Molecular identification of DNA barcoding of Leguminous toxic species and quantitative analysis by ELISA kits

Wang Jie,Wang Shuangyu,Sun Fenglin,Liu Chang,Zhao Jinquan,Yu Hongwei,Lv Xiaojing,Liu Ze,Bu Shuhua,Yu Weisen 한국식물생명공학회 2024 Plant biotechnology reports Vol.18 No.2

Some edible Leguminous are toxic when raw, and the Chinese are particularly fond of beans, so Leguminous poisoning is very common in China. Rapid and accurate identification of poisoned species and determination of their toxic components would better assist physicians in treating patients. However, traditional morphology-based identification methods possess many limitations. DNA barcoding technique is a new species identification technique developed in recent years, which is expected to make up for the shortcomings of traditional morphological identification. In this study, a comprehensive evaluation system based on DNA barcoding and ELISA kits was attempted. A total of 30 Leguminous toxic plants were collected, involving 9 genera and 10 species. We used simulated gastric fluid (SGF) to simulate the human gastric environment. Three markers (rbcL, trnH-psbA, and ITS) were amplified and sequenced for all untreated and 15 mock-digested samples. The validity of DNA barcoding for species identification was assessed using the Basic Local Alignment Search Tool (BLAST) method and the tree construction method. The levels of three toxic components (saponin, phytoagglutin and trasylol) were determined in all samples using ELISA kits. The amplification success rate of all three regions was high (rbcL 96.67%, trnH-psbA 100%, and ITS 100%), but the sequencing of the trnH-psbA region was less satisfactory (66.67%), and SGF had a significant impact on the sequencing of the ITS region (After 40 min of SGF treatment, the sequencing success rate decreased by 46.67%). The samples from different species and origins contained different levels of toxic components, and the levels of all three sub- stances decreased significantly after undergoing SGF digestion. After 1 h of SGF treatment, the saponin content decreased to 0–8.60% in untreated content (PHA decreased to 8.62–36.88%, trasylol decreased to 4.70–47.06%). The current results suggest that DNA barcoding has great potential for rapid identification of Leguminous poisoning in clinical settings. Toxins are probably not detectable in the patient for longer periods of poisoning. We recommend DNA barcoding technology as a first step for rapid screening and combined with toxin analysis for clinical diagnosis.

Selective Adsorption and Reusability for Pb<SUP>2+</SUP> of Chitosan-based Microporous Polymer

Jie Yu,Jidong Zheng,Quanfang Lu,Xing Wang,Xiaomin Zhang,Qizhao Wang,Wu Yang 한국고분자학회 2017 폴리머 Vol.41 No.3

A green and simple glow-discharge electrolysis plasma (GDEP) technique was used to synthesize the chitosan/attapulgite/poly(acrylic acid-co-2-acrylamido-2-methyl-1-propanesulfonic acid) (CS/ATP/P(AA-co-AMPS)) microporous polymer. The results showed that the optimum pH for the adsorption of Pb<SUP>2+</SUP>, Cd<SUP>2+</SUP>, Co<SUP>2+</SUP>, and Cu<SUP>2+</SUP> is 4.8, and time of adsorption equilibrium is 60 min. Adsorption kinetics fits well to the pseudo-second-order model. The maximum adsorption capacities for Pb<SUP>2+</SUP>, Cd<SUP>2+</SUP>, Cu<SUP>2+</SUP> and Co<SUP>2+</SUP> are 500.0, 301.7, 180.0 and 151.7 mg g<SUP>-1</SUP>, respectively. The microporous polymer has higher adsorption selectivity toward Pb<SUP>2+</SUP> with the coexistence of Cd<SUP>2+</SUP>, Co<SUP>2+</SUP>, and Cu<SUP>2+</SUP>. The CS/ATP/P(AAco-AMPS) possesses promising regeneration and reusability using 2.0 g L–1 ethylenediamine tetraacetic acid tetrasodium salt (EDTA-4Na) solution as eluent. CS/ATP/P(AA-co-AMPS) can be used as a very promising absorbent for the separation purification and selective recovery of Pb<SUP>2+</SUP> in aqueous solution containing Cd<SUP>2+</SUP>, Co<SUP>2+</SUP>, and Cu<SUP>2+</SUP> ions.

Preliminary Proteomic Analysis of Indomethacin’s Effect on TumorTransplanted with Colorectal Cancer Cell in Nude Mice

Yu-Jie Wang,Hong-Mei Wang,Gui-Ying Zhang,Zhi-Qiang Xiao,Zhu-Chu Chen 한국생화학분자생물학회 2006 BMB Reports Vol.39 No.2

Blood-Brain Barrier Disruption and Perivascular Beta-Amyloid Accumulation in the Brain of Aged Rats with Spontaneous Hypertension: Evaluation with Dynamic Contrast-Enhanced Magnetic Resonance Imaging

Yu Wang,Ruzhi Zhang,Chuanyuan Tao,Ziqian Xu,Wei Chen,Chunhua Wang,Li Song,Jie Zheng,Fabao Gao 대한영상의학회 2018 Korean Journal of Radiology Vol.19 No.3

Objective: Whether blood-brain barrier (BBB) disruption induced by chronic spontaneous hypertension is associated with beta-amyloid (Aβ) accumulation in the brain remains poorly understood. The purpose of this study was to investigate the relationship between BBB disruption and Aβ influx and accumulation in the brain of aged rats with chronic spontaneous hypertension. Materials and Methods: Five aged spontaneously hypertensive rats (SHRs) and five age-matched normotensive Wistar-Kyoto (WKY) rats were studied. The volume transfer constant (Ktrans) obtained from dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was used to evaluate BBB permeability in the hippocampus and cortex in vivo. The BBB tight junctions, immunoglobulin G (IgG), Aβ, and amyloid precursor protein (APP) in the hippocampus and cortex were examined with immunohistochemistry. Results: As compared with WKY rats, the Ktrans values in the hippocampus and cortex of the SHRs increased remarkably (0.316 ± 0.027 min-1 vs. 0.084 ± 0.017 min-1, p < 0.001 for hippocampus; 0.302 ± 0.072 min-1 vs. 0.052 ± 0.047 min-1, p < 0.001 for cortex). Dramatic occludin and zonula occludens-1 losses were detected in the hippocampus and cortex of SHRs, and obvious IgG exudation was found there. Dramatic Aβ accumulation was found and limited to the area surrounding the BBB, without extension to other parenchyma regions in the hippocampus and cortex of aged SHRs. Alternatively, differences in APP expression in the hippocampus and cortex were not significant. Conclusion: Blood-brain barrier disruption is associated with Aβ influx and accumulation in the brain of aged rats with chronic spontaneous hypertension. DCE-MRI can be used as an effective method to investigated BBB damage.

Genetic diversity and population structure of the amylolytic yeast Saccharomycopsis fibuligera associated with Baijiu fermentation in China

Wang Ju-Wei,Han Pei-Jie,Han Da-Yong,Zhou Sen,Li Kuan,He Peng-Yu,Zhen Pan,Yu Hui-Xin,Liang Zhen-Rong,Wang Xue-Wei,Bai Feng-Yan 한국미생물학회 2021 The journal of microbiology Vol.59 No.8

The amylolytic yeast Saccharomycopsis fibuligera is a predominant species in starters and the early fermentation stage of Chinese liquor (Baijiu). However, the genetic diversity of the species remains largely unknown. Here we sequenced the genomes of 97 S. fibuligera strains from different Chinese Baijiu companies. The genetic diversity and population structure of the strains were analyzed based on 1,133 orthologous genes and the whole genome single nucleotide polymorphisms (SNPs). Four main lineages were recognized. One lineage contains 60 Chinese strains which are exclusively homozygous with relatively small genome sizes (18.55–18.72 Mb) and low sequence diversity. The strains clustered in the other three lineages are heterozygous with larger genomes (21.85–23.72 Mb) and higher sequence diversity. The genomes of the homozygous strains showed nearly 100% coverage with the genome of the reference strain KPH12 and the sub-genome A of the hybrid strain KJJ81 at the above 98% sequence identity level. The genomes of the heterozygous strains showed nearly 80% coverage with both the sub-genome A and the whole genome of KJJ81, suggesting that the Chinese heterozygous strains are also hybrids with nearly 20% genomes from an unidentified source. Eighty-three genes were found to show significant copy number variation between different lineages. However, remarkable lineage specific variations in glucoamylase and α-amylase activities and growth profiles in different carbon sources and under different environmental conditions were not observed, though strains exhibiting relatively high glucoamylase activity were mainly found from the homozygous lineage.

Metabolism and excretion of novel pulmonary-targeting docetaxel liposome in rabbits

Wang, Jie,Zhang, Li,Wang, Lijuan,Liu, Zhonghong,Yu, Yu The Korean Society of Pharmacology 2017 The Korean Journal of Physiology & Pharmacology Vol.21 No.1

Our study aims to determine the metabolism and excretion of novel pulmonary-targeting docetaxel liposome (DTX-LP) using the in vitro and in vivo animal experimental models. The metabolism and excretion of DTX-LP and intravenous DTX (DTX-IN) in New Zealand rabbits were determined with ultra-performance liquid chromatography tandem mass spectrometry. We found DTX-LP and DTX-IN were similarly degraded in vitro by liver homogenates and microsomes, but not metabolized by lung homogenates. Ultra-performance liquid chromatography tandem mass spectrometry identified two shared DTX metabolites. The unconfirmed metabolite $M_{un}$ differed structurally from all DTX metabolites identified to date. DTX-LP likewise had a similar in vivo metabolism to DTX-IN. Conversely, DTX-LP showed significantly diminished excretion in rabbit feces or urine, approximately halving the cumulative excretion rates compared to DTX-IN. Liposomal delivery of DTX did not alter the in vitro or in vivo drug metabolism. Delayed excretion of pulmonary-targeting DTX-LP may greatly enhance the therapeutic efficacy and reduce the systemic toxicity in the chemotherapy of non-small cell lung cancer. The identification of $M_{un}$ may further suggest an alternative species-specific metabolic pathway.

Dissecting Causal Relationships Between Gut Microbiota, Blood Metabolites, and Stroke: A Mendelian Randomization Study

Wang Qi,Dai Huajie,Hou Tianzhichao,Hou Yanan,Wang Tiange,Lin Hong,Zhao Zhiyun,Li Mian,Zheng Ruizhi,Wang Shuangyuan,Lu Jieli,Xu Yu,Liu Ruixin,Ning Guang,Wang Weiqing,Bi Yufang,Zheng Jie,Xu Min 대한뇌졸중학회 2023 Journal of stroke Vol.25 No.3

Background and Purpose We investigated the causal relationships between the gut microbiota (GM), stroke, and potential metabolite mediators using Mendelian randomization (MR). Methods We leveraged the summary statistics of GM (n=18,340 in the MiBioGen consortium), blood metabolites (n=115,078 in the UK Biobank), and stroke (cases n=60,176 and controls n=1,310,725 in the Global Biobank Meta-Analysis Initiative) from the largest genome-wide association studies to date. We performed bidirectional MR analyses to explore the causal relationships between the GM and stroke, and two mediation analyses, two-step MR and multivariable MR, to discover potential mediating metabolites. Results Ten taxa were causally associated with stroke, and stroke led to changes in 27 taxa. In the two-step MR, <i>Bifidobacteriales</i> order, <i>Bifidobacteriaceae</i> family, <i>Desulfovibrio</i> genus, apolipoprotein A1 (ApoA1), phospholipids in high-density lipoprotein (HDL_PL), and the ratio of apolipoprotein B to ApoA1 (ApoB/ApoA1) were causally associated with stroke (all <i>P</i><0.044). The causal associations between <i>Bifidobacteriales</i> order, <i>Bifidobacteriaceae</i> family and stroke were validated using the weighted median method in an independent cohort. The three GM taxa were all positively associated with ApoA1 and HDL_PL, whereas <i>Desulfovibrio</i> genus was negatively associated with ApoB/ApoA1 (all <i>P</i><0.010). Additionally, the causal associations between the three GM taxa and ApoA1 remained significant after correcting for the false discovery rate (all q-values <0.027). Multivariable MR showed that the associations between <i>Bifidobacteriales</i> order, <i>Bifidobacteriaceae</i> family and stroke were mediated by ApoA1 and HDL_PL, each accounting for 6.5% (<i>P</i>=0.028) and 4.6% (<i>P</i>=0.033); the association between <i>Desulfovibrio</i> genus and stroke was mediated by ApoA1, HDL_PL, and ApoB/ApoA1, with mediated proportions of 7.6% (<i>P</i>=0.019), 4.2% (<i>P</i>=0.035), and 9.1% (<i>P</i>=0.013), respectively. Conclusion The current MR study provides evidence supporting the causal relationships between several specific GM taxa and stroke and potential mediating metabolites.

Intravenous Tenecteplase for Acute Ischemic Stroke Within 4.5–24 Hours of Onset (ROSE-TNK): A Phase 2, Randomized, Multicenter Study

Wang Lu,Dai Ying-Jie,Cui Yu,Zhang Hong,Jiang Chang-Hao,Duan Ying-Jie,Zhao Yong,Feng Ye-Fang,Geng Shi-Mei,Zhang Zai-Hui,Lu Jiang,Zhang Ping,Zhao Li-Wei,Zhao Hang,Ma Yu-Tong,Song Cheng-Guang,Zhang Yi,Ch 대한뇌졸중학회 2023 Journal of stroke Vol.25 No.3

Background and Purpose Intravenous tenecteplase (TNK) efficacy has not been well demonstrated in acute ischemic stroke (AIS) beyond 4.5 hours after onset. This study aimed to determine the effect of intravenous TNK for AIS within 4.5 to 24 hours of onset. Methods In this pilot trial, eligible AIS patients with diffusion-weighted imaging (DWI)-fluid attenuated inversion recovery (FLAIR) mismatch were randomly allocated to intravenous TNK (0.25 mg/kg) or standard care within 4.5–24 hours of onset. The primary endpoint was excellent functional outcome at 90 days (modified Rankin Scale [mRS] score of 0–1). The primary safety endpoint was symptomatic intracranial hemorrhage (sICH). Results Of the randomly assigned 80 patients, the primary endpoint occurred in 52.5% (21/40) of TNK group and 50.0% (20/40) of control group, with no significant difference (unadjusted odds ratio, 1.11; 95% confidence interval 0.46–2.66; <i>P</i>=0.82). More early neurological improvement occurred in TNK group than in control group (11 vs. 3, <i>P</i>=0.03), but no significant differences were found in other secondary endpoints, such as mRS 0–2 at 90 days, shift analysis of mRS at 90 days, and change in National Institutes of Health Stroke Scale score at 24 hours and 7 days. There were no cases of sICH in this trial; however, asymptomatic intracranial hemorrhage occurred in 3 of the 40 patients (7.5%) in the TNK group. Conclusion This phase 2, randomized, multicenter study suggests that intravenous TNK within 4.5–24 hours of onset may be safe and feasible in AIS patients with a DWI-FLAIR mismatch.

Clinical Application of Transcatheter Arterial Chemoembolization Combined with Synchronous C-arm Cone-Beam CT Guided Radiofrequency Ablation in treatment of Large Hepatocellular Carcinoma

Wang, Zhi-Jun,Wang, Mao-Qiang,Duan, Feng,Song, Peng,Liu, Feng-Yong,Wang, Yan,Yan, Jie-Yu,Li, Kai,Yuan, Kai Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.3

Objective: This work aimed to evaluate the safety and clinical efficacy of transcatheter arterial chemoembolization (TACE) combined with c-arm cone-beam CT guided synchronous radiofrequency ablation (RFA) in treatment of large hepatocellular carcinoma (HCC). Methods: 21 patients with large HCC were studied from January 2010 to March 2012. TACE combined with synchronous C-arm cone-beam CT guided RFA were performed on a total of 25 lesions. Conventional imaging examination (CEUS, enhanced CT or MRI) and AFP detection were regularly conducted to evaluate the technical success rate of combined treatment, complications, treatment response, time without disease recurrence and survival rate. Results: The technical success rate of combined treatment was 100%, without any significant complication. After 1 month, there were 19 cases with complete response and 2 cases with partial response, with an complete response rate of 90.4% (19/21) and a clinical effective rate of 100% (21/21). The complete response rates of single nodular lesions (100%, 17/17) was significantly higher than that of multiple nodular lesions (50%, 2/4) (P<0. 05). During 2 to 28 months of follow-up, in 19 cases with complete response, the average time without disease recurrence was $10.8{\pm}6$ months. The total survival rates of 6, 12 and 18 months in 21 patients were 100%, respectively. Conclusion: TACE combined with synchronous C-arm CT guided RFA is safe and effective for treatment of large HCC. The treatment efficacy for single nodular lesion is better than that for multiple nodular lesions.

Temperature dependent development of black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae) from Yangtze River Delta region of China

Wang Yinghui,Zhang Yanan,Wang Jiangfeng,Kang Chengtao,Hu Gengwang,Guo Yi,Chen Jie,Yang Lijun,Wang Yu 한국응용곤충학회 2023 Journal of Asia-Pacific Entomology Vol.26 No.4

The black soldier fly, Hermetia illucens (L.) (Diptera: Stratiomyidae), isomnivorous andwidely distributed in the world. Black soldier flies usually colonize corpse at a relatively late stage and have a longer developmental time compared to other flies. It can serve as a good supplementary indicator for estimating a longer minimum postmortem interval (PMI min ) in forensic investigations. In this paper, the development of H. illucens was investigated at six constant temperatures between 19 and 34 ◦ C. Under temperatures of 19, 22, 25, 28, 31, and 34 ◦ C, the mean developmental time from eggs to adults was 90.96, 58.29, 54.61, 42.33, 32.33 and 34.52 days, respectively. Developmental time and accumulated degree days results were used to construct thermal sum mation model and isomorphen diagram. The calculated values of lower developmental thresholds and thermal summation constant estimated by a linear model were 11.88 ◦ C and 650.89 degree days, respectively. Through a nonlinear model, the upper lethal developmental threshold temperature, intrinsic optimum temperature and lower developmental thresholds were estimated to be 35.18, 20.50 and 13.17 ◦ C, respectively. A growth curve, an equation and isomegalen diagram based on them show the relationship between developmental time and larval body length. The larval body length increased with increased developmental time and eventually pla teaued, revealing an S-shape growth curve. These results provide important basic developmental data for H. illucens, which can be used to estimate the PMI min .