Objective:To study the inductive effect of recombinant MUC1-MBP fusion protein combined with R848 on the immune activity of T cells in the mice,and to provide experimental basis for searching the suitable adjuvants of recombinant MUC1-MBP fusion protein.Methods:A total of 21 C57BL/6 mice were randomly divided into control group(treat with normal saline),MUC1-MBP+R848 group (treated with MUC1-MBP+R848),and MUC1-MBP+baillus calmette guerin(BCG) group(treated with MUC1-MBP+BCG)(n=7).After immunized for 4-7 d,the spleen tissue was taken and the spleen indexes of the mice in various groups were measured;the stimmulus index(SI) of the mice was detected by lymphocyte proliferation response;the levels of tumor necrosis factor-γ(TNF-γ) and interleukin-4(IL-4) were detected by ELISA method;the proprotions of T lymphocyte subsets in spleen cells were analyzed by flow cytometry.Results:Compared with control group,the spleen indexes,SI,and TNF-γ levels of the mice in MUC1-MBP+ R848 and MUC1-MBP+BCG groups were significantly increased (P<0.05 or P<0.01);the indexes metioned above of the mice in MUC1-MBP+ R848 were higher than those in MUC1-MBP+BCG group;the IL-4 levels had no significant difference(P>0.05).Compared with control group,the proprotions of CD3+T,CD4+T,and CD8+ T cells of the mice in MUC1-MBP+ R848 group and MUC1-MBP+BCG group were significantly increased(P<0.05 or P<0.01).Conclusion:Both MUC1-MBP fusion protein combined with R848 and BCG can induce the Th1 type of immune activity in the mice,and R848 is the potential candidate adjuvant for MUC1-MBP.

Objective:To explore the association between HLA-DR,DQ allele polymorphisms and onset of epidemic hemorrhagic fever(EHF)among Han Nationality in Zunyi area.Methods:Using group study,HLA-DR and DQ genotyping was conducted in 100 EHF cases and 100 controls among Han Nationality in Zunyi area with polymerase chain reaction-sequence specific primer(PCR-SSP),GF(gene frequency)and RR(relative risk)were calculated and compared.Results:The frequency of HLA-DRB1 16 in patients with EHF was higher than in the control group(RR=3.58,?2=4.916,P=0.0266

Objective: To investigate the MUC1 specific immune response enhanced by thmosinal (Tα1) using MUC1-MBP as the specific antigen, and to discuss the feasibility of MUC1-MBP as an adjuvant. Methods: The C57BL/6 mice were randomly divided into normal saline group, MUC1-MBP + BCG group and MUC1-MBP + Tα1 group. The T cellular immune activities, MUC1 special antibody and subclass, anti-tumor effect of Tα1 combined with MUC1-MBP were detected. 4-7 d after the 3rd immunization, the spleen indexes of the mice in various groups were measured; the lymphocyte proliferation response was used to detect the stimulate index (SI) of the mice in various groups; the levels of specific cytokines IFN-γ, IL-2, IL-4 and IL-10 in the supernatant of spleen cells of the mice were detected by ELISA; the level of serum MUC1-specific antibody was detected by ELISA. The C57BL/6 mice were inoculated with B16-MUC1 7 d after the last immunization and the survival of the mice was observed. Results: Compared with normal saline group, the spleen index and SI of the mice in MUCl-MBP+Tα1 and MUC1-MBP+BCG groups were significantly increased (P< 0.05 or P<0.01); the specific SI of MUC1 were significantly increased (P< 0.05). Compared with normal saline group, the levels of IFN-y and IL-2 in the supernatant of spleen cells of the mice in MUC1-MBP+BCG group were obviously increased (P<0.05); the levels of IL-4 and IL-10 were slightly increased, but there were no significant differences (P>0.05); compared with normal saline group, the level of IL-4 in MUCl-MBP+Tal group was obviously increased (P< 0.01); the levels of IFN-γ, IL-2 and IL-10 were slightly increased, but there were no significant differences (P>0.05). The titer of MUC1 specific antibody was increased with the increase of concentration of Tα1. The antibody subtype detection results showed that compared with normal saline group, the level of IgG1 in MUCl-MBP + BCG group was significantly increased (P< 0.05 or P< 0.01), and the level of IgG2a had no obvious change. The tumor prevention experiment results showed that compared with normal saline group, the survival rates of the mice in MUC1-MBP+BCG group and MUCl-MBP + Tal group had no significant differences. Conclusion: MUC1-MBP combined with Tα1 prefers to Th2 immune responses in the mice. It indicates that Tα1 can be used as an adjuvanted preventive vaccines, but not suitable for therapeutic vaccines.

Objective To enhance clinicians' intention to the importance of early diagnosis,early therapy and follow-up of type Ⅱ citrullinemia.Methods The clinical data of one adult-onset type Ⅱ citrullinemia pedigree were collected. The gene mutation type of SLC25A13 of proband and her daughter were determined by PCR and direct gene sequencing.Results The patient was a 27 years-old female,who complained of repeated dizziness, vomiting for more than 2 years and recurrent attacks of altered consciousness for about one and a half year.An abdominal ultrasonogram,liver magnetic resonance imaging and liver histology obtained by needle biopsy all determined the liver pathological changes of liver cirrhosis.Electroencephalogram showed sharp waves. The plasma amino acid showed a marked elevation of blood citrulline.Laboratory findings revealed a highly increased concentration of plasma ammonia during every episode. Mutation analysis of the SLC25A13 gene identified a homozygote of 851del4 in the patient,and heterozygote of 851del4 in her daughter. Conclusions For adults,unexplained dizziness,vomiting,but liver function still in the compensation,especially accompanied by neuropsychologic symptoms are highly suggestive of adult-onset type Ⅱ citrullinemia.SLC25A13 gene analysis contributes to the diagnosis of this disease,avoids invasive investigations and early confirmation of this disease means long-term dietary advice,genetic counseling,medical surveillance and early preparation for liver transplantation if is necessary.

Objective: To examine the effects of ethyl pyruvate (EP) on mitochondrial dynamics and cell apoptosis in lipopolysaccharide (LPS)-induced human kidney-2 (HK-2) cells. Methods: HK-2 cells were divided into three groups: HK-2 cells were challenged with LPS (800 μg/L) for 24 hours as LPS group, or LPS mixed with EP (0.25 mmol/L) for 24 hours as EP group. Cells were incubated with normal saline for 24 hours as control group. The levels of malondialdehyde (MDA), superoxide dismutase (SOD), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and intracellular adenosine triphosphate (ATP) were detected by enzyme linked immunosorbent assay (ELISA). JC-1 staining and Annexin V-fluorescein isothiocyanate/propidium iodide (FITC/PI) assays were used to evaluate mitochondrial membrane potential and cell apoptosis, respectively. Western Blot was used to evaluate the protein expressions of mitochondrial dynamics, including death-associated protein kinase 2 (DAPK-2), mitofusin (Mfn-1 and Mfn-2), and apoptotic associated biomarkers, including caspase-3, caspase-9, Bcl-2, Bcl-xL, cytochrome C (Cyt C), and DNA repair enzyme poly ADP-ribose polymerase (PARP). Results: Compared with the NC group, MDA, IL-6, TNF-α of LPS group were significantly increased, the expression of SOD, mitochondrial membrane potential and ATP level were significantly decreased, the expression of mitochondrial fission protein DAPK-2 was significantly increased, and mitochondrial fusion proteins Mfn-1 and Mfn-2 were significantly decreased, cell apoptosis and apoptotic protein caspase-3, caspase-9 and Cyt C were increased, and anti-apoptotic protein Bcl-2, Bcl-xL, PARP were significantly decreased. Compared with the LPS group, the oxidative activities and inflammatory factors above were inhibited in EP group [MDA (μmol/L): 12.35±2.21 vs. 45.95±1.76, SOD (kU/L): 54.68±1.42 vs. 40.73±1.60, IL-6 (ng/L): 67.87±2.61 vs. 338.92±20.91, TNF-α (ng/L): 19.23±1.80 vs. 180.69±6.51], mitochondrial membrane potential and ATP level were significantly increased [mitochondrial membrane potential: (99.43±0.25)% vs. (69.40±0.75)%, ATP (×10⁶ RLU): 0.19±0.01 vs. 0.12±0.05], the expression of mitochondrial fission protein was significantly decreased (DAPK-2/β-actin: 0.03±0.01 vs. 0.61±0.02), mitochondrial fusion proteins were significantly increased (Mfn-1/β-actin: 0.43±0.04 vs. 0.17±0.01, Mfn-2/β-actin: 0.201±0.004 vs. 0.001±0.001), percentage of cell apoptosis was significantly decreased [(5.25±0.17)% vs. (34.42±0.64)%], the expressions of apoptotic proteins were significantly decreased (caspase-3/β-actin: 0.25±0.15 vs. 1.76±0.01, caspase-9/β-actin: 0.09±0.02 vs. 1.52±0.12, Cyt C/β-actin: 0.001±0.001 vs. 0.350±0.030), and the expressions of anti-apoptotic proteins and PARP were significantly increased (Bcl-2/β-actin: 0.500±0.010 vs. 0.009±0.004, Bcl-xL/β-actin: 0.550±0.010 vs. 0.009±0.001, PARP/β-actin: 0.94±0.01 vs. 0.16±0.13), with statistically significant differences (all P < 0.05). Conclusions There are enhanced mitochondrial fission and diminished mitochondrial fusion in LPS-induced HK-2 cells. EP can protect mitochondria functions by regulate mitochondrial dynamics, and reducethe apoptosis of LPS-induced HK-2 cells.

BACKGROUNDMany studies have shown that continuous renal replacement therapy (CRRT) could clean lactate and treat the hyper-lactatemia. On the contrary, some other studies found that filter lactate clearance only accounted for a very small part of total lactate clearance and the hemofilter's contribution to the overall lactate clearance was negligible. The objective of this study was to evaluate the effects of various doses of continuous veno-venous hemofiltration (CVVH) on plasma lactate elimination in critically ill patients.

METHODSPatients were divided into three groups according to their incipient plasma lactate concentration. Group A: lactate ≤ 2 mmol/L, group B: lactate 2-5 mmol/L, group C: lactate ≥ 5 mmol/L. Three different doses (20 ml × kg(-1)× h(-1), 35 ml × kg(-1)× h(-1) and 45 ml × kg(-1)× h(-1)) of CVVH were applied to critically ill patients who experiencing CVVH. The concentrations of plasma lactate in pre-(A), post-dialyzer (V) sites and ultrafiltrate were measured after each dosage of CVVH was carried out for 30 minutes. Rate of lactate clearance by the filter (RLC) and filter lactate clearance (FLC) and Lactate-Sieving Coefficient (LSC) were calculated under different circumstances, including different doses of CVVH and different incipient lactate levels.

RESULTSFifteen patients were enrolled and 104 blood samples were drawn and lactate concentrations were measured in this study. RLC was found increased ((9.36 ± 9.73) mmol/h, (13.92 ± 12.56) mmol/h and (16.52 ± 12.71) mmol/h, P < 0.05 respectively) with the dose of CVVH increased. RLC was also increased ((3.46 ± 1.46), (10.38 ± 5.50) and (24.53 ± 14.69) mmol/h, P < 0.05 respectively) with the incipient lactate increased. FLC was increased ((1.95 ± 0.63), (2.95 ± 0.74) and (3.45 ± 0.54) L/h, P < 0.05 respectively) with the dose of CVVH increased. There was no significant difference of LSC in different doses of CVVH and different incipient lactate levels.

CONCLUSIONSPlasma lactate can be eliminated by CVVH and different doses of CVVH affect the rate of lactate clearance in critically ill patients.

Adult ; Aged ; Aged, 80 and over ; Critical Illness ; Female ; Hemofiltration ; Humans ; Lactic Acid ; blood ; Male ; Middle Aged