Batch fermentation of lycopene by recombinant E. c ol i ZYL-2 at various temperatures ranging between 23℃～37℃ were studied in 7L fermentor. At 33℃, cell specific growth rate in earlier stage of culture was higher, and the time obtained maximum cell dry weight was shorter at other tempe rature. while after 9 h, lycopene specific production rate was higher at 28℃. Based on these results, a two-stage temperature control strategy was developed in which 33℃ was used for fermentation for the first 9 h and the temperaturew as switched to 28℃ after 9h. Using this temperature-shift strategy, the maxim al lycopene content and productivity reached 605.25?g L-1 and 28.82?g L-1h-1. The lycopene fermentation level obtained by the strategy was higher than those in single temperature-control experiments.

Objective To investigate the effectiveness and safety of combined heparin and acutobin in the treatment of acute inferior vena cava thrombus in rabbits. Methods The inferior vena cava thrombus model was established in 72 rabbits and they were randomly divided into three groups; heparin group(A) , group for combination of urokinase and heparin (B), group for combination of acutobin and heparin (C) ,each group including 24 rabbits. Drugs were administrated 3 days after thrombosis. Coagulation indexes were tested to assess their safety, and Doppler ultrasound was used to assess their effectiveness, on day 3, day 7, and day 10. Results The prolongation of prothrombin time ( PT) in group C was shorter than that in group B( P < 0. 05 ) , the fibrinogen ( FBG) value in group C was lower than that in group B (P < 0. 05 ) , the prolongation of PT in group B and group C was longer than that in group A (P < 0. 01), the FBG value of group B and C were higher than that in group A ( P < 0. 01 ), D-dimer ( D-D) value in group B and C gradually returned to normal range. There was no difference between the two groups (P > 0. 05). The thrombolytic effect in group B and C were better than that in group A, statistical difference was reached between groups B and A (P <0. 01), and the difference was statistically significant between groups C and A 10 days after administration (P < 0. 01). Thrombolytic effect was not different statistically between groups B and C (P > 0. 05). Conclusion Acutobin combined with heparin in the treatment of acute inferior vena cava thrombus in rabbits was effective and safe.

Background Human leucocyte antigen (HLA)-B27-associated uveitis is one of the most common causes of non-infectious uveitis.T helper 17 (Th17) cells play an important role in human autoimmune diseases,but the pathology research on the production of Th17 cells in acute anterior uveitis patients positive for HLA-B27 was rarely reported.Objective The aim of this study was to investigate the expression and significance of the peripheral blood T helper cell subsets (Th1,Th2,Th17) in acute anterior uveitis patients positive for HLA-B27.Methods This study meets the criteria of the Helsinki Declaration.Informed consent was obtained from all the participants.A prospective cohort design was used in this study.Twenty-two patients with acute anterior uveitis positive for HLA-B27 were enrolled from Affiliated Second Hospital of Nanjing Medical University,and 16 normal healthy subjects with matched gender and age were enrolled as controls.The expression of interferon-γ (IFN-γ),interleukin-4 (IL-4) and IL-17 on lymphocytes (CD4+) in blood were assessed by flow cytometry,and immunoturbidimetry was used to detect the C reactive protein (CRP) level in blood.The degree of the severity of disease was evaluated by clinical scoring.The correlations between the percentage of IFN-γ+Th1,IL-4+Th2,or IL-17+Th17 with clinical factors and CRP were analyzed.Results The percentages of IFN-γ+Th1 and IL-17+Th17 in the peripheral blood were (23.11 ±9.69) ％ and (3.96±2.92) ％ in the patient group,showing a significant increase in comparison with (16.00±4.26)％ and (1.68±0.60) ％ in the control group (P=0.041,P=0.002).However,the IL-4+Th2 level was not significantly different between the patient group (0.33％ ±0.36％) and the control group (0.56％ ±0.34％) (P=0.122).No significant correlations were found between the percentage of IFN-γ+ Th1 with disease severity and CRP (r =0.197,P =0.500 ; r =0.253,P =0.383),between the percentage of IL-4+ Th2 with disease severity and CRP (r =0.068,P =0.817 ; r =0.439,P =0.116) as well as between the percentage of IL-17 + Th17 with CRP (r =0.226,P =0.436).However,a positive correlation was seen between the percentage of IL-17+ Th17 with disease severity (r =0.805,P =0.001).Conclusions IFN-γ and IL-17 in human CD4+T cells are significantly elevated in the blood of HLA-B27-related acute anterior uveitis patients.Disease severity is associated with the percentage of IL-17 +Th17,suggesting that Th1 cells together with Th17 cells participate in the pathogenesis of the disease and Th17 cells might play a dominant role in the disease.

Mesenchymal stem cells (MSCs) are promising candidates to develop new cell-based therapeutic strategies and bone marrow represents the main source of MSCs for experimental and clinical application currently. How to obtain enough MSCs is the biggest challenge faced by the experimental and clinical application. The difference of morphology, growth pattern, immunophenotype and multilineage differentiation of MSCs derived from human umbilical cord vein and their multiple differentiation capacity were investigated. The results showed that MSCs derived from human umbilical cord vein and normal adult bone marrow were similar in biological characteristics. They not only grew in fibroblast-like cells mode but had great expansion potency and multilineage differentiation ability in vitro. The MSCs derived from human umbilical cord vein may be an excellent alternative source for experimental and clinical application.

The recurrence rate of primary hepatocellularcarcinoma is very high,and repetitive resection is still the besttreatment for operable recurrent hepatocellular carcinoma.Indocyanine green(ICG) has been widely used andpopularized in resection of hepatocellular carcinoma,providesmore possibilities and diversity for the surgical treatment ofrecurrent hepatocellular carcinoma. Combined withpreoperative imaging,three-dimensional imaging technologyand intraoperative ultrasound,ICG can effectively improve theradical and accuracy of the operation for recurrent hepatocellular carcinoma. ICG-guided hepatectomy isappropriate for the new concept and technical system of liversurgery advocated by modern medical model, and realizesfunctional and anatomical hepatectomy for recurrenthepatocellular carcinoma.

Aviation ; Humans ; Physical Fitness ; Resistance Training ; Schools ; Students

OBJECTIVETo investigate the clinicopathologic features of reactive hyperplasia of lymph nodes in maxillofacial regions.

METHODSThirty-two patients with lymph node reactive hyperplasia (LRH) were analyzed clinicopathologically including pathomorphologic manifestation and immunohistochemical expression.

RESULTSHistopathology of 32 patients showed significant hyperplasia in folliculus lymphaticus, marginal zone lymph, paracortical area lymph, and seldom mixed hyperplasia. Immunohistochemical study showed that the hyperplastic lymphocytes were mainly composed of CD20 positive B cells positive in 23 cases(72%), CD3 positive T cell in 7 cases (22%), CD20 positive T cell and CD3 positive B cell partially as well as histiocytes in 2 cases (6%). Atypical hyperplasia was found in 10 cases.

CONCLUSIONHyperplastic lymphocytes in LRH of maxillofacial regions are mainly composed of B cells. Malignant transformation may occur in these patients with atypical hyperplasia. Regularity follow up is necessary for these patients.

Adult ; B-Lymphocytes ; Female ; Humans ; Hyperplasia ; Lymph Nodes

OBJECTIVETo investigate the role of T cell and its subsets in the induction of insulitis and type 1 diabetes mellitus (T1DM) in BALB/c mice.

METHODSAutoimmune diabetes mellitus was developed by intraperitoneal injection of 40 mg/kg streptozotocin (STZ) daily for 5 consecutive days in BALB/c mice as sources of donor cells. Spleen cells from diabetic mice were then cultured for 7 days in the stimulation of interleukin-2 (IL-2) to harvest diabetogenic T cells, which were subsequently transferred into normal BALB/c mice recipients. MTT, ELISA, and HE staining were used to analyze the lymphocyte proliferation, cytokine (IL-2, interferon-gamma, IL-4, and IL-10) levels, and pathological changes in pancreatic islets.

RESULTSAs few as 3 x 10(6) diabetogenic T cells successfully induced diabetes mellitus in recipients pretreated with STZ twice, whereas transfer of equal amount of normal splenocytes, T cell-depleted diabetogenic splenocytes, or diabetogenic CD4+ T cells alone in recipients receiving STZ twice pretreatment was proved not to induce diabetes mellitus either. A markedly increased lymphocyte proliferation, high levels of interferon-gamma and IL-2 in the supernatants of diabetogenic T cells were observed. In addition, a markedly enhanced lymphocyte proliferation, a high level of interferon-gamma secretion in serum, and numerous lymphocytes infiltration in pancreatic islets were detected in the diabetic mice induced by diabetogenic T cells transfer.

CONCLUSIONSA novel T1DM murine model is established in STZ-pretreated BALB/c mice by adoptive transfer of diabetogenic T cells. CD4+ T cells with interferon-gamma may promote the onset of diabetes mellitus.

Animals ; Blood Glucose ; metabolism ; Cytokines ; immunology ; Diabetes Mellitus, Experimental ; metabolism ; pathology ; Diabetes Mellitus, Type 1 ; immunology ; pathology ; Disease Models, Animal ; Islets of Langerhans ; cytology ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; cytology ; immunology

OBJECTIVETo investigate the apoptosis effect of isothiocyanates (ITCS) on human liver cancer cells HepG-2, and its mechanism.

METHODHepG-2 cells were treated with different concentrations of ITCS. MTT assay was used to evaluate the influence of ITCS on cell proliferation. Flow cytometry was used to test ROS levels, intracellular mitochondrial transmembrane potential (deltapsim) , and hypodiploid apoptosis peak in HepG-2 cells.

RESULTITCS obviously inhibited proliferation of HepG-2 cells. When treated with 15, 30, 60, 120, 240 microg x mL(-1) of ITCS for 24 h, ROS levels were (23.1+/-1. 8)%, (53.3+/-3.3)%, (57.9+/-2.0)%, (79.9+/-3.4)%, (93.4+/-2. 6)% respectively; and deltapsim were (94.8+/-5.5)%, (91.8+/-5.4)%, (66.0+/-5.6)%, (65. 5+/-6.6)% and (44.3+/-2.7)% respectively; when treated with 60, 120, 240 microg x mL(-1) of ITCS for 48 h, cell apoptotic rates were (16.6+/-2.8)%, (21.9+/-4.4) % and (70.2+/-5.3) % respectively.

CONCLUSIONITCS generates ROS in gastric cancer HepG-2 cells, which causes mitochondrial membrane permeabilization and deltapsim decrease, therefore, leads to apoptosis of HepG-2 cells.

Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Brassica ; chemistry ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Isothiocyanates ; isolation & purification ; pharmacology ; Liver Neoplasms ; metabolism ; pathology ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria, Liver ; drug effects ; physiology ; Plants, Medicinal ; chemistry ; Reactive Oxygen Species ; metabolism

To explore the new approach to prevent graft versus host disease (GVHD) by purging ex vivo T lymphocytes of bone marrow graft through Fas-FasL way, FasL-cDNA was transfected into BALB/c mouse bon e marrow cells by liposome ex vivo. The transfected cells were cultured together with BAC (BALB/c x C57BL/6) mouse bone marrow graft. The mixing bone marrow graft was infused into BALB/c mouse recipients after 60Co-gamma irradiation. The mortality, manifestation and pathologic change of GVHD in recipient mice were observed. The CFU-S and Y chromosome from donor mice were detected. The results showed that compared with control group, the mortality in 60 days of the recipients in the experimental group decreased (20% vs 70%, P < 0.01) and the morbidity of GVHD lowered (40% vs 100%, P < 0.01). The CFU-S counts for all groups were at normal level on 20 days after transplantation. The Y chromosome from donor mice was discovered in 70% bone marrow nucleated cells of recipient mice survived over 2 months in the experimental group. It is concluded that mFasL-cDNA transfected mouse bone marrow cells prevent GVHD after culturing together with bone marrow graft, and accelerate hematopoietic reconstitution in recipient mice.
Animals ; Bone Marrow Cells ; metabolism ; Bone Marrow Purging ; Bone Marrow Transplantation ; Fas Ligand Protein ; Female ; Genetic Therapy ; Graft vs Host Disease ; Male ; Membrane Glycoproteins ; genetics ; Mice ; Mice, Inbred BALB C ; Transfection