Objective To establish mice models of Kawasaki disease by Lcatobacillus casei cell wall extract(LCWE) ,in order to provide experimental materials for follow-up study .Methods LCWE group was given LCWE 0 .1 mL per mouse(containing 200 mg LCWE) via abdominal intramuscular injection .PBS group was given 0 .1 mL PBS per mouse .Orbital blood samples were collected respectively at the 1st ,3rd and 28th day after administration .The heart ,lung ,liver ,kidney ,spleen and other organs tissue samples were collected for pathological section ,HE staining .The histopathological changes were investigated ,and the routine blood test was proceeded .Results The WBC of LCWE group showed a trend of rising at the 1st ,3rd and 28th day after administration .PLT and MPV of LCWE group increased at the 3rd day after administration ,which returned to normal levels at the 28th day after adminis-tration .The pathological section showed the blood vessel walls of heart tissue enlargement ,surrounded and infiltrated by inflamma-tory cells infiltration ,the atheromatous plaque in blood vessels occasionally .Conclusion The study established mice model of Ka-wasaki disease successfully .

Objective: To investigate the possible mechanisms in acupuncture analgesia by interaction of &opioid receptor with neurotransmitter transport proteins or the Na+-K+pump. Methods: Microinjection of respective heterologous cRNA into the Xenopus oocytes as a model system, and measurement of steady-state currents under two-electrode voltage clamp. Results: The co-expression of the δ-opioid receptor with GAT1, EAAC1 or the sodium pump resulted in reducing activity of the respective transporter. Opioid receptor activation affected transporter activity in different ways: 1) GAT1 was further inhibited; 2) EAAC1 was stimulated; 3) Na+-K+ pump activity interfered with agonist sensitivity of DOR. Pump inhibition led to higher sensitivity for DPDPE. Conclusion: GABA transporter inhibition and glutamate transporter stimulation may counteract pain sensation by affecting the neurotransmitter concentration in the synaptic cleft and, therefore, may contribute synergistically to pain suppression by acupuncture. Sodium pump inhibition by endogenous ouabain may amplify these effects. These synergistic effects may be the molecular mechanism of inhibiting pain sense and/or acupuncture analgesia.

Objective To evaluate the role of vascular endothelial growth factor-D(VEGF-D)in lymphangiogenesis of breast cancer,and investigate its relationship with some clinicopathological parameters and prognosis. Methods VEGF-D expression was detected in 85 cases with primary breast carcinoma by immunohistochemistry,and VEGF-D mRNA was detected by RT-PCR.Podoplanin monoclonal antibody was used to mark lymphatic endothelial cell and lymphatic vessel density(LVD) was counted.The relationship among the aboved parameters,clinicopathological parameters and prognosis was analysed. Results It was revealed by immunohistochemistry that VEGF-D expression was ranked by 5 levels: negative,n=5(5.88%);"+",n=17(20%);"++",n=34(40%);"+++",n=22(25.88%),and "++++",n=7(8.24%).VEGF-D expression was associated with tumor lymph node metastasis and TNM clinical stage(P

To analyse the familial aggregation and genetic predisposition of Shen-yin deficiency syndrome (SYDS) in families with diabetes mellitus type 2 (DM2). Methods One hundred and forty-one DM2 patients were collected from 32 family lines in Nanjin area, in which the probands were differentiated as DM2 with SYDS. On them, genetic analysis on the characteristics of SYDS was conducted using pedigree analysis, morbidity and heritability of the first-degree relatives of the probands were calculated, and the action of familial SYDS factor on the genesis of the syndrome was assessed by multiple factors regression analysis. Results The morbidity rate of SYDS in the first-degree relatives of the probands was 33.71%, and the heritability, calculated by Falconer formula, was 80.6%. The fitting result of regression analysis showed that familial factor played an important role in SYDS genesis (OR = 5.61, P = 0.001), but DM2 itself is not an independent risk factor for it. Conclusion DM2 with SYDS shows the tendency of familial aggregation and genetic predisposition, genetic factor is associated with the genesis of the syndrome. Pedigree research is a good method for exploring the relationship between syndrome and genetic factor.
Adult ; Diabetes Mellitus, Type 2 ; genetics ; Diagnosis, Differential ; Female ; Genetic Predisposition to Disease ; Humans ; Male ; Medicine, Chinese Traditional ; Pedigree ; Yin Deficiency ; genetics

BACKGROUND & OBJECTIVE: Dendritic cells (DCs) or DClike cells had been successfully induced in vitro from leukemia cells, which may provide a promising immunotherapeutic protocol for leukemia. This study was designed to investigate the efficiency of in vitro generation of dendritic cells from CD14+ acute myelomonocytic (M4) or monocytic (M5) leukemia cells and their ability of stimulating specific antileukemia T-cell response.METHODS: Bone marrow mononuclear cells (BMMNCs) were isolated from 5M4/M5 leukemia patients with high CD14 expression, and then divided into 3groups: adherent leukemia cells, nonadherent blasts, and total unfractioned blasts. CD14 expression of the 3 groups was evaluated by flow cytometry (FCM). When cultured with or without granulocyte-macrophage colonystimulating factor (GM-CSF), interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-α) for 7-10 days, monocytic leukemia cell-derived dendritic cells (Mo-LDCs) were identified through morphologic observation and immunophenotype analysis using FCM. The immune function of Mo-LDCs was detected through allogeneic mixed lymphocyte reaction (Allo-MLR) and cytotoxicity assay of antileukemia cytotoxic T lymphocytes (CTLs). The leukemic origin of Mo-LDCs was confirmed by chromosomal karyotype analysis combined with the aberrant expression of myeloid antigens. RESULTS: The amount of CD14+ cells, which could differentiate into CD83+ mature DCs under induction of the cytokine combination, was higher in adherent leukemia cells than in nonadherent blasts and total unfractioned blasts. Regarding each 3 cell groups of the same patient or the unfractioned blasts of various patients, initial CD14 expression was positively related to the yield of CD83 + DCs after induction (r=0.967, P=0.007). Mo-LDCs exhibited typical morphology and phenotype as mature DCs, induced potent proliferation of homogeneous T cells in Allo-MLR, stimulated the expansion of leukemia-specific CTLs, and continued to possess the cytogenetic abnormalities of the original leukemia, as well as the aberrant expression of myeloid antigens. CONCLUSIONS: In M4/M5 subtype of AML, CD14+ cells could differentiate into immune-competent Mo-LDCs under the induction of the cytokine combination. CD14 expression level may predict the DCs differentiation ability of monocytic leukemia. MoLDCs, which possess the classical phenotype and function of DCs, as well as the abnormal leukemic antigens, may be useful for the immunotherapy of M4/M5 AML.

OBJECTIVETo compare the outcomes of intracytoplasmic sperm injection (ICSI) with retrieved epididymal and testicular sperm for obstructive azoospermia and with ejaculated sperm for severe oligozoospermia and asthenospermia.

METHODSWe retrospectively analyzed 431 ICSI cycles, which were divided according to sperm sources into Groups A (n=287 in patients with severe oligozoospermia or asthenospermia using ejaculated sperm), B (n=109 in obstructive azoospermia patients with sperm retrieved by percutaneous epididymal sperm aspiration, PESA) and C (n=35 in obstructive azoospermia patients with sperm retrieved by testicular sperm extraction, TESE). Comparisons were made among the three groups in the rates of embryo implantation, fertilization, pregnancy, cleavage, and miscarriage.

RESULTSGroup A showed statistically significant differences from Groups B and C in the rates of embryo implantation and pregnancy (18.46% vs. 25.23% and 28.76%, 31.23% vs. 42.16% and 39.39%, P < 0.05). But no significant differences were seen in the rates of fertilization, cleavage and miscarriage among the three groups (P > 0.05).

CONCLUSIONThe rates of embryo implantation and clinical pregnancy are higher in patients with obstructive azoospermia than in those with severe oligozoospermia or asthenospermia after ICSI with ejaculated sperm.

Azoospermia ; therapy ; Epididymis ; cytology ; physiopathology ; Female ; Humans ; Male ; Oligospermia ; therapy ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; methods ; Spermatozoa ; Testis ; cytology ; physiopathology

OBJECTIVETo evaluate the feasibility and efficacy of intraperitoneal chemotherapy for malignant ascites caused by different types of abdominal cancers guided by chemo-sensitivity methyl tetrojolium coloremetric (MTT) assay in vitro.

METHODSCancer cells in the malignant ascites were collected for MTT assay to determine the chemo-sensitivity. The drug producing the highest or the second highest inhibition rate was selected for intraperitoneal chemotherapy. The correlation between the results of MTT assay and the response of malignant ascites, the clinical features, Karnofsky performance score (KPS) and prognosis were analyzed.

RESULTSMTT assay indicated that Taxotere (TXT) and Hydroxycamptothecin (HCPT) were the most effective to cancer cells in malignant ascites, and HCPT was mostly frequently used for intraperitoneal chemotherapy (56.9%). Twenty-four patients showed response by intraperitoneal chemotherapy (complete response: 7; partial response: 17) with a slightly significant correlation between the results of MTT assay and response of malignant ascites (P = 0. 014). The KPS of the responders was improved significantly (P < 0.001), and the response of malignant ascites to intraperitoneal chemotherapy was demostrated as an independent prognostic factor by multi-variate analysis in this series.

CONCLUSIONIn vitro chemo-sensitivity MTT assay guided intraperitoneal chemotherapy for malignant ascites is simple, effective and safe, which can improve the KPS and prognosis of the responders.

Adenocarcinoma ; drug therapy ; pathology ; Adult ; Aged ; Antineoplastic Agents ; administration & dosage ; pharmacology ; therapeutic use ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; therapeutic use ; Ascites ; drug therapy ; pathology ; Camptothecin ; administration & dosage ; analogs & derivatives ; pharmacology ; therapeutic use ; Cell Survival ; drug effects ; Colorectal Neoplasms ; drug therapy ; pathology ; Female ; Humans ; Injections, Intraperitoneal ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Pancreatic Neoplasms ; drug therapy ; pathology ; Stomach Neoplasms ; drug therapy ; pathology ; Taxoids ; administration & dosage ; pharmacology ; therapeutic use ; Tumor Cells, Cultured

To study the effects of tyrosine-kinase inhibitor STI571 on the adhesion of RPMI8226 cells to fibronectin (FN), cell adhesion mediated adriamycin-resistance and the Rac1 mRNA expression, the adhesion of RPMI8226 cells to fibronectin and drug resistance mediated by cell adhesion were determined by means of crystal violet staining and MTT assays respectively, Rac1 mRNA levels in RPMI8226 cells were examined by semi-quantitative RT-PCR. The results showed that STI571 could inhibit the adhesion of RPMI8226 cells to fibronectin. When RPMI8226 cells had been adhered to FN or BSA-coated wells for 1, 6 and 12 hours, the adhesion rates were (43.71 +/- 2.18)%, (55.63 +/- 1.56)%, and (63.42 +/- 2.46)% respectively. After treatment with STI571 20 micromol/L, the adhesion rates decreased to (15.12 +/- 1.04)%, (17.58 +/- 1.32)% and (17.24 +/- 1.59)% respectively (P < 0.05). The experiment revealed that growth of RPMI8226 cells adhered to FN-coated plates had a significant advantage over growth on BSA-coated plates when exposed to adriamycin (Adr) for 1 hour followed by a 24-hour culture period, and the mean IC(50) value for FN-adhered cells was (1.46 +/- 0.04) micromol/L while mean IC(50) value for BSA control was (0.78 +/- 0.03) micromol/L (P < 0.05). Following treatment with 20 micromol/L STI571, the mean IC50 values for FN and BSA adhered cells were (0.81 +/- 0.05) micromol/L, (0.74 +/- 0.02) micromol/L respectively, there was no significant difference between them (P > 0.05). RT-PCR demonstrated that the relative Rac1 mRNA level (Rac1/GAPDH) in RPMI8226 cells was downregulated following being treated with 20 micromol/L STI571. It is concluded that STI571 can inhibit the adhesion of RPMI8226 cells to fibronectin, reverse cell adhesion mediated adriamycin-resistance, and downregulate Rac1 mRNA level.
Benzamides ; Cell Adhesion ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; Fibronectins ; metabolism ; Humans ; Imatinib Mesylate ; Multiple Myeloma ; metabolism ; pathology ; Piperazines ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; Pyrimidines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Tumor Cells, Cultured ; rac1 GTP-Binding Protein ; biosynthesis ; genetics

OBJECTIVETo compare the efficacy and safety of standard or reduced doses of bortezomib combined with adriamycin and dexamethasone (PAD) in patients with multiple myeloma (MM).

METHODSEighty-two newly diagnosed or refractory/relapsed patients received bortezomib [either 1.2-1.3 mg/m(2) (standard dose) or 1.0-1.1 mg/m(2) (reduced dose) on day 1, 4, 8 and 11], and adriamycin (10 mg/m(2)) plus dexamethasone (40 mg/m(2)) on day 1-4 at 3-week intervals for 1 to 6 courses. The International Myeloma Working Group Criteria were used to evaluate the response. Adverse events were graded according to the National Cancer Institute Common Toxicity Criteria (Version 3.0).

RESULTSTwo courses of standard dose of PAD resulted in a similar response rate of partial and very good partial complete remissions (PR) compared with reduced dose (80.0% vs 80.8%, P=0.728). Grade III- Ⅳ neutropenia and thrombocytopenia were higher with standard dose than that with reduced doses of PAD (21.1% vs11.1%, P=0.270; 10.5% vs 6.3%, P=0.619, respectively). Grade III-Ⅳ bortezomib-induced peripheral neuropathy, herpes zoster, fatigue or abdominal distention were significantly higher with standard dose than that with reduced dose of PAD (15.8% vs 1.6%, P=0.037; 26.3% vs 6.3%, P=0.028; 36.8% vs 14.3%, P=0.046; 15.8% vs 1.6%, P=0.037, respectively).

CONCLUSIONReduced dose of PAD appears to result in a similar overall response rate, but a better tolerance and safety compared with standard dose.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Boronic Acids ; administration & dosage ; adverse effects ; therapeutic use ; Bortezomib ; Dexamethasone ; administration & dosage ; adverse effects ; Doxorubicin ; administration & dosage ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; Pyrazines ; administration & dosage ; adverse effects ; therapeutic use ; Treatment Outcome

Cytomegalovirus (CMV) infection is a dangerous complication in patients with chronic graft versus host disease (cGVHD). CMV-specific immunity depends on the activity of T cells. This study was aimed to investigate the effect of CMV pp65 gene modified dendritic cells (DCs) on activation of autologous T cells. Lentivirus system was utilized to introduce the CMV full-length pp65 gene into mouse DCs; CpG-DNA was used to induce mature DCs; flow cytometry and immunofluorescence were used to determine the expression of antigen and IFNgamma in T lymphocytes. The results showed that the DCs were infected with lentivirus at a multiplicity of infection (MOI) of 50 with optimal infectious efficiency of 30%-40%; mature DCs expressing pp65 gene could stimulate autologous naive T cells to express CD69 specifically; mature DCs expressing PP65 could stimulate autologous CD4+ or CD8+ T cells to produce IFNgamma. It is concluded that CMV pp65-modified and CpG-DNA-induced mature DCs can activate CMV-specific T lymphocytes in vitro.
Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, T-Lymphocyte ; genetics ; metabolism ; Antigens, Viral ; immunology ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; CpG Islands ; genetics ; Cytomegalovirus ; immunology ; DNA ; genetics ; Dendritic Cells ; cytology ; immunology ; metabolism ; Humans ; Interferon-gamma ; genetics ; metabolism ; Lectins, C-Type ; Lentivirus ; genetics ; metabolism ; Mice ; Phosphoproteins ; genetics ; metabolism ; Viral Matrix Proteins ; genetics ; metabolism