Luciferase Assay to Screen Tumour-specific Promoters in Lung Cancer

Xu, Rong,Guo, Long-Jiang,Xin, Jun,Li, Wen-Mao,Gao, Yan,Zheng, You-Xian,Guo, You-Hong,Lin, Yang-Jun,Xie, Yong-Hua,Wu, Ya-Qing,Xu, Rui-An Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11

Objective: Specific promoters could improve efficiency and ensure the safety of gene therapy. The aim of our study was to screen examples for lung cancer. Methods: The firefly luciferase gene was used as a reporter, and promoters based on serum markers of lung cancer were cloned. The activity and specificity of seven promoters, comprising CEACAM5 (carcinoembryonic antigen, CEA), GRP (Gastrin-Releasing Peptide), KRT19 (cytokeratin 19, KRT), SFTPB (surfactant protein B, SP-B), SERPINB3 (Squamous Cell Carcinoma Antigen, SCCA), SELP (Selectin P, Granule Membrane Protein 140kDa, Antigen CD62, GMP) and DKK1 (Dickkopf-1) promoters were compared in lung cancer cells to obtain cancer-specific examples with strong activity. Results: The CEACAM5, DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB promoters were cloned. Furthermore, we successfully constructed recombinant vector pGL-CEACAM5 (DKK1, GRP, SELP, KRT19, SERPINB3 and SFTPB) contained the target gene. After cells were transfectedwith recombinant plasmids, we found that the order of promoter activity from high to low was SERPINB3, DKK1, SFTPB, KRT19, CEACAM5, SELP and GRP and the order for promoters regarding specificity and high potential were SERPINB3, DKK1, SELP, SFTPB, CEACAM5, KRT19 and GRP. Conclusion: The approach adopted is feasible to screen for new tumour specific promoters with biomarkers. In addition, the screened lung-specific promoters might have potential for use in lung cancer targeted gene therapy research.

miR-181b as a Potential Molecular Target for Anticancer Therapy of Gastric Neoplasms

Guo, Jian-Xin,Tao, Qing-Song,Lou, Peng-Rong,Chen, Xiao-Chun,Chen, Jun,Yuan, Guang-Bo Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.5

Objective: MicroRNAs (miRNAs) play important roles in carcinogenesis. The aim of the present study was to explore the effects of miR-181b on gastric cancer. Methods: The expression level of miR-181b was quantified by qRT-PCR. MTT, flow cytometry and matrigel invasion assays were used to test proliferation, apoptosis and invasion of miR-181b stable transfected gastric cancer cells. Results: miR-181b was aberrantly overexpressed in gastric cancer cells and primary gastric cancer tissues. Further experiments demonstrated inducible expression of miR-181b by Helicobacter pylori treatment. Cell proliferation, migration and invasion in the gastric cancer cells were significantly increased after miR-181b transfection and apoptotic cells were also increased. Furthermore, overexpression of miR-181b downregulated the protein level of tissue inhibitor of metalloproteinase 3 (TIMP3). Conclusion: The upregulation of miR-181b may play an important role in the progress of gastric cancer and miR-181b maybe a potential molecular target for anticancer therapeutics of gastric cancer.

Acute Normovolemic Hemodilution Effects on Perioperative Coagulation in Elderly Patients Undergoing Hepatic Carcinectomy

Guo, Jian-Rong,Jin, Xiao-Ju,Yu, Jun,Xu, Feng,Zhang, Yi-Wei,Shen, Hua-Chun,Shao, Yi Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.8

Background: Acute normovolemic hemodilution (ANH) has been widely used to prevent the massive blood loss during hepatic carcinoma. The influences of ANH on coagulation function are still controversy, especially in elderly patients. The study observed ANH effects on coagulation function and fibrinolysis in elderly patients undergoing the disease. Materials and Methods: Thirty elderly patients (aged 60-70 yr) with liver cancer (ASA I or II) taken hepatic carcinectomy from February 2007 to February 2008 were randomly divided into ANH group (n=15) and control group (n=15). After tracheal intubation, patients in ANH group and control group were infused with 6% hydroxyethyl starch (130/0.4) and Ringer's solution, respectively. Blood samples were drawn from patients in both groups at five different time points: before anesthesia induction (T1), 30 min after ANH (T2), 1 h after start of operation (T3), immediately after operation (T4), and 24 h after operation (T5). Then coagulation function, soluble fibrin monomer complex (SFMC), prothrombin fragment (F1+2), and platelet membrane glycoprotein (CD62P and activated GP IIb/GP IIIa) were measured. Results: The perioperative blood loss and allogeneic blood transfusion were recorded during the surgery. The perioperative blood loss was not significantly different between two groups (p>0.05), but the volume of allogeneic blood transfusion in ANH group was significantly less than in control group ($350.0{\pm}70.7$) mL vs. ($457.0{\pm}181.3$) mL (p<0.01). Compared with the data of T1, the prothrombin time (PT) and activated partial thromboplastin time (APTT) measured after T3 were significantly longer (p<0.05) in both groups, but within normal range. There were no significant changes of thrombin time (TT) and D-dimer between two groups at different time points (p>0.05). SFMC and F1+2 increased in both groups, but were not statistically significant. PAC-1-positive cells and CD62P expressions in patients of ANH group were significantly lower than those at T1 (p<0.05) and T2-T5 (p>0.05). Conclusions: ANH has no obvious impact on fibrinolysis and coagulation function in elderly patients undergoing resection of liver cancer. The study suggested that ANH is safe to use in elderly patients and it could reduce allogeneic blood transfusion.

Prevalence of Spina Bifida Occulta and Its Relationship With Overactive Bladder in Middle-Aged and Elderly Chinese People

Jun Wei Wu,Yu Rong Xing,Yi Bo Wen,Tian Fang Li,Quan De Feng,Xiao Ping Shang,Yun Long Li,Jin Jin Feng,Xin Xin Wang,Rong Qun Zhai,Xiang Fei He,Tao Chen,Xin Jian Liu,Jian Guo Wen 대한배뇨장애요실금학회 2016 International Neurourology Journal Vol.20 No.2

Purpose: To investigate the prevalence of spina bifida occulta (SBO) and its relationship with the presence of overactive bladder (OAB) in middle-aged and elderly people in China. Methods: A cross-sectional community-based survey was carried out at 7 communities in Zhengzhou City, China from December 15, 2013 to June 10, 2014, where residents aged over 40 years were randomly selected to participate. All of the participants underwent lumbosacral radiographic analysis and relevant laboratory tests. A questionnaire including basic information, past medical history and present illness, and the OAB symptom score was filled out by all participants. Chi-square tests and logistic regression were used for data analysis with a P-value of <0.05 denoting statistical significance. Results: A total of 1,061 subjects were qualified for the final statistical analysis (58.8±11.7 years; male, 471 [44.4%]; female, 590 [55.6%]). The overall prevalence of SBO was 15.1% (160 of 1,061): 18.3% (86 of 471) in men and 12.5% (74 of 590) in women. Among these subjects, 13.7% (145 of 1,061) had OAB: 13.2% (62 of 471) in men and 14.1% (83 of 590) in women. The results of logistic regression showed that age, SBO, history of cerebral infarction (HCI), and constipation were risk factors for OAB (P<0.05), while sex, history of childhood enuresis (HCE), body mass index (BMI), and diabetes mellitus (DM) were not (P>0.05). In men, age, SBO, and constipation were risk factors for OAB (P<0.05), while HCE, BMI, DM, HCI, and benign prostate hyperplasia were not (P>0.05). In women, age, SBO, and HCI were risk factors for OAB (P<0.05), while HCE, BMI, DM, vaginal delivery, and constipation were not (P>0.05). Conclusions: The prevalence of SBO is high and it is related to OAB in middle-aged and elderly people in China.

Effects of Stellate Ganglion Block on the Peri-operative Vasomotor Cytokine Content and Intrapulmonary Shunt in Patients with Esophagus Cancer

Guo, Wei,Jin, Xiao-Ju,Yu, Jun,Liu, Yang,Zhang, Jian-Ping,Yang, Da-Wei,Zhang, Lei,Guo, Jiang-Rong Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.21

Objective: To investigate the effects of stellate ganglion block (SGB) on the peri-operative vasomotor cytokine content and intrapulmonary shunt in patients with esophagus cancer who underwent thoracotomy. Materials and Methods: Forty patients undergoing elective resection of esophageal cancer patients who had I~II American Society of Anesthesiologist (ASA) were randomly divided into total intravenous anesthesia group (group N, n=20) and total intravenous anesthesia combined with SGB group (group S, n=20, 0.12 mL/kg 1% lidocaine was used for SGB 10 min before induction). Heart rate (HR), mean arterial pressure (MAP), central venous pressure (CVP), mean pulmonary arterial pressure (MPAP) and continuous cardiac output (CCO) were continuously monitored. The blood from internal jugular vein was drawn respectively before induction ($T_0$), and 30 min ($T_1$), 60 min ($T_2$) and 120 min ($T_3$) after one-lung ventilation (OLV), and 30 min (T4) after two-lung ventilation. The contents of plasma endothelin (ET), nitric oxide (NO) and calcitonin gene-related peptide (CGRP) were detected with enzyme linked immunosorbent assay (ELISA). Meanwhile, arterial and mixed venous blood samples were collected for determination of blood gas and calculation of intrapulmonary shunt fraction (Qs/Qt). Results: During OLV, ET contents were increased significantly in two groups (P<0.05), and no significant difference was presented (P>0.05). NO content in group S was obviously higher than in group N at T3 (P<0.05), whereas CGRP content in group N was markedly lower than in group S at each time point (P<0.05). Qs/Qt was significantly increased in both groups after OLV, but there was no statistical significant regarding the Qs/Qt at each time point between two groups. Conclusions: Total intravenous anesthesia combined with SGB is conducive to regulation of perioperative vasomotor cytokines in thoracotomy, and has little effect on intrapulmonary shunt at the time of OLV.

Effects of Low Light Intensity on Photosynthetic Capacity, Photosynthetic Nitrogen Utilization Efficiency and Nitrogen Partition in Non-Heading Chinese Cabbage Leaves

Jun Huang,Zhen Wu,Shi-Rong Guo 한국원예학회 2006 한국원예학회 기타간행물 Vol.- No.-

Three Dimensional Reconstruction of Bone-Cartilage Transitional Structures Based on Semi-Automatic Registration and Automatic Segmentation of Serial Sections

Hua Guo,Zheng-Wei Xu,Bao-Rong He,Ding-Jun Hao,Wei-Guo Bian 한국조직공학과 재생의학회 2014 조직공학과 재생의학 Vol.11 No.5

A well-defined three-dimensional (3-D) reconstruction of bone-cartilage transitional structures is crucial for the osteochondral restoration. This paper presents an accurate, computationally efficient and semi-automated algorithm for the alignment and segmentation of two-dimensional (2-D) serial to construct the 3-D model of bonecartilage transitional structures. Entire system includes the following five components: (1) image harvest, (2) image registration, (3) image segmentation, (4) 3-D reconstruction and visualization, and (5) evaluation. A computer program was developed in the environment of Matlab for the semi-automatic alignment and automatic segmentation of serial sections. Semi-automatic alignment algorithm based on the position’s cross-correlation of the anatomical characteristic feature points of two sequential sections. A method combining an automatic segmentation and an image threshold processing was applied to capture the regions and structures of interest. SEM micrograph and 3-D model reconstructed directly in digital microscope were used to evaluate the reliability and accuracy of this strategy. The morphology of 3-D model constructed by serial sections is consistent with the results of SEM micrograph and 3-D model of digital microscope.

Changes in Milk Production and Metabolic Parameters by Feeding Lactating Cows Based on Different Ratios of Corn Silage: Alfalfa Hay with Addition of Extruded Soybeans

Yana, Rong,Zhang, Ruizhong,Zhang, Xian,Jiang, Chao,Han, Jian-Guo,Zhang, Ying-Jun Asian Australasian Association of Animal Productio 2011 Animal Bioscience Vol.24 No.6

The objective of this study was to investigate the effects of different ratios of corn silage (CS): alfalfa hay (AH), and extruded soybeans (ESB) on milk yield, milk composition, blood metabolites, and fatty acids in milk fat and plasma. Ninety multiparous Holstein cows were arranged in a randomized block design experiment which lasted 14 weeks. Treatments were arranged as a $3{\times}3$ factorial with 0%, 5% or 10% ESB (dry matter basis) and three forage treatments: I) 30% CS, 10% AH and 10% Leymus chinense hay (LC); ii) 20% corn silage, 20% alfalfa hay and 10% LC; iii) 10% CS, 30% AH and 10% LC. Cows were allowed to consume a total mixed ration ad libitum. There was no change of dry matter intake when cows were fed the experimental diets. As more AH was added to the diets, milk yield, milk protein content and yield, and trans9, cis11-conjugated linoleic acids (CLA) concentrations in milk fat and plasma increased. When ESB were supplemented to the diets, milk yield, and trans9, cis11-CLA concentration in milk fat and plasma increased. When 10% ESB was added to the diet containing 30% AH the trans9, cis11-CLA content (1.46 g/100 g of total fatty acids) in milk was the highest among all treatments. These results suggests that AH could replace part of a CS diet and be a good forage source of diet for dairy cows to improve milk yield and milk composition. Meanwhile, ESB could be included in the diet with high AH to improve production performance of dairy cows.

Cryptotanshinone Induces Inhibition of Breast Tumor Growth by Cytotoxic CD4+ T Cells through the JAK2/STAT4/ Perforin Pathway

Zhou, Jun,Xu, Xiao-Zhen,Hu, Yao-Ren,Hu, Ai-Rong,Zhu, Cheng-Liang,Gao, Guo-Sheng Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.6

Cryptotanshinone (CPT), is a quinoid diterpene isolated from the root of the Asian medicinal plant, Salvia miotiorrhiza bunge. Numerous researchers have found that it could work as a potent antitumor agent to inhibit tumor growth in vitro, buith there has been much less emphasis on its in vivo role against breast tumors. Using a mouse tumor model of MCF7 cells, we showed that CPT strongly inhibited MCF7 cell growth in vivo with polarization of immune reactions toward Th1-type responses, stimulation of naive CD4+ T cell proliferation, and also increased IFN-${\gamma}$ and perforin production of CD4+ T cells in response to tumor-activated splenocytes. Furthermore, data revealed that the cytotoxic activity of CD4+ T cells induced by CPT was markedly abrogated by concanamycin A(CMA), a perforin inhibitor, but not IFN-${\gamma}$ Ab. On the other hand, after depletion of CD4+ T cells or blocked perforin with CMA in a tumor-bearing model, CPT could not effectively suppress tumor growth, but this phenomenon could be reversed by injecting naive CD4+ T cells. Thus, our results suggested that CPT mainly inhibited breast tumor growth through inducing cytotoxic CD4+ T cells to secrete perforin. We further found that CPT enhanced perforin production of CD4+ T cells by up-regulating JAK2 and STAT4 phosphorylation. These findings suggest a novel potential therapeutic role for CPT in tumor therapy, and demonstrate that CPT performs its antitumor functions through cytotoxic CD4+ T cells.

Microbiological Purification of L-Arabitol from Xylitol Mother Liquor

( Ming Guo Jiang ),( Ben Wang ),( Li Fang Yang ),( Shuang Jun Lin ),( Hai Rong Cheng ) 한국미생물 · 생명공학회 2011 Journal of microbiology and biotechnology Vol.21 No.1

As a rare sugar alcohol, L-arabitol can be used in food and can prevent extra fat deposits in the intestinal tract. Commercially, L-arabitol is prepared from pure L-arabinose by hydrogenation, which needs a high temperature and high pressure, leading to a high production cost for Larabitol. Therefore, this study describes a novel L-arabitol production method based on biological purification from the xylitol mother liquor, a cheap and readily available raw material that contains a high concentration of Larabitol. First, a novel Bacillus megaterium strain was screened that can utilize xylitol, sorbitol, and mannitol, yet not L-arabitol. The isolated strain was inoculated into a medium containing the xylitol mother liquor under formulated culture conditions, where a high L-arabitol yield (95%) and high purity (80%) were obtained when the medium was supplemented with 50 g/l of xylitol mother liquor. Upon further purification of the fermentation broth by ion exchange and decolorization, L-arabitol was crystallized with a purity of 98.5%.