2010 Vol. 23, No. 4

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Proteome Analysis of Neisseria meningitidis Serogroup Strains C Associated with Outbreaks in China
YUAN HU, ZHU-JUN SHAO, XIAO-MEI YAN, BO-QING LI, FEI ZHAO, DI XIAO, JUN REN, MING-HUAN ZHENG, CHUN-XIANG FAN, LI-HUA HE, LI XU, YI-XIN GU, HAI JIANG, FENG-HUA GUO, ZHEN-WEI DAI, HONG-YU REN, MEI-JUAN LU, XIA CHEN, QING-HUA ZOU, FAN-LIANG MENG, JIAN-ZHONG ZHANG
2010, 23(4): 251-258.
Objective During 2003-2005, an outbreak of meningitis due to Neisseria meningitidis serogroup C occurred in China.With the aim to find strain clues result in the final epidemics, the ancestral strain 053442, a clinical isolate, and a carrier strain 053426 with different gene type were analyzed. Methods Clinical strain 053442 and carrier strain 053426 were cultured on GC agar plates under the same condition. Two-dimensional electrophoresis was performed using the pH 3-10 nonlinear IPG strips of 24 cm length, and all the protein spots were identified by matrix-assisted laser desorption/ionization time of flight spectrometry. Results 502 and 380 protein spots were identified in 053426 and 053442 respectively, relating to 266 and 202 different genes covering a wide range of cellular functions. The express volume and number of proteins involved in energy metabolism, protein synthesis and amino acid biosynthesis in 053426 were higher than in 053442. Virulence factor Opa, Opc and a series of proteins involved in pilus assembly and retraction were identified in 053442, which appear to be of primary importance in colonization and invasion of human cells. Compared to 053442, virulence protein species were less in 053426,with lower express volumes too. No Opa and Ope were detected in 053426. Conclusion The different protein expression profiles of the clinical strain 053442 and carrier strain 053426 in the present study provide some clues of the different pathogenicity of the two strains, which may account for result in the final epidemics.
Gene Expression Profiles Comparison between 2009 Pandemic and Seasonal H1N1 Influenza Viruses in A549 Cells
XIAO-XING YANG, NING DU, JIAN-FANG ZHOU, ZI LI, MIN WANG, JUN-FENG GUO, DA-YAN WANG, YUE-LONG SHU
2010, 23(4): 259-266.
Objective To perform gene expression profiles comparison so that to identify and understand the potential differences in pathogenesis between the pandemic and seasonal A (H1N1) influenza viruses. Methods A549 cells were infected with A/California/07/09 (H1N 1) and A/GuangdongBaoan/51/08 (H1N1) respectively at the same MOI of 2 and collected at 2, 4, 8,and 24 h post infection (p.i.). Gene expression profiles of A549 cells were obtained using the 22 K Human Genome Oligo Array,and differentially expressed genes were analyzed at selected time points. Results Microarrays results indicated that both of the viruses suppressed host immune response related pathways including cytokine production while pandemic H1N1 virus displayed weaker suppression of host immune response than seasonal H1N1 virus. Observation on similar anti-apoptotic events such as activation of apoptosis inhibitor and down-regulation of key genes of apoptosis pathways in both infections showed that activities of promoting apoptosis were different in later stage of infection. Conclusion The immuno-suppression and anti-apoptosis events of pandemic H1N1 virus were similar to those seen by seasonal H1N1 virus. The pandemic H1N1 virus had an ability to inhibit biological pathways associated with cytokine responses, NK activation and macrophage recognition.
Expressions of Nucleoside Diphosphate Kinase (nm23)in Tumor Tissues are Related with Metastasis and Length of Survival of Patients with Hepatocellular Carcinoma
RUN AN, JIE MENG, QI SHI, XIAO-XIA DAI, JING-HONG CHEN, YAN-JUN LEI, BING SHAN, CHEN GAO, YONG-LIE CHU, XIAO-PING DONG
2010, 23(4): 267-272.
Objective To evaluate the relationship of expressions of nucleoside diphosphate kinase (nm23) and proliferating cell nuclear antigen (PCNA), as well as apoptosis, with the prognosis of HCC patients by analyzing their pathological and clinical data. Methods The expressions of nm23 and PCNA were analyzed by immunohistochemistry and the apoptotic phenomena were detected by TUNEL technique in the liver samples from 43 HCC tissues, 39 para-neoplastic tissues, and 10 normal tissues.The mean apoptosis index (AI) and proliferative index (PI) in individual sample were calculated. Results As shown by the detection, 32.6% of carcinomas had negative nm23 signal in tumor tissues, whereas all para-neoplastic and normal tissues had positive nm23. The AI in nm23 positive HCC was significantly higher than that in nm23 negative one, with statistical difference (P<0.05). Furthermore, the expressions of nm23, and the values of AI and PI were contrastively analyzed with some main pathological and clinical data of HCC. It revealed that HCC with extrahepatic metastasis showed remarkable correlation with the negative nm2.3 (P=0.013) and higher PI values of HCC (P=0.015). The disease-free survival in HCC patients with negative nm23 expression was significantly poorer than that in patients with positive nm23 expression. Conclusion These data suggest that expressions of nm23 protein in rumor tissues are correlated with occurrences of metastasis and length of survival of the HCC patients, which may be an indicator for their prognosis.
CagA+ H. pylori Induces Akt1 Phosphorylation and Inhibits Transcription of p21WAF1/CIP1 and p27KIP1 via PI3K/Akt1 Pathway
SHU-PING LI, XUE-JUN CHEN, AI-HUA SUN, JIN-FANG ZHAO, JIE YAN
2010, 23(4): 273-278.
Objective Cytotoxin-associated protein (CagA) of H. pylori has been confirmed to be closely associated with gastric inflammation and tumorigenesis, but the mechanism behind it is little understood. In this study, we try to determine roles of CagA+ strain in activating PI3K/Aktl signaling pathway, and affecting expression of p21WAF1/CIP1 and p27KIP1, and also in releasing IL-8 in host cells. Methods Akt1 phosphorylation and IL-8 levels of CagA- and CagA- strain infected AGS cells were detected by ELISAs. Two quantitative RT-PCRs were established to measure p21WAF1/CIP1 and p27KIP1 mRNA levels in the CagA+ and CagA- strain infected cells. LY294002, an inhibitor of PI3K/Akt pathway, was used to define effect of the pathway in IL-8 release. Results CagA+ strain could induce an obvious elevation ofAktl phosphorylation in the infected AGS cells while CagA- strain failed to do so. The CagA+ H. pylori strain infected AGS cells showed significant drops both in p21WAF1/CIP1and p27KIP1 mRNA levels, whereas the CagA- H. pylori strain caused a remarkable increase in p21WAF1/CIP1 mRNA without affecting p27KIP1 gene transcription in the AGS cells. Both the CagA+ and CagA- H. pylori strains enabled AGS cells to produce close elevated levels of IL-8, and the LY294002 block resulted in unexpected elevations of IL-8 levels. Conclusion CagA can activate PI3K/Aktl pathway that plays an inhibitory role in IL-8 release in H. pylori infected AGS cells. Activation of PI3K/Aktl pathway and subsequent negative regulation of p21WAF1/CIP1 and p27KIPt expression might be involved in CagA-associated carcinogenesis.
Living Conditions and Palliative Care Needs among End-of-Life Former Commercial Plasma Donors Affected with HlV/AIDS in Rural Henan of China
YU SHENG, ZE-QI QIU, YUN HE, NAOMI JUNIPER, YAN ZHANG
2010, 23(4): 279-286.
Objective China began providing antiretroviral therapy to people living with HIV/AIDS (PLWHA) in 2003. This study was to investigate the living conditions, including quality of life and happiness, and need for palliative care of end-of-life PLWHA in rural Henan. Methods One hundred end-of-life AIDS patients were selected from Weishi, Zhenping and Tanghe counties in Henan, using convenience sampling. The World Health Organization Quality of Life for HIV (WHOQOL-HIV)BREF Chinese Version was used to measure the quality of life and the Memorial University of Newfoundland Scale of Happiness (MUNSH) was employed to measure subjective welfare. Qualitative interviews and focus group discussions were undertaken to learn about the palliative care provided and the specific needs of the end-of-life patients. Results Patients,overall quality of life was moderate (12.62±1.97). Highest scores were in the spirituality/religion/personal beliefs, higher than the average scores in the Chinese population (P<0.01), while psychological (13.58±2.06) and environment (12.50±3.28) domain scores were similar to the latter (P>0.05). Both independence (12.15±2.15) and physiological (14.04+3.16) domain scores were lower than the average of the people living with HIV/AIDS in other studies (P<0.01); however, all were in the moderate range.The average MUNSH score was 21.00±6.20, which was also moderate. The in-depth interviews indicated that the Henan Provincial Government's policy of treatment and care had a beneficial impact on end-of-life AIDS patients, although the care components could be improved. Conclusions Living conditions of the end-of-life AIDS patients were moderate, and the HIV/AIDS palliative care model used was beneficial to them. Care could be improved by assisting the family unit as a whole.
Estimation of Probability of Unprotected Heterosexual Vaginal Transmission of HIV-1 from Clients to Female Sex Workers in Kaiyuan,Yunnan Province,China
WEN-JUAN MA, JUN-JIE WANG, KATHLEEN-H. REILLY, AI-MEI BI, GUO-WEI DING, KUMI SMITH, NING WANG
2010, 23(4): 287-292.
Objective To estimate the probability of unprotected heterosexual vaginal transmission of HIV-1 from clients to Female Sex Workers (FSWs) in Kaiyuan County, Yunnan province, China, and analyze factors which impact the transmission probability. Methods An open cohort research of FSWs in Kaiyuan was created from surveys conducted in April 2008,October 2008, and April 2009. Two cross-sectional surveys of local clients were also carried out in May 2008 and November 2008. A model was developed to estimate the probability of unprotected heterosexual vaginal transmission of HIV- 1 from clients to FSWs. Results The transmission probability from clients to FSWs was estimated as 0.0023 [95% CI 0.0014-0.0032] per unprotected heterosexual act. Conclusion The transmission probability among this group engaging in commercial sex in Yunnan province differs from that found in studies from other countries, and future studies should estimate the influence factors of HIV- 1 transmission probability, such as stage of HIV infection, co-morbid sexually transmitted infections (STIs).
Gender Differences in Pulmonary Inflammation Following Systemic Cadmium Administration in Rats
JELENA STOSIC, IVANA MIRKOV, SANDRA BELIJ, MIROSLAV NIKOLIC, ALEKSANDRA POPOV, DRAGAN KATARANOVSKI, MILENA KATARANOVSKI
2010, 23(4): 293-299.
Objective To examine the presence of gender differences in pulmonary inflammation evoked by acute systemic cadmium administration in rats. Methods Presence of basic indicators of lung inflammation (inflammatory cytokine lung content, leukocyte infiltration and activity of cells recovered from lungs by enzyme digestion) was analyzed and compared in animals of the two sexes. Results Intraperitoneal administration of cadmium (1.0 mg/kg) resulted in higher cadmium content in lungs of female rats. Higher tumor necrosis factor (TNF) content was noted in lung homogenates of male rats, while interleukin-6 (IL-6) content was slightly, but signifaicantly greater in lungs of female rats. Increased leukocyte infiltration was observed in lungs of male rats, mainly due to neutrophils. Increased responsiveness to phorbol myristate acetate (PMA)stimulation was noted in cells recovered from lungs of male rats. Rise in intracellulgr content of myeloperoxidase (MPO) was noted in lung cells from cadmium-treated rats of both sexes, but higher in cells from male rats. Conclusion Presented data documented a more intense pulmonary inflammatory response to systemic cadmium administration in males, with higher IL-6levels in lungs of female individuals. These sex differences in proinflamatory activity of cadmium in lungs should be taken into consideration in studying the remote toxicity of this heavy metal.
Early Results of Urothelial Carcinoma Screening in a Risk Population of Coke Workers: Urothelial Carcinoma among Coke Workers
GIBERTI C, GALLO F, SCHENONE M, GENOVA A
2010, 23(4): 300-304.
Objective To present the protocol and the early results of a urothelial carcinoma (UC) screening analysis performed in a risk population of coke workers. Methods Between June 2006 and October 2008, 171 male workers (mean age 43 years),employed in a Ligurian coke plant (Italiana Coke S.r.l) and exposed to polycyclic aromatic hydrocarbons (PAHs) for a median period of 16 years, underwent screening for UC. Urological evaluation included medical history, physical examination, routine laboratory tests, urine analysis, urinary cytology and uCyt+ assay. In the event of signs and symptoms suggestive of UC or positive urinary tests, the workers were also subjected to urinary ultrasonography and cystoscopy with biopsy of any suspicious lesions. Results Regarding the laboratory tests, 19/171 (11%) uCyt+ samples were considered inadequate and were excluded from the outcomes assessment. Overall, urine analysis, cytology and uCyt+ were positive in 18/152 (12%) subjects who showed no evidence of UC at the scheduled check-ups. No significant association was identified between marker positivity and occupational activity. Conclusions Our results fail to show an increased risk of UC among the coke workers evaluated.However, they will need to be confirmed in the future by a larger enrollment and a longer follow-up in order to assess the definitive risk for UC after exposure to coke.
Vascular and Morphogenetic Abnormalities Associated with Exposure of Cigarette Smoke Condensate during Chicken and Murine Embryogenesis
SOHAIL EJAZ, AHMED EJAZ, AMARA SOHAIL, CHAN WOONG LIM
2010, 23(4): 305-311.
Objective Embryonic movements (EM) and angiogenesis pathways are evolutionarily conserved mechanisms which are essential for proper embryonic development. Deviations in these processes by exposure to cigarette smoke condensate (CSC)may cause vascular and morphogenetic disorders. Methods Using chicken and mouse embryos, we have demonstrated the in vivo effects of CSC on EM, vascular development, and organogenesis. Results Examination of the CSC exposed chicken embryos revealed a significant reduction in EM, stunted growth, deviated pattern of blood vessels, hemorrhages, and localized necrosis. Likewise, mouse embryos that were exposed to CSC at E8.5 and E9.5 died between El 1.5 and E12.5, respectively.These mouse embryos showed defects in morphogenesis and remodeling of the embryonic vasculature, while littermate controls showed normal development.Conclusion Cigarette smoking during pregnancy is fatal for growing embryos. CSC may induce the remodeling of embryonic vasculature, leading to various pathologies.
Metabolomic Study on Vitamins B1, B2, and PP Supplementation to Improve Serum Metabolic Profiles in Mice under Acute Hypoxia Based on 1H NMR Analysis
JIN LIU, JIAN-QUAN WU, JI-JUN YANG, JING-YU WEI, WEI-NA GAO, CHANG-JIANG GUO
2010, 23(4): 312-318.
Objective To explore metabolic changes after acute hypoxia and modulating effect of vitamins B1,B2, and PP supplementation in mice exposed to acute hypoxia. Methods Fifty male Kunming mice were randomly divided into 5 groups: normal, acute hypoxia, acute hypoxia with 2, 4 and 8 time- vitamins B1, B2, and PP supplementation. All mice were fed with corresponding diets for two weeks and then were exposed to a simulated altitude of 6 000 meters for 8 h, except for the normal group. Nuclear magnetic resonance analysis was used to identify the changes of serum metabolic profiles. Results There were significant changes in some serum metabolites under induced acute hypoxia, essentially relative increase in the concentrations of lactate, sugar and lipids and decrease in ethanol. The serum levels of choline, succinate, taurine, alanine, and glutamine also increased and phosphocholine decreased in the acute hypoxia group. After vitamins B1, B2, and PP supplementation, all these metabolic changes gradually recovered. Conclusion Significant changes in serum metabolic profile were observed by metabolomics in mice exposed to acute hypoxia, and vitamins B1, B2, and PP supplementation proved to be beneficial to improving some metabolic pathways. It is suggested that the dietary intakes of vitamins B1, B2, and PP should be increased under hypoxia condition.
A Study on Pollen Allergens in China
ZHI-GANG LIU, JUAN-JUAN SONG, XIAO-LI KONG
2010, 23(4): 319-322.
Objective Allergic disease caused by airborne pollen is a major health problem in China. Intensive study on pollen allergens can be of great help for preventing and treating pollinosis. Four aspects of the study on pollen allergens in China including major allergic pollen in our country, analysis and purification of pollen allergen composition, recombinant pollen allergens and clinical application of pollen allergens are described in this paper.
A(H1N1)Influenza Pneumonia with Acute Disseminated Encephalomyelitis: A Case Report
JUN YANG, YU-GUANG WANG, YUN-LIANG XU, XIAN-LING REN, YU MAO, XING-WANG LI
2010, 23(4): 323-326.
An Association between Immunosenescence and CD4+CD25+ Regulatory T Cells: A Systematic Review
LING WANG, YAN XIE, LI-JING ZHU, TING-TING CHANG, YAN-QING MAO, JIE LI
2010, 23(4): 327-332.
Objective Age-related increment of the prevalence of CD4+CD25+ regulatory T (Treg) cells were described controversially, and whether such changes explain immune dysfunction in the elderly is still unclear. The aim of this systematic review is to evaluate the role of the Tregs in immunosenescence. Methods Medline and manual searches were performed to identify all published epidemiological and animal studies investigating the efficacy of the association between immunosenescence and Treg cells. Results It was founded that the frequency, phenotypic characteristics, and number/function of Tregs were altered significantly with aging. Medical conditions in individuals with advanced ageas well as apoptosis intensity of Treg cells had an impact on the accumulation of Tregs which in turn could deteriorate cytotoxic activity of CD8+ T and NK cells and production of IL-2. The range of immune cells that could be suppressed by Treg cells was quite wide and covered CD4+CD25+ T cells, NK cells, dendritic cells and even monocyrtes. These changes were observed both in humans and experimental animals. Besides, it was believed that frequency of Tregs increased with age and was accompanied by intensified suppressive activity for Tregs in patients, for example, with Alzheimer disease (AD) and Parkinson disease (PD).The impaired condition of CD4+ T cells, so-called immunosenescence, rendered transplant recipients less responsive to an allogeneic kidney graft, an effect that was limited to transplant recipients who were aged over 60 years. Conclusions Treg cells are associated with immunosenescence. All these changes contribute to the aging-related decline of immune responses and lead to the higher risk of immune-mediated diseases, cancer or infections in aged individuals.
Biomedical and Environmental Sciences
2010, 23(4)