Objective　The aim of this paper was to investigate the changes in cyclic 3'5'-guanosine monophosphate (cGMP) before and after hemodialysis to estimate the value of cGMP to the dry boby-weight. Methods: Plasma cGMP levels (by radioimmunoassay), cardiothoracic ratio (CTR), and the body weight (BW) before and after hemodialysis were determined in chronic hemodialysis patients and clinical signs and symptoms were observed at the same time. Results　① The predialytic cGMP value of the patients was significantly higher than that of healthy controls (P<0.05). ② The postdialytic cGMP level was significantly lower than the predialytic cGMP level (P<0.01). ③ Postdialytic CTR and BW values were significantly lower than predialytic values (P<0.01). ④Compared to those of predialysis, postdialytic clinic signs and symptoms of the patients were significantly relieved. Conclusions　① The plasma cGMP level can sensitively reflect the hydration state and is a reliable marker for dry body-weight estimation.②The measurement of plasma cGMP combined with clinical parameters and radiological indexes permit a more accurate dry body-weight estimation.

Objective To indentify the effect of hemodialysis(HD) and continuous veno-venous hemofiltration (CVVH) on serum hepatocyte growth factor (HGF) in the patients with acute renal failure (ARF). Methods Of 28 patients with ARF, 19 received HD and 9 underwent CVVH. HGF concentrations in patients serum and ultrafiltrate were measured by ELISA. Results Compared with the controls, serum HGF concentration increased significantly in ARF patients (P

Objective:To determine the characteristics and differences in bleomycin-induced lung ifbrosis model by repeated low-dose intravenous injection and single dose intratracheal instillation of bleomycin.
Methods:Forty male ICR (Institute for Cancer Research) mice were randomly divided into a model group I, a model group II, and 2 control groups (10 mice in each group). In model group I, bleomycin was injected intravenously at 10 mg/(kg·d) for 14 consecutive days;and in model group II, bleomycin was instilled intratracheally at 5 mg/kg. The 2 control groups were given isotonic saline solution. At the 28th day, the mice were sacrificed and the bronchoalveolar lavage lfuid (BALF) was collected. The total cells and proteins in the BALF, pulmonary coeffcient, and hydroxyproline (HYP) content were determined. The pathological changes were observed by the eosin staining and Masson's trichrome staining.
Results:1) Both intravenous injection and intratracheal instillation of bleomycin resulted in severe and extensive inlfammation and ifbrosis in the lungs. The total cells and proteins in the BALF, HYP content, pulmonary coeffcient and the pathological score of pulmonary ifbrosis were all signiifcantly increased in the 2 model groups (P<0.01). 2) Fibrosis was mainly under the pleura or around the vessel in model group I, and it was located near the bronehia and bronchioles in model group II. 3) The death rate was higher in the model group II than that in the model group I. 4) Proteins in the BALF were significantly higher in model group II than that in model group I (P<0.05). There was no difference in the total cells in the BALF, the pulmonary coefficient, the HYP content, and the pathological score of pulmonary ifbrosis between the 2 groups (P>0.05).
Conclusion:The pulmonary fibrosis model can be successfully established by intravenous injection or intratracheal instillation of bleomycin, but the sites of pulmonary ifbrosis are different. The histological changes caused by the repeated low-dose intravenous injection of bleomycin is more similar to idiopathic pulmonary ifbrosis than that by the single dose intratracheal instillation.

0.05). Conclusions One mechanism of SF pretreatment cardioprotective effect is mediated by bradykinin. The combined use of SF and CP doesn′t result in significant improvement, and thenefore is not advocated.

Objective To evaluate immune protective effects of Treponema pallidum(Tp)pcD/Tp92 DNA vaccine delivered through different inoculation routes against Tp-induced skin infection in New Zealand rabbits. Methods A total of 108 New Zealand rabbits were randomly and equally divided into 6 groups:A1 and A2 groups treated with intramuscular injection of empty plasmids pcD and pcD/Tp92 DNA vaccine respectively for 2 sessions, B1, B2, C1 and C2 groups firstly treated with intramuscular injection of the pcD/Tp92 DNA vaccine for 1 session for primary immunization, then receiving nasogastric feeding with pcD/Tp92 DNA vaccine, pcD/Tp92 DNA vaccine+cytosine-phosphate-guanine(CpG)oligodeoxynucleotide (ODN), and recombinant Tp92 protein, and recombinant Tp92 protein+CpG ODN respectively for booster immunization. Enzyme-linked immunosorbent assay(ELISA)was conducted to detect the serum level of anti-Tp92 IgG antibody at week 0, 2, 4, 6, 8 after immunization, the SIgA level in the nasopharyngeal region and vaginal mucosa at week 8 after immunization, as well as levels of interleukin-2(IL-2)and interferon-γ (IFN-γ)in the culture supernatant of rabbit spleen cells at week 8 after immunization, and methyl thiazolyl tetrazolium(MTT)assay was performed to estimate proliferative activity of rabbit splenic lymphocytes in three rabbits from each group. At week 10 after immunization, other 15 rabbits from each group were subcutaneously inoculated with Tp standard strain, and changes of skin lesions at the inoculation site during early-stage infection were observed and recorded. Results At week 8 after immunization, the C2 group showed significantly higher serum level of anti-Tp92 IgG antibody(1.825 ± 0.175), supernatant levels of IL-2 (154.7 ± 14.6)and IFN-γ(277.4 ± 24.4), and proliferative activity of T cells(3.57 ± 0.24)compared with the A2(1.372 ± 0.322, 112.3 ± 13.4, 232.8 ± 25.3, 3.08 ± 0.22, respectively, all P<0.05), B1(0.893 ± 0.297, 76.6 ± 21.5, 165.7 ± 22.6, 2.12 ± 0.14, respectively, all P<0.05)and B2(1.294 ± 0.124, 97.3 ± 18.7, 211.3 ± 24.6, 2.88 ± 0.18, respectively, all P<0.05)groups. In addition, effective immunoprotection was achieved in the C2 group with more production of mucosa-specific SIgA antibody, as well as the lowest Tp-positive rate (6.67%) and ulcer formation rate (6.67%) in skin lesions at the inoculation sites. Conclusion The effective vaccination strategy, namely intramuscular injection of the pcD/Tp92 DNA vaccine for primary immunization followed by nasogastric feeding with mucosal adjuvant CpG ODN combined with recombinant Tp92 protein for booster immunization, can induce the strongest mucosal immune responses and immune protective effects.

Objective:To establish a method to determine the metabolites in rat kidney tissues by gas chromatography-mass spectrometry (GC-MS) combined with chemometric techniques. Methods:Metabolites were separated and identiifed on HP-5MS column (30 m × 0.25 μm × 0.25 mm). The initial column temperature was 100℃lasting 3 min, and then programmed at 8℃/min to 300℃, maintaining at this temperature for 6 min. hTe internal standard was heptadecanoic acid. hTe grinded kidney tissue was exacted by methanol. hTe supernatant was dried by nitrogen. Atfer the oximation and derivation, the supernatant was analyzed by GC-MS. hTe overlapped peaks were resolved into pure chromatogram and mass spectra with chemometric techniques. Qualitative analysis was performed by comparing the obtained pure mass spectra with those in NIST mass spectra database and certiifcated by the standards and the references. hTe internal method was used for semi-quantitation. Results:A total of 53 compounds were identiifed. hTe main constitutions in the kidney tissue were amino acids, saccharides, fatty acids and urea. Conclusion:hTe combination of methods is rapid and accurate for the analysis of metabolites in the kidney tissue, which provides more information for further study of metabonomics in kidney tissues.

Objective To investigate the expression of p21 and p53 in renal interstitial fibrosis rats and the effect of enalapril on it. Methods Sprague-Dawley rats were randomly divided into 3 groups, shame operation rats group, unilateral urethral obstruction and enalapril treatment group. Histological chan-ges were observed by Masson stain. The expression of p21 mRNA and p53 mRNA was detected by RT-PCR. Results With degree of interstitial fibrosis aggravating, the expression of p21 and p53 increasing,p21 and p53 expression of UUO rats at every time point were positive correlative. Enalapril can inhibit the expression of p21 and p53. Concinsion p21 and p53 expression increased in UUO rats renal cortex and enalapril can significantly inhibit its expression, p21 may participate in the pathogenesis of renal tubule-in-terstitial fibrosis through p53 pathway.

AIM: To investigate clonal expansion and specific cytotoxicity of TCR V? subfamily T cells from acute myelogenous leukemia M_ 2a subtype (AML-M_ 2a)patients and normal individuals induced by AML-M_ 2a cells, respectively. METHODS: Complementarity determining region 3(CDR3) of TCR ? with variable region genes was amplified in autologous or allogeneic T cells from mixed lymphocyte and tumor culture (MLTC) using RT-PCR. The positive products were further analyzed to identify the clonality of T cells by genescan. The specific cytotoxicity of T cells was analyzed by MTT. RESULTS: T cells from both M_ 2a patients and normal individuals after MLTC showed high response to M_ 2a cells with 4-17 TCR V? subfamily dominant utilization, one or two clonal expansion of T cells were identified in some predominant TCR V? subfamilies. Difference of distribution and clonal expansion of TCR V? subfamily T cells were related to source of T cells and the phase during MLTC. Compared with LAK cell, most of T cells from MLTC were CD3+CD8+T cells with higher and more specific cytoxicity to the induced cells, M_ 2a cells, but not HL60 or K562 cell line. CONCLUSION: Clonal expansion of TCR V? subfamily T cells stimulated selectively by M_ 2a cells may be a specific immune response of autologous and allogeneic T cells to M_ 2a cells associated antigen. The T cells induced by M_ 2a cells have the ability of specific cytoxicity to the AML-M_ 2a cells.

AIM: To investigate the role of Cx43 in the myocardialization of the proximal outflow tract(OFT) septum in the mouse heart.METHODS: C57/BL6 mice of ED11.5 to 1 day after birth were used in this study,which included Cx43 knockout homozygotes((Cx43-/-)),heterozygotes((Cx43+/-)) and wildtypes((Cx43+/+)).Pathohistological analysis was used to examine the structure of the hearts.The expression of alpha-sarcomeric actin(?-SCA),active caspase-3 and activator protein-2(AP-2) were detected by immunohistochemistry.RESULTS: Most(Cx43-/-) mice died within 24 h after birth with a swelling and blockage of the conotruncal region,which led to the obstruction of OFT and enlargement of right ventricle.HE staining showed plenty of abnormal tissues in this region forming many pouches.No apparent malformations were observed in(Cx43+/-) and(Cx43+/+) mice.The expression of ?-SCA in the proximal OFT septum was delayed obviously in(Cx43-/-).The apoptotic cells existed in the proximal OFT septum of(Cx43+/+) mostly during ED12.5 to ED15.5.However,there were less apoptotic cells observed in(Cx43+/-),and few in(Cx43-/-).The expression of AP-2,marker of neural crest cells,was increased in (Cx43-/-) and abnormally located in the proximal OFT septum.CONCLUSIONS: Cx43 KO mice are characterized by hyperplasia in conotruncal region,which may be associated with the delayed myocardialization of OFT septum.The decreased apoptosis and the abnormal distribution of cardiac neural crest cells are likely to contribute to the abnormal myocardialization in mice with Cx43 defects.

AIM: To construct recombinant adenovirus containing transcription factor T-bet (T-box expressed in T cells),and induce type-1 T-helper differentiation of lymphocytes. METHODS: T-bet gene was cloned from total RNA of lymphocyte stimulated with IFN-? with RT-PCR methods,then subcloned into transfer vector pAdtrack-CMV in BgIII/SaII sites. The new transfer vector pAdtrack-CMV. T-bet was digested with Pme I,subsequently cotransformed into BJ5183 cells with adenoviral backbone plasmid pAdEasy-1. The resultant plasmid pAd. T-bet was linearized by Pac I and transfected into 293 cells with liposome LIPOFECTAMINE 2000 for producing Ad.T-bet. The recombined adenovirus Ad.T-bet was identified through RT-PCR and Western blotting methods. Lymphocytes purified from patients suffering from liver cancer was infected with liposome and Ad.T-bet with multiplicity of infection (m.o.i) 5000,and the concentration of IFN-? in culture media was evaluated with ELISA methods. RESULTS: T-bet gene was successfully cloned from lymphocytes and incorporated into recombinant adenovirus Ad.T-bet. Lymphocytes infected with Ad. T-bet constantly and strongly secreted Th1 cytokine IFN-?. CONCLUSION: Recombinant adenovirus Ad.T-bet effectively induces type-1 T-helper differentiation,which is a promising method for restoration of patients' immune reaction against cancer.