Lysins are efficient bacteria cell wall digesting enzymes encoded by DNA bacteriophage. Gram-positive bacteriophage lysins feature similar domain structure, high lytic efficiency, synergic antibacterial effect with antibiotics, rare neutralization by antibodies, less chance of developing drug-resistant strains, et al. The past decade has seen a considerable amount of research worldwidely focused on lysin, and lysins have been used successfully in a variety of animal models to control pathogenic antibiotic resistant bacteria found on mucosal surfaces and infected tissues. The great potential of lysins as an anti-infective agent prompted this review.

The objective of this work was to investigate the effect of six individual strains of fungi on the reduction of gossypol levels and nutritional value during solid substrate fermentation of cottonseed meal (CSM). Six groups of disinfected CSM substrate were incubated for 48 h after inoculation with either of the fungi C. capsuligena ZD-1, C. tropicalis ZD-3, S. cerevisae ZD-5, A. terricola ZD-6, A. oryzae ZD-7, or A. niger ZD-8. One not inoculated group (substrate) was used as a control. Levels of initial and final free gossypol (FG), crude protein (CP), amino acids (AA) and in vitro digestibility were assayed. The experiment was done in triplicate. The experimental results indicated that microbial fermentation could greatly decrease (P<0.05) FG levels in CSM. The detoxification efficiency differed between the species of microorganisms applied. From the perspective of reducing CSM potential toxicity, C. tropicalis ZD-3 was most successful followed by S. cerevisae ZD-5 and A. niger ZD8. They could reduce FG levels of CSM to 29.8, 63.07 and 81.50 mg/kg based on DM (dry matter), respectively, and their detoxification rate were 94.57%, 88.51% and 85.16%, respectively. If crude protein, amino acids content and their in vitro digestibility were also taken into account, A. niger ZD-8 may be the best choice. The CP content of CSM substrate fermented by C. tropicalis ZD-3 and A. niger ZD-8 were improved by 10.76% and 22.24%; the TAA (total amino acids) contents were increased by 7.06% and 11.46%, and the EAA (essential amino acids) were raised by 7.77% and 12.64%, respectively. Especially, the levels of methionine, lysine and threonine were improved greatly (P<0.05). The in vitro CP digestibility of CSM fermented by C. tropicalis ZD-3 and A. niger ZD-8 was improved by 13.42% and 18.22%, the TAA were increased by 17.75% and 22.88%, and the EAA by 16.61% and 21.01%, respectively. In addition, the in vitro digestibility of methionine, lysine and threonine was also improved greatly (P<0.05).
Amino Acids ; analysis ; Cottonseed Oil ; chemistry ; metabolism ; Digestion ; Fermentation ; Fungi ; metabolism ; Gossypol ; analysis ; Nutritive Value ; Plant Proteins ; analysis

Objective To study the osteogenic effects of induced endothelial cell(EC)on bone marrow stromal cells(BMSCs)of rabbits in co-culture condition.Methods BMSCs were obtained from rabbits by density gradient centrifugation.The adhesive ceils were preserved to passage in culture.The cultured cells were divided into four groups:group A(BMSCs),group B(BMSCs osteogenic induction),group C(EC induction)and group D (co-culture of induced BMSCs and EC).The cell morphology,immunofluorescence,cell proliferation,alkaline pbosphatase(ALP)activity,osteocalcin synthesis were observed to determine the effects of induced autologous EC on the osteogenic potential and cellular compatibility of BMSCs.Results The immunochemical staining showed that the BMSCs were induced into EC in group C.The cellular compatibility of BMSCs and EC was good.The ALP activity and osteocalcin content were obviously higher in group D than in any other groups(P＜0.05).The cell proliferation difference was not obvious between groups(P＞0.05).Conclusions The cellular compatibility of induced osteoblasts and induced EC is perfect.The ECs can significantly increase the viability and ALP activity of induced osteoblasts.

This study was purposed to investigate the amplification of CD3(-)CD56(+)NK cells in umbilical cord blood and their change of immunophenotype and cytotoxicity after stimulation with IL-2 and IL-15. Mononuclear cells were isolated from umbilical cord blood and cultured in serum-free medium supplemented with IL-2 or (and) IL-15 for 14 d. The subset level of CD3(-)CD56(+)NK cells and expression of CD16, CD62L, NKG2A, NKG2D, NCR44, NCR46, granzyme B and perforin were analyzed by flow cytometry. The cytotoxicity of NK cells to K562 was detected by WST-1 method. The results showed that NK cells stimulated with IL-2, IL-15 and IL-2/IL-15 were amplified by 10.78 ± 2.51, 10.42 ± 3.72, and 10.54 ± 6.24 times respectively after 14 d, there was no statistically significant difference between these three groups. The expression of CD16 decreased obviously in NK cells after amplification; there was significant difference between IL-2 and IL-15 groups. The expression of CD62L was not changed statistically after stimulation with cytokines, the IL-2 down-regulated the expressions of NKG2A and NCR46, while IL-15 showed the opposite effect. IL-2 or IL-15 displayed upregulation effect on the expression of NKG2D, perforin and NCR44, but there was statistically significant difference between effects of these two cytokines. IL-15 up-regulated the expression of granzyme B on NK cells. The cytotoxicity of NK cells stimulated and amplified by cytokines significantly increased, but there was no statistically significant difference between IL-2 and IL-15. It is concluded that IL-2 or IL-15 can effectively amplify umbilical cord blood NK cells under serum-free conditions. Although the immunophenotype associated with NK cells function showed different characteristics between them, however, cytotoxicity of NK cells increased obviously after amplification and there is no statistically significant difference between effect of these two cytokines, their synergistic effect is not obvious. The cytotoxicity of NK cells is the result from combined effect of all active molecules.
Cells, Cultured ; Cytotoxicity, Immunologic ; drug effects ; Fetal Blood ; cytology ; Flow Cytometry ; Humans ; Immunophenotyping ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Killer Cells, Natural ; drug effects

T lymphocytes are an integral part of the effective immune response against various tumors, but they are frequently functionally unresponsive in tumor-bearing patients. This study was aimed to investigate the T lymphocyte function of patients with acute myeloid leukemia (AML) in different status through analyzing intracellular cytokine characteristics of T lymphocytes in AML patients. The T lymphocytes from 18 de nuevo AML patients in different status and 10 healthy controls were stimulated with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of monensin, and were stained with fluorescent McAbs CD4-PITC, CD8-FITC and IFNgamma-PE, ILA-PE, then their T lymphocytes were analyzed by flow cytometry. The results showed that IFN-gamma level in CD4+ and CD8+ T cells of de novo AML patients was significantly lower as compared with healthy controls (p<0.05), while IL-4 level in CD4+ and CD8+ T cells was low too, there was no significant difference between de novo AML patients and healthy controls. In AML patients in clinical remission, IFNgamma level in CD8+ T cells was significantly higher than that in de novo AML patients (p<0.05), but there was no significant difference as compared with healthy controls (p>0.05). In relapsed AML patients, IFNgamma level in CD4+ and CD8+ T cells was significantly lower than that in healthy controls and in AML patients with CR (p<0.05), while IL-4 level was significantly higher than that in healthy controls and de nuevo AML patients (p<0.05). It is concluded that the cytokine secretion in T cell subsets of AML patients in different status is changed. Correspondingly, the Th1/Tc1 level is low after stimulating CD4+ and CD8+ T cells in peripheral blood of de novo AML patients, but not different from healthy controls. Though the Th1 level in T cells of AML patients in complete remission is low, but Tc1 response is even enhanced as high as the healthy controls. The Th2/Tc2-like response of CD4+ and CD8+ T cells in relapsed AML patients obviously increases when compared with Th1/Tc1-like response.
CD4-Positive T-Lymphocytes ; metabolism ; CD8-Positive T-Lymphocytes ; metabolism ; Humans ; Interferon-gamma ; secretion ; Interleukin-4 ; secretion ; Leukemia, Myeloid, Acute ; immunology

Objective： The current stady was designed to investigate the reconstitution of T cell receptor repertoire in the patients with chronic Graft-versus-host disease (cGVHD). Methods： The TCR repertoire in peripheral blood mononuclear cells from 5 cGVHD patients was examined after PCR amplification of 24 Vβ gene subfamilies. Results： Only 2－8 Vβ subfamily T cells were found in the samples from these patients, and there were different demonstrations in different patients. We found Vβ T cells proliferated in 4 patients. Conclusion： The TCR repertoire complexities was abnormal in all patients,Vβ may be associated with cGVHD, and the method may be demonstrated the reconstitution of T cell after transplantation.

OBJECTIVETo formulate the seed quality grading standard of Glycyrrhiza uralensis.

METHODThousand-grain weight, seed moisture, germination rate, purity of G. uralensis seed samples from 24 regions were tested. Through statistical analysis, the key indicator and the reference indicators for seed quality grading were defined.

RESULTGermination percentage was the primary indicator of seed quality grading, thousand-grain weight, cleanliness and moisture content were important reference indicators.

CONCLUSIONThe seed quality of each grade should reach the following requirements: for first grade seeds, germination percentage > or = 85% , purity > or = 92%, thousand-grain weight > or = 13 g, seed moisture < or = 11%; for second grade seeds, germination percentage 75%-85%, purity 83%-92%, thousand-grain weight 11-13 g, seed moisture < or = 11%; for third grade seeds, germination percentage 65%-75%, purity 74%-83%, thousand-grain weight 9-11 g, seed moisture < or = 11%.

Germination ; Glycyrrhiza uralensis ; chemistry ; classification ; physiology ; Quality Control ; Seeds ; chemistry ; classification ; physiology

OBJECTIVETo observe the effects of astragalus membranacaus injection on the activity of the intestinal mucosal mast cells (IMMC) and inflammatory response after hemorrahagic shock-reperfusion in rats.

METHODThirty-two Wistar rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with astragalus membranacaus 10 g kg(-1)) and high dosage group (treated with astragalus membranacaus 20 g kg(-1)). Models of hemorrhage shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology, ultrastructure of IMMC, and expression of tryptase were observed. The levels of MDA, TNF-a, histamine, and SOD activity of intestinal were detected, and the number of IMMC was counted.

RESULTThe degranulation of IMMC was seen in model group and was attenuated by astragalus membranacaus treatment. Chiu's score of model group was higher than that of the other groups. Astragalus membranacaus could attenuate the up-regulation of the Chiu' s score, the levels of MDA and TNF-alpha, expression of tryptase, and the down-regulation of SOD activity and histamine concentration. The Chiu's score and MDA content were negatively, while SOD activity was positively correlated to the histamine concentration respectively in the four groups.

CONCLUSIONAstragalus membranacaus can reduce small intestine mucosal damage by inhibiting the activity of IMMC after hemorrhage shock reperfusion.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Injections, Intravenous ; Intestinal Mucosa ; metabolism ; pathology ; Intestine, Small ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mast Cells ; drug effects ; metabolism ; ultrastructure ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; pathology ; Shock, Hemorrhagic ; metabolism ; pathology ; Tryptases ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the expression of TGF-betaR I, Smad2, Smad3 and Smad7 in keloids, normal scars and normal skins. Discuss the significance of these proteins in the course of keloid.

METHODSImmunohistochemistry method was used to detect the expression intensity and distribution of these proteins in above 3 kinds of different tissues in 44 cases. Statistics was used to analyze the data.

RESULTSThe expression of TGF-betaR I were much stronger in keloid than in the other two tissues. The expression of Smad7 were lower in keloids. The increase expression of Smad2,3 were not obvious, but they were found to accumulate in the nucleus.

CONCLUSIONSThe results indicate that over-expression of TGF-betaR I, low-expression of Smad7 and accumulation of Smad2,3 may be one of the etiological factors of keloids. This research may provide a new idea to prevent and treat keloids or other fibrosis diseases in the future.

Adolescent ; Adult ; Female ; Humans ; Keloid ; metabolism ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; metabolism ; Receptors, Transforming Growth Factor beta ; metabolism ; Smad2 Protein ; metabolism ; Smad3 Protein ; metabolism ; Smad7 Protein ; metabolism ; Young Adult

BACKGROUNDCurrently, various calculation methods for evaluating blood-loss in patients with total knee arthroplasty (TKA) are applied in clinical practice. However, different methods may yield different results. The purpose of this study was to determine the most reliable method for calculating blood-loss after primary TKA.

METHODSWe compared blood-loss in 245 patients who underwent primary unilateral TKA from February 2010 to August 2011. We calculated blood-loss using four methods: Gross equation, hemoglobin (Hb) balance, the Orthopedic Surgery Transfusion Hemoglobin European Overview (OSTHEO) formula, and Hb-dilution. We determined Pearson's correlation coefficients for the four methods.

RESULTSThere were large differences in the calculated blood-loss obtained by the four methods. In descending order of combined correlation coefficient based on calculated blood-loss, the methods were Hb-balance, OSTHEO formula, Hb-dilution, and Gross equation.

CONCLUSIONSThe Hb-balance method may be the most reliable method of estimating blood-loss after TKA.

Aged ; Arthroplasty, Replacement, Knee ; adverse effects ; Blood Loss, Surgical ; statistics & numerical data ; Female ; Humans ; Male ; Middle Aged ; Models, Theoretical ; Retrospective Studies