Objective To study the specific anti-tumor immunity of dendritic cell(DC) modified by HSP70-tumor peptide complexes and sCD40L in vitro and in vivo.Methods The murine marrow cells were cultured for 7 d,and then randomized to 4 groups: control,only HSP70-H22 peptide complexes,sCD40L,and HSP70-H22 peptide complexes combined with sCD40L.After intervention for 24 h,IL-12 and IL-10 in the supernatant were detected by ELISA assay,CD40 and CD80 of DC by FACS,and the proliferation of spleen lymphocytes by MRL.The ability of spleen lymphocytes activated by DC to H22 cells in vitro was investigated by MTT.The models of murine hepatoma were established,and then randomized to 5 groups(Ⅰ,Ⅱ,Ⅲ,Ⅳ and Ⅴ),followed by interference with normal saline and DC respectively.At 21 d,mice were sacrificed and the weight of tumor was measured.The levels of IL-10 and IFN-? in blood serum were detected by ELISA assay.Results Compared with that in the groups of control,only sCD40L,and HSP70-H22 peptide complexes,the level of IL-10 in the group stimulated by HSP70-H22 peptide complexes combined with sCD40L decreased significantly,but other indexes in this group increased significantly(P

ZHANG Lin-hua, GAO Rui-chang, XU Ming-li (School of Chemical Engineering, Tianjin University, Tianjin 300072, China)

Objective To study the therapeutic effect and mechanism of Dicliptera chinensis polysaccharide ( DCP) on liver injury induced by antituberculosis drugs. Methods Sixty mice were randomly divided into six groups, namely normal control group, model group, glucurolactone group (in the dosage of 200 mg·kg-1·d-1), and high-, middle- and low-dose DCP groups ( in the dosage of 600, 400, 200 mg·kg-1·d-1, respectively). Except for the normal control group, the rats in the other groups were given intragastric administration of isoniazid and rifampicin ( 100 mg/kg) to induce liver injury model, and were simultaneously treated with corresponding agents, once a day. On the experiment day 30, the blood and liver tissue were sampled. The serum levels of alanine aminotransferase ( ALT) , aspartate aminotransferase ( AST) , alkaline phosphatase ( AKP) and microsomal nitric oxide ( NO) were detected by biochemical method. The contents of tumor necrosis factor alpha (TNF-α) and interleukin 6 (IL-6) in liver tissue were determined by enzyme-linked immunosorbent assay ( ELISA) , and the hepatic histopathology was observed after HE staining. Results In DCP groups, the hepatic pathological changes of the mice were improved, the number of the inflammatory cells was reduced, and the activities of serum ALT, AST and AKP as well as the contents of hepatic TNF-α, IL-6 and NO were reduced ( P<0.05 or P<0.01 compared with those in the model group). Conclusion Dicliptera chinensis polysaccharide is effective for liver injury induced by antituberculosis drug, and the mechanism may be associated with its anti-inflammatory action.

BACKGROUND: The occurrenceand development of rheumatoid arthritis were strongly associated with unbalance proliferation and apoptosis of synovial cells and lymphocytes. Some synovial cell apoptosis was abnormal. OBJECTIVE: To observe effects of heterogenic double umbilical cord blood stem cell transplantation on Bcl-2 and Bax expression in mice with type Ⅱcollagen-induced arthritis. METHODS: C57BL/6(H-2b) mice were induced with Freund's complete adjuvant and type Ⅱcollagen to establish mouse models of type Ⅱcollagen-induced arthritis.At 2days after secondary immunity incubation, mice were injected with saline via tail vein in the model and normal control groups. Umbilical cord blood hemopoietic stem cells were injected into mouse tail vein in the UBSCs transplantation groups (single dose:2×106/50 g; double dose: 1×106/50 g each, totally 2×106/50 g). Mice were intragastrically administrated methotrexate in the methotrexate positive control group, 0.017 5 g/kg once, once every 5 days, totally six times. RESULTS AND CONCLUSION: Joint tissue below knee and elbow was obtained at42 days following transplantation. Histopathology displayed that smooth and glossy articular surface, no inflammatory cell infiltration in the synovial layer and normal chondrocytes in the normal control group. Hyperplasia, a lot of inflammatory cell infiltration and damaged cartilage surface were visible in the model group. Slight hyperplasia and a little inflammatory cell infiltration were detectable in the methotrexate positive controlgroup and single UBSCs transplantation group. Double UBSCs transplantation group exhibited smooth and glossy articular cartilage surface, no damage, a little inflammatory cell infiltration. Immunohistochemistry demonstrated that Bcl-2 and Bax expression was lower in the double UBSCs transplantation group compared with single UBSCs transplantation group (P < 0.05). Compared with normal control group, no significant difference was detected (P > 0.05). Results suggest that double umbilical cord blood stem cells can induce synovial cell apoptosis in mice with type Ⅱcollagen-induced arthritis in a certain number and action time, and protect synovial membrane against damage.

BACKGROUND: Matrix metalloproteinase (MMP) degrades extracellular matrix, which is a necessity of joint destruction in rheumatoid arthritis patients. MMP-2 and MMP-9 can evaluate rheumatoid arthritis and serve as an index to predict progressive destruction of the joint. OBJECTIVE: To observe heterogenous allogeneic umbilical cord blood stem cell (UBSC) transplantation on MMP-2 and MMP-9 expression in the spleen of mice with type Ⅱ collagen-induced arthritis. METHODS: Fetus cord blood was sterilely obtained and cord blood stem cells were separated. The C57BL/6(H-2b) mice were assigned to five groups (n=10). Except normal control group, models of collagen-induced arthritis were established using complete Freund's adjuvant + type Ⅱ collagen. Mice from the methopterin group were intragastrically administered methopterin suspension 0.017 5 g/kg, once every 5 days. Other groups used caudal vein injection. Mice from the model and normal control groups were injected with saline. Mice from the mono-UBSCs group and double-UBSCs group were injected with 2×106/kg UBSCs from one and two parents. At 42 days following injection, animals were sacrificed and the ankle joint was obtained for histopathological detection. MMP-2 and MMP-9 mRNA expression in the spleen was examined using reverse transcription-polymerase chain reaction. RESULTS AND CONCLUSION: Double-UBSC transplantation could significantly inhibit inflammatory cell infiltration in synovial tissue of mice with type Ⅱ collagen-induced arthritis, repaired impaired cartilage tissue. The repair effect was better than that in methopterin group and mono-UBSCs group. MMP-2 and MMP-9 mRNA expression in the spleen was significantly lower in the double-UBSCs group than the mono-UBSCs group (P < 0.01). These suggest that heterogenous allogeneic double-UBSCs transplantation participated in pathological changes in rheumatoid arthritis cartilage and in synthesis of cartilage extracellular matrix and effectively treated rheumatoid arthritis by regulating MMP-2 and MMP-9 mRNA expression.

Aim To study the effects of Dicliptera chinensis polysaccharide(DCP)on alcoholic fatty liver disease(AFLD)in rats based on anti-inflammation and antioxidation.Methods 60 rats were randomly divid-ed into six groups:control group,model group,silybin group and DCP of high,medium and low dose groups. The control group was fed with normal diet, other groups were fed with high sugar and high fat diet,and given 5% alcohol 5 mL·kg-1 by gavage.The alcohol consistency increased 5%every week until AFLD mod-els in rats were made after 7 weeks.Except control group,other groups were fed with high sugar and high fat diet,and given 35% alcohol 5 mL · kg-1 and DCP.All rats were killed after five weeks,and blood and liver tissues were collected.The activity of alanine aminotransaminase (ALT),aspartate aminotransferase (AST ), alkaline phosphatase (AKP ), triglyceride (TG),total cholesterol (TC ),low-density lipoprotein cholesterol(LDL-C)and high-density lipoprotein cho-lesterol(HDL-C)in serum were detected by using bio-chemical method. The contents of malondialdehyde (MDA),superoxide dismutase (SOD),reduced gluta-thione(GSH)in liver tissues were detected.The con-tents of tumor necrosis factor-α(TNF-α),interleukin-6 (IL-6 )and transforming growth factor-β1 (TGF-β1 ) were determined by enzyme-linked immunosorbent as-say(ELISA)in liver tissues.The liver tissues were ob-tained and histologic analysis was done through HE. Results DCP reduced the activity or content of ALT, AST,AKP,TG,TC,LDL-C,HDL-C,TNF-α,IL-6, TGF-β1 in serum and liver tissues of rats(P<0.05 ), and increased the activity or content of HDL-C,SOD and GSH (P<0.05 ).DCP could remarkably inhibit the NF-κB expression in liver tissues(P<0.01 ).The pathological examination indicated that DCP could ob-viously alleviate the inflammation and fat denaturation of the liver cells.Conclusion DCP can inhibit the de-velopment of AFLD.The mechanism may be related to antioxidation,free radical scavenging, inhibition of lipidperoxidation,anti-inflammation,and inhibition of the TGF-β1 and NF-κB expression.

Objective To develop the Assessment Scale for Patient Safety Culture in ICU and do psychometric test.Methods The Assessment Scale for Patient Safety Culture in ICU was developed based on literature review,qualitative interview,and two-round Delphi consultation.The reliability and validity were tested among 235 chnical nurses and doctors in ICU.Results A 45-item questionnaire was originally developed and eleven of them were deleted after subjective screening,project analysis and exploratory factor analysis.Seven factors were extracted with cumulative contribution rate of 59.347％.The scale of total Cronbach's alpha coefficient was 0.909; the split half reliability was 0.832.Each dimension of Cronbach's alpha coefficient was between 0.481～0.909,and the split half reliability was 0.481～0.866.All had significance in statistics above.Conclusions The Assessment Scale for Patient Safety Culture in ICU has good reliability and validity,which can be used to evaluate the ICU patient safety culture atmosphere domestically.

Objective To evaluate the potential differences in serum proteomic profiles between patients with esophageal squamous cell carcinoma(ESCC)and precancerous lesions in order to establish proteomic pattern model for diagnosis of ESCC and precancerous lesions in high risk area,and to investigate its value in screening ESCC.Methods The serum and endoscopic biopsy samples were obtained from 38 normal controls,63 patients with atypical hyperplasia(class Ⅰ 26 cases,class Ⅱ 26 cases,class Ⅲ 11 cases)and 36 patients with advanced esophageal carcinoma.The serum proteomic patterns were examined using surface enhanced laser desorption/ionization time of flight mass spectrometry(SELDI-TOF-MS)and CM10 protein chip.The data was analyzed and disease diagnostic models were established using support vector machine(SVM).The diagnostic model was evaluated and validated by leave one cross validation.Results ①The diagnostic model could differentiate advanced esophageal carcinoma from normal controls with a specificity of 89.47%and a sensitivity of 83.33%.②The results delivered 92.31%,80.77% and 90.91%specificity,and 80.56%,83.33%and 94.44%sensitivity for discrimination of atypical hyperplasia Ⅰ,Ⅱand Ⅲ,respectively,using diagnostic models.③Four(4291,5644,5664,8775)m/z peaks observed repeatedly using diagnostic models.Conclusions The SELDI-TOF-MS and SVM provide a new approach for discrimination of ESCC and precancerous lesions in high risk area.Four(4291,5644,5664,8775)m/z peaks may considered as potential biomarkers which related to the ESCC and esophageal precancerous lesions.

Objective: To evaluate the effect of dexmedetomidine on hypoxia-inducible factor-1alpha (HIF-1 α) signaling pathway during hypoxia in mice with lung cancer. Methods: Eighteen clean-grade healthy adult male BALB/c nude mice, aged 8 weeks, weighing 20-30 g, were divided into 3 groups (n=6 each) using a random number table method: lung cancer group (group L), hypoxia+ lung cancer group (group HL), and hypoxia plus lung cancer plus dexmedetomidine group (group HLD). Human lung adenocarcinoma A549 cell suspension 1×10⁶/150 μl was injected via the tail vein to establish the mouse model of lung cancer.After the model was established successfully, chronic intermittent hypoxia was performed as follows: the mice were placed in air-tight modular incubation chambers, and the atmosphere was controlled by a constant gas flow containing 10% O₂ for 3 h once a day for 3 weeks.The mice in group HL were exposed to hypoxia.After the end of hypoxia exposure, the animals in group HLD were intraperitoneally injected with dexmedetomidine 25 μg/kg 3 times a week for 3 weeks in total.The mice in group L were placed in air-tight modular incubation chambers and the atmosphere was controlled by a constant gas flow containing 21% O₂ for 3 h once a day for 3 weeks, and the equal volume of normal saline was injected intraperitoneally.The mice were sacrificed by cervical dislocation, and the lung tissues were removed for determination of lung cancer nodules count, HIF-1α expression (by immunohistochemistry), expression of HIF-1α, survivin and X chromosome linked inhibitor of apoptosis protein (XIAP) (by Western blot), and expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 (by real-time polymerase chain reaction). Results: Compared with group L, the number of lung cancer nodules was significantly increased, and the expression of HIF-1α, survivin, XIAP, MMP-2 and MMP-9 was up-regulated in the other two groups (P<0.05). Compared with group HL, the number of lung cancer nodules was significantly increased, and the expression of HIF-1α, survivin, XIAP, MMP-2 and MMP-9 was up-regulated in group HLD (P<0.05). A small number of HIF-1α positive cells were found in group L, a medium number of HIF-1α positive cells in group HL, and a large number of HIF-1α positive cells in group HAD. Conclusion Dexmedetomidine can promote proliferation of tumor cells in mice with lung cancer during hypoxia, and the mechanism is related to activating HIF-1α signaling pathway.

To evaluate bioequivalence and safety of two kinds of metformin hydrochloride sustained-release tablets (test preparation vs reference preparation) under the condition of fed and single administration.A single center, randomized, open, single-dose, two-period, two-sequence, and double-crossover design was used.32 healthy subjects took 0.5 g of test preparation or reference preparation under fed and single-dose administration.4 mL of venous blood was collected from before administration (0 h) to 1, 3, 4, 4.5, 5, 5.5, 6, 7, 8, 9, 10, 12, 15, 24, 36 and 48 h after administration.The concentration of metformin in plasma samples was detected, and then the pharmacokinetic parameters were calculated by WinNonlin 7.0 software.When the 90% confidence intervals of cmax, AUC0-t and AUC0-∞ geometric mean ratio of test preparation and reference preparation were within 80.00%-125.00% equivalent intervals respectively, the bioequivalence of the two preparations was proved.One subject fell off due to adverse events.The main pharmacokinetic parameters of test preparation and reference preparation as follows: cmax were (0.68 ± 0.14) and (0.65 ± 0.11) mg/L, AUC0-t were (7.33 ± 1.65) and (7.00 ± 1.89) h·mg/L, AUC0-∞ were (7.39 ± 1.67) and (7.06 ± 1.91) h·mg/L, respectively.The 90% confidence intervals of the geometric mean ratio of the two main pharmacokinetic parameters were 101.45%-109.14%, 100.08%-112.32% and 100.24%-112.28%, respectively, which fell within the bioequivalence interval of 80.00%-125.00%.There were no serious adverse events and unexpected adverse events during the trial.The results show that test preparation and reference preparation are bioequivalent under fed and single-dose administration, safe and well tolerated in healthy subjects.