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Design of stable adsorbents for selective gold recovery with large capacity and fast adsorption kinetics is of great challenge, but significant for the economy and the environment. Herein, we show the design and preparation of an irreversible amide-linked covalent organic framework (COF) JNU-1 via a building block exchange strategy for efficient recovery of gold. JNU-1 was synthesized through the exchange of 4,4'-biphenyldicarboxaldehyde (BA) in mother COF TzBA consisting of 4,4',4''-(1,3,5-triazine-2,4,6-triyl)trianiline (Tz) and BA with terephthaloyl chloride. The irreversible amide linked JNU-1 gave good stability, unprecedented fast kinetics, excellent selectivity and outstanding adsorption capacity for gold recovery. X-ray photoelectron spectroscopy along with thermodynamic study and quantum mechanics calculation reveals that the excellent performance of JNU-1 for gold recovery results from the formation of hydrogen bonds C(N)-H⋅⋅⋅Cl and coordinate interaction of O and Au. The rational design of irreversible bonds as both inherent linkage and functional groups in COFs is a promising way to prepare stable COFs for diverse applications., (© 2020 Wiley-VCH GmbH.)

Inspired by nature, many functional surfaces have been developed with special structures in biology, chemistry, and materials. Many research studies have been focused on the preparation of surfaces with static structure. Achieving dynamical manipulation of surface structure is desired but still a great challenge. Herein, a polyelectrolyte film capable of regional and reversible changes in the microporous structure is presented. Our proposal is based on the combination of azobenzene (Azo) π-π stacking and electrostatic interaction, which could be affected respectively by ultraviolet (UV) irradiation and water plasticization, to tune the mobility of polyelectrolyte chains. The porous patterns can be obtained after regional ultraviolet irradiation and acid treatment. Owing to the reversibility of Azo π-π stacking and electrostatic interaction, the patterns can be repeatedly created and erased in the polyelectrolyte film made by layer-by-layer (LbL) self-assembly of poly(ethyleneimine)-azo and poly(acrylic acid). Furthermore, through two rounds of porous pattern formation and erasure, different functional species can be loaded separately and confined regionally within the film, showing potential applications in the functional surface. This work highlights the coordination of two noncovalent interactions in thin films for regional and reversible controlling its structure, opening a window for more in-depth development of functional surfaces.

Development of efficient adsorbents for nitrate removal is vital for tackling increasing nitrate contamination. We report a free-radical polymerization strategy to prepare a cationic surfactant-modified covalent organic framework (DhaTab-S) for removing nitrate ions from aqueous solution. DhaTab-S was prepared by grafting a cationic surfactant diallyldimethylammonium chloride solution on vinyl-containing COF (DhaTab-V). The adsorption capacity for nitrate was pH-dependent with the maximum at pH 6, and the adsorption process was largely influenced by ionic strength. The adsorption equilibrium for nitrate on DhaTab-S was reached within 40 minutes and followed pseudo-second-order kinetics. DhaTab-S showed a nitrate adsorption capacity of 108.8 mg g -1 , which is about 15 times that of COF before surfactant modification. The large adsorption capacity makes DhaTab-S a promising candidate for nitrate removal from aqueous media., (© 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Herein, we have reported the facile room-temperature synthesis of a chiral covalent organic framework (CCOF) for the enantioselective adsorption of amino acids. The prepared CCOF provides various stereoscopic interactions with amino acids for highly selective adsorption of their enantiomers., Competing Interests: There are no conflicts to declare., (This journal is © The Royal Society of Chemistry.)

We report a thiol-ene click strategy for the preparation of a novel phenylboronic acid-functionalized covalent organic framework (COF) for selective removal of catechol in aqueous solution. Vinyl-functionalized 2,5-diallyloxyterephthalaldehyde (Da-V) was prepared as a building ligand. Da-V was then condensed with 1,3,5-tris(4-aminophenyl)benzene (Tab) to give a vinyl-functionalized COF DhaTab-V. Subsequently, 4-mercaptophenylboronic acid (4-MPBA) was covalently linked on DhaTab-V via thiol-ene click reaction to give phenylboronic acid-functionalized COF DhaTab-PBA. The adsorption isotherms, energetics and kinetics, and reusability of DhaTab-PBA for the adsorption and removal of catechol from aqueous solution were investigated in detail. This phenylboronic acid-functionalized COF is promising as sorbent for selective removal of catechol from aqueous medium with large adsorption capacity and good reusability.

Gold nanoparticles (AuNPs) without surface capping agents are easily aggregated owing to their high surface energy, leading to an unexpected decrease of their catalytic activity. Herein, we report gold nanoparticles decorated covalent organic framework (COF) as mimetic peroxidase for colorimetric detection of mercury. 1,3,5-Tris-(4-formyl-phenyl)triazine (PT) and 4, 4'-azodianiline (Azo) was employed as the monomers to prepare a novel imine based COF PTAzo under solvothermal conditions. AuNPs are further decorated into the PTAzo to form COF-AuNPs hybrid via a citrate reducing method. The COF-AuNPs show high stability, and exhibits enhanced peroxidase mimetic activity in the presence of Hg 2+ . The Hg 2+ concentration dependent peroxidase mimetic activity of COF-AuNPs enables the development of a sensitive and selective method for detection of Hg 2+ in aqueous solution. The developed method gives good linearity in the range of 5-300 nM with the limit of detection of 0.75 nM. The results show that Hg 2+ -enhanced peroxidase mimetic activity of COF-AuNPs offers great potential for the detection of Hg 2+ in real samples., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Aflatoxin B 1 (AFB 1 ) contamination is one of the most critical global issues in food safety. The high carcinogenic nature necessitates rapid and specific methods for the determination of AFB 1 in foodstuffs at ultratrace levels. Here, we report an enhanced bienzymatic chemiluminescence competitive immunoassay for ultrasensitive and high-throughput determination of AFB 1 . In this assay, protein G was first coated on the wells of a microplate for recognizing the Fc fragment of anti-AFB 1 mAbs to reduce the antibody dosage and guarantee high immunological reaction efficiency. The target AFB 1 competed with glucose oxidase labeled AFB 1 for the limited anti-AFB 1 mAbs in the wells of the microplate. p -Bromophenol was employed as an enhancer to obtain intense and long-lasting chemiluminescence. The utilization of an enhancer and bienzymatic catalysts effectively improved the detection sensitivity. The developed method offered a good linearity over 5 orders of magnitude, a detection limit of 5 pg L -1 , and a relative standard deviation of 1.9% for AFB 1 . The application of the developed method to the analysis of grain samples gave quantitative recoveries from 94.0% to 97.0%. The developed method provides a universal platform for high-throughput, ultrasensitive, and high specific detection of pollutants or nutrients in foods.

Covalent organic framework TpBD was grafted on stainless steel wire with polydopamine as a linker. The fabricated TpBD bonded stainless steel wire was used as the solid-phase microextraction fiber to extract sixteen polycyclic aromatic hydrocarbons (PAHs) for subsequent GC-MS/MS determination in grilled meat samples. The developed method gave the limits of detection (S/N = 3) from 0.02 (pyrene)-1.66 (naphthalene) ng L -1 and enhancement factors from 1069 (naphthalene)-10879 (benz(a)anthracene). The relative standard deviations (RSDs) for intra-day and inter-day study are in the range of 2.6%-8.5% and 4.5%-9.4%, respectively. The fiber-to-fiber RSDs for three parallel prepared fibers were 5.3%-10.0%. One TpBD bonded fiber can stand at least 200 cycles without significant loss of extraction efficiency. The developed method was successfully applied for the determination of trace PAHs in grilled meat samples with recoveries from 85.1% to 102.8%., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Surface-mediated transfection has drawn tremendous interest for gene therapy due to its localized gene delivery characteristic and promising perspective for combination devices in clinical applications. However, a method for the controllable load of genetic agents and tunable transfection efficiency to primary cells remains unsatisfactory. Herein, we present a polymeric spongy film with modification of polydopamine (PDA) for controlling load of plasmid DNAs and improving transfection to primary endothelial cells. We demonstrate that, via wicking action, the loading of DNA into the film is simple, rapid, and highly controllable while easily reaching ∼95 μg/cm 2 by only a one-shot loading process. Meanwhile, PDA endows the spongy films with a very good photothermal conversion capability. Consequently, we obtain an enhanced transfection up to ∼85% to hard-to-transfect primary endothelial cells upon NIR irradiation. Furthermore, we realize a spatial cell transfection through NIR irradiation in the defined area, suggesting a high potential for precise gene therapy. This photothermal spongy film could serve as a robust platform for surface-mediated gene therapy, and extend the paradigm of a light enhanced delivery system.

Background: AT-rich interactive domain-containing protein 1A (ARID1A) is a subunit of the mammary SWI/SNF chromatin remodeling complex and a tumor suppressor protein. The loss of ARID1A been observed in several types of human cancers and associated with poor patient prognosis. Previously, we have reported that ARID1A protein was rapidly ubiquitinated and destructed in gastric cancer cells during DNA damage response. However, the ubiquitin e3 ligase that mediated this process remains unclear., Materials and Methods: The interaction between ARID1A and β-TRCP was verified by co-immunoprecipitation (Co-IP) assay. The degron site of ARID1A protein was analyzed by bioinformatics assay. Short hairpin RNAs (shRNAs) were used to knockdown (KD) gene expression., Results: Here we show that DNA damage promotes ARID1A ubiquitination and subsequent destruction via the ubiquitin E3 ligase complex SCFβ-TRCP. β-TRCP recognizes ARID1A through a canonical degron site (DSGXXS) after its phosphorylation in response to DNA damage. Notably, genetic inactivation of the Ataxia Telangiectasia Mutated (ATM) kinase impaired DNA damage-induced ARID1A destruction., Conclusions: Our studies provide a novel molecular mechanism for the negative regulation of ARID1A by β-TRCP and ATM in DNA damaged gastric cancer cells., Competing Interests: Competing interestsThe authors declare that they have no competing interests.

This Article contains an error in Fig. 1, in which the structures of '(+)-Ac-L-Ta' and 'CTp' are drawn incorrectly. The correct version of Fig. 1 is shown below. The error has not been fixed in the original PDF and HTML versions of the Article.

Objective: To explore the related factors of death from severe heat stroke in Shanghai from 2013 to 2017. Methods: The data of 1 152 patients with severe heat stroke who were divided into survival ( n= 1 037) and death ( n= 115) groups including gender, age and heat stroke type (heat cramp, heat exhaustion, heat apoplexy and the mixed type) were collected from meteorological bureau and case report system for high temperature heat stroke in Shanghai from 2013 to 2017. Meanwhile, the meteorological data of the onset date of severe heat stroke cases were collected, including maximum temperature, minimum temperature, daily temperature, relative humidity, air pressure, precipitation and wind speed. The differences of individual and meteorological factors between the two groups were compared, and multivariate logistic regression model was used to analyze the related factors of death from severe heat stroke. Results: Among 1 152 cases, the mean± SD of age was (56.29±18.95) years old, 843 (73.18%)were male, 962 (83.51%) were in the heat wave period; 322 cases (27.95%) were heat cramp, 170 cases (14.76%) were heat exhaustion, 533 cases (46.27%) were heat apoplexy and 114 cases (9.90%) were the mixed type. Daily average temperature ((32.81±1.99) ℃), daily maximum and minimum temperatures ((38.20±2.24) ℃ and (29.22±1.94) ℃) in survival group were lower than those in death group (all P values<0.001), which were (33.76±1.17) ℃, (39.19±1.31) ℃ and (29.72±1.66) ℃. Daily average relative humidity ((60.36±9.75)%) and daily minimum relative humidity ((41.26±9.71)%) in survival group were higher than those in death group(all P values <0.05), which were (54.59±6.89)% and (35.60±7.24)%. The results of logistic regression analysis suggested that compared with the cases with daily average humidity <60% and a mixed type heat stroke, the death OR (95 %CI ) values of cases with daily average humidity >60%, heat cramp, heat exhaustion and heat apoplexy were 0.31 (0.18,0.54), 0.13 (0.05,0.34), 0.68 (0.58,2.30) and 0.87 (0.48,1.58). Conclusion: The temperature, relative humidity and the type of heat stroke were the main related factors affecting the prognosis of severe heat stroke.

A layer-by-layer approach was developed to prepare 3D covalent organic framework COF-300/silica composites (COF-300@SiO2) with uniform morphology as the stationary phase for high performance liquid chromatography separation of position isomers with high efficiency, selectivity and precision.

Objective: To study the effects of metastasis associated 1 (MTA1) on biological characteristics such as migration, invasion and proliferation of gastric cancer (GC) cells. Methods: pSilencer3.1-MTA1-siRNA vector was used to establish human gastric cancer BGC-823 cell lines with constitutive MTA1-knockdown. Boyden, wound healing, clony forming assay and 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2H tetrazolium bromide (MTT) assay were performed to identify the effects of MTA1-deficiency on the biological behaviors of BGC-823 cells in vitro. Simultaneously, MTA1 overexpressed BGC-823 cell line was established by pcDNA3-MTA1 plasmid transfection for reverse verification. In addition, the role of MTA1 in the tumorigenicity of gastric cancer BGC823 cells in vivo was examined by subcutaneous injection of BGC-823 cells expressing different MTA1 levels into nude mice. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot were used to detect the expression levels of integrin β1, cyclin D1 and uPAR in pSilencer3.1-MTA1-siRNA, pcDNA3-MTA1 transfected cells and control cells. Results: MTA1 knocked down or upregulated BGC-823 cell lines were successfully generated by transfecting pSilencer3.1-MTA1-siRNA or pcDNA3-MTA1 vector with lipofectamine 2000, respectively. The Boyden and wound healing experiments showed metastasis and invasion ability in MTA1 knocked down cells (25±2, 12±1) were significantly decreased when compared with those of control (78±2, 50±2) and MTA1-overexpressed groups (218±2, 269±3; P <0.05). The results of MTT assay and colony forming assay were significantly decreased when compared with those of showed that MTA1 overexpressed cells grew more rapidly and formed more colonies in vitro and induced worse malignant tumors in vivo, while MTA1 knocked down cells presented the reversed phenotype[control group (1 482.41±511.90) mm(3,) (1.39±0.29)g; MTA1 overexpressed group [(3 158.73±1 823.22) mm(3,) (2.23±0.51)g; MTA1-downregulated group (711.32±284.30)mm(3,) (0.87±0.21) g ; P <0.05)]. In addition, RT-PCR result showed that the expression level of MTA1 was positively correlated with the known metastasis-related genes (integrinβ1, cyclinD1, uPAR). Conclusions: MTA1 promotes the invasion, migration and proliferation of human gastric cancer BGC-823 cells. On the contrary, down-regulation of MTA1 significantly inhibits tumorigenicity of BGC-823 cells and induces favorable phenotypes. MTA1 may promote the malignant phenotype of BGC-23 cells via regulating the expressions of integrinβ1, cyclinD1 and uPAR.

With the development of radiotherapeutic oncology, computer technology and medical imaging technology, radiation therapy has made great progress. Research on the impact and the specific mechanism of radiation on tumors has become a central topic in cancer therapy. According to the traditional view, radiation can directly affect the structure of the DNA double helix, which in turn activates DNA damage sensors to induce apoptosis, necrosis, and aging or affects normal mitosis events and ultimately rewires various biological characteristics of neoplasm cells. In addition, irradiation damages subcellular structures, such as the cytoplasmic membrane, endoplasmic reticulum, ribosome, mitochondria, and lysosome of cancer cells to regulate various biological activities of tumor cells. Recent studies have shown that radiation can also change the tumor cell phenotype, immunogenicity and microenvironment, thereby globally altering the biological behavior of cancer cells. In this review, we focus on the effects of therapeutic radiation on the biological features of tumor cells to provide a theoretical basis for combinational therapy and inaugurate a new era in oncology.

Aim: To determine the efficacy and safety of transarterial embolization and low-dose continuous hepatic arterial infusion chemotherapy with oxaliplatin and raltitrexed in hepatocellular carcinoma (HCC) with major portal vein tumor thrombus (MPVTT)., Methods: Eighty-six patients with MPVTT accepted routine embolization. The catheter was kept in the hepatic artery and oxaliplatin (50 mg in 250 mL of glucose) was infused by pump for 4 h, followed by raltitrexed (2 mg in 100 mL of 0.9% saline) infusion by pump for the next 1 h. The efficacy and safety were evaluated after the transarterial chemoembolization (TACE)., Results: Full or partial embolization was achieved in 86 cases, where all the cases received low dose continuous hepatic arterial infusion chemotherapy. Complete responses (CRs), partial responses (PRs), stable disease (SD), and disease progression (PD) for intrahepatic disease were observed in 0, 45, 20, and 21 patients, respectively. The 1-, 2-and 3-year overall survival rates of the 86 patients were 40.7%, 22.1%, and 8.1% respectively, and the median survival time was 8.7 mo. Complication was limited., Conclusion: TACE with low dose continuous hepatic arterial infusion of oxaliplatin and raltitrexed could be an option in MPVTT patient; it was shown to be effective in patients with advanced HCC with MPVTT with less toxicity., Competing Interests: Conflict-of-interest statement: The authors declare, to the best of our knowledge, no conflict of interest exists.

Covalent organic frameworks (COFs) are a new class of multifunctional crystalline organic polymer constructed with organic monomers via robust covalent bonds. The unique properties such as convenient modification, low densities, large specific surface areas, good stability and permanent porosity make COFs great potential in separation science. This review shows the state-of-the art for the application of COFs and their composites in analytical separation science. COFs and their composites have been explored as promising sorbents for solid phase extraction, potential coatings for solid phase microextraction, and novel stationary phases for gas chromatography, high-performance liquid chromatography and capillary electrochromatography. The prospects of COFs for separation science are also presented, which can offer an outlook and reference for further study on the applications of COFs., (Copyright © 2018 Elsevier B.V. All rights reserved.)

High crystallinity of covalent organic frameworks (COFs) with dual fluorescence emissions has not been reported so far. Here, we show the rational design and preparation of high-crystallinity COF TzDa via the synergetic interaction of docking sites and hydrogen bonds: 4,4',4″-(1,3,5-Triazine-2,4,6-triyl)trianiline (Tz) with the docking site and 2,5-dihydroxyterephthalaldehyde (Da) with the OH group are employed to synthesize the imine-linked two-dimensional high-crystallinity layered structure TzDa. The prepared mesoporous TzDa (ca. 36 Å) exhibits high thermal and chemical stability. The intramolecular charge transfer (ICT) and excited-state intramolecular proton transfer (ESIPT) effects bring TzDa two main fluorescence emissions around 500 and 590 nm. Water molecules can interfere with the ICT and ESIPT effects, allowing the development of a ratiometric fluorescent sensor for water in organic solvents. The proposed sensor shows high sensitivity to trace water in conventional organic solvents. The high stability of TzDa allows its recyclable uses for trace water detection. This work not only offers a platform for the construction of high-crystallinity COFs, but also provides a rational design of COFs with dual fluorescence emissions for ratiometric sensing applications.

Objective: To study the acute effects of compound ambient air pollution on small airway lung functions among school children in Shanghai. Method: A longitudinal survey on lung functions was conducted among 233 school-children from three schools (A, B and C, located in innerring, mid-ring and outer-ring areas). Lung function test was performed once a week for 3 times respectively, among children in school A and B in Dec. 2013 and in school C in Dec. 2014. The fourth lung function test was tested in Jun. 2014 and May 2015 in the respective schools. Results: from the lung function would include items as: forced mid-expiratory flow at 25% of forced vital capacity (MEF(25%)), mid-expiratory flow at 50% of forced vital capacity (MEF(50%)), mid-expiratory flow at 75% of forced vital capacity (MEF(75%)) and mid-expiratory flow between 25% and 75% of the forced vital capacity (FEF(25%-75%)). Data regarding the daily air quality real-time of PM(2.5), PM(10), SO(2) and NO(2) in Dec. 2013, Dec. 2014, Jun. 2014 and May. 2015 from the three environmental monitoring spots and meteorological data from the Shanghai Meteorological Service system which were physically close to the three schools, were collected simultaneously. Linear mixed effect model was used to examine the levels of correlation between lung function indicators and ambient air pollutants. Results When confounding factors on meteorology and individuals were controlled, the lag effects and accumulated lag effects were found to have existed between the internal quarter rang (IQR) concentration of PM(2.5) and PM(10) in lag2 day and lag02 days, IQR concentration of SO(2) in lag02 day and IQR concentration of NO(2) lag0 day, when small airway lung functions like MEF(25%), MEF(50%), MEF(75%) and FEF(25%-75%)( P <0.05) were inspected. Results from the two air pollutants model analysis showed that SO(2) and NO(2) presenting interactive effects with PM(2.5), PM(10) and lag effects more significant than the individual SO(2) and NO(2), respectively ( P <0.05). Conclusion: Contents on the ambient air pollutants as PM(2.5), PM(10), SO(2) and NO(2) were negatively associated with the lung functions in the small airways of children, in Shanghai.

Covalent organic frameworks (COFs) are a novel class of porous materials, and offer great potential for various applications. However, the applications of COFs in chiral separation and chiral catalysis are largely underexplored due to the very limited chiral COFs available and their challenging synthesis. Here we show a bottom-up strategy to construct chiral COFs and an in situ growth approach to fabricate chiral COF-bound capillary columns for chiral gas chromatography. We incorporate the chiral centres into one of the organic ligands for the synthesis of the chiral COFs. We subsequently in situ prepare the COF-bound capillary columns. The prepared chiral COFs and their bound capillary columns give high resolution for the separation of enantiomers with excellent repeatability and reproducibility. The proposed strategy provides a promising platform for the synthesis of chiral COFs and their chiral separation application.

Aggressive angiomyxoma (AAM) and angiomyofibroblastoma (AMFB) are two rare types of mesenchymal tumors with overlapping clinicopathological features. In certain cases, the differential diagnosis between the two tumors is difficult even for experienced pathologists. The present study reported the case of a well-circumscribed soft tissue mass on the anterior wall of the vagina in a 25-year-old woman. The mass was initially removed without disturbance to the adjacent tissues. The histopathological features included spindle cells in inconspicuous myxoid stroma and a well-demarcated mass without evidence of invasion, which prompted the initial diagnosis of AMFB. After 2 years, a mass returned in the same area and a wide tumor excision was performed. The histopathological examination confirmed the final diagnosis of AAM upon review.

In May 2015, Professor Xiao Yang authored a review on the development of CRISPR-Cas9 techniques in the journal of Military Medical Research. This review provided a valuable overview of this major scientific advance. It has been four years since the first publication of the CRISPR-Cas9 breakthrough (Science. 2012; 337: 816-21). The use of this technique has expanded into various scientific areas and is being developed into a systematic technical platform that may contribute to many bioengineering fields involving DNA sequence editing.

Objective: The gene product of the AT-rich interactive domain 1A (SWI-like) gene (ARID1A) is a member of the SWI/SNF adenosine triphosphate-dependent chromatin-remodeling complexes, which plays an essential role in controlling gene expression and is also involved in cancer development. ARID1A is frequently mutated in a wild variety of cancers and function as a tumor suppressor in several kinds of cancers. ARID1A was down-regulated in gastric cancer, and associated poor patient prognosis. However, how ARID1A protein is regulated in gastric cancer remains largely unknown., Materials and Methods: Here, we show that ARID1A protein is rapidly ubiquitinated and degradated in gastric cancer cells in response to DNA damage treatment., Results: Using genetic and pharmacologic Cullin inactivation coupled with in vitro ubiquitination assay, we demonstrate that ARID1A is a substrate of the Cullin-SKP1-F-box protein (SCF) complexes. Moreover, gastric cancer cells with forced expression of ARID1A showed an increased sensitivity to DNA damage reagents. Thus, our data uncovered a previous unknown posttranscriptional regulation of ARID1A by SCF E3 ligase in gastric cancer cells in DNA damage response., Conclusions: These findings suggest ARID1A might be a promising drug target in gastric cancer treatment.

Background: Tuberculosis remains one of the deadliest infectious diseases in humans. It has caused more than 100 million deaths since its discovery in 1882. Currently, more than 5 million people are infected with TB bacterium each year. The cell wall of Mycobacterium tuberculosis plays an important role in maintaining the ability of mycobacteria to survive in a hostile environment. Therefore, we report a virtual screening (VS) study aiming to identify novel inhibitors that simultaneously target RmlB and RmlC, which are two essential enzymes for the synthesis of the cell wall of M. tuberculosis., Methods: A hybrid VS method that combines drug-likeness prediction, pharmacophore modeling and molecular docking studies was used to indentify inhibitors targeting RmlB and RmlC., Results: The pharmacophore models HypoB and HypoC of RmlB inhibitors and RmlC inhibitors, respectively, were developed based on ligands complexing with their corresponding receptors. In total, 20 compounds with good absorption, distribution, metabolism, excretion, and toxicity properties were carefully selected using the hybird VS method., Discussion: We have established a hybrid VS method to discover novel inhibitors with new scaffolds. The molecular interactions of the selected potential inhibitors with the active-site residues are discussed in detail. These compounds will be further evaluated using biological activity assays and deserve consideration for further structure-activity relationship studies., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Objective: To observe the therapeutic effect of different schemes of mild moxibustion for treatment of (yang-deficiency induced cold-accumulation type)knee osteoarthritis (KOA)., Methods: Fifty-nine KOA patients were chosen and randomly divided into control group (n =30) and test group (n =29). Patients of the control group were treated by mild moxi- bustion of Neixiyan (EX-HE4) and Waixiyan (ST 35) for 30 min, once daily for 14 days (two courses), and those of the test group were treated by mild moxibustion of EX-HE 4 and ST 35 for 30 mini once daily for 7 days (the same to control group), followed by moxibustion of Yaoyangguan (GV 3) and Mingmen (GV 4) once daily for next 7 days more. In addition, patients of the two groups were also treated by routine acupuncture stimulation of EX-HE 4, ST 35, Yanglingquan (GB 34), Kunlun (BL 60), etc. The interval between two therapeutic courses was one day. The Lysholm Knee Score Scale (LKSS) was used to evaluate the therapeutic effect. Visual analogue scale (VAS) was employed to assess the patient's knee-joint pain severity (arthralgia), and scores of morning stiffness, arthrocele, and walking restraint degree of the knee-joint were also evaluated before and after the treatment., Results: After the treatment, the scores of VAS, morning stiffness, arthrocele and walking restraint degree of the knee-joint of both groups were significantly decreased (P<0. 05), and the scores of the test group were obviously lower than those of the con- trol group (P<0. 05). The effective rate of the test group was 89.66% (26/29)which was obviously higher than that (70.00%, 21/30) of the control group (P<0. 05)., Conclusion: Mild moxibustion of Neixiyan (EX-HE 4) and Waixiyan (ST 35, local acu- points), and Yaoyangguan (GV 3) and Mingmen (GV 4) has a better therapeutic effect for KOA patients than moxibustion of local acupoints only.

Insulin-like growth factor binding protein-3 (IGFBP-3) exerts anti-proliferative or pro-apoptotic effects through IGF-dependent as well as IGF-independent mechanisms in vitro. The purpose of this study was to examine the association between genetic variants in IGFBP-3 (rs2270628) and the risk of esophageal squamous cell carcinoma (ESCC) in a Chinese Han population. Five hundred ESCC cases and 500 cancer-free controls of the Chinese Han population were involved in this study. The IGFBP-3 single-nucleotide polymorphism (SNP) rs2270628 was genotyped and the estimated adjusted odds ratios (ORs) and 95% confidence intervals (CIs) for its association with the risk of ESCC were determined using unconditional logistic regression analysis. Compared with the rs2270628 CC genotype, TT genotype was associated with a significantly increased ESCC risk with OR (95%CI) of 2.07 (1.05-4.09), but CT genotype was not (OR = 1.25, 95%CI =0.94-1.66). IGFBP-3 SNP rs2270628 may contribute to the risk of ESCC in the Chinese Han population.

Objective: To explore the incidence, clinical characteristics, diagnosis and treatment strategies, prognosis of patients with malignancy-associated hypercalcemia (MAH)., Methods: The data of 115 patients with MAH who were treated at the Medical Oncology Department of Chinese PLA General Hospital from Jan., 2001 to Dec., 2010 was retrospectively reviewed. Survival analysis was performed using the Kaplan-Meier method and the Cox proportional hazard model with statistic software SPSS 18.0., Results: The patients had blood calcium levels ranging from 2.77 to 4.87 mmol/L. Except for 9 cases who died or were discharged within 5 days after admission, all other patients recovered to normal blood calcium level after treatment with bisphosphonates or intravenous hydration and diuretics; their survival after occurrence of MAH was from 1 day to 4,051 days, and the median survival time was only 50 days. In the log-rank test, the male, renal metastasis, central nervous system symptoms and hypercalcemia occurring over 140 days after cancer diagnosis were predictors of poor survival (P=0.002, P=0.046, P=0.000, P=0.009). In the COX analysis, being male, central nervous system symptoms and hypercalcemia lasting over 140 days after cancer diagnosis were independent prognostic factors for survival time (RR=2.131, P=0.027; RR=3.054, P=0.002; RR=2.403, P=0.001). According to these factors, a score system was established to predict the patient prognosis and adjust the treatment., Conclusion: Cancer patients with MAH have an extremely poor median survival. Some independent factors indicate poor prognosis, including male gender, central nervous system symptoms and hypercalcemia lasting over 140 days after cancer diagnosis. The prognostic score can serve as a reference for MAH prognosis and treatment, worthy of further investigation.

Objective: To compare the outcome difference between the heat-sensitive and non-sensitive moxibustion stimulation of Yifeng (TE 17) in the treatment of peripheral facial palsy., Methods: A total of 43 patients with peripheral facial paralysis were divided into heat-sensitive moxibustion (n = 21) and non-sensitive moxibustion (n = 22) groups in accordance with their reactions to moxa-heat stimulation. Mild moxibustion was applied to bilateral Yifeng (TE 17) for 45 minutes, followed by mild acupuncture stimulation of Cuanzhu (BL 2), Yangbai (GB 14), Sibai (ST 2), Hegu (LI 4), etc. Moxibustion treatment was conducted once daily for 10 days, and acupuncture treatment given once daily for 20 days (with 2 days interval between every 10 days). Following moxibustion, if the patient felt regional heat penetrating to the deep tissue, extending peripherally, or propagating to other part of the body, or felt mild warm in the stimulated region but warmer in the slightly distant part, or felt mild warm on the skin surface but warmer in the deep tissue, it was considered to be heat-sensitivity. Patients with occurrence of heat-sensitivity being equal to and more than 3 times during the 10 sessions of treatment were assigned to heat-sensitive group, and those with occurrence of heat-sensitivity being equal to or less than 2 times assigned to non-sensitive moxibustion group. According to Portmann Scale (including movement and resting posture) for the voluntary movement state of the face, forehead, winkles eye closure, open mouth smile, snarl, and pucker; points 0, 1, 2 and 3 indicate no muscular movement, marked asynersis, asynersis and normal, respectively. In the light of resting posture of the eye, the nasolabial fold and mouth corner, 0, 1, and 2 points indicate severe, mild dissymmetry and normal, separately. In evaluation of the therapeutic effect, twenty points (at most) indicate cured, 17-19 points marked improvement; 14- 16 points improved, and < or = 13 points failure., Results: Compared with pre-treatment, Portmann scores of both heat-sensitive moxibustion and non-sensitive moxibustion groups were significantly increased after the treatment (P < 0.001), and the score of the heat-sensitive moxibustion group was markedly higher than that of the non-sensitive moxibustion group (P < 0.05). Of the 21 and 22 facial palsy patients in the heat-sensitive moxibustion and non-sensitive moxibustion groups, 8 and 5 cases were cured, 10 and 7 were markedly effective, 3 and 10 were improved, with the markedly effective rates being 85.71% and 54.55%, respectively. The therapeutic effect of the heat-sensitive moxibustion group was statistically better (P < 0.05)., Conclusion: Heat-sensitive moxibustion is significantly superior to non-sensitive moxibustion in improving symptoms of peripheral facial palsy patients, suggesting a necessity for paying attention to patients' reactions during moxibustion treatment.

Aim: To detect the expression of huCdc7 in colorectal cancer., Methods: The mRNA and protein expression of huCdc7 in 39 colorectal cancer tissue specimens and matched tumor-adjacent normal colorectal tissue specimens was detected by reverse transcription-polymerase chain reaction and immunohistochemistry, respectively., Results: The relative expression level of huCdc7 mRNA in colorectal cancer was significantly higher than that in tumor-adjacent normal colorectal tissues (0.03675 ± 1.00 vs 0.01199 ± 0.44, P < 0.05). huCdc7-positive cells displayed brown granules in the nucleus. Tumor tissues contained many huCdc7-positive cells, whereas normal colorectal tissues contained very few positive cells., Conclusion: huCdc7 may play an important role in the development and progression of colorectal cancer.

Objective: To investigate changes of protein expression profiles between human choriocarcinoma JeG-3 cell line and its floxuridine (FUDR)-resistant sub-line., Methods: The differentially expressed proteins were identified by using two dimension difference gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) approaches. Gene ontology (GO) analysis and Pathway analysis were used to screen the candidate proteins. The levels of the proteins in chemo-resistant sub-lines were validated by western blot. Ribonucleic acid interference (RNAi) was used to knockdown the expression of calreticulin (CALR) and (or) protein disulfide-isomerase A3 (PDIA3) respectively., Results: Forty-six proteins spots were found to be significantly different in spot intensity by statistical analysis between chemo-resistance sub-line and parent cell line, of which 31 proteins were identified by MALDI-TOF-MS. Comparing to the parent cell lines, three endoplasmic reticulum (ER) protein folding molecular chaperones: CALR, PDIA3 and 78 000 glucose-regulated protein (GRP78) screened out were increased significantly in floxuridine-resistant sub-line and were verified by western blot. The resistance index decreased by knockdown the CALR and/or PDIA3 expression in FUDR-resistant sub-line 76.3% (36.7 ± 2.0 vs. 8.7 ± 3.1, P < 0.05) and 51.4% (36.7 ± 2.0 vs. 17.8 ± 1.2, P < 0.05) respectively., Conclusion: These ER protein folding molecular chaperones, CALR, PDIA3 and GRP78, may involved in the mechanism of FUDR-resistance choriocarcinoma.

Objective: To explore the expression and significance of estrogen receptor (ER), progestrone receptor (PR), vascular endothelial growth factor (VEGF), CA15-3, CA125 and carcinoma embryonic antigen (CEA) expression in judging the prognosis of breast cancer., Materials and Methods: Sixty-five patients with breast cancer undergoing operations in the general surgery department were considered as the observation group, and 50 healthy outpatients of our hospital as the control group. Cubital venous blood was drawn in the morning from fasting patients in the two groups and chemiluminescence immunoassays were used to detect the levels of CA15-3, CA125 and CEA in serum. The follow-up duration was from 4 months to 2 years, and change in levels of the indicators was detected by dynamically drawing blood. After surgery, cancer tissue samples of patients in observation group remained on file (the non-recurrent patients were biopsied). Immunohistochemistry was applied to determine the expression of ER, PR and VEGF in tissue., Results: The effective rate of 12 patients with negative ER and PR expression was 33.3% in the observation group, being associated with prognosis to varying extents. Serum CA15-3, CA125 and CEA in the observation group were all significantly higher than in control group (p<0.01). With increase in pathological staging, levels of serum CA15-3, CA125 and CEA gradually increased (p<0.01). Levels in patients with lymph node metastasis were markedly higher than in those without (p<0.01). In addition, values with distal lymph node metastasis were notably higher than with adjacent lymph node metastasis (p<0.01). The postoperative follow-up results revealed that positive VEGF and levels of serum VEGF, CA15-3, CA125 and CEA in recurrence group were obviously higher than in non-recurrence group (p<0.01)., Conclusions: Joint detection of ER and PR expression as well as levels of serum VEGF, CA15-3, CA125 and CEA is meaningful and can guide the diagnosis and treatment for breast cancer.

Objective: To investigate the impact of the application of chemical pesticides on the distribution of Anopheles anthropophagus in rice fields and the malaria incidence., Method: Twenty-four villages from 16 counties in the provinces of Zhejiang, Sichuan and Guangxi were chosen for the surveys in the period of 1983-1987. For the survey of An. anthropophagus, indoor human bait trapping until midnight and catching the mosquitoes in all the nets in early morning were carried out to get the density and population ratio of the mosquitoes. Historical data on Anopheles spp., malaria incidence, acreage of the single or double season cropping of rice and on the quantity of chemical pesticide used in rice fields were collected from the study areas., Results: In Hang-jia-hu region of Zhejiang Province, double season rice cropping was performed at that time, the quantity of pesticides used in 1973 was 45 kg/hm2, which was as high as 50 times than that in the 1950s. The density of An. anthropophagus decreased yearly, no An. anthropophagus could be found at 11 survey points in late 1980s. The malaria incidence dropped to less than 1 per 10000. In Leshan and Yibin areas of Sichuan Province, the major cultivation was single cropping, pesticides were applied in paddy fields since 1960s, and the average quantity of pesticides used was 8.6 kg/hm2 during 1970s-1980s. No significant difference on the density of An. anthropophagus was revealed between 1980s (86.2%) and 1960s (82.2%) (chi2=0.63, P>0.05). After mid-1980s, pesticide use gradually increased, and reached to 18.18 kg/hm2 in average in the years after 2000. The density of An. anthropophagus decreased, no An. anthropophagus was found in 2010 in the area surveyed and no malaria cases were reported as well. With double season cropping in Huanjiang County of Guangxi, the pesticide amount consumed was 1.79kg/hm2, 25.13 kg/hm2 and 7.68 kg/hm2 in paddy fields in 1960s, 1970s, and 1980s, respectively. The proportion of An. anthropophagus in anophelines was 52%(1 747/3 392) in the beginning of the 1980s. After the year 2000, the average pesticide use increased to 20.38 kg/hm2 in paddy fields. It was difficult to find An. anthropophagus in human dwellings after 2008. The average annual malaria incidence dropped to 0.14 per 10 000., Conclusion: Change of farming activities and especially use of chemical pesticides in high quantity at the rice fields undermine the breeding environments of An. anthropophagus, greatly reduce the mosquito population and therefore the malaria incidence.

Uric acid is released from damaged cells and serves as a danger signal that alerts the immune system to potential threats, even in the absence of microbial infection. Uric acid modulation of innate immune responses has been extensively studied, but the impact of this damage-associated molecular pattern on adaptive responses remains largely unknown. In this study, we report that, in the presence of NF-κB signaling, uric acid crystals were capable of stimulating dendritic cells to promote the release of cytokines associated with Th17 polarization. Accordingly, naive CD4(+) T cells cocultured with uric acid-treated dendritic cells differentiated toward the Th17 lineage. Th17 differentiation required the inflammasome-dependent cytokines IL-1α/β and IL-18 in both in vitro and in vivo models, and the inflammasome adaptor protein ASC and caspase-1 were essential for Th17 responses. Collectively, our findings indicate a novel role for the danger signal uric acid, in cooperation with NF-κB activation, in driving proinflammatory Th17 differentiation. Our data indicate that sterile inflammation shapes adaptive immunity, in addition to influencing early innate responses.

Objective: To investigate the relationship between metastasis-associated gene 1 (MTA1) expression and invasive and metastatic ability of cervical cancer cell., Methods: Three kinds of plasmids pcDNA3 (control group), pcDNA3-MTA1 (MTA1 group) and pSilencer3.1-MTA1-siRNA (MTA1-siRNA group) were transfected into human cervical cancer cell line CaSki cells. Reverse transcription (RT)-PCR and western blot were used to detected MTA1 mRNA and protein expressions. The effects of MTA1 expression on CaSki cell growth and proliferation, cell migration, adhesion and invasion, and cell cycles were tested by methyl thiazolyl tetrazolium (MTT), clone formation experiment, wound-healing assay, transwell assay, adhesion assay and flow cytometry, respectively. In animal experiment, three groups of cells were inoculated to BALB/c nude mouse subcutaneously to observe tumor formation ability., Results: Compared with control group, MTA1 mRNA and protein were significantly overexpressed in MTA1 group, while MTA1-siRNA group showed lower MTA1 expression. Compared with control group, MTA1 group showed significantly accelerated cell growth; while MTA1-siRNA group showed decreased cell growth since the second day (P < 0.05). Clone formation number in control, MTA1 and MTA1-siRNA group were 133 ± 6, 169 ± 10 and 57 ± 5, respectively. MTA1 group showed accelerated cell formation, while MTA1-siRNA group showed the reverse effect compared with that in control group (P < 0.05). At 24, 48 and 72 hours after wounding, the healing ability of MTA1-siRNA group significantly lagged behind that in the control group, while MTA1 group showed accelerated cell healing ability. The adhesion rate of control, MTA1 and MTA1-siRNA group were (69.3 ± 3.6)%, (80.4 ± 5.6)% and (39.2 ± 7.4)% separately at 90 minutes after cell seeding. In contrast with control group, MTA1 group promoted the adhesion of CaSki cell to matrigel matrix, while MTA1-siRNA group inhibited the adhesion process (P < 0.05). In the migration assay, the number of cells migrated to the bottom side of the membrane in control, MTA1 and MTA1-siRNA group were 153 ± 17, 247 ± 38 and 82 ± 10, respectively. The number of cells in the invasion assay were 231 ± 19, 354 ± 36 and 76 ± 7, respectively. Compared with the control group, MTA1 group significantly increased the migration and invasion ability, while MTA1-siRNA group showed lower cell migration and invasion ability (P < 0.05). In cell cycle experiment, no significant differences of cell proportions including G(1), S and G(2) stage were found among three groups (P > 0.05). In animal experiment, compared with control group, MTA1 group showed accelerated tumor formation and growth, while the MTA1-siRNA group showed the reverse effect (P < 0.05)., Conclusions: MTA1 may play its roles to promote cervical cancer cell invasion, migration, adhesion, as well as cell growth and colony formation, while RNA interference against MTA1 may decrease the malignant phenotypes. This study shows that it will be an effective beginning to explore metastasis mechanisms and cancer gene therapy strategy targeting MTA1 in cervical cancer.

NLRs are cytoplasmic proteins that sense cellular stress and intracellular damage resulting from pathogen uptake. To date, the role of NLRs has been studied using combinations of NLR and TLR agonists, but the interplay between two different NLRs remains uncharacterized. In this study, we employed microarrays to investigate in DCs the regulation of gene transcription mediated by activation of NOD2 and NLRP3 pathways using MDP and MSU. MDP and MSU co-stimulation of murine BMDCs up-regulated the expression of genes encoding molecules for antigen presentation and co-stimulation (MHC class II, CD80, CD86), integrins (ITGB3, ITGAV), cytokines (IL-1α, IL-1β, IL-6, IL-2, IL-23p19, IL-12p40), and chemokines (CXCL1, CXCL2). Transcription of the cytokine genes induced by MDP and MSU partially depended on NOD2 but was independent of NLRP3. Finally, we showed that ERK1 and c-JUN activation increased upon MDP and MSU co-stimulation. As a whole, the results indicate that two different NLR activators synergize at the transcriptional level, leading to unique differential expression of genes involved in the innate immune response.

Objective: To construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20 positive primary chronic lymphocytic leukemia (CLL) cells and provide a promising tool for tumor adoptive immunotherapy., Methods: The recombinant vectors were transduced into PA 317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection for one week. Then transduced T lymphocytes and primary CLL cells were co-cultured. The status of primary chronic lymphocytic leukemia cells were observed by microscope. The level of IL-2 and IFN-gamma in the culture medium were measured., Results: Primary T cells expressing anti-CD20scFv/IgGFc/CD80/CD28/zeta could be constructed successfully. These T cells were able to lyse CD20+ targets and secrete high levels of IL-2 (1301.00 pg/ml) and IFN-gamma (602.18 pg/ml) in vitro., Conclusion: (1) Recombinant gene modified T cells can be constructed successfully. (2) Recombinant gene modified T cells can specially kill CD20 positive primary CLL cells in vitro.

Objective: To investigate the effects of hydroquinone (HQ) on expression of topoisomerase IIα (TOPOIIα) in human bone marrow mononuclear cells, and to explore the role and possible regulatory mechanism of TOPOIIα involved in toxicity of HQ to hematopoietic cells., Methods: After human bone marrow mononuclear cells were exposed to 50 µmol/L HQ (used the cells which were exposed to sterile distilled water as control); the activity of TOPOII was measured by TOPOII assay kit; the expression levels of TOPOIIα mRNA and protein were detected by RT-PCR technique and Western blotting method respectively; the chromatin immunoprecipitation (ChIP) assay was carried out to study the possible mechanism of TOPOIIα expression changes., Results: (1) The activity of TOPOII was inhibited obviously; the protein and mRNA expression of TOPOIIα were 0.017 ± 0.029 and 0.610 ± 0.128, significantly lower than that in the control with the significant difference (P < 0.01) after treated with HQ for 10 h; (2) The decreased content of TOPOIIα was associated with descended level of histone H4 acetylation than in the control, from 1.198 ± 0.056 to 0.324 ± 0.229, with the significant difference (P < 0.01), without accompanied descended level of histone H3 acetylation, from 1.253 ± 0.045 to 1.177 ± 0.025 (P > 0.05); (3) TOPOIIα mRNA expression decreased gradually after HQ processing, and the chemical modification (histone H4 acetylation) of TOPOIIα promoter happened prior to the mRNA expression., Conclusion: HQ could repress the expression of TOPOIIα in human bone marrow mononuclear cells; the change of histone chemical modification plays an important role in the benzene's hematopoietic toxicity.

This study identified differences in expression of angiopoietin-1 (Ang-1) and angiopoietin-2 (Ang-2) in the endometrium of 33 women with abnormal bleeding induced by an intra-uterine device (IUD; 23 proliferative phase, 10 secretory phase) compared with 28 control samples from normal endometria in women without an IUD fitted (12 proliferative phase, 16 secretory phase). Expression of Ang-1, Ang-2 and endothelial cell-specific tyrosine kinase receptor-2 (Tie-2) mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR), and Ang-1 and Ang-2 protein levels were detected by immunohistochemistry. The RT-PCR results showed significantly decreased Ang-1 expression in the endometrium from IUD patients during both phases compared with the control women. Expression of Tie-2 mRNA and the Ang-1:Ang-2 mRNA ratio were also significantly decreased in endometria from IUD patients in the secretory phase compared with the control women. Immunohistochemical analysis showed elevated Ang-2 protein levels in secretory phase endometrium from IUD patients compared with the control women. These results suggest that the angiopoietin/Tie-2 system promotes vascular remodelling in the endometrium and that changes in the expression of Ang-1, Ang-2 and Tie-2 may contribute to abnormal uterine bleeding in some IUD users.

Highly pathogenic avian influenza (HPAI) virus of the H5N1 subtype has caused devastating damage to poultry flocks and sporadic human H5N1 infections. There is concern that this virus subtype may gain transmissibility and become pandemic. Rapid diagnosis and surveillance for H5N1 subtype viruses are critical for the control of H5N1 infection. In this study, we report a robust antigen-capture enzyme-linked immunosorbent assay (AC-ELISA) based on H5- and N1-specific monoclonal antibodies (MAbs) for the rapid detection of H5N1 subtype viruses. The H5 hemagglutinin (HA)-specific MAb (2D9) targets a conformational epitope which recognized multiple clades of H5N1 viruses, including clades 0, 1, 2.1, 2.2, 2.3, 4, 7, and 8. The N1 neuraminidase (NA)-specific MAb (8H12) recognized a linear epitope comprising the sequence AELPF. This epitope was 99% conserved in the NA of 708 analyzed H5N1 viruses, while the epitope was absent in NAs of subtypes N2 through N9. The specificity of the AC-ELISA was examined by using 41 H5N1 HPAI strains from multiple clades, 36 non-H5N1 viruses, and 4 influenza B viruses. No cross-reactivity was observed for any of the non-H5N1 viruses tested. The estimated detection limit was 1 to 2 HA titers. It is concluded that this H5N1 AC-ELISA can simultaneously detect H5 and N1 subtype antigens, eliminating the need for secondary testing for the NA subtype. Implementation of this assay in ELISA-like formats suitable for field use, such as dot ELISA, immunofiltration, or electrochemical biosensor technologies, would provide dual on-site detection of H5 and N1 in clinical or environmental specimens.

This study investigated human cytomegalovirus (HCMV) glycoprotein genotypes in the genital tract tissue of 125 tubal pregnancy patients. The HCMV glycoprotein-B N-terminus (gBn, 54 - 485 NT), gB endoprotease cleavage site (gBclv, 1284 - 1600 NT) and glycoproteinH (gH, -58 - 213 NT) gene fragments were amplified by nested polymerase chain reaction and sequenced to identify gB and gH genotypes. Of 16 gBn-positive samples, four were gBn1, one was gBn2 and 11 were the gBn3 genotype. Of 13 positive gBclv samples, seven were gBclv1, two were gBclv2 and four were the gBclv3 genotype. Of 20 positive gH samples, 10 were gH1, six were gH2 and four were a combined gH1/gH2 genotype. In 10 of the samples that were positive for the gBn and gBclv genotypes, the gBn and gBclv genotypes were not consistent (four were gBclv1-gBn3). This study showed that: (i) HCMV infection with the gB1 - gB3 glycoprotein genotypes is present in tubal pregnancy; (ii) the gBclv and gBn genotypes are not strictly consistent; and (iii) intragenetic variability within the gB gene due to homologous recombination occurs frequently.

Aim: To construct recombinant retroviruses expressing anti-CD20 scFv/CD80/CD28/zeta gene and detect its expression in Jurkat cells., Method: CD28-zetacDNA were amplified from plasmids pBULLET and inserted into pLNCX vector that contained anti-CD20 scFv/CD80 gene. The recombinant plasmids were transfected into PA 317 cells. Retroviruses were harvested from culture medium of PA 317 cells. Then NIH 3T3 were transfected with retroviruses. Objective gene expression was determined by PCR and FACS. Jurkat cells were transfected with high titer of retroviruses and resistant clones were obtained by G418 selection. Objective mRNA was determined by RT- PCR., Results: The recombinant eukaryotic vector was constructed successfully by PCR and enzyme digestion analysis and objective gene was amplified from NIH 3T3 cells transfected with retroviruses by PCR; FACS showed that objective protein could be expressed in NIH 3T3 cells. Objective gene was amplified from Jurkat cells transfected with retroviruses by RT-PCR., Conclusion: Recombinant retrovirus expressing anti-CD20 scFv/CD80/CD28/zeta gene was successfully constructed and objective protein could be expressed in Jurkat cells.

Objective: To explore the effects of adenovirus delivered tissue inhibitor of metalloproteinases-3 (Ad-TIMP-3) on the biological behaviors of cervical cancer cell lines and to evaluate its potential application in cervical cancer gene therapy., Methods: We transferred Ad-TIMP-3 into cervical cancer cells. The TIMP-3 mRNA expression was assessed by RT-PCR, and the TIMP-3 and p53 protein expressions were assessed with Western blot. The apoptotic changes of cells were illustrated with morphology and DAPI staining. The viability of cells was determined with MTT assay. The abilities of in vitro invasion and adhesion were evaluated by the invasion and adhesion assays respectively., Results: After infection, the TIMP-3 mRNA and protein were significantly upregulated in a time-dependent manner. Overexpression of TIMP-3 markedly increased p53 protein level in spite of the backgrounds of p53 gene in cells. Ad-TIMP-3 infection induced massive apoptosis of cervical cancer cells with a marked bystander effect. The abilities of in vitro invasion and adhesion were inhibited significantly (P < 0.01). The cytotoxicity of Ad-TIMP-3 was significantly stronger than that of Ad-p53 (P < 0.05, P < 0.01)., Conclusions: Ad-TIMP-3 infection has cytotoxic effects on cervical cancer cells and can inhibit the expressions of these malignant phenotypes. Ad-TIMP-3 may be a potentially useful agent for cervical cancer gene therapy.

Objective: To construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20+ lymphoma cells and provide a probably new approach to tumor adoptive immunotherapy., Methods: CD28-zeta cDNA were amplified from vector pBULLET and inserted into pLNCX vector that contained anti-CD20scFv/CD80 gene. The recombinant vectors were transduced into PA317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection at one week. Then transduced T lymphocytes and lymphoma cell lines Daudi Raji were cocultured. The cytotoxicity and cytokine production of transduced T cells were determined by non-radio-activation cytotoxicity assay and ELISA respectively., Results: The recombinant eukaryotic vector was constructed successfully as proved by enzyme digestion analysis and sequencing. These T cells were able to lyse CD20+ target cells and secrete high levels of IL-2 and IFN-gamma in vitro., Conclusion: Recombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.

Objective: To investigate the effects of adenovirus-delivered tissue inhibitor of metalloproteinases-3 ( Ad-TIMP-3) on sensitivity of cervical cancer cells to cisplatin and evaluate the potential application of this combined scheme in cervical cancer treatment., Methods: Cells of cervical cancer CaSKi cell line were infected with Ad-TIMP-3 in vitro. Apoptotic effect, cell cycle changes and p53 protein expression were detected. After combined treatment of those cells with cisplatin, colony formation test was performed and cytotoxicity was detected by MTT. The growth curve and tumor growth inhibition in vivo were evaluated., Results: The expressions of TIMP-3 mRNA and protein were significantly upregulated after transfection. As a result, massive apoptosis was induced and the cells were arrested at G2/M phase. Exogenous overexpression of TIMP-3 increased p53 protein level markedly in spite of the backgrounds of p53 gene in cells. Combined with cisplatin treatment, the cloning efficiency was decreased. A synergism was observed by isobolic method ( D < 1 ) in vitro and tumor growth was significantly inhibited in vivo., Conclusion: Ad-TIMP-3 is a powerful proapoptotic agent. It increases sensitivity of the cells to cisplatin and the Ad-TIMP-3 gene therapy in combination with cisplatin could be a promising alternative in cervical cancer treatment.

Objective: To explore the feasibility and mechanism of recombinant adenovirus Ad-pl4ARF in cancer gene therapy., Methods: The proliferation of different liver cancer cells was assessed by morphology and trypan blue assay. Cell apoptosis was confirmed by detecting phosphatidylserine (PS) externalization with Annexin V/PI double staining. The expression of related proteins was analyzed by Western bloting. Nude mouse model bearing subcutaneous transplanted BEL7402 tumor was established to study the therapeutic ability of Ad-pl4ARF., Results: Ad-pl4ARF suppressed cell growth and proliferation, and promoted cell apoptosis of cancer cell lines with different genetic background. Ad-pl4ARF inhibited growth of liver cancer cells ( HepG2, BEL7402) in a dose-dependent manner. Ad-pl4ARF lead to overexpression of Bax and p21, the downstream regulating genes of p53. In the experimental therapy on nude mice bearing subcutaneous transplanted BEL7402 tumor, Ad-pl4ARF suppressed tumor growth significantly., Conclusion: pl4ARF is a short gene and with powerful function, which are consistent with the requirements for tumor suppressor genes used in gene therapy. It may play an important role in gene therapy against malignancies in the future.

Objective: Based on two sets of data from occupational epidemiology, Benchmark dose (BMD) was applied to estimate biological exposure limit (BEL)., Methods: Cadmium exposed workers were selected from a cadmium smelting and a zinc products factory and control group was selected from doctors or nurses and staff from shops living in the same area; Urinary cadmium (UCd) was used as exposure biomarker and urinary beta(2) microglobulin (UBM), NAG (UNAG) and albumin (UALB) were as effect biomarkers. All urine parameters were adjusted by urinary creatinine. Software of BMDS (Version 1.3.2, EPA.U.S) was used to calculate BMD., Results: Calculated abnormal prevalence was based on the upper limit of 95% of effect biomarkers in control group; There are significant dose response relationship between the prevalence of effect biomarkers (UBM, UNAG and UALB) and exposure biomarker (UCd); BEL was 5 microg/g creatinine for UBM as effect biomarker, It consists with the recommendation of WHO; BEL was 3 microg/g creatinine for UNAG as effect biomarker; BEL can be estimated by using the method of BMD; the more sensitive biomarker would used, the more occupational people would protected., Conclusion: The application of BMD in estimating biological exposure limit (BEL) is proper. UNAG is suggested as most sensitive biomarker to be used to estimate BEL for cadmium exposure.

Objective: To estimate the benchmark dose for osteoporosis caused by cadmium exposure in a Chinese general population with an epidemiological study., Methods: The inhabitants living in both cadmium polluted and non-polluted areas served as the exposure group and the control group. Urinary cadmium (UCd) and Blood cadmium (BCd) were used as exposure biomarkers while the Z score was used as effect biomarker for the osteoporosis., Results: The UCd and BCd in the habitants of the polluted areas were significantly higher than those in the habitants of the control area on average (P < 0.05) and the UCd and BCd in the habitants of the highly polluted areas were significantly higher than those in the habitants of the moderately polluted area on average (P < 0.05). The bone mineral density was significantly decreased in the groups of the highest UCd and BCd level compared with the 5 microg/g Cr group with the significant difference (P < 0.05). The morbidity of the osteoporosis would increase significantly with the increase of the cadmium exposure (P < 0.05) with the linear correlation (P < 0.05). BMDs were calculated using BMDS Version l.3.2 software and BMDLs were also determined. The BMDL of UCd for cadmium-induced osteoporosis was higher than those representing cadmium-induced renal dysfunction., Conclusion: High level of cadmium exposure can induce osteoporosis, which occurs later than renal damage related to cadmium exposure. The BMD is a practical method.

Aim: To characterize p62 expression and define the relationship between p62 expression and cell proliferation in primary carcinomas of the digestive system., Methods: p62 expression was characterized in surgically resected tumor specimens from 60 patients with primary carcinomas of the digestive tract (including 22 esophageal carcinomas, 17 gastric carcinomas, and 21 colorectal carcinomas) and 40 patients with hepatocellular carcinoma (HCC) by immunohistochemistry (IHC). The cell proliferation was determined by IHC of Ki-67 in 40 patients with HCC., Results: Twenty-two cases of esophageal carcinoma were histopathologically diagnosed as squamous cell carcinoma. We combined the gastric and colorectal carcinomas based on the equivalent histology. The 38 tumors in the combined groups, consisted of 17 well-differentiated, eight moderately differentiated, nine poorly differentiated carcinomas, and four mucinous adenocarcinomas. According to the criteria of Edmondson and Steiner, 40 patients with HCC were graded (2 grade I, 17 grade II and 21 grade III). p62 expression in primary carcinomas of the gastrointestinal tract (60/60,100%) was higher than that (27/40, 67.5%) of HCC (P<0.01, chi(2) = 19.63). High expression levels of p62 were positively correlated with histological grades in gastric and colorectal carcinomas (P<0.0001) and inversely associated with those in HCC (P = 0.0322). No significant correlations were observed for esophageal carcinomas (P = 0.8246). p62 expression was also detected in the cytoplasm of morphologically normal columnar epithelial cells adjacent to the cancer foci of gastric and colorectal carcinomas. In 40 HCC specimens, the mean Ki-67 labeling index (LI) was (19.6+/-16.0)%. It was (28.3+/-18.73)% in 12 cases with high p62 expression (+++), (7.53+/-14.83)% in 13 cases without p62 expression(-). Patients with a high p62 expression showed a significantly higher level of Ki-67 staining than those without p62 expression (P<0.05, t = 2.069)., Conclusion: p62 expression is common in carcinomas of the digestive system and higher in carcinomas of the gastrointestinal tract than in primary HCC. p62 is a cellular differentiation-related protein. Cancer cells with a high p62 expression exhibited highgrowth fractions in HCC.