Objective To discuss the related factors of the malignant tumor complicated with venous throm-boembolism(VTE),which may provide the clinical evidence for prevention,diagnosis and treatment of this disease. Methods Retrospectively reviewed the clinical data of patients with malignant tumor who were diagnosed with VTE from June 2012 to June 2015 in our hospital.Results Among 74 cases with tumor merger VTE,female with thrombosis was higher than the male.67.57% of the tumor with VTE patients was 45 -80 years old.Lung cancer patient with VTE was the highest almost 36.49%.Patients with deep vein catheterization,a history of surgery or merging of hyper-tension were easy occurred VTE,those were 14 cases (18.92%),12 cases (16.22%)and 11 cases (14.86%) respectively.57 VTE patients underwent radiotherapy or chemotherapy treatment.The platelet count,D -dimer, fibrinogen and CRP were associated with thrombosis.Among 74 cases with VTE,67 cases (90.54%)accepted anticoagulation therapy.Conclusion Sex,tumor disease,chemotherapy or radiotherapy treatment,medical history and coagluative function index were associated with VTE.Early diagnosis and treatment is the key to reduce mortality and improve prognosis of malignant tumor with VTE.

Objective To monitor the changes of T lymphocyte subsets during chemotherapy in patients with non-small cell lung cancer(NSCLC).Furthermore,to analyze the relationship between chemotherapy curative effect and the T lymphocyte subsets.Methods 42 cases with NSCLC were selected as the subjects,and 28 people undergoing health examination were selected as control group.All NSCLC patients just received NSCLC standard chemotherapy,then respectively determined T lymphocyte subsets at pre-chemotherapy and after 2,4 courses of chemotherapy.Every two cycles of chemotherapy we would evaluate the effect of chemotherapy.Results Before chemotherapy,the patients′ CD+3(t=3.222,P<0.01),CD+4 (t=9.655,P<0.01),CD+4/CD+8 (t=10.657,P<0.01) and NK lymphocytes (t=5.310,P<0.01) were significantly decreased than those of the control group,and CD+8 lymphocytes (t=7.926,P<0.01) increased significantly.There were no significant differences in patients' gender,age,but Ⅰ-Ⅱ period patients′ CD+4(t=2.097,P<0.05),CD+4/CD+8(t=2.045,P<0.05) immune function were better than Ⅲ-Ⅳ period patients.In patients with adenocarcinoma,CD+8(t=2.015,P<0.05),CD+4/CD+8(t=2.315,P<0.05) immune function were better than squamous carcinoma.The patients received chemotherapy after 2,4 courses,the T lymphocyte subgroup was comprehensively improved (P<0.01).The curative effect of chemotherapy was positively correlated with T lymphocyte function,PR>SD>PD.Conclusion Chemotherapy can improve the NSCLC patients′ T lymphocyte subgroup,and improve the situation related to the curative effect of chemotherapy.

ObjectiveTo study the effects of small-dose glucocorticoid (GC) on glucocorticoid receptor (GR) and cellular immune function in critical patients.MethodsForty ICU critical patients admitted in Shanghai Changzheng Hospital from March 2007 to March 2009 were enrolled in the study and were divided into GC group and non-GC group according to the use or absence of GC.Blood samples were collected at days 1,7 and 10 after GC treatment to detect GR binding affinity of mononuclear leukocytes (MNLs) and polymorphonuclear leukocytes (PMLs) in the peripheral blood and the CD4/CD8 ratio in the T lymphocytes.The method of GC use was that the hydrocortisone was given intravenously at a dose of 100 mg every eight hours.ResultsGR binding capacity of MNLs at day 1 and 7 showed no statistical difference between the GC and non-GC groups.GR binding capacity of MNLs in the GC group was lower at day 1 and was much lower at day 7 (P ＜ 0.05 ).However,in the non-GC group,it was lower at day 1,but showed significant improvement at day 7 ( P ＜ 0.05 ).The change of GR binding capacity of PMLs was similar to that of MNLs.There was no significant difference of CD4/CD8 ratio between the GC and non-GC group at day 1.The ratio of CD4/CD8 in the non-GC group was significantly higher than that in the GC group at day 10 (P ＜0.05).CD4/CD8 ratio in the GC group showed a slight reduction at day 10,with no significant difference from that at day 1.While,the non-GC group showed a significant increase of CD4/CD8 ratio at day 10 as compared with that at day 1 (P ＜ 0.05 ).ConclusionLow-dose GC plays some role in the negative feedback regulation of GR binding capacity of peripheral blood leukocytes and in the inhibition of cellular immune function.

Objective To investigate the efficacy of transplantation of induced bone marrow mesenchymal stem cells(MSCs)via various route for treatment of chronic liver injury.Methods MSCs were isolated and expanded by density gradient centrifugation combined with adherent culture.MSCs were induced to differentiate into hepatocyte-like cells in vitro.The markers of hepatocyte lineage were examined by RT-PCR and immunocytochemistry.The changes of ultramicrostructure of MSCs were observed by transmission electron microscope.One hundred and fourteen Wistar rats were treated with mixture of 40%CCl4 and peanut oil to establish chronic liver injury model and another 6 rats were served as control.Twelve rats in model group were died within 8 weeks,and the rest rats were divided into portal vein,spleen and tail vein transplantation groups.The rats were transplanted with DAPI-labelled MSCs which were induced for 14 days.The distribution of the DAPI-labelled cells were observed by fluorescence microscopy.The liver function and pathology were examined.Results AFP mRNA expression were detected at 7th and 14th day after induction,expressions of ALB mRNA,CK18 and hepatocyte antigen were found at 14th and 2lth day.The ultrastructure examination revealed that MSCs were enlarged with a lot of endoplasmic reticulum,mitochondrion,lysosome and glycogen granule.DAPI labelled cells were found in the spleen and liver transplantion groups.The pathological changes of liver were alleviated in all treated groups,especially in intraspleenic transplantion group(P＜0.05).Conclusions Induced bone marrow MSCs can ameliorate liver function of chronic hepatic injury after transplantation.The intraspleenic transplantation is better than others.

Objective: To observe the influence of epidermal growth factor tyrosine kinase inhibitors (EGFR-TKIs) on peripheral blood T lymphocyte subsets, and its efficacy and safety in the treatment of non-small cell lung cancer (NSCLC) patients.Methods: A total of 35 cases with NSCLC in our hospital were selected as the observation group and 28 healthy persons undergoing health examination at the same time were used as the control group.All the NSCLC patients were with EGFR mutations and accepted the EGFR-TKIs therapy.Flow cytometry was employed to detect T lymphocyte subsets in peripherial blood (including CD3+,CD4+,CD8+,CD4+/CD8+ and NK lymphocy-tes)in one week before the treatment, at the 1st, 2nd and 4th cycle after the EGFR-TKIs treatment, and the curative effect and safety were evaluated as well.Results: Before the chemotherapy, the levels of CD3+, CD4+, CD4+/CD8+ and NK lymphocytes in the NSCLC patients were significantly lower than those in the control group (P<0.01), and CD8+ lymphocytes increased significantly (P<0.01).After the treatment of 1, 2 and 4 cycles, the levels of CD4+and CD4+/CD8+ increased in varying degrees when compared with those before the treatment (P<0.01), and CD8+ decreased significantly when compared with that before the treatment (P<0.01).After the treatment of 2 and 4 cycles, NK cells increased in varying degrees when compared with those before the treatment (P<0.05).After the treatment of 4 cycles, the level of CD3+lymphocytes was significantly higher than that before the treatment (P<0.05).The T lymphocyte immune function in the patients with different therapeutic effects showed statistically significant difference (P<0.05).The order of lymphocyte subgroup improvement was PR>SD>PD.The adverse reactions caused by EGFR-TKIs were slight, and all could be improved markedly after the symptomatic treatment.Conclusion: EGFR-TKIs can significantly regulate the expression of T lymphocyte subsets in NSCLC patients and improve the immune function of the patients with promising safety.

Parkinson's disease(PD)is a common neurodegenerative disease mainly affecting old persons.The pathogenesis of PD is still unclear, although it has been studied for many years.Recently, more and more evidences show that the dysfunction of autophagy plays a pivotal role in the pathogenesis of PD.Substantial progress in the genetic research of PD has showed that several pathogenic genes were identified correlated with autophagy, in particular, playing an important role in sporadic cases of PD.Here, we will discuss pathogenic genes-correlated dysfunctional autophagy-lysosomes system, so as to specify the pathogenesis of PD and provide a clue for its treatment.

Objective To investigate the effects of rhubarb on the expression of glucocorticoids receptor (GR)and peripheral blood lymphocytes in burning-induced septic rats. Methods Sixty-six male healthy Sprague-Dawley(SD)rats were randomly divided into sham operated control group(n=18),sepsis model group(n=24) and rhubarb treatment group(n=24),each group was further randomly divided into 12,24 and 72 hours subgroups according to different time points. The model of scald sepsis was replicated by scald injury induced by boiling water at the rat back accounting for 30% total body surface area(Ⅲ grade of scald),and administration of endotoxin (5 mg/kg)into the peritoneal cavity 12 hours after scald injury. After the successful establishment of septic models, the rats in the rhubarb treatment group were immediately infused with 50 mg/kg rhubarb powder dissolved in 1 mL saline through a gastric tube,while the rats in sham operated control group and sepsis model group received saline by the same way as a substitute for rhubarb. The the binding capacity of GR of peripheral blood leucocyte and binding activity of GR of hepatocyte were analyzed by radiation ligands binding assay. The CD4+,CD8+as well as CD4+/CD8+ ratio in peripheral blood lymphocytes were detected by flow cytometer. Results The binding capacity of GR of peripheral blood leucocyte and binding activity of GR of hepatocyte were significantly decreased in a time-dependent manner in sepsis model group compared to those of the sham operated control group,while in the rhubarb treatment group they were increased in a time-dependent manner after interference of rhubarb, and they were higher than those in the model group at the same time points〔leukocyte GR binding capacity (locus/cell)at 12,24,72 hours :1 515.38±300.44,1 859.63±258.26,1 890.50±307.88 vs. 1 122.63±225.39, 1 008.88±150.41,724.38±91.19;hepatocyte GR binding capacity(fmol/mg):210.19±26.26,258.01±20.98, 283.38±38.21 vs. 153.11±30.07, 129.83±26.89, 94.08±14.30, all P<0.01〕. Compared with the sham operated control group,the CD4+ and CD8+ were decreased in various degrees at 12 hours and 24 hours in the septic group, at 24 hours the differences being statistically significant (P<0.01 and P<0.05). CD4+/CD8+ratios were decreased significantly at all time points,the differences were statistically significant at 24 hours and 72 hours(both P<0.01). The CD4+ T cell and CD4+/CD8+ ratio at all the time points were increased at various degrees in the rhubarb treatment group,and the differences from those in the sepsis model group at 24 hours and 72 hours were statistically significant (1.58±0.69, 1.56±0.49 vs. 1.02±0.41, 1.01±1.68, both P<0.01). Conclusion Rhubarb can modulate the binding capacity of GR of peripheral blood leucocyte and the binding activity of GR of hepatocyte,and via its influence on the number of peripheral leucocytes,the immune dysfunction in the sepsis processes is improved.

Objective:To observe the effect of protoscolex on Th subsets and correlative cytokine in mice spleen cells in vitro.Methods:Co-culture spleen cells from BALB/c mice with protoscolices,then IL-4,IFN-γand TGF-βproduction in cell culture supernatants were analyzed by ELISA.The percentage of Th subsets were detected by Flow Cytometry analysis.Results:Secretion levels of IL-4 and TGF-βwere significantly increased in spleen cells at different time point in co-culture system with protoscolices.Ratios of Th2 and Treg cells were also significantly increased in co-culture system at different time points than the control groups.However,there was no statistical significance for ratio of Th1 cells at different time points.Conclusion:The protoscolex can increase the ratios of Th2 cells and Treg cells from spleen cells.Secretion levels of IL-4 and TGF-βwere also increased in spleen cells co-cultured with protosco-lices.The results suggest that these Th cell subsets play a role in the immune escape of the hydatid disease.

Objective:To investigate the role of growth arrest specific protein 6 (Gas6) in regulating macrophage polarization in wound healing.Methods:Clean male B6 mice were randomly(random number) divided into the normal group, skin defect group, skin defect group + normal saline group (PBS group), skin defect + Gas6 (1 μg) group, skin defect + Gas6 (5 μg) group, and skin defect + Gas6 (10 μg) group. Ten mice in each group were used to observe the healing of skin wounds. Macrophages were isolated from the wound tissues of the remaining 6 mice on the fifth day after modeling. The levels of IL-6 and IL-10 were detected by enzyme-linked immunosorbent assay (ELISA), the mRNA expression levels of arginase-1 (Arg-1) and inducible nitric oxide synthase (iNOS) were detected by RT-PCR, and flow cytometry was used to detect the expression of M1 marker CD197, M2 marker CD163 and F4/80. HE staining was used to detect the pathological changes of skin wounds. Masson staining was used to analyze the granulation tissue and collagen deposition.Results:Scab began to form on the surface of the wound on the third day after the skin defect model was established. The wound area of the Gas6 treatment group was smaller than that of the PBS group, and the wound healing was better than that of the PBS group. Compared with the normal group, the proportion of CD197 in macrophages of the skin defect group was significantly increased ( P=0.00 49), the proportion of CD163 and F4/80 double positive was significantly decreased ( P=0.00 86), the level of IL-6 was significantly increased ( P=0.00 13), the level of IL-10 was significantly increased ( P=0.00 14), the level of iNOS mRNA was significantly increased ( P=0.00 8), and Arg-1 was significantly increased in the skin defect group The mRNA level was significantly decreased ( P=0.01 21), and the inflammatory infiltration was aggravated. Compared with the PBS group, the proportion of CD197 in the Gas6 treatment group was significantly decreased ( P=0.00 0), the double positive rates of CD163 and F4/80 were significantly increased ( P = 0.00 0), the level of IL-6 was significantly decreased (P = 0.00 0), the level of IL-10 was significantly increased ( P=0.00 03), the level of iNOS mRNA was significantly decreased ( P=0.00 18), the level of Arg-1 mRNA was significantly increased ( P=0.00 1), and the number of inflammatory cells and the number of collagen fibers were increased. Conclusions:Gas6 can promote the transformation of macrophages from M1 to M2 in mice with skin defect, which is beneficial to the wound healing of skin defect.

Objective: To investigate the innate immunity of cultured human umbilical vein endothelial cells (HUVEC) after recombinant hemagglutinin (HA) protein of influenza virus innoculation. Methods: HUVECs were divided into four groups: control, H9N2 viral particle, recombinant H3HA and recombinant H5HA. After 24 h of innoculation, the supernatant of the culture was extracted and the HUVECs were lysed. The levels of cytokine in HUVEC were measured by real-time PCR and ELISA. Results: The mRNA and protein expression of IFNB1 and the mRNA expression of CCL5 and IFITM3 in HUVECs of HA (H3HA and H9HA) innoculated group did not show significant difference compared to control (IFNB1 mRNA and protein expression of H3HA group, P=0.051, 0.839; IFNB1 mRNA and protein expression of H9HA group, P=0.127, 0.561; CCL5 mRNA expression of H3HA and H9HA croups, P=0.314, 0.429; IFITM3 mRNA expression of H3HA and H9HA croups, P=0.530, 0.269). The mRNA expression of CCL5 and IFITM3 in HUVECs of H9N2 viral particle innoculated group increased significantly compared to control (P=0.022, 0.047). Conclusions These results suggest that recombinant HA alone did not trigger innate immunity in HUVEC, but viral particle could activate endothelial innate immunity.