tandem mass spectrometry method was established to identify the metabolites of PPD in human plasma and urine following oral administration in phase IIa clinical trial. Results: A total of 40 metabolites in human plasma and urine were identified using this method. Four metabolites identified were isolated from rat feces, and two of them were analyzed by NMR to elucidate the exact structures. The structures of isolated compounds were confirmed as (20S,24S)-epoxydammarane-12,23,25-triol-3-one and (20S,24S)-epoxydammarane-3,12,23,25-tetrol. Both compounds were found as metabolites in human for the first time. Upon comparing our findings with the findings of the in vitro study of PPD metabolism in human liver microsomes and human hepatocytes, metabolites with m/z 475.3783 and phase II metabolites were not found in our study whereas metabolites with m/z 505.3530, 523.3641, and 525.3788 were exclusively detected in our experiments. Conclusion: The metabolites identified using ultra-performance liquid chromatography coupled with triple quadrupole time-of-flight mass spectrometry in our study were mostly hydroxylated metabolites. This indicated that PPD was metabolized in human body mainly through phase I hepatic metabolism. The main metabolites are in 20,24-oxide form with multiple hydroxylation sites. Finally, the metabolic pathways of PPD in vivo (human) were proposed based on structural analysis.

Does HBV Infection Increase Risk of Endometrial Carcinoma?

Jiang, Xue-Feng,Tang, Qiong-Lan,Zou, Yuan,Xu, Ling,Zeng, Hua,Chi, Cong,Jiang, Jing-Ru,Zhang, Bin-Zhong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.2

Objective: Connections between chronic inflammation and tumor development and progression are now generally accepted. Recent evidence indicates that hepatitis B is associated with several types of cancer, but whether endometrial carcinoma (EC) is included has not been reported. Methods: We analyzed HBV serum marker status in 398 patients with endometrial cancer, comparing them to 788 control women undergoing health examination. Results: The total prevalence of HBsAg tested positive in cancer group was significantly higher than the control group (12.8% vs 6.0%, P=0.001), while positive HBsAb was significantly lower (41.2% vs 68.5%, P=0.001). Hepatitis B carriers in endometrial cancer group were also more frequent than in the control group (9.3% vs 5.5%, P=0.013). Interestingly, in the endometrial cancer group, 147 cases were HBV serum marker negative, which was also higher than in the control group (36.9% vs 15.6%, P=0.001). Conclusion: There may be a correlation between HBV infection and endometrial carcinoma.

Metabolic engineering of Corynebacterium glutamicum for increasing the production of L-ornithine by increasing NADPH availability.

Jiang, Ling-Yan,Zhang, Yuan-Yuan,Li, Zhen,Liu, Jian-Zhong Published by Stockton Press on behalf of the Socie 2013 Journal of industrial microbiology & biotechnology Vol.40 No.10

<P>The experiments presented here were based on the conclusions of our previous proteomic analysis. Increasing the availability of glutamate by overexpression of the genes encoding enzymes in the L-ornithine biosynthesis pathway upstream of glutamate and disruption of speE, which encodes spermidine synthase, improved L-ornithine production by Corynebacterium glutamicum. Production of L-ornithine requires 2 moles of NADPH per mole of L-ornithine. Thus, the effect of NADPH availability on L-ornithine production was also investigated. Expression of Clostridium acetobutylicum gapC, which encodes NADP-dependent glyceraldehyde-3-phosphate dehydrogenase, and Bacillus subtilis rocG, which encodes NAD-dependent glutamate dehydrogenase, led to an increase of L-ornithine concentration caused by greater availability of NADPH. Quantitative real-time PCR analysis demonstrates that the increased levels of NADPH resulted from the expression of the gapC or rocG gene rather than that of genes (gnd, icd, and ppnK) involved in NADPH biosynthesis. The resulting strain, C. glutamicum δAPRE::rocG, produced 14.84 g l?1 of L-ornithine. This strategy of overexpression of gapC and rocG will be useful for improving production of target compounds using NADPH as reducing equivalent within their synthetic pathways.</P>

Improved kinetics of LiNi_1/3Mn_1/3Co_1/3O₂ cathode material through reduced graphene oxide networks

Jiang, Ke-Cheng,Xin, Sen,Lee, Jong-Sook,Kim, Jaekook,Xiao, Xiao-Ling,Guo, Yu-Guo The Royal Society of Chemistry 2012 Physical chemistry chemical physics Vol.14 No.8

<P>An electronically conducting 3D network of reduced graphene oxide (RGO) was introduced into LiNi<SUB>1/3</SUB>Mn<SUB>1/3</SUB>Co<SUB>1/3</SUB>O<SUB>2</SUB> (LNMC) cathode material in a special nano/micro hierarchical structure. The rate test and cycling measurement showed that the hierarchical networks remarkably improve the high rate performance of LNMC electrode for lithium-ion batteries. The effect of RGO conducting networks on kinetic property was investigated by electrochemical impedance spectroscopy (EIS) and potentiostatic intermittent titration (PITT). The EIS results reveal that the RGO network greatly decreases the resistance of lithium batteries, especially the charge transfer resistance which can be attributed to the significantly improved conducting networks. The enhancement of apparent diffusion coefficient by the RGO conducting networks is shown by PITT. The power performance was found to be limited by the electrical conduction in the two-phase region, which can be greatly facilitated by the hierarchical RGO network together with carbon black. The as-obtained LNMC/RGO cathode exhibits an outstanding electrochemical property supporting the design idea of electronically conducting 3D networks for the high-energy and high-power lithium-ion batteries.</P> <P>Graphic Abstract</P><P>A 3D conducting network of reduced graphene oxide nanosheets is introduced into LiNi<SUB>1/3</SUB>Mn<SUB>1/3</SUB>Co<SUB>1/3</SUB>O<SUB>2</SUB> and is found to significantly promote the kinetics for Li storage. <IMG SRC='http://pubs.rsc.org/services/images/RSCpubs.ePlatform.Service.FreeContent.ImageService.svc/ImageService/image/GA?id=c2cp23363k'> </P>