The purpose of this study is to obtain effective and stable bacteriocin-like substance from lactic acid bacteria. Lactobacillus buchneri KLDS1.0364,which was isolated from fermented cream,a traditional dairy product in Inner Mongolia,could produce bacteriocin-like substance. The bacteriocin-like substance produced by KLDS1.0364 was partially purified and the characteristics were studied. The bacteriocin-like substance was purified by SP-Sepharose fast flow cation exchange chromatography. The molecular weight of the bacteriocin-like substance was 21.6kD,as determined by tricine-SDS-PAGE. The bacteriocin-like substance remained stable after 30 min at 121℃ and after 2 h of incubation during pH 2～10. Complete inactivation of antimicrobial activity was observed after treatment of the bacteriocin-like substance with several proteinases. Treatment with catalase and ?-amylase did not result in any changes of antimicrobial activity. The mode of action of the bacteriocin-like substance is bactericidal. It exhibited a broad spectrum of antagonistic activity against various species of Gram-positive bacteria,Gram-negative bacteria and fungi.

ObjectiveTo investigate mechanisms for IL-27 induced proliferation and differentiation of peripheral blood CD4+ T cells in primary biliary cirrhosis (PBC).MethodsPeriperal blood CD4+ T cells were isolated from patients with PBC,chonic hepatitis B (CHB) and health controls (HCs).After IL-27 stimulation,proliferation ability of CD4+ T cells was evaluated by CCK-8 kit,and cytokines were analyzed by ELISA.Real-time PCR was employed to assay mRNA expression of T-bet and GATA3 in CD4+ T cells.p-STAT-1 and pSTAT-3 expression in CD4+ T cells were detected by Western blot.ResultsEnhanced proliferation of CD4+ T cells was found in all subjects after IL-27 stimulation.However,the proliferation ability in patients with PBC was greater than that in CHB and HCs ( P＜0.001 ).Levels of IL-2 and IFN-γ in supernatant from IL-27-incubated PBC blood CD4+ T cells were higher than that from CHB and HCs (P＜0.001 ).In normal situation,T-bet mRNA of CD4+ T cells in PBC group was higher than that in CHB group (P=0.007).Furthermore,after IL-27 stimulation,elevated T-bet mRNA expression and GATA3 inhibition were found in patients with PBC.High expression of p-STAT-1 and p-STAT-3 in blood CD4+ T cells were found in PBC,CHB and HCs after stimulation by IL-27.But their expression in patients with PBC were higher than those in patients with CHB and HCs.ConclusionProliferation of blood CD4+ T cells could be induced by IL-27 in patients with PBC.The signaling pathways of p-STAT-1,p-STAT-3 were involved to induce Th1 immune response and related cytokines expression.This study implicated that IL-27 may play important roles in early inflammation damage in PBC.

Objective To systematically investigate the molecular epidemiological profiles of human papillomavirus (HPV) in patients with condyloma acuminata(CA). Methods Two hundred and one samples of HPV DNA isolated from CA were PCR amplified by the PGMY09/11 primer system. The PCR products were simultaneously hybridized to 37 specific HPV probes immobilized on a nylon strip and then genotyped. All DNA templates were further PCR amplified using HPV 6 and 11 type specific primers for verification. Results All samples were HPV DNA positive consisting of totally 31 genotypes, the types of which were type 11(53.7%, 108/201), 6(43.8%, 88/201), 16(6.5%, 13/201), 52(6.0%, 12/201), 33(5.5%, 11/201), cp6108 (5.5%, 11/201) and 42 (5.0%, 10/201). The samples infected with a single and mixed types of HPV accounted for 60.2% (121/201) and 39.8% (80/201) respectively. Consistent results were found with the detection of HPV6 and 11 between hybridization assay and type-specific PCR. Conclusions At least 31 HPV genotypes are associated with CA. HPV 11 predominates while 68, 40, 54, 67, 73, 82, 35, 64 and 83 are rare in CA. Type cp6108 is detected in CA for the first time with a high prevalence. HPV26, 69, 70, 71,72 and IS39 might be not associated with CA. CA infected with a single and mixed HPV types accounts for 60.2% and 39.8%, respectively.

AIMTo explore the effect of chronic hypoxic hypercapnia on learning-memory and the possible mechanisms involved.

METHODSFifty-eight male SD rats were randomly divided into three groups: Normal control group (NC, n=18), 2-week (2HH, n=18), and 4-week hypoxic hypercapnia (4HH, n=20) group. The rats, spatial learning-memory tasks were assessed by the Morris water maze. The expression of NMDAR1mRNA was determined by hybridization in situ.

RESULTSCompared with NC group, rats exposed to chronic hypoxic hypercapnia displayed significant impairment in their performance assessed by two measures: mean escape latencies (2HH: 38.59 +/- 8.35 s, 4HH: 60.59 +/- 17.28 s) and swim path distances(2HH: 9893.45 +/- 1958.16 mm, 4HH: 18077.57 +/- 6878.85 mm). The expression level of NMDAR1mRNA in the hippocampus and cortex were lower than those in the NC group, especially, the NMDAR1mRNA expression of hippocampus CA1 in 4HH decreased by 21.4% (P < 0.01).

CONCLUSIONChronic hypoxic hypercapnia could impair the rat spatial learning-memory and the decrease in expression of NMDAR1mRNA might be involved in.

Animals ; Hypercapnia ; metabolism ; Hypoxia ; metabolism ; Male ; Maze Learning ; Memory ; Rats ; Rats, Sprague-Dawley ; Receptors, N-Methyl-D-Aspartate ; metabolism

OBJECTIVETo investigate the protective effect of exogenous inhibitor of matrix metalloproteinases-9 (MMP-9), batimastat, in the lung injury induced by cardiopulmonary bypass (CPB) in dogs.

METHODSThirty healthy mongrel puppies were randomly divided into 3 groups: control group, low-dose group [batimastat 10 mg/(kg.d) for 3 days before operation] and high-dose group [batimastat 30 mg/(kg.d) for 3 days before operation]. The off-pump puppies' model of acute lung injury was established, and hemodynamic and respiratory parameters were monitored. The preoperative and postoperative alveolar-arterial oxygen difference (A-aDO(2)) and respiratory index (RI) were calculated. From the beginning of surgery, blood samples were taken at the time 0, 60, 120, and 270 min. Plasma concentrations of MMP-9 were measured by ELISA, and blood MMP-9 mRNA expressions were determined by RT-PCR. The myeloperoxidase (MPO) activity of centrifugal bronchoalveolar lavage fluid were measured by Colorimetry. And MMP-9 activity was determined by Gelatin zymography. Light and electronic microscope were used to observe the morphological changes of lung tissue. A small piece of left lung tissue was taken, weighed and baked to calculate the wet weight (W/D) index.

RESULTSAfter cardiopulmonary bypass, the concentrations of MMP-9 and mRNA expressions of the control group were increased significantly, and lung injury was apparent. At 270 min, the MMP-9 plasma concentration of high-dose group (17.36 +/- 1.18) microg/L was significant reducing than control group (30.47 +/- 2.22) microg/L (P < 0.05). After operation, A-aDO(2) and RI of high-dose group were significantly improved than control group (P < 0.05). The W/D index of the high-dose group (2.8 +/- 0.48) was significantly lower than that of control group (4.7 +/- 0.6) (P < 0.05). And the pathological changes of lung tissue were significantly improved in the high-dose group. However, there was no significant difference in the MMP-9 mRNA expression in three groups.

CONCLUSIONSBatimastat plays a role in the protection of the lung injury of CBP by reducing the concentration and activity of MMP-9, the degradation of the cell membrane and pulmonary neutrophil infiltration and reduction of pulmonary edema.

Acute Lung Injury ; etiology ; prevention & control ; Animals ; Cardiopulmonary Bypass ; Disease Models, Animal ; Dogs ; Lung ; pathology ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinase Inhibitors ; Phenylalanine ; analogs & derivatives ; pharmacology ; Postoperative Complications ; prevention & control ; Thiophenes ; pharmacology

Membrane (M) protein genes of 20 infectious bronchitis virus (IBV) strains isolated in China between 1995 and 2004 were sequenced and analyzed. The M genes of twenty isolates were composed of 672 to 681 nucleotides, encoding polypeptides of 223 to 226 amino acid residues. Variations of the deduced amino acids of M gene mainly occurred at positions 2 to 17 and 221 to 233, comparing with that of the IBV strain LX4. There were deletions or insertions in the M gene of Chinese isolates at amino acid position 2 to 6, leading to the loss or gain of a glycosylation site. Phylogenetic tree based on amino acid sequences of M genes from 20 Chinese isolates and 34 reference strains showed that they were classified into five distinct clusters. Most of the Chinese IBV strains were included in clusters II and IV, forming distinct groups. The isolates in cluster II showed a close evolutionary relationship with Taiwan isolates. Furthermore, recombination especially the recombination between field isolates and vaccine strains had been observed while comparing the phylogeny of M genes with those of S1 and N genes.
Amino Acid Sequence ; China ; Genetic Variation ; Infectious bronchitis virus ; classification ; genetics ; isolation & purification ; Molecular Sequence Data ; Phylogeny ; Sequence Homology, Amino Acid ; Viral Matrix Proteins ; genetics

Adolescent ; Adult ; China ; HIV Infections ; prevention & control ; Humans ; Male ; Rural Population ; Sexual Behavior ; Single Person ; Young Adult

OBJECTIVETo observe the effect of compound Puerarin on collagen IV of streptozotocin-induced diabetic rats.

METHODSDiabetic nephropathy rats were induced by intraperitoneal injection of streptozotocin (STZ). Rats were allocated randomly to control group (10), diabetes model group (10), Vitamin C group (10), Puerarin group (10), vitamin C plus Puerarin group (10). The study period lasted for 12 weeks. During and after the treatment, the general state, blood glucose levels, glycosylated hemoglobin, blood urea nitrogen, serum collagen IV, blood urea nitrogen, serum creatinine, urinary albumin excretion rate of the 24-hour, and clearance rate of creatinine collagen IV protein were determined by immunohistochemistoche analysis as well as type the gene expression of collagen IV alpha 1 mRNA were determined by in situ hybridization analysis in the kidney tissue of different groups.

RESULTS(1) Diabetes mellitus and renal function lesion occurred in the four groups. (2) Vitamin C and Puerarin could improve the general conditions of diabetic Rats, decrease blood urea nitrogen [(8.68 +/- 0.43), (7.98 +/- 0.47) and (5.76 +/- 0.82) micromol/L, serum creatinine [(74.68 +/- 8.20), (75.52 +/- 7.98) and (58.66 +/- 6.65) mmol/L], and urinary albumin excretion rate of the 24-hour [(18.40 +/- 0.37), (17.24 +/- 0.30) and (9.97 +/- 1.27) mg/24 h x 10(-3)]; increase clearance rate of creatinine [(0.59 +/- 0.21), (0.61 +/- 0.14) and (0.69 +/- 0.32) ml/min], the expression of collage IV absorbance [(111.56 +/- 14.61), (110.78 +/- 9.69) and (95.44 +/- 9.97) ] in the diabetic Rats were significantly inhibited at the same time.

CONCLUSIONThe compound Puerarin might have some functions on preventing ren by inhibiting expression of type IV collagen.

Animals ; Collagen Type IV ; antagonists & inhibitors ; biosynthesis ; Diabetes Mellitus, Experimental ; drug therapy ; metabolism ; Diabetic Nephropathies ; drug therapy ; metabolism ; Isoflavones ; pharmacology ; therapeutic use ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

To study the impact of the enterovirus 71(EV71) on the nuclear transport mechanism,The pGFP-NLS vector with nuclear location signal(NLS) was constructed, RD cells transfected by the pGFP-NLS vector were inoculated with the EV71 or cotransfected by EV71-2A vector. The results showed that GFP protein with NLS was expressed in the cytoplasm due to the inhibition of nuclear transport. In order to further study the mechanism of the EV71 to prevent nuclear transport,Nup62 was detected by Western blotting after RD cells were infected with EV71 or transfected by EV71-2A vector. The results showed that decreased expression of Nup62 could be detected after infection with EV71 and transfection by EV71-2A vector. This study demonstrates that the cleavage of Nup62 by EV71 2A protease may be the mechanism of nuclear transport inhibition.
Active Transport, Cell Nucleus ; Cell Line, Tumor ; Cell Nucleus ; metabolism ; Enterovirus A, Human ; enzymology ; genetics ; metabolism ; Enterovirus Infections ; virology ; Gene Expression Regulation, Viral ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Membrane Glycoproteins ; metabolism ; Nuclear Localization Signals ; metabolism ; Nuclear Pore Complex Proteins ; metabolism ; Peptide Hydrolases ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Transfection

OBJECTIVETo explore the clinical and laboratory characteristics of two myelodysplastic syndromes (MDS) patients with double isochromosome 20q- anomaly.

METHODSBone marrow cell chromosome preparations were made with both direct method and short-term culture. Karyotype analysis was performed by R-banding technique, and dual-color FISH (fluorescence in situ hybridization) by using a 20q telomeric probe and a sequence-specific probe for 20q12.

RESULTSThe clinical and hematological findings were comparable with diagnosis of MDS. Karyotype analysis showed that both patients had double isochromosome 20q- anomaly: case 1 is 46, XX, der(20)? i(20q-) [6]/46, idem, der (6) i (6p) [1]/47, idem, +der (20)? i (20q-) [3]/47, idem, der(6)i (6p), +der(20)? i (20q-) [20]; case 2 is 45, XY, -7, der (20)? i (20q-) [17]/46, idem, +der(20) ? i(20q-) [3]. Two derivative chromosomes 20 were proved 20q isochromosomes with interstitial deletions by dual-color FISH in one patient.

CONCLUSIONSDouble isochromosome 20q- anomaly is a rare recurrent karyotype abnormality in MDS, and signals a poor prognosis.

Chromosome Banding ; Chromosomes, Human, Pair 20 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Isochromosomes ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics