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Construction of lentiviral vector carrying Rab9 gene and its expression in mouse brain

Youguo HAO, Min ZHANG, Jinzhi XU, Bitao BU, Jiajun WEI

《医学前沿（英文）》 2009年 第3卷 第2期   页码 141-147 doi: 10.1007/s11684-009-0041-6

摘要： Rab proteins and their effectors facilitate vesicular transport by tethering donor vesicles to their respective target membranes. Rab9 mediates late endosome-to- -Golgi-network trafficking. To explore the possibility of Rab9-related gene therapy for neurodegenerative diseases, we packed Lentivirus encoding Rab9. The expressing plasmid pCDH1-MCF1-Rab9-EF1-copGFP was constructed by using molecular biological techniques. The Lentivirus encoding Rab9 cDNA was packed by Lifectamine-2000 mediated co-transfection of the plasmid pPACKH1- , pPACKH1- and pVSV- into 293T cells. DNA sequencing proved the successful construction of pCDH1-MCF1-Rab9-EF1-copGFP. After 72 hours, the expression of GFP could be detected in BV-2 cells. Western blotting revealed that the Rab9 gene expression in BALB/c mice brain was up-regulated significantly 4 weeks after injection with Lentivirus encoding Rab9, which evidenced a satisfactory increasing effect of this virus. Administration of Lenti-Rab9 to postnatal day 3 Niemann-Pick disease type C (NPC) mice reduced motor defects and prevented the weight loss associated with female NPC mice, as well as modulating the death rate of Purkinje neurons. It is concluded that the packaging of Lentivirus encoding Rab9 was successful. Lentivirus encoding Rab9 can increase the expression of Rab9 gene effectively, which might offer a novel means for the treatment of neurodegenerative diseases.

关键词： Rab9     lentivirus     gene therapy     gene transfer    

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Acetylated HOXB9 at lysine 27 is of differential diagnostic value in patients with pancreatic ductal

Xiaoran Sun, Jiagui Song, Jing Zhang, Jun Zhan, Weigang Fang, Hongquan Zhang

《医学前沿（英文）》 2020年 第14卷 第1期   页码 91-100 doi: 10.1007/s11684-019-0696-6

摘要： Pancreatic ductal adenocarcinoma (PDAC) is the ninth most common human malignancy and the sixth leading cause of cancer-related death in China. AcK27-HOXB9 is a newly identified HOXB9 post-transcriptional modification that can predict the outcome in lung adenocarcinoma and colon cancer well. However, the role of AcK27-HOXB9 in PDAC is unclear. The present study aims to investigate the differential diagnostic role of patients with AcK27-HOXB9 PDAC. Tissue microarrays consisting of 162 pancreatic tumor tissue samples from patients with PDAC and paired normal subjects were used to examine HOXB9 and AcK27-HOXB9 levels and localizations by immunohistochemical analysis and Western blot assay, respectively. HOXB9 was upregulated ( <0.0001), and AcK27-HOXB9 ( =0.0023) was downregulated in patients with PDAC. HOXB9 promoted ( =0.0115), while AcK27-HOXB9 ( =0.0279) inhibited PDAC progression. AcK27-HOXB9 predicted favorable outcome in patients with PDAC ( =0.0412). AcK27-HOXB9 also suppressed PDAC cell migration in a cell migration assay. The results of this study showed that HOXB9 promoted and AcK27-HOXB9 suppressed PDAC progression. The determination of ratio between HOXB9 and AcK27-HOXB9 exhibited potential diagnostic value in patients with PDAC.

关键词： HOXB9     AcK27-HOXB9     PDAC    

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662 A/G gene variation in human tumor necrosis factor receptor superfamily, member 9 (TNFRSF9)

QU Yanchun, YANG Ze, SUN Liang, JI Linong

《医学前沿（英文）》 2008年 第2卷 第3期   页码 283-285 doi: 10.1007/s11684-008-0053-7

摘要： The aim of this paper is to report a new coding variance of the gene, a candidate for autoimmune diseases. We found the variation in two families with type 2 diabetes mellitus by D-HPLC mutation screening method and confirmed our results by direct sequencing and PCR-RFLP. Although without changing the amino acid coding, the variance may have an effect on codon usage and play a role in disease development, such as type 2 diabetes mellitus. However, we cannot define the role of this variance because the frequency of the minor allele is low in the Chinese population and no homozygote of the variance was found. More research in multiple populations will be necessary to define the role of this variance.

关键词： D-HPLC mutation     development     autoimmune     PCR-RFLP     candidate    

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The 9th China Engineering Management Forum Guangzhou Consensus

《工程管理前沿（英文）》 2015年 第2卷 第2期   页码 105-107 doi: 10.15302/J-FEM-2015019

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Mobile CRISPR-Cas9 based anti-phage system in E. coli

《化学科学与工程前沿（英文）》 2022年 第16卷 第8期   页码 1281-1289 doi: 10.1007/s11705-022-2141-7

摘要： Escherichia coli is one of the most important microbial cell factories, but infection by bacteriophages in the environment may have a huge impact on its application in industrial production. Here, we developed a mobile CRISPR-Cas9 based anti-phage system for bacteriophages defense in E. coli. Two conjugative plasmids pGM1 (phosphoglucomutase 1) and pGM2 carrying one and two guide RNAs, respectively, were designed to defend against a filamentous phage. The results showed that the pGM1 and pGM2 could decrease the phage infection rate to 1.6% and 0.2% respectively in infected cells. For preventing phage infection in E. coli, the pGM2 decreased the phage infection rate to 0.1%, while pGM1 failed to block phage infection. Sequence verification revealed that point mutations in protospacer or protospacer adjacent motif sequences of the phage genome caused loss of the defense function. These results support the potential application of MCBAS in E. coli cell factories to defend against phage infections.

关键词： phage infections     anti-phage     CRISPR-Cas9     conjugative transfer     synthetic biology    

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REDUCTION OF NON-POINT SOURCE POLLUTION IN THE YONG’AN RIVER BY CONSTRUCTED WETLAND BASED ON 9 YEARS

《农业科学与工程前沿（英文）》 2023年 第10卷 第4期   页码 627-638 doi: 10.15302/J-FASE-2023516

摘要：

The agricultural and livestock activities surrounding the rivers flowing into the lakes have caused non-point source pollution, leading to excessive amounts of nutrient salts in downstream rivers. Introducing river water into constructed wetlands along river course has proven to be an effective solution for decreasing nitrogen (N) and phosphorus (P) loads. This paper reports 9 years of monitoring the Yong’an River and its surrounding constructed wetlands in the upper reaches of Erhai Lake, located in Yunnan Province, China. This study analyzed the main types of pollutants in the river, and evaluated the removal efficiency of pollutants by the constructed wetlands. The findings indicate that total nitrogen (TN) and nitrate nitrogen (NO3–-N) are the primary pollutants in the Yong’an River, which exhibit variation throughout the year corresponding to the alternating wet and dry seasons. Although constructed wetlands are effective in removing NO3–-N and P, their efficacy in removing ammonium nitrogen (NH4+-N) and organic pollutants is limited. This limitation can be attributed to the lack of timely disposal of aquatic plant residues. This research contributes to the understanding of the potential issues that may arise during the extended use of constructed wetlands and provides solutions to address them.

关键词： inflowing rivers     surface-flow constructed wetland     nutrients     long-term monitoring    

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CRISPR-Cas9 mediated LAG-3 disruption in CAR-T cells

null

《医学前沿（英文）》 2017年 第11卷 第4期   页码 554-562 doi: 10.1007/s11684-017-0543-6

摘要：

T cells engineered with chimeric antigen receptor (CAR) have been successfully applied to treat advanced refractory B cell malignancy. However, many challenges remain in extending its application toward the treatment of solid tumors. The immunosuppressive nature of tumor microenvironment is considered one of the key factors limiting CAR-T efficacy. One negative regulator of T cell activity is lymphocyte activation gene-3 (LAG-3). We successfully generated LAG-3 knockout T and CAR-T cells with high efficiency using CRISPR-Cas9 mediated gene editing and found that the viability and immune phenotype were not dramatically changed during in vitro culture. LAG-3 knockout CAR-T cells displayed robust antigen-specific antitumor activity in cell culture and in murine xenograft model, which is comparable to standard CAR-T cells. Our study demonstrates an efficient approach to silence immune checkpoint in CAR-T cells via gene editing.

关键词： CAR-T     CRISPR-Cas9     LAG-3    

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Compiling of comprehensive data of human infections with novel influenza A (H7N9) virus

null

《医学前沿（英文）》 2013年 第7卷 第3期   页码 275-276 doi: 10.1007/s11684-013-0285-z

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Research progress on human infection with avian influenza H7N9

Xiaoxin Wu, Lanlan Xiao, Lanjuan Li

《医学前沿（英文）》 2020年 第14卷 第1期   页码 8-20 doi: 10.1007/s11684-020-0739-z

摘要： Since the first case of novel H7N9 infection was reported, China has experienced five epidemics of H7N9. During the fifth wave, a highly pathogenic H7N9 strain emerged. Meanwhile, the H7N9 virus continues to accumulate mutations, and its affinity for the human respiratory epithelial sialic acid 2-6 receptor has increased. Therefore, a pandemic is still possible. In the past 6 years, we have accumulated rich experience in dealing with H7N9, especially in terms of virus tracing, epidemiological research, key site mutation monitoring, critical disease mechanisms, clinical treatment, and vaccine development. In the research fields above, significant progress has been made to effectively control the spread of the epidemic and reduce the fatality rate. To fully document the research progress concerning H7N9, we reviewed the clinical and epidemiological characteristics of H7N9, the key gene mutations of the virus, and H7N9 vaccine, thus providing a scientific basis for further monitoring and prevention of H7N9 influenza epidemics.

关键词： H7N9     pandemic     epidemiology     mutations     vaccine     influenza    

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Frequency domain a9ctive vibration control of a flexible plate based on neural networks

Jinxin LIU, Xuefeng CHEN, Zhengjia HE

《机械工程前沿（英文）》 2013年 第8卷 第2期   页码 109-117 doi: 10.1007/s11465-013-0252-z

摘要：

A neural-network (NN)-based active control system was proposed to reduce the low frequency noise radiation of the simply supported flexible plate. Feedback control system was built, in which neural network controller (NNC) and neural network identifier (NNI) were applied. Multi-frequency control in frequency domain was achieved by simulation through the NN-based control systems. A pre-testing experiment of the control system on a real simply supported plate was conducted. The NN-based control algorithm was shown to perform effectively. These works lay a solid foundation for the active vibration control of mechanical structures.

关键词： active vibration control (AVC)     neural network (NN)     low frequency noise     frequency domain control     multi-frequency control    

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Zr66Al9Cu16Ni9块体非晶合金制备工艺研究

阮许平,阮宏伟

《中国工程科学》 2004年 第6卷 第5期   页码 76-78

摘要：

运用真空电弧炉熔炼合金制备试样，采用三种不同的方法对金属熔体进行处理，试样的X射线衍射晶化相分析结果表明，B2O3和Na2SiO3的混合物与所选组分的熔体发生了反应，不适合作Zr基合金的包裹物；组分为CaF2(45%)，AlF3(50%)，CaO(5%)以及微量的TiO2的氟盐，由于该净化剂与所选的组分发生了反应，不适合Zr基合金的净化；设备的高真空度、原材料成分的高纯度是促进合金形成非晶态的重要因素。

关键词： 块体非晶合金     制备工艺     玻璃     熔盐     真空度    

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The first avian influenza A (H7N9) viral infection in humans in Zhejiang Province, China: a death report

null

《医学前沿（英文）》 2013年 第7卷 第3期   页码 333-344 doi: 10.1007/s11684-013-0275-1

摘要：

This study reports the first death caused by a novel avian influenza A (H7N9) virus in Zhejiang Province, China. The patient had chronic hepatitis B and history of exposure to poultry. The patient initially complained of diarrhea and influenza-like symptoms on March 7 and 14 respectively. The disease progressed to severe pneumonia, sustained hypoxia, and coagulation abnormalities. The patient died on March 27 because of respiratory failure, multiple organ failure, and disseminated intravascular coagulation without oseltamivir treatment. This H7N9 virus from Zhejiang is highly similar to isolates obtained from Shanghai, Jiangsu, Anhui, etc. Analysis of hemagglutinin, neuramidinase, and matrix genes indicated that the isolates share the same avian origin, have low virulence, and are sensitive to oseltamivir, but are resistant to adamantine. Only the isolate that caused the fatality exhibited substitution of Q226I in the HA gene, which indicates a potentially enhanced human affinity. The secondary transmission rate was 1.6% (2/125). Only two health workers presented with influenza-like symptoms, and they subsequently tested negative for H7N9 RNA. In conclusion, underlying disease, late diagnosis, and untimely antiviral treatment are possible high-risk factors for infections and death caused by the low-pathogenicity avian influenza A (H7N9). Person-to-person transmission of the H7N9 virus was not detected among close contacts, but such transmission should be investigated in the future. Expanding and enhancing surveillance will help in the early discovery and diagnosis of suspected cases, which will reduce the number of severe cases and deaths.

关键词： avian influenza A (H7N9) virus     epidemiology     contacts     person-to-person transmission    

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Serum carbohydrate antigen (CA) 19-9 as a prognostic factor in cholangiocarcinoma: A meta-analysis

Shang-Long LIU, Zi-Fang SONG, Qing-Gang HU, Shao-Bo HU, Jun LI, Qi-Chang ZHENG, Duo SHAN,

《医学前沿（英文）》 2010年 第4卷 第4期   页码 457-462 doi: 10.1007/s11684-010-0240-1

摘要： This study was performed to determine the prognostic role of preoperative serum carbohydrate antigen (CA) 19-9 levels in the survival of patients with cholangiocarcinoma. Articles published up to June 1, 2010 that evaluated preoperative CA19-9 levels and the prognosis of cholangiocarcinoma were collected for meta-analysis. The required information for calculating individual relative risk (RR) was extracted from the studies, and a combined overall RR was estimated. Nine eligible studies were included. One study dealt with extra-hepatic cholangiocarcinoma, while the other eight studies analyzed intra-hepatic cholangiocarcinoma. The mean methodological quality score was 74.1%, ranging from 65.5% to 82.5%. The overall RR for the nine studies was 1.28 (95% confidence interval= 1.10–1.46), and the Z-score for overall effect was 13.83 (<0.001). The association between serum CA19-9 level and lymph node involvement was also assessed. The combined RR was 1.471 (95% confidence interval= 0.411–5.264) and Z-score for overall effect was 0.59 ( = 0.553). CA19-9 levels were associated significantly with the prognosis of patients with cholangiocarcinoma. This meta-analysis shows that elevation of preoperative CA19-9 levels is correlated with a poor prognosis of patients with cholangiocarcinoma. However, larger scale and randomized studies are needed to draw a more substantive conclusion.

关键词： cholangiocarcinoma     prognostic factors     serum carbohydrate antigen 19-9    

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Characterization of polyclonal antibodies against nonstructural protein 9 from the porcine reproductive

Mengmeng ZHAO,Juanjuan QIAN,Jiexiong XIE,Tiantian CUI,Songling FENG,Guoqiang WANG,Ruining WANG,Guihong ZHANG

《农业科学与工程前沿（英文）》 2016年 第3卷 第2期   页码 153-160 doi: 10.15302/J-FASE-2016097

摘要： Porcine reproductive and respiratory syndrome (PRRS) is considered to be one of the most important infectious diseases impacting the swine industry and is characterized by reproductive failure in late term gestation in sows and respiratory disease in pigs of all ages. The nonstructural protein 9 gene, , encoding the RNA-dependent RNA polymerase, is generally regarded as fairly conserved when compared to other viral proteins. Antibodies against 9 will be of great importance for the diagnosis and treatment of the causal agent, PRRS virus. A study was undertaken to generate polyclonal antibodies against the immunodominant 9. For this purpose, the 9 was expressed in and subsequently used as an antigen to immunize New Zealand rabbits. Antiserum was identified via an indirect ELISA, and then verified based on the ability to react with both naturally and artificially expressed 9. Results of virus neutralization test showed that this antiserum could not neutralize the PRRSV. Nevertheless, this antiserum as a diagnostic core reagent should prove invaluable for further investigations into the mechanism of PRRS pathogenesis.

关键词： PRRSV     Nsp9     expression     antibody     neutralize    

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Lymphatic metastasis is related to the epithelial-mesenchymal transition and expressions of VEGF, MMP-9,

Lihui WANG, Lianhong LI, Shen LV, Shujun FAN, Li ZHAN, Bo WANG, Zhong ZHANG

《医学前沿（英文）》 2009年 第3卷 第2期   页码 164-170 doi: 10.1007/s11684-009-0038-1

摘要： The invasion and metastasis of breast cancer are supposed to involve several stages in which epithelial-mesenchymal transition (EMT) is regarded as the mechanistic basis for the behavior of cancer cells. A series of factors related to EMT are apparently involved in such process. The current study aimed to investigate the contributions of EMT and related factors in lymph node metastasis of breast cancer. The expressions of E-cadherin (E-Cad), N-cadherin (N-Cad), vascular endothelial cell growth factor (VEGF), matrix metalloproteinase-9 (MMP-9), cyclooxygenase-2 (COX-2), and CD34 were examined in 74 cases of breast cancer, including 39 cases with lymph node metastasis and 35 cases without lymph node metastasis by immunohistochemistry. Multivariable Cox proportional hazards model was used to analyze the patients’ prognosis. The expressions of N-Cad, VEGF, MMP-9, and COX-2 in cases with lymph node metastasis were significantly higher than those without lymph node metastasis ( <0.05), while the E-Cad level was inversely related to status of lymph node metastasis ( <0.05). The metastasis rate of lymph node in the cases with EMT (lower E-Cad expression and higher N-Cad expression) was 78.3%, while that without EMT (higher E-Cad expression and lower N-Cad expression) was 11.1%. There was a statistical difference in the expression of COX-2 protein between histological grade I and grade II or III, respectively ( <0.05). In the cases with higher grade, the expression of E-Cad was decreased, while that of N-Cad was increased. Higher microvascular density (MVD) was also found to be significantly associated with lymphatic metastasis ( <0.05), and the cases with higher MVD had shorter survival time. This study indicates that EMT and expressions of VEGF, MMP-9 and COX-2, and MVD value are strongly correlated with lymph node metastasis in breast cancer.

关键词： epithelial-mesenchymal transition     vascular endothelial cell growth factor     matrix metalloproteinase-9     cyclooxygenase-2     higher microvascular density     breast cancer    

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标题 作者 时间 类型 操作

Construction of lentiviral vector carrying Rab9 gene and its expression in mouse brain

Youguo HAO, Min ZHANG, Jinzhi XU, Bitao BU, Jiajun WEI

期刊论文

Acetylated HOXB9 at lysine 27 is of differential diagnostic value in patients with pancreatic ductal

Xiaoran Sun, Jiagui Song, Jing Zhang, Jun Zhan, Weigang Fang, Hongquan Zhang

期刊论文

662 A/G gene variation in human tumor necrosis factor receptor superfamily, member 9 (TNFRSF9)

QU Yanchun, YANG Ze, SUN Liang, JI Linong

期刊论文

The 9th China Engineering Management Forum Guangzhou Consensus

期刊论文

Mobile CRISPR-Cas9 based anti-phage system in E. coli

期刊论文

REDUCTION OF NON-POINT SOURCE POLLUTION IN THE YONG’AN RIVER BY CONSTRUCTED WETLAND BASED ON 9 YEARS

期刊论文

CRISPR-Cas9 mediated LAG-3 disruption in CAR-T cells

null

期刊论文

Compiling of comprehensive data of human infections with novel influenza A (H7N9) virus

null

期刊论文

Research progress on human infection with avian influenza H7N9

Xiaoxin Wu, Lanlan Xiao, Lanjuan Li

期刊论文

Frequency domain a9ctive vibration control of a flexible plate based on neural networks

Jinxin LIU, Xuefeng CHEN, Zhengjia HE

期刊论文

Zr66Al9Cu16Ni9块体非晶合金制备工艺研究

阮许平,阮宏伟

期刊论文

The first avian influenza A (H7N9) viral infection in humans in Zhejiang Province, China: a death report

null

期刊论文

Serum carbohydrate antigen (CA) 19-9 as a prognostic factor in cholangiocarcinoma: A meta-analysis

Shang-Long LIU, Zi-Fang SONG, Qing-Gang HU, Shao-Bo HU, Jun LI, Qi-Chang ZHENG, Duo SHAN,

期刊论文

Characterization of polyclonal antibodies against nonstructural protein 9 from the porcine reproductive

Mengmeng ZHAO,Juanjuan QIAN,Jiexiong XIE,Tiantian CUI,Songling FENG,Guoqiang WANG,Ruining WANG,Guihong ZHANG

期刊论文

Lymphatic metastasis is related to the epithelial-mesenchymal transition and expressions of VEGF, MMP-9,

Lihui WANG, Lianhong LI, Shen LV, Shujun FAN, Li ZHAN, Bo WANG, Zhong ZHANG

期刊论文