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author:("Liu, liaodong")
1.  Preparation of γ-Fe2O3/ZnFe2O4 nanoparticles by enhancement of surface modification with NaOH 
During liquid-phase synthesis of γ-Fe2O3 nanoparticles by chemically induced transition in FeCl2 solution, enhancement of surface modification by adding ZnCl2 was attempted by using NaOH. By using transmission electron microscopy, X-ray diffraction, X-ray photoelectron spectroscopy, energy-dispersive X-ray spectrometry, and vibrating sample magnetometry, the dependence of the synthesis on the amount of additional NaOH was studied.
The experimental results show that the surface of the γ-Fe2O3 nanoparticles could be modified by adding ZnCl2 to form composite nanoparticles with γ-Fe2O3/ZnFe2O4 ferrite core coated with Zn(OH)2 and adsorbed FeCl3, and that modification could be enhanced by adding NaOH.
In the experimental conditions, when the concentration of additional NaOH was below 0.70 M, the amounts of ZnFe2O4 and Zn(OH)2 phases increased slightly and that of adsorbed FeCl3 was unchanged. When the concentration of NaOH exceeded 0.70 M, the amount of FeCl3, ZnFe2O4, and Zn(OH)2 increased.
PMCID: PMC4077838  PMID: 24991231
Nanoparticles; Composite; Surface modification; γ-Fe2O3; ZnFe2O4
2.  A rice calcium-dependent protein kinase OsCPK9 positively regulates drought stress tolerance and spikelet fertility 
BMC Plant Biology  2014;14:133.
In plants, calcium-dependent protein kinases (CDPKs) are involved in tolerance to abiotic stresses and in plant seed development. However, the functions of only a few rice CDPKs have been clarified. At present, it is unclear whether CDPKs also play a role in regulating spikelet fertility.
We cloned and characterized the rice CDPK gene, OsCPK9. OsCPK9 transcription was induced by abscisic acid (ABA), PEG6000, and NaCl treatments. The results of OsCPK9 overexpression (OsCPK9-OX) and OsCPK9 RNA interference (OsCPK9-RNAi) analyses revealed that OsCPK9 plays a positive role in drought stress tolerance and spikelet fertility. Physiological analyses revealed that OsCPK9 improves drought stress tolerance by enhancing stomatal closure and by improving the osmotic adjustment ability of the plant. It also improves pollen viability, thereby increasing spikelet fertility. In OsCPK9-OX plants, shoot and root elongation showed enhanced sensitivity to ABA, compared with that of wild-type. Overexpression and RNA interference of OsCPK9 affected the transcript levels of ABA- and stress-responsive genes.
Our results demonstrated that OsCPK9 is a positive regulator of abiotic stress tolerance, spikelet fertility, and ABA sensitivity.
PMCID: PMC4036088  PMID: 24884869
Abscisic acid (ABA) signaling; Abiotic stresses; Calcium-dependent protein kinase (CDPK); Drought stress tolerance; Rice; Spikelet fertility
3.  Molecular Epidemiology of Japanese Encephalitis Virus in Mosquitoes during an Outbreak in China, 2013 
Scientific Reports  2014;4:4908.
Japanese encephalitis virus (JEV) can cause serious encephalitis and Culex mosquitoes are the primary vector. In 2013, a JE outbreak occurred in Shandong Province, China with 407 confirmed cases, including 11 deaths. An investigation on JEV in mosquitoes during the outbreak was conducted. A total of 14,719 mosquitoes were collected at 3 sites. For the 12,695 Culex tritaeniorhynchus mosquitoes, 88/201 pooled samples were positive by RT-PCR for the presence of the pre-membrane or envelope protein coding genes. The maximum likelihood estimates of JEV positive individuals per 1,000 vectors were 12.0, 7.2, and 6.0 in the 3 sites respectively with an overall estimate of 9.1. Phylogenetic analysis on these pre-membrane (n = 72) and envelope (n = 26) sequences with those of reference strains revealed they belonged to genotype I. This study describes the molecular epidemiology of JEV and suggests the high infection rate in mosquitoes is an important factor for the outbreak.
PMCID: PMC4014896  PMID: 24809635
4.  Risk Intelligence: Making Profit from Uncertainty in Data Processing System 
The Scientific World Journal  2014;2014:398235.
In extreme scale data processing systems, fault tolerance is an essential and indispensable part. Proactive fault tolerance scheme (such as the speculative execution in MapReduce framework) is introduced to dramatically improve the response time of job executions when the failure becomes a norm rather than an exception. Efficient proactive fault tolerance schemes require precise knowledge on the task executions, which has been an open challenge for decades. To well address the issue, in this paper we design and implement RiskI, a profile-based prediction algorithm in conjunction with a riskaware task assignment algorithm, to accelerate task executions, taking the uncertainty nature of tasks into account. Our design demonstrates that the nature uncertainty brings not only great challenges, but also new opportunities. With a careful design, we can benefit from such uncertainties. We implement the idea in Hadoop 0.21.0 systems and the experimental results show that, compared with the traditional LATE algorithm, the response time can be improved by 46% with the same system throughput.
PMCID: PMC4030500  PMID: 24883392
5.  Synergistic killing of lung cancer cells by cisplatin and radiation via autophagy and apoptosis 
Oncology Letters  2014;7(6):1903-1910.
Cisplatin is a commonly used drug for chemotherapy, however, whether it may be used synergistically with radiotherapy remains unclear. The present study investigated the underlying mechanisms of synergistic killing by radiosensitization and cisplatin, with a focus on the growth inhibition, apoptosis and autophagy of non-small cell human lung cancer cells in vitro and in a tumor xenograft in vivo. A549 cells were used for the in vitro experiments and divided into the following four treatment groups: Sham-irradiated; conventional radiotherapy (CRT) of five doses of 2 Gy every day; hyperfractionated radiotherapy of five doses of 2 Gy (1 Gy twice a day at 4 h intervals) every day; and CRT plus cisplatin. A xenograft tumor-bearing C57BL/6 model was established for the in vivo experiments and the above-mentioned treatments were administered. MTT and colony formation assays were used to detect cell viability and western blotting was performed to detect the levels of protein expression. Monodansylcadaverine staining and the immunofluorescence technique were used to analyze the autophagy rate, while flow cytometry and immunohistochemistry were performed to detect the expression levels of the genes associated with apoptosis and autophagy, including microtubule-associated protein 1 light chain 3 (MAPLC3)-II, phosphoinositide 3-kinase (PI3K) III, Beclin1, phosphorylated protein kinase B (p-AKT), damage-regulated autophagy modulator (DRAM), B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein, caspase-3 and p21. The MTT assay demonstrated that cisplatin exhibits a dose-dependent cytotoxicity in A549 cells and synergizes with radiation to promote the cell-killing effect of radiation. In the xenograft mouse model of Lewis cells, cisplatin plus ionizing radiation (IR) (five doses of 2 Gy) yielded the most significant tumor suppression. The autophagic vacuoles, the ratio of MAPLC3-II to MAPLC3-I (LC3-II/LC3-I) and the levels of Beclin1 were found to increase in all treatment groups, with the most marked upregulation observed in the CRT plus cisplatin treatment group. In addition, caspase-3 processing was enhanced in the group treated with the combination of cisplatin with radiation, compared with the group treated with radiation alone. Fractionated IR resulted in a significant increase in p21 expression, which was further enhanced when combined with cisplatin. Furthermore, treatment with cisplatin and fractionated IR resulted in a significant elevation of the expression of the autophagy-related genes, PI3KIII, Beclin1 and DRAM1. However, the levels of p-AKT were observed to decline following exposure to fractionated IR in the presence or absence of cisplatin. As for the apoptosis signaling genes, the combination of cisplatin and fractionated IR therapy resulted in a significant decrease in Bcl-2 expression and a marked upregulation of p21 expression. The current study offers strong evidence that the combination of cisplatin with radiation strengthens the killing effect of radiation via pro-apoptotic and pro-autophagic cell death.
PMCID: PMC4049698  PMID: 24932256
cisplatin; radiosensitivity; synergistic killing; lung cancer; autophagy; apoptosis
6.  The Tumor Suppressor, p53, Contributes to Radiosensitivity of Lung Cancer Cells by Regulating Autophagy and Apoptosis 
Cell death is one of the most important endpoints of radiosensitivity. The tumor suppressor p53 participates not only in regulation of apoptosis, but also in autophagy mechanism. In this study, H1299-P53 (with wild-type p53) and H1299-175H (with mutant 175H) were used, and the effects of p53 on radiosensitivity were analyzed.
Cell models with different p53 status were established by gene engineering, and cell viability was examined by colony formation assay, and cell counting kit-8 (CCK-8), 3-Methyladenine, and Z-VAD were used to block autophagy and apoptosis, respectively. Western blot was used to detect protein expression; monodansylcadaverine (MDC) staining was used to analyze autophagy rate; DAPI/Propidium Iodide (PI) staining and flow cytometry were used to assess apoptosis and necrosis.
In parental H1299, H1299-P53, and H1299-175H cells, radiosensitivity exhibited different by colony formation and CCK-8 assay (D0: 1.764 Gy, 1.407 Gy and 1.695 Gy; Dq: 2.977 Gy, 1.199 Gy and 2.312 Gy in turn). The radiosensitization of p53 was associated with the increase of MDM2 and P21 expression. The ionizing radiation (IR)-induced apoptosis was significant in H1299-P53 compared with in H1299 and H199-175H (p<0.05) by flow cytometry, and the expression of cleaved-caspase3 was increased in H1299-P53 cells. While the IR-induced autophagy was significant in H1299 cells (p<0.01) and decreased in H1299-P53 and H1299-175H cells (p<0.01) by MDC staining, the expression of MAPLC3II and Beclin-1 increased in H1299, but not in H1299-p53 and H199-175H cells. The IR-induced cell survival was significantly increased by Z-VAD-FMK and decreased by 3MA in H1299-P53 cells; IR- induced autophagy was significantly increased by Z-VAD-FMK in H1299-P53 cells (p<0.01), but not changed in H1299 cells.
p53 could regulate radiosensitivity by inhibiting autophagy and activating apoptosis; autophagy provides a prosurvival mechanism, and p53 potently abrogated the IR-induced autophagy, while mutant 175H shown no effect on radiosensitivity, suggesting that individual treatment strategies should be based on p53 status in patients.
PMCID: PMC3589888  PMID: 23268708
p53; apoptosis; autophagy; lung cancer; radiosensitivity
7.  Stat3 Inhibition Attenuates Mechanical Allodynia through Transcriptional Regulation of Chemokine Expression in Spinal Astrocytes 
PLoS ONE  2013;8(10):e75804.
Signal transducer and activator of transcription 3 (Stat3) is known to induce cell proliferation and inflammation by regulating gene transcription. Recent studies showed that Stat3 modulates nociceptive transmission by reducing spinal astrocyte proliferation. However, it is unclear whether Stat3 also contributes to the modulation of nociceptive transmission by regulating inflammatory response in spinal astrocytes. This study aimed at investigating the role of Stat3 on neuroinflammation during development of pain in rats after intrathecal injection of lipopolysaccharide (LPS).
Stat3 specific siRNA oligo and synthetic selective inhibitor (Stattic) were applied to block the activity of Stat3 in primary astrocytes or rat spinal cord, respectively. LPS was used to induce the expression of proinflammatory genes in all studies. Immunofluorescence staining of cells and slices of spinal cord was performed to monitor Stat3 activation. The impact of Stat3 inhibition on proinflammatory genes expression was determined by cytokine antibody array, enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Mechanical allodynia, as determined by the threshold pressure that could induce hind paw withdrawal after application of standardized von Frey filaments, was used to detect the effects of Stat3 inhibition after pain development with intrathecal LPS injection.
Intrathecal injection of LPS activated Stat3 in reactive spinal astrocytes. Blockade of Stat3 activity attenuated mechanical allodynia significantly and was correlated with a lower number of reactive astrocytes in the spinal dorsal horn. In vitro study demonstrated that Stat3 modulated inflammatory response in primary astrocytes by transcriptional regulation of chemokine expression including Cx3cl1, Cxcl5, Cxcl10 and Ccl20. Similarly, inhibition of Stat3 reversed the expression of these chemokines in the spinal dorsal horn.
Stat3 acted as a transcriptional regulator of reactive astrocytes by modulating chemokine expression. Stat3 regulated inflammatory response in astrocytes and contributed to pain modulation. Blockade of Stat3 represents a new target for pain control.
PMCID: PMC3789727  PMID: 24098399
9.  Bone Selective Protective Effect of a Novel Bone-seeking Estrogen on Trabecular Bone in Ovariectomized Rats 
Calcified Tissue International  2013;93(2):172-183.
The drawbacks of estrogen restrict the clinical use of hormone replacement therapy, and it would be most helpful to explore new estrogenic substances that could prevent bone loss and be free from any adverse effects. We synthesized a new compound named bone-seeking estrogen (SE2) by combining 17β-estradiol (E2) with iminodiacetic acid through the Mannich reaction. E2 and SE2 were labeled with isotope 3H, and the tissue distribution tests of E2-3H and SE2-3H were analyzed by the radioactivity. The specific nuclear binding of E2 and SE2 in osteoblasts was measured. SE2 exhibited significantly greater affinity for bone but lower affinity for ovary and uterus than did E2, and SE2 maintained a high affinity for the estrogen receptor alpha similar to that of E2. SE2 administration did not induce uterine hypertrophy. Body weight increase was significantly suppressed by treatment with E2 but not by SE2 after ovariectomy (OVX). SE2 decreased bone turnover as E2 after OVX detected by serum biochemical markers. Bone histology and micro-CT analysis revealed that SE2 administration, similar to E2, could improve bone mass and trabecular architecture after OVX. Biomechanical analyses showed that SE2 treatment effectively increased mechanical properties after OVX. The results suggested that SE2 was effective in preventing OVX-induced bone loss and exhibited few side effects on body weight and uterine hypertrophy, which was beneficial in reducing the adverse effects caused by E2. SE2 may be a better choice than E2 for the prevention of postmenopausal osteoporosis.
PMCID: PMC3717164  PMID: 23780350
Bone-seeking estrogen; Estrogen; Postmenopausal osteoporosis; Side effects; Trabecular architecture
10.  Knockdown of FRAT1 Expression by RNA Interference Inhibits Human Glioblastoma Cell Growth, Migration and Invasion 
PLoS ONE  2013;8(4):e61206.
FRAT1 positively regulates the Wnt/β-catenin signaling pathway by inhibiting GSK-3-mediated phosphorylation of β-catenin. It was originally characterized as a protein frequently rearranged in advanced T cell lymphoma, but has recently also been identified as a proto-oncogene involved in tumorigenesis. Our previous studies showed that FRAT1 was dramatically overexpressed in gliomas and its expression level was significantly increased along with clinicopathological grades.
In the current study, we used RT-PCR and Western blotting to assess the mRNA and protein levels of FRAT1 in three glioma cell lines. In addition, to evaluate its functional role in gliomas, we examined the effects of FRAT1 knockdown on proliferation, migration and invasion in vitro and tumor growth in vivo using glioblastoma U251 cells and RNAi.
FRAT1 was highly expressed in all three glioma cell lines. RNAi-mediated down-regulation of endogenous FRAT1 in human glioblastoma U251 cells resulted in suppression of cell proliferation, arrest of cell cycle, inhibition of cell migration and invasion in vitro. Moreover, FRAT1 depletion significantly impaired tumor xenograft growth in nude mice.
Our results highlight the potential role of FRAT1 in tumorigenesis and progression of glioblastoma. These findings provide a biological basis for FRAT1 as a potential molecular marker for improved pathological grading and as a novel candidate therapeutic target for glioblastoma management.
PMCID: PMC3629175  PMID: 23613813
11.  Preliminary evidence for a 1000-year-old tsunami in the South China Sea 
Scientific Reports  2013;3:1655.
The risk of large, devastating tsunamis in the South China Sea and its surrounding coastal region is commonly underestimated or unrecognized due to the difficulty of differentiating tsunami from storm deposits. As a consequence, few convincing records have documented tsunami deposits in this region. Here we report preliminary evidence from Xisha Islands in the South China Sea for a large tsunami around AD 1024. Sand layers in lake sediment cores and their geochemical characteristics indicate a sudden deposition event around AD 1024, temporally consistent with a written record of a disastrous event characterized by high waves in AD 1076. Heavy coral and shell fossils, which are older than AD 1024, deposited more than 200 meters into the island, further support the occurrence of a high-energy event such as a tsunami or an unusually large storm. Our results underscore the importance of acknowledging and understanding the tsunami hazard in this area.
PMCID: PMC3622915  PMID: 23575432
12.  The Novel Function of Advanced Glycation End Products in Regulation of MMP-9 Production 
The Journal of surgical research  2010;171(2):871-876.
Advanced glycation end products (AGEs), formed from proteins and peptides by nonenzymatic glycoxidation after contact with aldose sugars, have been implicated in the pathogenesis of age-related cardiac and vascular dysfunction. Our previous study demonstrated significantly elevated levels of AGE and the receptor for AGE (RAGE) in human abdominal aortic aneurysm (AAA) tissues. Inhibition of AGE signaling by targeted gene deletion of RAGE markedly reduced the development of aneurysm in a mouse model of AAA. We also showed that AGE may stimulate aneurysm formation by promoting metalloproteinase (MMP)-9 expression. In this study, we investigated the molecular mechanism underlying this novel function of AGE.
The murine macrophage cell line RAW 264.7 was pretreated with AGE, TGF-β, and MAPK inhibitors. The protein was collected for Western blot analysis. Culture supernatants were collected to determine MMP-9 activity by gelatin zymography.
We found that AGE induced the production of MMP-9 in macrophages in a dose-dependent manner. This induction of MMP-9 was markedly diminished by pretreatment with TGF-β. To delineate the underlying molecular mechanism, we showed that AGE increased phosphorylation of p44/42 ERK, p38, JNK, and PI3K in macrophages. Moreover, AGE induced active p65 subunit of NF-κB. Inhibition of ERK (UO126) or p38 (SB203580), but not PI3K (LY294002 or wortmannin), blocked AGE-induced MMP-9 expression. In contrast, inhibition of JNK (SP-600125) significantly enhanced the stimulatory effect of AGE on MMP-9. Furthermore, TGF-β suppressed AGE-induced expression of the active p65 subunit of NF-κB.
Our data indicate that AGE induces MMP-9 through activation of ERK, p38 mitogen-activated protein and NF-κB, a pathway that is antagonized by TGF-β. This finding in conjunction with previously reported AGE functions in inflammation suggests that anti-AGE therapies could be effective in the prevention of human AAA development and progression.
PMCID: PMC3623272  PMID: 20638679
MAP kinases; abdominal aortic aneurysm; signaling; TGF-beta
13.  Two cases of non-intervention-related vascular intense spasm following stent implantation in the coronary artery 
The clinical occurrence of non-intervention-related vascular spasm following coronary stenting is rare. In the present study, 2 cases are reported. One patient developed continuous spasms in the proximal segment of the left anterior descending (LAD) and left circumflex (LCX) arteries following LAD artery stenting. The second patient developed an intense spasm in the right coronary artery (RCA) following LAD artery stenting. Clinical course and prognosis are dangerous. The main treatment for this condition is a combination of repeated injections of nitroglycerin into the coronary artery and the administration of calcium antagonists. In the clinic, intervention-related vascular spasms are common in percutaneous coronary intervention (PCI) due to the mechanical stimulation caused by balloon dilatation or stent expansion. Injections of a vasodilator into the coronary artery are able to mitigate the spasms and the consequent prognosis is good.
PMCID: PMC3702719  PMID: 23837043
vascular intense spasm; stent implantation; coronary artery
14.  The synergistic effects of traditional Chinese herbs and radiotherapy for cancer treatment 
Oncology Letters  2013;5(5):1439-1447.
Traditional Chinese medicine (TCM) has been demonstrated to have potent cytotoxic activity against certain malignant tumors. Ionizing radiation (IR) is one of the most effective methods used in the clinical treatment of cancer. The drawback of a single formula is that it limits the treatment efficacy for cancer, while comprehensive strategies require additional theoretical support. However, a combination of different antitumor treatment modalities is advantageous in restricting the non-specific toxicity often observed with an extremely high dose of a single regimen. The induction of apoptotic cell death is a significant process in tumor cells following radiotherapy or chemotherapy, and resistance to these treatments has been linked to a low propensity for apoptosis. Autophagy is a response of cancer cells to IR or chemotherapy, and involves the prominent formation of autophagic vacuoles in the cytoplasm. In this review, the synergistic effects of TCM and radiotherapy are summarized and the underlying mechanisms are illustrated, providing new therapeutic strategies for cancer.
PMCID: PMC3678704  PMID: 23760551
traditional Chinese medicine; apoptosis; autophagy; radiosensitization; cancer treatment
15.  Pathology of Bursae of Fabricius in Methionine-Deficient Broiler Chickens 
Nutrients  2013;5(3):877-886.
The purpose of this 42-day study was to investigate the effects of methionine (Met) deficiency on immune function by determining the relative weight, morphological and ultrastructural changes of bursae of Fabricius, cell cycle, and apoptosis of bursa cells. One hundred and twenty one-day-old avian broilers were randomly divided into two groups and fed on a control diet (starter diet, Met 0.50%; grower diet, Met 0.40%) and Met-deficient diet (starter diet, Met 0.26%; grower diet, Met 0.28%) for six weeks. The relative weight of bursae was decreased with Met deficiency when compared to that of the control group. Lesions were also observed in the Met-deficient group. Histopathologically, the numbers of lymphocytes in the follicles were decreased. Ultrastructurally, the mitochondria of lymphocytes were swollen in the Met-deficient group. As measured by flow cytometry, bursal cells in the G0G1 phase were significantly higher (P < 0.01), and bursal cells in the S, G2M phases and proliferating index were obviously lower (P < 0.01) with Met deficiency than in the control group. Moreover, the percentage of apoptotic cells in the bursae were significantly increased in Met-deficient birds (P < 0.01). It was concluded that Met deficiency restrained the development of the bursae of Fabricius and affected the humoral immunity of the chickens.
PMCID: PMC3705324  PMID: 23486195
broilers; bursa of Fabricius; flow cytometry (FCM); methionine deficiency; pathology
16.  Risk Perceptions for Avian Influenza Virus Infection among Poultry Workers, China  
Emerging Infectious Diseases  2013;19(2):313-316.
To determine risk for avian influenza virus infection, we conducted serologic surveillance for H5 and H9 subtypes among poultry workers in Beijing, China, 2009–2010, and assessed workers’ understanding of avian influenza. We found that poultry workers had considerable risk for infection with H9 subtypes. Increasing their knowledge could prevent future infections.
PMCID: PMC3563274  PMID: 23343592
Avian influenza; influenza; viruses; poultry workers; serologic survey; knowledge; attitudes; practices
17.  Enzyme-inhibitor-like tuning of Ca2+ channel connectivity with calmodulin 
Nature  2010;463(7283):968-972.
Ca2+ channels and calmodulin are two prominent signaling hubs1 that synergistically impact functions as diverse as cardiac excitability2, synaptic plasticity3, and gene transcription4. It is thereby fitting that these hubs are in some sense coordinated, as the opening of CaV1-2 Ca2+ channels are regulated by a single calmodulin (CaM) constitutively complexed with channels5. The Ca2+-free form of CaM (apoCaM) is already preassociated with the IQ domain on the channel carboxy terminus, and subsequent Ca2+ binding to this ‘resident’ CaM drives conformational changes that then trigger regulation of channel opening6. Another potential avenue for channel-CaM coordination could arise from the absence of Ca2+ regulation in channels lacking a preassociated CaM6,7. Natural fluctuations in CaM levels might then influence the fraction of regulatable channels, and thereby the overall strength of Ca2+ feedback. However, the prevailing view has been that the ultra-strong affinity of channels for apoCaM ensures their saturation with CaM8, yielding a significant form of concentration independence between Ca2+ channels and CaM. Here, we reveal significant exceptions to this autonomy, by combining electrophysiology to characterize channel regulation, with optical FRET sensor determination of free apoCaM concentration in live cells9. This approach translates quantitative CaM biochemistry from the traditional test-tube context, into the realm of functioning holochannels within intact cells. From this perspective, we find that long splice forms of CaV1.3 and CaV1.4 channels include a distal carboxy tail10-12 that resembles an enzyme competitive inhibitor, which retunes channel affinity for apoCaM so that natural CaM variations affect the strength of Ca2+ feedback modulation. Given the ubiquity of these channels13,14, the connection between ambient CaM levels and Ca2+ entry via channels is broadly significant for Ca2+ homeostasis. Strategies like ours promise key advances for the in situ analysis of signaling molecules resistant to in vitro reconstitution, such as Ca2+ channels.
PMCID: PMC3553577  PMID: 20139964
18.  Autophagy inhibition plays the synergetic killing roles with radiation in the multi-drug resistant SKVCR ovarian cancer cells 
Autophagy has attracted attentions as a novel mechanism for tumor development. In this study Human ovarian carcinoma cell line SKOV3 and multidrug-resistant phenotype SKVCR cells were used and the roles of autophagy in radiation-induced cell death were analyzed.
Methods and materials
Cell viability was examined by colony formation and cell counting kit-8 (CCK-8) assay, 3MA and ZVAD were used to block autophagy and apoptosis, respectively. Quantitative real-time PCR was used to detect mRNA level and Western blot was used to detect protein expression, monodansylcadaverine (MDC) staining and flow cytometery were used for autophagy, apoptosis and cell cycle dynamics, respectively.
(1) The radiosensitivity exhibited differently in SKOV3 and SKVCR cells (SKOV3: D0=3.37, SKVCR: D0= 4.18); compared with SKOV3 the constitutive expression of MAPLC3 in SKVCR was higher, but no change of Caspase-3 and cleaved Caspase-3. (2) The ionizing radiation (IR)- induced apoptosis and autophagy were significant in both cells (P<0.05); inhibition of apoptosis with ZVAD showed no impact on survival of SKOV3 and SKVCR cells after radiation, while inhibition of autophagy significantly decreased viability in SKVCR cells, for SKVO3 cells only low level of radiation (2 Gy and 4 Gy) could decrease the viability(P<0.05). (3) ZVAD inhibited apoptosis and autophagy in both cells, 3MA inhibit apoptosis in SKOV3, and promote apoptosis in SKVCR, together with inhibition of autophagy. (4) G2/M arrest was induced by radiation in both cells; the accumulation of G2/M was more significant in SKOV3, 3MA attenuated the radiation-induced S phase delay in SKVCR.
IR-induced autophagy provides a self-protective mechanism against radiotherapy in SKVCR cells, the use of autophagy inhibitor, 3MA, increases the killing effects of radiation by inhibiting autophagy and radiation- induced S phase delay, also by the increase of apoptosis, which suggests a better therapeutic strategy in drug- resistant SKVCR ovarian cancer cells.
PMCID: PMC3552756  PMID: 23244773
Autophagy; Radiosensitivity; Multidrug resistance; Ovarian cancer; Apoptosis
19.  Virulent Avian Infectious Bronchitis Virus, People’s Republic of China 
Emerging Infectious Diseases  2012;18(12):1994-2001.
YN strain shows severe pathogenicity in chickens.
A virulent avian infectious bronchitis virus (IBV) was isolated from 30-day-old broiler chickens that exhibited respiratory symptoms, nephropathologic lesions, and a high proportion of deaths in the People’s Republic of China during 2005. The strain, designated YN, was genetically and pathologically characterized. Phylogenetic analysis showed that YN and most of the previously characterized IBV isolates found in China were phylogenetically classified into 2 main genetic clusters. The YN isolate caused severe lesions and resulted in deaths of 65% in experimental infections of 30-day-old specific-pathogen–free chickens. Tracheal and severe kidney lesions developed in all infected birds, confirming the ability of YN strain to induce both respiratory and renal disease. IBV antigens were detected by immunohistochemical analysis in the trachea, lung, kidney, and bursa, consistent with histopathologic observations, virus isolation, and reverse transcription PCR detection. We showed that YN IBV exhibits severe pathogenicity in chickens, and that similar viruses are prevalent in China.
PMCID: PMC3557894  PMID: 23171741
Avian infectious bronchitis virus; virulence; pathogenicity; tissue tropism; phylogenetic analysis; viruses; People’s Republic of China; chickens
20.  The dependence of Ni-Fe bioxide composites nanoparticles on the FeCl2 solution used 
Ni2O3- γ-Fe2O3 composite nanoparticles coated with a layer of 2FeCl3·5H2O can be prepared by co-precipitation and processing in FeCl2 solution. Using vibrating sample magnetometer (VSM), X-ray diffraction (XRD), transmission electron microscopy (TEM) and X-ray photoelectron spectroscopy (XPS) diffraction techniques, the dependence of the preparation on the concentration of the FeCl2 treatment solution is revealed.
The magnetization of the as-prepared products varied non-monotonically as the FeCl2 concentration increased from 0.020 M to 1.000 M. The Experimental results show that for the composite nanoparticles, the size of the γ-Fe2O3 phase is constant at about 8 nm, the Ni2O3 phase decreased and the 2FeCl3·5H2O phase increased with increasing concentration of FeCl2 solution. The magnetization of the as-prepared products mainly results from the γ-Fe2O3 core, and the competition between the reduction of the Ni2O3 phase with the increase of the 2FeCl3·5H2O phase resulted in the apparent magnetization varying non-monotonically.
When the concentration of FeCl2 treatment solution did not exceed 0.100 M, the products are spherical nanoparticles of size about 11 nm; their magnetization increased monotonically with increasing the concentration of FeCl2 solution due to the decreasing proportion of Ni2O3 phase.
PMCID: PMC3532291  PMID: 23110795
Composite; Nanoparticles; FeCl2 solution; Concentration
21.  Multiple linear epitopes (B-cell, CTL and Th) of JEV expressed in recombinant MVA as multiple epitope vaccine induces a protective immune response 
Virology Journal  2012;9:204.
Epitope-based vaccination might play an important role in the protective immunity against Japanese encephalitis virus (JEV) infection. The purpose of the study is to evaluate the immune characteristics of recombinant MVA carrying multi-epitope gene of JEV (rMVA-mep). The synthetic gene containing critical epitopes (B-cell, CTL and Th) of JEV was cloned into the eukaryotic expression vector pGEM-K1L, and the rMVA-mep was prepared. BALB/c mice were immunized with different dosages of purified rMVA-mep and the immune responses were determined in the form of protective response against JEV, antibodies titers (IgG1 and IgG2a), spleen cell lymphocyte proliferation, and the levels of interferon-γ and interleukin-4 cytokines. The results showed that live rMVA-mep elicited strongly immune responses in dose-dependent manner, and the highest level of immune responses was observed from the groups immunized with 107 TCID50 rMVA-mep among the experimental three concentrations. There were almost no difference of cytokines and neutralizing antibody titers among 107 TCID50 rMVA-mep, recombinant ED3 and inactivated JEV vaccine. It was noteworthy that rMVA-mep vaccination potentiates the Th1 and Th2-type immune responses in dose-dependent manner, and was sufficient to protect the mice survival against lethal JEV challenge. These findings demonstrated that rMVA-mep can produce adequate humoral and cellular immune responses, and protection in mice, which suggested that rMVA-mep might be an attractive candidate vaccine for preventing JEV infection.
PMCID: PMC3511265  PMID: 22985466
Japanese encephalitis virus; rMVA-mep; Immune response; Protection response
22.  Interplay between the Westerlies and Asian monsoon recorded in Lake Qinghai sediments since 32 ka 
Scientific Reports  2012;2:619.
Two atmospheric circulation systems, the mid-latitude Westerlies and the Asian summer monsoon (ASM), play key roles in northern-hemisphere climatic changes. However, the variability of the Westerlies in Asia and their relationship to the ASM remain unclear. Here, we present the longest and highest-resolution drill core from Lake Qinghai on the northeastern Tibetan Plateau (TP), which uniquely records the variability of both the Westerlies and the ASM since 32 ka, reflecting the interplay of these two systems. These records document the anti-phase relationship of the Westerlies and the ASM for both glacial-interglacial and glacial millennial timescales. During the last glaciation, the influence of the Westerlies dominated; prominent dust-rich intervals, correlated with Heinrich events, reflect intensified Westerlies linked to northern high-latitude climate. During the Holocene, the dominant ASM circulation, punctuated by weak events, indicates linkages of the ASM to orbital forcing, North Atlantic abrupt events, and perhaps solar activity changes.
PMCID: PMC3431539  PMID: 22943005
23.  The Role of Affect in Attentional Functioning for Younger and Older Adults 
Although previous research has shown that positive affect (PA) and negative affect (NA) modulate attentional functioning in distinct ways, few studies have considered whether the links between affect and attentional functioning may vary as a function of age. Using the Attention Network Test (Fan et al., 2002), we tested whether participants’ current state of PA and NA influenced distinct attentional functions (i.e., alerting, orienting, and executive attention) and how the relationships between affective states and attentional functioning differ in younger (18–25 years) and older (60–85 years) age groups. While there were age differences in alerting efficiency, these age differences were mediated by PA, indicating that the higher state PA found in older adults may contribute to age differences in alerting. Furthermore, age group moderated the relationship between PA and orienting as well as NA and orienting. That is, higher levels of PA and lower levels of NA were associated with enhanced orienting efficiency in older adults. Neither PA nor NA had any influence on executive attention. The current results suggest that PA and NA may influence attentional functioning in distinct ways, but that these patterns may depend on age groups.
PMCID: PMC3431793  PMID: 22969741
affect; age differences; attentional networks; individual differences; attention
24.  Deoxycytidine kinase regulates the G2/M checkpoint through interaction with cyclin-dependent kinase 1 in response to DNA damage 
Nucleic Acids Research  2012;40(19):9621-9632.
Deoxycytidine kinase (dCK) is a rate limiting enzyme critical for phosphorylation of endogenous deoxynucleosides for DNA synthesis and exogenous nucleoside analogues for anticancer and antiviral drug actions. dCK is activated in response to DNA damage; however, how it functions in the DNA damage response is largely unknown. Here, we report that dCK is required for the G2/M checkpoint in response to DNA damage induced by ionizing radiation (IR). We demonstrate that the ataxia–telangiectasia-mutated (ATM) kinase phosphorylates dCK on Serine 74 to activate it in response to DNA damage. We further demonstrate that Serine 74 phosphorylation is required for initiation of the G2/M checkpoint. Using mass spectrometry, we identified a protein complex associated with dCK in response to DNA damage. We demonstrate that dCK interacts with cyclin-dependent kinase 1 (Cdk1) after IR and that the interaction inhibits Cdk1 activity both in vitro and in vivo. Together, our results highlight the novel function of dCK and provide molecular insights into the G2/M checkpoint regulation in response to DNA damage.
PMCID: PMC3479177  PMID: 22850745
25.  Temporal trend and climate factors of hemorrhagic fever with renal syndrome epidemic in Shenyang City, China 
BMC Infectious Diseases  2011;11:331.
Hemorrhagic fever with renal syndrome (HFRS) is an important infectious disease caused by different species of hantaviruses. As a rodent-borne disease with a seasonal distribution, external environmental factors including climate factors may play a significant role in its transmission. The city of Shenyang is one of the most seriously endemic areas for HFRS. Here, we characterized the dynamic temporal trend of HFRS, and identified climate-related risk factors and their roles in HFRS transmission in Shenyang, China.
The annual and monthly cumulative numbers of HFRS cases from 2004 to 2009 were calculated and plotted to show the annual and seasonal fluctuation in Shenyang. Cross-correlation and autocorrelation analyses were performed to detect the lagged effect of climate factors on HFRS transmission and the autocorrelation of monthly HFRS cases. Principal component analysis was constructed by using climate data from 2004 to 2009 to extract principal components of climate factors to reduce co-linearity. The extracted principal components and autocorrelation terms of monthly HFRS cases were added into a multiple regression model called principal components regression model (PCR) to quantify the relationship between climate factors, autocorrelation terms and transmission of HFRS. The PCR model was compared to a general multiple regression model conducted only with climate factors as independent variables.
A distinctly declining temporal trend of annual HFRS incidence was identified. HFRS cases were reported every month, and the two peak periods occurred in spring (March to May) and winter (November to January), during which, nearly 75% of the HFRS cases were reported. Three principal components were extracted with a cumulative contribution rate of 86.06%. Component 1 represented MinRH0, MT1, RH1, and MWV1; component 2 represented RH2, MaxT3, and MAP3; and component 3 represented MaxT2, MAP2, and MWV2. The PCR model was composed of three principal components and two autocorrelation terms. The association between HFRS epidemics and climate factors was better explained in the PCR model (F = 446.452, P < 0.001, adjusted R2 = 0.75) than in the general multiple regression model (F = 223.670, P < 0.000, adjusted R2 = 0.51).
The temporal distribution of HFRS in Shenyang varied in different years with a distinctly declining trend. The monthly trends of HFRS were significantly associated with local temperature, relative humidity, precipitation, air pressure, and wind velocity of the different previous months. The model conducted in this study will make HFRS surveillance simpler and the control of HFRS more targeted in Shenyang.
PMCID: PMC3247297  PMID: 22133347

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