1.Isolation and Identification of Marine Actinomycetes with Quorum Sensing Inhibitory Activity
Wei-Shan WANG ; Shan-Shan ZOU ; Shou-Liang YIN ; Qian-Hong GONG ; Wen-Gong YU ;
Microbiology 2008;0(09):-
Quorum sensing systems of pathogens are central regulators for the expression of virulence factors. Increasing evidence implies that targeting the quorum sensing system of many pathogenic bacteria is a promising therapeutic approach to control infections. In this work,we isolated 47 strains of actinomycetes from the mud sample of Jiaozhou Bay. Quorum sensing inhibitory activity was monitored by Chromobacterium violaceum CV026. As a result,the culture broth extract of actinomycetes WA-7 was found to have significant quorum sensing inhibitory activity. This strain was assigned to the genus Streptomyces based on its 16S rDNA sequence. Further investigation revealed that the extract could inhibit the quorum sensing-controlled violacein and proteases production of C. violaceum in a concentration-dependent manner.
3.Low-dose radiation induces endoplasmic reticulum stress and activates PERK-CHOP signaling pathway in mouse testicular cells.
Fang FANG ; Ping-Sheng GONG ; Xiang-Fu SONG ; Shou-Liang GONG ; Zhi-Cheng WANG
National Journal of Andrology 2012;18(9):777-782
OBJECTIVETo explore the correlation of low-dose radiation with endoplasmic reticulum stress and the activation of the PERK-CHOP signaling pathway in mouse testicular cells.
METHODSHealthy Kunming mice were randomly assigned to time-effect (0, 3, 6, 12 and 24 h of irradiation at 75 mGy) and dose-effect (12 h of irradiation at 0, 50, 75, 100 and 200 mGy) groups. The contents of H202 and MDA were measured by colorimetry with the agent kits, the expressions of GRP78, PERK and CHOP mRNA detected by quantitative RT-PCR, and the levels of GRP7B, PERK, phosphorylated PERK (pho-PERK) and CHOP proteins determined by Western blotting and image analysis.
RESULTSAfter whole-body irradiation of the mice with 75 mGy, the content of H2 02 in the testis tissue was increased with time prolongation, while that of MDA decreased slightly at 3 and 6 h and then increased with the lengthening of time, both increased significantly at 12 and 24 h as compared with those at 0 h (P < 0. 05, P < 0. 01). Apart from reduced levels of GRP78 mRNA at 3 and 24 h and GRP78 protein at 6 h after irradiation, significant increases were found in the mRNA expressions of GRP78 at 12 h, PERK at 3,6, 12 and 24 hand CHOP at 12 and 24 h (P < 0.05, P < 0.01), as well as in the protein levels of GRP78 at 12 and 24 h, pho-PERK at 3, 12 and 24 h and CHOP at 3, 6, 12 and 24 h in comparison with those at 0 h (P < 0. 05, P < 0. 01). No obvious regularity was observed in the change of the PERK protein expression. After 12 h of whole-body irradiation, the content of H202 was increased at 50, 75 and 100 mGy, but decreased slightly at 200 mGy, while that of MDA was increased with dose increasing, with significant increases in the content of H2 02 at 75 and 100 mCy and in that of MDA at 75, 100 and 200 mGy as compared with the 0 mGy group. Apart from the reduced levels of GRP78 mRNA at 50 and 200 mCy, significant increases were found in the mRNA expressions of PERK at 75, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 (P c 0. 05, P < 0.01) as well as in the protein levels of GRP78 at 100 and 200 mGy, pho-PERK at 50, 100 and 200 mGy and CHOP at 50, 75, 100 and 200 mCy as compared with those at 0 mGy (P < 0. 05, P < 0. 01). There were differences in the changes of different protein expressions, but no obvious regularity was seen in the change of the PERK protein expression.
CONCLUSIONLow-dose radiation can induce endoplasmic reticulum stress in mouse testicular cells, and activate the PERK-CHOP signaling pathway.
Animals ; Endoplasmic Reticulum Stress ; radiation effects ; Heat-Shock Proteins ; metabolism ; Male ; Mice ; Mice, Inbred Strains ; Radiation Dosage ; Radiation, Ionizing ; Signal Transduction ; radiation effects ; Testis ; cytology ; metabolism ; radiation effects ; Transcription Factor CHOP ; metabolism ; Whole-Body Irradiation ; eIF-2 Kinase ; metabolism
4.Influence of preoperative nutritional support on surgical outcomes of chronic radiation enteritis patients complicated with intestinal obstruction.
Liang ZHANG ; Jian-feng GONG ; Ling NI ; Qi-yi CHEN ; Zhen GUO ; Wei-ming ZHU ; Ning LI ; Jie-shou LI
Chinese Journal of Gastrointestinal Surgery 2013;16(4):340-344
OBJECTIVETo investigate the effect of preoperative nutritional support in the management of patients with chronic radiation enteritis (CRE) with intestinal obstruction undergoing resectional surgery.
METHODSClinical data of 158 CRE patients undergoing diseased bowel resection from 2001 to 2011 were analyzed retrospectively. A total of 130 patients received preoperative nutritional support, including 28 patients with enteral nutrition support, 60 patients with total parenteral nutrition support, and 42 patients with combined nutritional support. The nutritional parameters, procedures, operation-related complications, and postoperative hospital stay were recorded.
RESULTSAfter aggressive nutritional support in 130 patients, patients nutritional index, such as serum prealbumin, transferrin, serum albumin improved significantly preoperatively, while the change of body mass index and hemoglobin was not significant. Compared to those without preoperative nutritional support, those who received preoperative nutritional support had lower stoma rate (31.5% vs. 53.6%, P=0.027), less postoperative infection rate (13.8% vs. 32.1%, P=0.019), shorter postoperative hospital stay [(14.1±7.3) d vs. (18.8±15.8) d, P=0.013). Enteral nutrition group had less postoperative infection rate (7.1% vs. 21.7%, P=0.017), lower stoma rate (28.6% vs. 48.3%, P=0.02), and shorter postoperative hospital stay [(15.5±9.6) d vs. (21.7±19.0) d, P=0.025) as compared to total parenteral nutrition group.
CONCLUSIONSPreoperative nutritional support can decrease the stoma rate, postoperative infection rate, and shorten hospital stay in CRE patients complicated with intestinal obstruction. If tolerated, enteral nutrition support should be chosen.
Adult ; Aged ; Aged, 80 and over ; Chronic Disease ; Enteritis ; etiology ; surgery ; Female ; Humans ; Intestinal Obstruction ; complications ; surgery ; Male ; Middle Aged ; Nutritional Support ; methods ; Preoperative Care ; Radiation Injuries ; complications ; Retrospective Studies ; Treatment Outcome ; Young Adult
6.Research progress of new antibacterial drugs that target bacterial quorum sensing systems.
Shou-Liang YIN ; Ya-Jing CHANG ; Su-Ping DENG ; Qing-Chi WANG ; Wen-Gong YU ; Qian-Hong GONG
Acta Pharmaceutica Sinica 2011;46(6):613-621
In recent years, antibiotic resistance of bacteria has become a global health crisis. Especially, the new class of "superbug" was found in South Asia, which is resistant to almost known antibiotics and causes worldwide alarm. Through the underlying mechanisms of bacterial pathogenecity, the expression of many pathogen virulence factors is regulated by the process of quorum sensing. Screening efficient quorum sensing inhibitors is an especially compelling approach to the future treatment of bacterial infections and antibiotic resistance. This article focuses on bacterial quorum sensing system, quorum sensing screening model for in vitro and evaluation of animal models in vivo, recent research of quorum sensing inhibitors and so on.
Animals
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Anti-Bacterial Agents
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pharmacology
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therapeutic use
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Bacterial Infections
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drug therapy
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Disease Models, Animal
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Drug Resistance, Bacterial
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Pseudomonas aeruginosa
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drug effects
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pathogenicity
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physiology
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Quorum Sensing
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drug effects
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physiology
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Virulence
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drug effects
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Virulence Factors
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metabolism
7.Mitochondrial modulation of apoptosis induced by low-dose radiation in mouse testicular cells.
Fang FANG ; Ping Sheng GONG ; Hong Guang ZHAO ; Yu Jing BI ; Gang ZHAO ; Shou Liang GONG ; Zhi Cheng WANG
Biomedical and Environmental Sciences 2013;26(10):820-830
OBJECTIVETo investigate whether apoptosis induced by low-dose radiation (LDR) is regulated by mitochondrial pathways in testicular cells.
METHODSMale mice were exposed to whole-body LDR, and changes in mitochondrial function and in expression of apoptotic factors were analyzed in the testicular cells as follows. Total nitric-oxide synthase (T-NOS) and Na+/K+ ATPase activities were biochemically assayed. Reactive oxygen species (ROS) and mitochondrial membrane potential (Δψm) were determined by flow cytometry using fluorescent probes. Levels of mRNAs encoding cytochrome c (Cyt c) and apoptosis-inducing factor (AIF) were quantified by real-time reverse-transcription PCR (RT-PCR). Expression of Cyt c, AIF, caspase-9, and caspase-3 at the protein level was assessed by western blotting and immunohistochemistry.
RESULTSLDR induced an increase in T-NOS activity and ROS levels, and a decrease in Na+/K+ ATPase activity and mitochondrial Δψm, in the testicular cells. The intensity of these effects increased with time after irradiation and with dose. The cells showed remarkable swelling and vacuolization of mitochondria, and displayed a time- and dose-dependent increase in the expression of Cyt c, AIF, procaspase-9, and procaspase-3. Activation of the two procaspases was confirmed by detection of the cleaved caspases. The changes in expression of the four apoptotic factors were mostly limited to spermatogonia and spermatocytes.
CONCLUSIONLDR can induce testicular cell apoptosis through mitochondrial signaling pathways.
Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspases ; Cytochromes c ; metabolism ; Membrane Potential, Mitochondrial ; drug effects ; Mice ; Mitochondria ; drug effects ; Reactive Oxygen Species ; metabolism
8.Effects of corticosterone, cAMP, cGMP, Ca2+, and protein kinase C on apoptosis of mouse thymocytes induced by X-ray irradiation.
Shou-Liang GONG ; Li-Hua DONG ; Guang-Wei LIU ; Ping-Sheng GONG ; Wen-Tian LU ; Hong-Guang ZHAO ; Xiao-Jing JIA ; Yong ZHAO
Biomedical and Environmental Sciences 2008;21(2):167-172
OBJECTIVETo observe the effects of signal factors of corticosterone (CS), cAMP, cGMP, Ca2+ andprotein kinase C (PKC) on lymphocyte apoptosis in mouse thymus induced by X-rays of 4 Gy in vitro.
METHODSThe DNA lytic rate for thymocytes was measured by fluorospectrophotometry.
RESULTSThe DNA lyric rate for thymocytes 4-8 hours after irradiation with 2-8 Gy was significantly higher than that in the control (P<0.01). As compared with the control, the DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.05-0.4 microg/mL ionomycin (Iono, P<0.05 or P<0.01) or 0.05-0.4 ng/mL phorbol myristate acetate (PMA, P<0.05 or P<0.01), respectively, was significantly increased, while the rate for thymocytes treated with 50 ng/mL cGMP was not significantly increased. The DNA lytic rate for thymocytes treated with 0.01 micromol/L CS (P<0.01), 50 ng/mL cAMP (P<0.01), 0.2 and 0.4 microg/mL Iono (P<0.05), and 0.2 and 0.4 ng/mL PMA (P<0.05) plus 4-Gy irradiation, respectively, was significantly higher than that treated with single 4-Gy irradiation, while the rate for thymocytes treated with 50 ng/mL cGMP plus 4-Gy irradiation was not increased. When both 0.4 microg/mL Iono and 0.4 ng/mL PMA acted on the thymocytes, the DNA lytic rate for thymocytes was significantly higher than that in the control (P<0.01), the DNA lytic rate for thymocytes treated with both 0.4 microg/mL Iono and 0.4 ng/mL PMA plus 4-Gy irradiation was significantly higher than that treated with single 4-Gy irradiation (P<0.05), but was not significantly higher than that treated with 0.4 microg/mL Iono plus 4-Gy irradiation or 0.4 ng/mL PMA plus 4-Gy irradiation.
CONCLUSIONCS, cAMP, Ca2+, and PKC signal factors can promote thymocyte apoptosis induced by larger dose X-rays.
Animals ; Apoptosis ; drug effects ; radiation effects ; Calcium ; pharmacology ; Corticosterone ; pharmacology ; Cyclic AMP ; pharmacology ; Cyclic GMP ; pharmacology ; Ionomycin ; pharmacology ; Male ; Mice ; Protein Kinase C ; metabolism ; Spectrometry, Fluorescence ; Tetradecanoylphorbol Acetate ; pharmacology ; Thymus Gland ; cytology ; drug effects ; X-Rays
9.Changes of cycle and apoptosis of spermatogenic cells and antioxidant capacity in male rats with diabetes mellitus.
Hong-guang ZHAO ; Guang-wei LIU ; Shu-chun LIU ; Zhi-cheng WANG ; Yang LIU ; Zhen-qi WANG ; Cai LI ; Lu CAI ; Shou-liang GONG
National Journal of Andrology 2005;11(10):735-739
OBJECTIVETo explore the changes of cycle and apoptosis of spermatogenic cells and antioxidant capacity of the serum and testis in male rats with diabetes mellitus.
METHODSThirty male rats were divided into two groups, 10 for normal control and 20 for the diabetes group. The rats were injected intraperitoneally with streptozocin (TZ) to develop diabetes, and 12 weeks later, their survival rate and testis weight were recorded. The percentage of G0/G, S and G2/M phases and apoptosis in spermatogenic cells were measured with flow cytometry (FCM). Malondialdehyde (MDA) and nitric oxide (NO) levels, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and NO synthase (NOS) activities in the serum and testis were measured with thiobarbituric acid reactive substances (TBARs), nitric acid reoxidized enzyme, xanthine oxidative enzyme, 5,5 Dithiobis (2,2 nitrobenzoate) (TNB) and visible light photometer methods, respectively.
RESULTSTwelve weeks after the male rats got diabetes, their survival rate, body weight and testis weight were significantly lower (p < 0.05), and the percentages of G0/G1 phases and apoptotic spermatogenic cells were obviously higher (P < 0.05) than the normal control. At the same time, the percentage of S and G2/M phases spermatogenic cells decreased. So the spermatogenic cells were arrested in G0/G1 phase. In the diabetic rat serum and testis, especially in the testis, MDA levels were distinctly higher and SOD activities were significantly lower than those in the control. Serum GSH-Px activities of the diabetic rats were significantly lower (p < 0.05), while testis GSH-Px activities were significantly higher than those in control group (P < 0.01). NO contents in the serum and testis of the diabetic rats (P < 0.01) increased significantly, particularly the former, while NOS activities in the serum decreased significantly as compared with the control (P < 0.5).
CONCLUSIONThe increase in testis and serum MDA levels and NO contents and the decrease in the antioxidant enzyme activity of the diabetic rats may be relevant to spermatogenic disorder caused by the increase of G0/G1 phases arrest and spermatogenic cells apoptosis.
Animals ; Apoptosis ; Cell Cycle ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Male ; Malondialdehyde ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism ; Testis ; cytology ; metabolism
10.Construction and immunogenicity analysis of recombinant replication-defective human adenovirus type 5 bearing the porcine circovirus type 2 Cap protein gene.
Ting GONG ; Shou-Feng ZHANG ; Ye LIU ; Cheng-Long SUN ; Yang YANG ; Qi CHEN ; Fang QIAN ; Bo-Hao LIU ; Rong-Liang HU
Chinese Journal of Virology 2013;29(1):26-31
To construct a recombinant replication-defective human adenovirus type 5 expressing Cap protein of PCV2 and test the immunological efficacy in mice. In this study, the recombinant replication-defective human adenovirus type 5, named as rAd5-Cap (wt-rAd5), was constructed through homologous recombination internally in the HEK293AD cells after co-transfection of the Pac I-linearized backbone plasmid and the shuttle plasmid pacAd5CMV-Cap containing the open reading frame (ORF2) of the porcine circovirus type 2 (PCV2) cap protein or pacAd5CMV without inserted fragment. Furthermore, the rAd5-Cap could induce the expression of PCV2 cap protein in the HEK293AD cells with high efficacy evaluated by the RT-PCR and indirect immunofluorescence assay (IFA). The virus titer of rAd5-Cap could reach up to 10(8.5) TCID50/mL similarly to that of wt-rAd5, indicating that there was little affect on the virus proliferation after the insertion of PCV2 cap protein gene. The humeral immune responses could be activated and detected 14 days after the inoculation of the mice with 10(7) TCID50 rAd5-Cap intramuscularly, and constantly in crease in another 14 days. These molecular biological and animal experiments results demonstrated that the PCV2 cap protein could be efficiently expressed by the recombinant adenovirus rAd5-Cap in eukaryotic cells and induce robust immune responses in mice, which laid a good foundation for the development of new type vaccine against porcine circovirus.
Adenoviruses, Human
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genetics
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Animals
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Antibodies, Viral
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blood
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Capsid Proteins
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genetics
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immunology
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Circovirus
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immunology
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Defective Viruses
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genetics
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HEK293 Cells
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Humans
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Mice
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Recombinant Proteins
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biosynthesis
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immunology
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Virus Replication