Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

Yu-Guo Yuan,Shao-Zheng Song,Meng-Ming Zhu,Zheng-Yi He,Rui Lu,Ting Zhang,Fei Mi,Jin-Yu Wang,Yong Cheng 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.8

Objective: To create genetically modified goat as a biopharming source of recombinant human lacotoferrin (hLF) with transcription activator-like effector nucleases. Methods: TALENs and targeting vector were transferred into cultured fibroblasts to insert hLF cDNA in the goat beta-lactoglobulin (BLG) locus with homology-directed repair. The gene targeted efficiency was checked using sequencing and TE7I assay. The bi-allelic gene targeted colonies were isolated and confirmed with polymerase chain reaction, and used as donor cells for somatic cell nuclear transfer (SCNT). Results: The targeted efficiency for BLG gene was approximately 10%. Among 12 Bi-allelic gene targeted colonies, five were used in first round SCNT and 4 recipients (23%) were confirmed pregnant at 30 d. In second round SCNT, 7 (53%), 4 (31%), and 3 (23%) recipients were confirmed to be pregnant by ultrasound on 30 d, 60 d, and 90 d. Conclusion: This finding signifies the combined use of TALENs and SCNT can generate bi-allelic knock-in fibroblasts that can be cloned in a fetus. Therefore, it might lay the foundation for transgenic hLF goat generation and possible use of their mammary gland as a bioreactor for large-scale production of recombinant hLF.

A Prediction Model Based on the Risk Factors Associated with Pathological Upgrading in Patients with Early-Stage Gastric Neoplasms Diagnosed by Endoscopic Forceps Biopsy

Zhao Yu Han,Zheng Yu,Sha Jie,Hua Hong Jin,Li Ke Dong,Lu Yu,Dang Yi Ni,Zhang Guo Xin 거트앤리버 소화기연관학회협의회 2023 Gut and Liver Vol.17 No.1

Background/Aims: The discrepancies between the diagnosis of preoperative endoscopic forceps biopsy (EFB) and endoscopic submucosal dissection (ESD) in patients with early gastric neoplasm (EGN) exist objectively. Among them, pathological upgrading directly influences the accuracy and appropriateness of clinical decisions. The aims of this study were to investigate the risk factors for the discrepancies, with a particular focus on pathological upgrading and to establish a prediction model for estimating the risk of pathological upgrading after EFB. Methods: We retrospectively collected the records of 978 patients who underwent ESD from December 1, 2017 to July 31, 2021 and who had a final histopathology determination of EGN. A nomogram to predict the risk of pathological upgrading was constructed after analyzing subgroup differences among the 901 lesions enrolled. Results: The ratio of pathological upgrading was 510 of 953 (53.5%). Clinical, laboratorial and endoscopic characteristics were analyzed using univariable and binary multivariable logistic regression analyses. A nomogram was constructed by including age, history of chronic atrophic gastritis, symptoms of digestive system, blood high density lipoprotein concentration, macroscopic type, pathological diagnosis of EFB, uneven surface, remarkable redness, and lesion size. The C-statistics were 0.804 (95% confidence interval, 0.774 to 0.834) and 0.748 (95% confidence interval, 0.664 to 0.832) in the training and validation set, respectively. We also built an online webserver based on the proposed nomogram for convenient clinical use. Conclusions: The clinical value of identifying the preoperative diagnosis of EGN lesions is limited when using EFB separately. We have developed a nomogram that can predict the probability of pathological upgrading with good calibration and discrimination value.

Comparison of Liver Transplantation and Liver Resection for Hepatocellular Carcinoma Patients with Portal Vein Tumor Thrombus Type I and Type II

Jia-Yu Lv,Ning-Ning Zhang,Ya-Wei Du,Ying Wu,Tian-Qiang Song,Ya-Min Zhang,Yan Qu,Yu-Xin Liu,Jie Gu,Ze-Yu Wang,Yi-Bo Qiu,Bing Yang,Da-Zhi Tian,Qing-Jun Guo,Li Zhang,Ji-San Sun,Yan Xie,Zheng-Lu Wang,Xin 연세대학교의과대학 2021 Yonsei medical journal Vol.62 No.1

Purpose: The aim of this study was to compare the efficacy of liver transplantation (LT) and liver resection (LR) for hepatocellularcarcinoma (HCC) patients with portal vein tumor thrombus (PVTT) and to investigate risk factors affecting prognosis. Materials and Methods: A total of 94 HCC patients with PVTT type I (segmental PVTT) and PVTT type II (lobar PVTT) were involvedand divided into LR (n=47) and LT groups (n=47). Recurrence-free survival (RFS) and overall survival (OS) were comparedbefore and after inverse probability of treatment weighting (IPTW). Prognostic factors for RFS and OS were explored. Results: Two treatment groups were well-balanced using IPTW. In the entire cohort, LT provided a better prognosis than LR. Among patients with PVTT type I, RFS was better with LT (p=0.039); OS was not different significantly between LT and LR(p=0.093). In subgroup analysis of PVTT type I patients with α-fetoprotein (AFP) levels >200 ng/mL, LT elicited significantly longermedian RFS (18.0 months vs. 2.1 months, p=0.022) and relatively longer median OS time (23.6 months vs. 9.8 months, p=0.065). Among patients with PVTT type II, no significant differences in RFS and OS were found between LT and LR (p=0.115 and 0.335,respectively). Multivariate analyses showed treatment allocation (LR), tumor size (>5 cm), AFP and aspartate aminotransferase(AST) levels to be risk factors of RFS and treatment allocation (LR), AFP and AST as risk factors for OS. Conclusion: LT appeared to afford a better prognosis for HCC with PVTT type I than LR, especially in patients with AFP levels>200 ng/mL.

The Biotypes, Wing-forms and the Immigration of Brown Planthopper, Nilaparvata lugens Stal, in Zhejiang Province, China

Yu, Xiaoping,Lu, Zhongxian,Wu, Guorui,Tao, Linyong,Chen, Jianming,Zheng, Xusong,Xu, Hongxing Korean Society of Applied Entomology 2001 Journal of Asia-Pacific Entomology Vol.4 No.2

The virulence change (biotype), wing form development and immigration of brown planthopper (BPH) Nilaparvata lugens Stal, population in Zhejiang province was studied as compared to these BPH populations collected from Guangxi and Yunnan province, China and Philippines. The results indicate that the Zhejiang population could successfully infest variety IR26 and Mudgo after 1989, and ASD7 and IR36 after 1998, showing the characters of BPH biotype 2 and biotypes 3. The BPH tropical populations such as Guangxi population, however, adapted the resistant varieties IR26, Mudgo and ASD7 earlier. In 1997, the nymphal survival indices of Guangxi and Yunnan populations on Rathu Heenati (Bph3) were up to 70.9 and 66.7 respectively, higher than Zhejiang population. The Zhejiang populations show low brachupterous rates and have a negative correlation with nymphal density, belonging to the temperate type. The Philippine populations are almost brachypterous, however, the brachypterous rates of BPH males raise with the increase of nymphal density. The Guangxi population has a high brachypterous rate, being similar to the tropical type. On the other hand, tests showed that the lowest survival rate, longer duration of nymphs and the lightest weight of brachypterous female occurred in the $2^{nd}$ generation of BPH after continuously feeding on resistant variety. Physiologically, the highest activities of superoxide dismutase (SOD) and catalase (CAT) were detected in the second generation of the BPH feeding on resistant rice variety.

Biosynthesis of rare 20(R)-protopanaxadiol/protopanaxatriol type ginsenosides through Escherichia coli engineered with uridine diphosphate glycosyltransferase genes

Lu Yu,Yuan Chen,Jie Shi,Ru-Feng Wang,Ying-Bo Yang,Li Yang,Shujuan Zhao,Zheng-Tao Wang 고려인삼학회 2019 Journal of Ginseng Research Vol.43 No.1

Background: Ginsenosides are known as the principal pharmacological active constituents in Panax medicinal plants such as Asian ginseng, American ginseng, and Notoginseng. Some ginsenosides, especially the 20(R) isomers, are found in trace amounts in natural sources and are difficult to chemically synthesize. The present study provides an approach to produce such trace ginsenosides applying biotransformation through Escherichia coli modified with relevant genes. Methods: Seven uridine diphosphate glycosyltransferase (UGT) genes originating from Panax notoginseng, Medicago sativa, and Bacillus subtilis were synthesized or cloned and constructed into pETM6, an ePathBrick vector, which were then introduced into E. coli BL21star (DE3) separately. 20(R)-Protopanaxadiol (PPD), 20(R)-protopanaxatriol (PPT), and 20(R)-type ginsenosides were used as substrates for biotransformation with recombinant E. coli modified with those UGT genes. Results: E. coli engineered with GT95syn selectively transfers a glucose moiety to the C20 hydroxyl of 20(R)-PPD and 20(R)-PPT to produce 20(R)-CK and 20(R)-F1, respectively. GTK1- and GTC1-modified E. coli glycosylated the C3eOH of 20(R)-PPD to form 20(R)-Rh2. Moreover, E. coli containing p2GT95synK1, a recreated two-step glycosylation pathway via the ePathBrich, implemented the successive glycosylation at C20eOH and C3eOH of 20(R)-PPD and yielded 20(R)-F2 in the biotransformation broth. Conclusion: This study demonstrates that rare 20(R)-ginsenosides can be produced through E. coli engineered with UTG genes.

Common genetic determinants of breast-cancer risk in East Asian women: a collaborative study of 23 637 breast cancer cases and 25 579 controls

Zheng, Wei,Zhang, Ben,Cai, Qiuyin,Sung, Hyuna,Michailidou, Kyriaki,Shi, Jiajun,Choi, Ji-Yeob,Long, Jirong,Dennis, Joe,Humphreys, Manjeet K.,Wang, Qin,Lu, Wei,Gao, Yu-Tang,Li, Chun,Cai, Hui,Park, Sue K Oxford University Press 2013 Human Molecular Genetics Vol.22 No.12

<P>In a consortium including 23 637 breast cancer patients and 25 579 controls of East Asian ancestry, we investigated 70 single-nucleotide polymorphisms (SNPs) in 67 independent breast cancer susceptibility loci recently identified by genome-wide association studies (GWASs) conducted primarily in European-ancestry populations. SNPs in 31 loci showed an association with breast cancer risk at <I>P</I> < 0.05 in a direction consistent with that reported previously. Twenty-one of them remained statistically significant after adjusting for multiple comparisons with the Bonferroni-corrected significance level of <0.0015. Eight of the 70 SNPs showed a significantly different association with breast cancer risk by estrogen receptor (ER) status at <I>P</I> < 0.05. With the exception of rs2046210 at 6q25.1, the seven other SNPs showed a stronger association with ER-positive than ER-negative cancer. This study replicated all five genetic risk variants initially identified in Asians and provided evidence for associations of breast cancer risk in the East Asian population with nearly half of the genetic risk variants initially reported in GWASs conducted in European descendants. Taken together, these common genetic risk variants explain ∼10% of excess familial risk of breast cancer in Asian populations.</P>

Prediction of Lung Cancer Based on Serum Biomarkers by Gene Expression Programming Methods

Yu, Zhuang,Chen, Xiao-Zheng,Cui, Lian-Hua,Si, Hong-Zong,Lu, Hai-Jiao,Liu, Shi-Hai Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

In diagnosis of lung cancer, rapid distinction between small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC) tumors is very important. Serum markers, including lactate dehydrogenase (LDH), C-reactive protein (CRP), carcino-embryonic antigen (CEA), neurone specific enolase (NSE) and Cyfra21-1, are reported to reflect lung cancer characteristics. In this study classification of lung tumors was made based on biomarkers (measured in 120 NSCLC and 60 SCLC patients) by setting up optimal biomarker joint models with a powerful computerized tool - gene expression programming (GEP). GEP is a learning algorithm that combines the advantages of genetic programming (GP) and genetic algorithms (GA). It specifically focuses on relationships between variables in sets of data and then builds models to explain these relationships, and has been successfully used in formula finding and function mining. As a basis for defining a GEP environment for SCLC and NSCLC prediction, three explicit predictive models were constructed. CEA and NSE are requentlyused lung cancer markers in clinical trials, CRP, LDH and Cyfra21-1 have significant meaning in lung cancer, basis on CEA and NSE we set up three GEP models-GEP 1(CEA, NSE, Cyfra21-1), GEP2 (CEA, NSE, LDH), GEP3 (CEA, NSE, CRP). The best classification result of GEP gained when CEA, NSE and Cyfra21-1 were combined: 128 of 135 subjects in the training set and 40 of 45 subjects in the test set were classified correctly, the accuracy rate is 94.8% in training set; on collection of samples for testing, the accuracy rate is 88.9%. With GEP2, the accuracy was significantly decreased by 1.5% and 6.6% in training set and test set, in GEP3 was 0.82% and 4.45% respectively. Serum Cyfra21-1 is a useful and sensitive serum biomarker in discriminating between NSCLC and SCLC. GEP modeling is a promising and excellent tool in diagnosis of lung cancer.

Development and Validation of a Prognostic Nomogram Based on Clinical and CT Features for Adverse Outcome Prediction in Patients with COVID-19

Zheng Yingyan,Xiao Anling,Yu Xiangrong,Zhao Yajing,Lu Yiping,Li Xuanxuan,Mei Nan,She Dejun,Wang Dongdong,Geng Daoying,Yin Bo 대한영상의학회 2020 Korean Journal of Radiology Vol.21 No.8

Objective: The purpose of our study was to investigate the predictive abilities of clinical and computed tomography (CT) features for outcome prediction in patients with coronavirus disease (COVID-19). Materials and Methods: The clinical and CT data of 238 patients with laboratory-confirmed COVID-19 in our two hospitals were retrospectively analyzed. One hundred sixty-six patients (103 males; age 43.8 ± 12.3 years) were allocated in the training cohort and 72 patients (38 males; age 45.1 ± 15.8 years) from another independent hospital were assigned in the validation cohort. The primary composite endpoint was admission to an intensive care unit, use of mechanical ventilation, or death. Univariate and multivariate Cox proportional hazard analyses were performed to identify independent predictors. A nomogram was constructed based on the combination of clinical and CT features, and its prognostic performance was externally tested in the validation group. The predictive value of the combined model was compared with models built on the clinical and radiological attributes alone. Results: Overall, 35 infected patients (21.1%) in the training cohort and 10 patients (13.9%) in the validation cohort experienced adverse outcomes. Underlying comorbidity (hazard ratio [HR], 3.35; 95% confidence interval [CI], 1.67–6.71; p < 0.001), lymphocyte count (HR, 0.12; 95% CI, 0.04–0.38; p < 0.001) and crazy-paving sign (HR, 2.15; 95% CI, 1.03–4.48; p = 0.042) were the independent factors. The nomogram displayed a concordance index (C-index) of 0.82 (95% CI, 0.76–0.88), and its prognostic value was confirmed in the validation cohort with a C-index of 0.89 (95% CI, 0.82–0.96). The combined model provided the best performance over the clinical or radiological model (p < 0.050). Conclusion: Underlying comorbidity, lymphocyte count and crazy-paving sign were independent predictors of adverse outcomes. The prognostic nomogram based on the combination of clinical and CT features could be a useful tool for predicting adverse outcomes of patients with COVID-19.

Dynamics of Yeast-like Symbiote and Its Relationship with the Virulence of Brown Planthopper, Nilaparvata lugens Stal, to Resistant Rice Varieties

Lu, Zhong-Xian,Yu, Xiao-Ping,Chen, Jian-Ming,Zheng, Xu-Song,Xu, Hong-Xing,Zhang, Jue-Feng,Chen, Lie-Zhong Korean Society of Applied Entomology 2004 Journal of Asia-Pacific Entomology Vol.7 No.3

Abundance of the yeast-like symbiote (YLS) in brown planthopper (BPH), Nilaparvata lugens Stal, collected from rice fields in China and obtained from International Rice Research Institute (IRRI), was monitored. The role of YLS in nymph survival rate, development and adult reproduction, and activities of transaminases in BPH on different resistant rice varieties were examined. Also, the relationship between YLS and virulence shifts of BPH to resistant rice varieties was discussed. The results indicated that the virulence of BPH populations to resistant rice varieties was strongly related to the abundance of YLS in 5th instar nymphs and brachypterous females of BPH field populations at Nanning in South China and Hangzhou m East China, showing the more abundance of YLS in BPH, the higher virulence of BPH to resistant rice varieties, while no obvious differences in YLS abundance were monitored between three BPH biotypes. Numbers of YLS reduced significantly in female adult of BPH biotype 1 fed on resistant rice varieties IR26, Mudgo and ASD7, compared with that fed on susceptible rice variety TN1. The lowest YLS abundance of BPH from field population was found in 2nd generation successively fed on resistant varieties, and the number of YLS was resumed at 4th generation when the BPH has adapted the resistant varieties. The markedly lower nymphal survival rate, adult fecundity and activities of alanine and aspartic transminase, and significantly longer nymph duration in the three aposymbiotic biotypes than in the symbiotic ones were recorded on both susceptible and resistant rice varieties. However, the much greater differences between aposymbiotic and symbiotic BPH were found on the inadaptable resistant rice varieties than that on the fitness resistant rice varieties. Those results implied that YLS in BPH should playa crucial role in the virulent shift of BPH populations to resistant rice varieties.

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.