Adult ; Aged ; Female ; Humans ; Male ; Manipulation, Orthopedic ; methods ; Middle Aged ; Shoulder Dislocation ; therapy

This study was purposed to investigate the proliferation and antitumor activity of rhG-CSF-mobilized peripheral blood mononuclear cells (G-PBMNCs) activated by interleukin 21 (IL-21) alone or in combination with interleukin 15 (IL-15)/interleukin 2 (IL-2) and to evaluate the feasibility and value of tumor immunotherapy with cytokine combinations. G-PBMNCs were activated by IL-21 alone or in combination with IL-15/IL-2 in vitro, and the proliferation of the activated G-PBMNCs was analyzed by CCK-8 assay. The cytotoxicity of the activated G-PBMNC to the K562 cells was studied by the test principle which is based on target cell labeling with 5-(6)-carboxy-fluorescein succinimidyl ester (CFSE) and subsequent DNA-labeling with propidium iodide (PI) for identification of target cells with compromised cell membranes. The phenotypes of the activated G-PBMNCs were assayed by flow cytometry. The results showed that the cytotoxicity of IL-21 group had no difference from which of IL-2 group. When G-PBMNCs were exposed to the combinations of IL21+IL15/IL21+IL15+IL2, the cytotoxicity was significantly enhanced at E:T ratio of 25:1, as compared with combination of IL21+IL2 (p<0.05). The cytotoxicity of the cytokines combinations was significantly higher than that in cytokine used alone at E:T ratio of 50:1 (p<0.05). The cryopreservative and resuscitative G-PBMNCs showed the same result with the fresh G-PBMNCs in cytotoxicity test. The proportions of CD3+ and CD8+ T cells were increased when G-PBMNCs were incubated with cytokines for 72 hours. CD4, CD3-56+ and CD3+56+ counts were significantly elevated when G-PBMNCs were exposed to IL21 + IL15 (p<0.05). It is concluded that IL-21 alone enhance the antitumor activity of G-PBMNCs, which further strengthens when IL-21 combinated with IL-15.
Cell Proliferation ; Cells, Cultured ; Drug Synergism ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukins ; pharmacology ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; immunology ; Recombinant Proteins

OBJECTIVESTo study the expression of nicotinamide phosphoribosyl transferase (Nampt) and vascular endothelial growth factor-A (VEGF-A) in gastric carcinoma and investigate their correlations to clinicopathologic features and prognostic significance.

METHODSThe proteins of Nampt and VEGF-A in 68 specimens of gastric carcinoma and 59 specimens normal gastric tissue were detected by immunohistochemistry during January 2000 to December 2004, and the 68 patients were followed up.

RESULTSNampt protein was detected in the cytoplasm of both tissues, and Nampt in gastric carcinoma (13 ± 5) were significantly higher than that in normal gastric tissue (6 ± 3) (t = 7.46, P < 0.01). The expression of Nampt was correlated to invasive depth (F = 4.693, P = 0.034), lymph node metastasis (F = 4.027, P = 0.049), clinical TNM stage (F = 9.979, P = 0.002), but not to gender, age, tumor location, tumor size, differentiation (P > 0.05). The expression of Nampt is correlated with survival of patients that underwent surgical resection for gastric cancer. The survival rate of patients in negative of Nampt was very higher than that of the positive patients, and its co-expression with VEGF-A showed a trend towards poorer survival. The positive correlation was found between the expression of Nampt and VEGF-A in gastric carcinoma (r = 0.293, P = 0.015).

CONCLUSIONSThe expression of Nampt is positively correlated to that of VEGF-A in gastric carcinoma. The correlation between the expression of Nampt and VEGF-A in gastric carcinoma plays an important role cooperatively in carcinogenesis, development and metastasis of gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; Cytokines ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Nicotinamide Phosphoribosyltransferase ; metabolism ; Prognosis ; Stomach Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism

This study was purposed to investigate the effect of G-CSF on the proliferation, differentiation, and cell cycle distribution of thymocytes in sublethally irradiated mice. Female BALB/c mice were exposed to 6.0 Gy γ-ray irradiation and then randomly divided into control and G-CSF treatment group. In the treatment group rhG-CSF 100 µg/(kg·d) was given subcutaneously for 14 continuous days and to make sure the first injection was given within 1 hour after irradiation. Cell cycle distribution and apoptosis of thymocytes were detected within 72 hours after irradiation. Subpopulations of CD4(-)CD8(-) cells and sequential changes in the distribution of CD4(+)CD8(+), CD8(+)CD4(-), CD8(-)CD4(+) cells were detected by a three-color flow cytometry during a four-weeks period after irradiation. The results showed that in G-CSF treatment group marked increase of cells in G(0)/G(1) phase (G-CSF vs control: 82.0 ± 5.0% vs 75.9 ± 2.8%) (p < 0.05) and a decrease of cells in S phase (G-CSF vs control: 10.2 ± 4.8% vs 15.7 ± 2.3%) (p < 0.05)could be observed as early as 6 hours after irradiation, but G-CSF seems have no evident effects on the cells in G(2)/M phase. G-CSF could also protect thymocytes against apoptosis. 6 and 12 hours after irradiation the apoptosis rates of thymic cells in G-CSF treatment group were 11.5 ± 2.4% and 15.5 ± 3.3% respectively, while in the control group the apoptosis rates were 16.5 ± 2.2% and 22.6 ± 0.7% respectively. Comparison between the two group demonstrated significant difference (p < 0.05). CD4(-)CD8(-) double negative thymocytes (DN)can be defined as DN1-4 according to their maturation. G-CSF treatment resulted in a significant increase in DN1 thymocytes and promoted their proliferation and differentiation to a more mature DN3 and DN4 stage. G-CSF could enhance the recovery of CD4(+)CD8(+) thymocytes and mitigate their relapse during reconstitution. The percentage of CD4(+)CD8(+) thymocytes in the G-CSF treatment group 28 days after irradiation was significantly higher than that of the control group (71.0 ± 6.3% vs 25.5 ± 6.3%) (p < 0.05). It is concluded that G-CSF has a positive effects on the thymic cell cycle distribution, proliferation and differentiation, which may contribute to the reconstitution of central immune system after acute irradiation.
Animals ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Differentiation ; drug effects ; Cells, Cultured ; Female ; Flow Cytometry ; Granulocyte Colony-Stimulating Factor ; pharmacology ; therapeutic use ; Lymphocyte Count ; Mice ; Mice, Inbred BALB C ; Radiation Injuries, Experimental ; therapy ; Thymus Gland ; cytology

Objective To investigate the genetic polymorphism of mec Ⅰ in the clinical isolates of methicillin-resistant Staphylococcus anreus(MRSA).Methods 40 isolates(MRSA)carrying mecA gene were selected randomly from the clinical isolates of Staphylococcus anreus from Jan,2005 to Aug,2006 in our hospital.The mec Ⅰ gene was detected by PCR followed with sequencing.Staphylococcal cassette chromosome mec(SCCmec)in MRSA were detected by multiplex-PCR.Agar dilution method was used for determining the MICs of oxacillin against MRSA.Results 35 of 40(87.5%)MRSA carried mec Ⅰ gene.All isolates carrying mec Ⅰ gene have mecI 202C→T substitution,which resulted in Gln at 68 aminophenol position replaced by stop condon.32 isolates carried single point mutation.3 isolates carried double-point mutation,including additonal A at 3 positon,A→C at 41 position and C→T at 142 position beside C→T at 202 position,respectively.Among 35 isolates carrying mec Ⅰ gene,there were 27 isolates of SCCmec Ⅲ, 7 isolates of SCCmec Ⅲ A and 1 isolate of SCCmec Ⅱ.Among 5 isolates with deletion of mec Ⅰ gene,there were 3 isolates of SCCmecⅣ,1 isolate of SCCmec Ⅰ and 1 isolate of non-known SCCmec tpye.The MICs of oxacillin were 256-512 ?g/ml,≥512 ?g/ml and 8-256 ?g/ml in 31 isolates with single point mutation at 202 position in mec Ⅰ gene,3 isolates with double-point mutation in mecI gene and 5 isolates with deletion of mec Ⅰ gene,respectively.1 isolate with single point mutation in mec Ⅰ gene had contrary result(MIC

Sarcopenia,osteoporosis and osteosarcoporosis,which belong to the same category of musculoskeletal unit disease,seriously impair the homeostasis of the musculoskeletal system in the aging process of the human body.In recent years,researchers have applied various emerging technologies to conduct multi-disciplinary,multi-level and multi-dimensional research on musculoskeletal decay disease,and have come to realized that the disease involves complex biochemical and mechanical crosstalk,and the intestinal microecology has also aroused great interest in this crosstalk because of its profound impact on musculoskeletal health.In view of this,this paper takes the close relationship between intestinal microecology and musculoskeletal decay disease as the starting point,and under the guidance of the overall concept,we deeply analyzes the mechanism and regulatory information behind this relationship,and on this basis,we excavates the potential therapeutic targets for musculoskeletal decay disease,so that we can provide new ideas and strategies for the management of the disease in the context of the musculoskeletal subunit.

OBJECTIVETo study the relationship between the changes of inflammation-associated factors, C-reactive protein (CRP), procalcitonin (PCT), erythrocyte sedimentation rate (ESR), white blood cell (WBC) and neutrophils, and the severity in children with Mycoplasma pneomoniae pneumonia (MPP).

METHODSNinety-two children with acute MPP consisting of 52 cases with concomitant systemic inflammation response syndrome (SIRS) and 40 cases without SIRS were enrolled in this study. The 52 cases with concomitant SIRS were classified into two groups based on the severity of SIRS: mild SIRS (n=25) and severe SIRS (n=27). CRP, PCT, ESR and WBC count and the percentage of neutrophils (NE%) were detected on admission and one week after anti-inflammation treatment.

RESULTSAll of patients showed increased serum CRP contents at admission. The serum CRP contents were the highest in the severe SIRS group, followed by the mild SIRS and non-SIRS groups on admission (P < 0.05 or 0.01). The serum CRP contents were reduced in all of patients after 1-week treatment. The severe SIRS group still demonstrated higher serum CRP contents than the non-SIRS and the mild SIRS groups (P < 0.01). The severe SIRS group had increased serum PCT contents on admission, which were significantly higher than those of the mild SIRS and non-SIRS groups (P < 0.01). After 1-week treatment, the serum PCT contents were reduced in the severe SIRS group but remained higher than in the mild SIRS and non-SIRS groups (P < 0.01). ESR increased significantly in the severe SIRS group than in the mild SIRS and non-SIRS groups on admission (P < 0.01). One-week treatment did not significantly decrease ESR in all three groups. The WBC count and NE% in the mild and severe SIRS groups were significantly higher than in the non-SIRS group and the severe SIRS group had higher WBC count and NE% than the mild SIRS group on admission (P < 0.05). The WBC count and NE% decreased after 1-week treatment in the mild and severe SIRS groups (P < 0.05). One inflammation-associated factor (only CRP) increase was predominant in the non-SIRS group (65%), 2 factors increase in the mild SIRS group (56%), and three or more factors increase in the severe SIRS group (70.4%).

CONCLUSIONSThe detection of inflammation-associated factors, CRP, PCT, ESR, WBC and neutrophils, are valuable to the evaluation of severity in MPP.

Adolescent ; Blood Sedimentation ; C-Reactive Protein ; analysis ; Calcitonin ; blood ; Calcitonin Gene-Related Peptide ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Leukocyte Count ; Male ; Pneumonia, Mycoplasma ; blood ; complications ; drug therapy ; Protein Precursors ; blood ; Systemic Inflammatory Response Syndrome ; blood ; drug therapy

This study was purposed to investigate the reconstitution of immune system in patients with acute lymphocyte leukemia (ALL) or acute myeloid leukemia (AML) after HLA-mismatched nonmyeloablative hematopoietic stem cell transplantation (NHSCT) and its relation with infection and GVHD. 6 ALL and 4 AML patients having HLA-mismatched related donors received the nonmyeloablative precondition regimen composed of fludarabine (Fln), ATG, Ara-C, CTX and total body irradiation (TBI) in dose 2 Gy. The GVHD was prevented and treated by CsA, anti-CD25 antibody and mycophenolic mofetil (MMF) before and after transplantation. The flow cytometry was used to detect the changes of total T cells, help/inducer T cells, suppressor/killer T cells, gamma/delta T cells, B cells, NK cells, NKT cells, regulatory T cells, activated T cells, naive T cells, memory T cells and ratio of CD4/CD8 in patients with remission resulting from chemotherapy before transplantation, and analyse the relation of immunofunctional cells to infection and GVHD after transplantation, compare the difference in recovery of immune system of ALL and AML patients. The results showed that the recovery of total lymphocytes and lymphocyte subsets displayed one's own regular pattern. As compared with patients without GVHD, the counts of lymphocyte subsets in patients with GVHD was higher, while the counts of gamma/delta T cells, regulatory T cells, NK cells, the counts of B cells, NK cells, naive cells and CD4/CD8 ratio as well as the counts of B cells, naive T cells and NK cells were lower at 1 month, 2 - 3 months and 6 - 8 months after transplantation respectively. The total T cells and subsets recovered slowly, but NK cells and NKT cells recovered rapidly in patients with infection at early period after transplantation, the B cells and naive B cells recovered rapidly at 3 months after transplantation. There was no difference in lymphocyte recovery between ALL and AML patients. It is concluded that the analysis of each lymphocyte subsets may indirectly show the recovery of thymus function in patients, the changes of NK cells, B cells and naive T cells have an important significance for identifying and forecasting the GVHD and infection.
Adolescent ; Adult ; Child ; Female ; Graft vs Host Disease ; etiology ; HLA Antigens ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia, Myeloid, Acute ; immunology ; Lymphocyte Subsets ; immunology ; Male ; Middle Aged ; Postoperative Period ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; immunology ; Young Adult

This study was aimed to investigate the expression difference of serum cytokines in 20 patients receiving HLA-identical nonmyeloablative allogeneic hematopoietic stem cell transplantation (iNAHSCT) and HLA-haploidentical nonmyeloablative allogeneic hematopoietic stem cell transplantation (hiNAHSCT). IL-2, IL-4, IL-6, IL-10, TNF-α, γ-IFN and IL-17 were detected by flow cytometric bead array before and on week 1, 2, 4 after transplantation respectively. The results showed that the IL-2 level was found to be up-regulated at week 1 and 2 after transplantation in iNAHSCT group and in hiNAHSCT group respectively, but there was no difference between these two groups (P > 0.05). The γ-IFN levels was up-regulated at week 4 after transplantation in above-mentioned two groups, but no difference was found between these two groups. The IL-4 level increased at week 2 and 1 after transplantation in iNAHSCT and hiNAHCT groups respectively, but the IL-4 level in iNAHSCT group was higher than that in hiNAHSCT group. The IL-6 level rose at week 1 and 2 after transplant in above mentioned groups respectively, and reached to peak level at week 4 after transplantation, but IL-6 level in hiNAHSCT was higher than that in iNAHSCT group (P < 0.02). The IL-10 level was up-regulated at week 1 and 2 in iNAHSCT and hiNAHSCT groups respectively, but the IL-10 level in iNAHSC was higher than that in hiNAHSCT group. The TNF-α level was up-regulated at week 1 in hiNAHSCT group, but at week 2 in iNAHSCT group after transplantation. The TNF-α level in hiNHASCT group was higher than that in iNAHSCT group (P < 0.01). The IL-17 level was up-regulated at week 1 and week 4 after transplantation in hiNAHSCT and iNAHSCT groups respectively, the IL-17 level in hiNAHSCT group was high as compared with that in iNAHSCT group. It is concluded that the serum cytokine levels are obviously up-regulated in iNAHSCT and hiNHASCT groups, and reach to peak level at week 4 after transplantation. The IL-6, TNF-α and IL-17 level up-regulated significantly in hiNAHSCT group, but the IL-4 and IL-10 level up-regulated significantly in iNAHSCT.
Adolescent ; Adult ; Child ; Cytokines ; blood ; Female ; HLA Antigens ; immunology ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Interleukin-10 ; blood ; Interleukin-17 ; blood ; Interleukin-4 ; blood ; Interleukin-6 ; blood ; Male ; Middle Aged ; Transplantation, Homologous ; Tumor Necrosis Factor-alpha ; blood ; Young Adult

OBJECTIVETo exploit the role of bone marrow (BM) and peripheral blood (PB) fluorescence in situ hybridization (FISH) in cytogenetic evaluation of myelodysplastic syndrome (MDS).

METHODSThe metaphase cytogenetics and BM interphase FISH were prospectively compared in 112 cases of de novo MDS. At the same time, comparison of BM and PB FISH was conducted in 56 cases.

RESULTSThe differences between metaphase cytogenetics and BM FISH were observed in 22 (54%) of 41 cases with clonal karyotypic abnormalities, most of differences were caused by the limitation of FISH probe panel which could not target all of the regions with aberrations. Only 6 (27%) of 22 differences were involved in our probe regions, the FISH results did not change their cytogenetic risk categories. BM FISH testing was abnormal in 15 (21%) of 71 cases with normal karyotypes, FISH testing was abnormal in 14/51 (27%) and 1/20 (5%) cases with fewer than 20 normal metaphases or more than 20 normal metaphases. Comparison of FISH results of PB and BM samples showed abnormal PB FISH results in 21 (72%) of 29 cases with abnormal BM FISH results, and in 1 (4%) of 27 cases with normal BM FISH results.

CONCLUSIONBM FISH should be used to MDS cases with fewer than 20 normal metaphases. Although PB FISH testing is limited by a relatively high false negative rate, it is a reasonable choice to cases with failure of BM aspiration.

Adolescent ; Adult ; Aged ; Female ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; blood ; genetics ; Prospective Studies ; Young Adult