OBJECTIVEGlutamine has protective effects against renal injuries. This study was designed to explore the possible mechanism underlying the protections by examining the effects of glutamine on extracellular signal regulated kinase (ERK) and p38MAPK expression in the kidney in rats with endotoxemia.

METHODSOne hundred and twenty-one 18-day-old Wistar rats were randomly injected with LPS (4 mg/kg; n=55), LPS (4 mg/kg)+glutamine (1 mL/kg) (n=55), or normal saline (control group; n=11). The two LPS groups were subdivided into five groups sacrificed at 2, 4, 6, 24 and 72 hrs after administration (n=11 each). ERK-2 and p38MAPK mRNA and protein expression in the kidney were measured by RT-PCR and immunohistochemistry.

RESULTSCompared to the control group, the mRNA and protein expression of both ERK-2 and p38MAPK in the LPS group significantly increased 2, 4, 6, 24 and 72 hrs after administration (P<0.01), and reached a peak at 6 hrs (P<0.01). In the LPS+glutamine group, the trend of ERK-2 and p38MAPK expression was similar to the LPS group but their expression levels were significantly lower than those in the LPS group at all time points (P<0.05 or 0.01).

CONCLUSIONSBoth ERK-2 and p38MAPK expression increased in young rats with LPS-induced endotoxemia. Glutamine alleviates renal injuries possibly by decreasing the expression of both.

Animals ; Endotoxemia ; enzymology ; pathology ; Extracellular Signal-Regulated MAP Kinases ; analysis ; genetics ; Female ; Glutamine ; pharmacology ; Immunohistochemistry ; Kidney ; enzymology ; Lipopolysaccharides ; toxicity ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; p38 Mitogen-Activated Protein Kinases ; analysis ; genetics

Objective To analyze the epidemic tendency of Microtus fascus plague during 2000-2008 in Sichuan province. Methods To investigate the plague each year according to "overall Plan of the Plague in the Whole Nation" and "Surveillance Program of Sichuan Province Plague". Results There were plague epidemic from 2000 to 2008, with the average density as 312.41/ha. 42.57% of the Microtus fuscus were infected by body Fleas. The Fleas Index was 0.88 and the Index for nest Fleas of Microtas fuscus was 55.89. Six kinds of animals were infected by not only the Microtus fuscus but also herd-dog, sand fox, Tibetan sheep, domestic cats and Cricetulus longicaulatus as well. The positive rate of live Microtus fuscus was 0.32% but 22.99% in the dead Microtus fuscus. The overall positive rate on serological test was 6.70%. There were 4 Sections, 11 species and 19 kinds Fleas identified and carrying 3 kinds of fleas, Callopsylla sparsilis, Amphipsylla tntua tutua and Rhadinopsylla dahurica vicina, with the overall infection rate as 0.054%. Conclusion Plague among Microtus fuscus showed a continuous epidemic in Sichuan province during 2000-2008.

Objective To analyzed the variant information on the indices regarding fleas from natural foci of Microtus plague in Sichuan epidemic area during 2000 to 2008.Methods Statistical and analytical methods were used on the surveillance data regarding Microtus fuscus plague.Results There were 19 flea species identified and the share of Callopsylla sparsilis was 62.79 percent while the share of Amphipsylla tuta tuta was 30.90 percent on Microtus fuscus plague.The infection rate of fleas and the flea index were the highest in October and the lowest in December and March.Species as Callopsylla sparsilis,Amphipsylla tuta tuta and Rhadinopsylla dahurica vicina could naturally infect the Yersinia pestis.Conclusion Microtus fuscus plague could become epidemic when animals and flea species were infected.We should emphasis on plague monitoring program so as to prevent the occurrence of the disease.

OBJECTIVETo construct a recombinant human CD59 gene containing intercellular adhesion molecule-2 promoter for high level endothelial-specific expression in xenotransplantation.

METHODSICAM-2 promotor fragment and CD59-intron 1 fragment were produced by PCR from the human blood genome, and then clone these fragments into a pcDNA3-CD59 eukaryotic expression vector which was followed by digestion with the specific restricted endonuclease (for example: EcoRI, Hind III). The ICAM-2 promoter and CD59-intron 1 fragments were identified by PCR, and sequencing. The recombinant was then transfected into pig aorta endothelial cells with Lipofection, and the expression was measured by flow cytometer.

RESULTSProducts of the sequences measured were in accord with the frames of the gene bank. The expression of the protein of this recombinant was positive.

CONCLUSIONThe CD59 recombinant gene is constructed successfully, providing a basis for transgenic research.

Animals ; Antigens, CD ; genetics ; CD59 Antigens ; biosynthesis ; genetics ; Cell Adhesion Molecules ; genetics ; Cloning, Molecular ; Endothelium, Vascular ; cytology ; metabolism ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation ; Humans ; Promoter Regions, Genetic ; Recombinant Proteins ; biosynthesis ; genetics ; Swine ; Transfection ; Transplantation, Heterologous

The aim of the study is to evaluate the effects of the single and mixed decoction of Thallus laminariae (kelp) and Glycyrrhiza glabra (licorice) on the metabolism and their difference. The mixed decoction of kelp and licorice and the single decoction were made and intragastrically administered to the SD rats. The effect on system metabolism, the toxicity of liver and kidney were assessed by GC-MS profiling of the endogenous molecules in serum, routine biochemical assays and histographic inspection of tissues from SD rats, separately. The mixed decoction of kelp and licorice induced more obvious pathological abnormalities in SD rats than a single decoction of kelp, while the extracts of licorice did not show any pathological change. Neither the mixed, nor the single decoction showed abnormal histopathology. After intragastric administration of extracts for 5 days, the mixed decoction induced a decrease of ALT (no significant change in the groups of single decoction) and an increase of BUN (so did the single decoction of kelp). Metabolomic profile of the molecules in serum revealed that the metabolic patterns were all obviously affected for the three groups, i.e., the mixed and single decoction of kelp and licorice. The rats given with the single decoction of kelp showed a similar pattern to that of the mixed decoction, indicating that the kelp primarily contributed the perturbation of metabolism for the mixed decoction. All three groups induced a decrease of branched chain amino acids, TCA cycle intermediates and glycolysis intermediates (e.g., pyruvic acid and lactic acid) and an increase of 3-hydroxybutyric acid. Kelp decoction showed stronger potential in reducing TCA cycle intermediates and glycolysis intermediates than the other two groups, while the levels of branched chain amino acids were the lowest after licorice extracts were given. These results suggested that the effect of the mixed decoction on metabolism was closely associated with both kelp and licorice. The continuous administration of single decoction of kelp and the mixed decoction of licorice and kelp resulted in pathological abnormalities in kidney of SD rats. The mixed decoction of kelp and licorice distinctly perturbed sera molecules and hence system metabolism, which showed associated with those of kelp and licorice. Although the metabolic effect was associated with both kelp and licorice, the results suggested kelp contributed to it primarily.
Animals ; Glycyrrhiza ; chemistry ; Kelp ; chemistry ; Kidney ; drug effects ; Liver ; drug effects ; Metabolomics ; Plant Preparations ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo evaluate the anticancer effects of exogenous human WT-PTEN overexpression on bladder transitional carcinoma cell line EJ.

METHODSThe plasmid containing WT-PTEN or mutant PTEN was separately transfected into bladder transitional carcinoma cell line EJ, and the protein expression of PTEN in the EJ cells was detected by Western blot. Cell morphological changes were observed under the inverted microscope and transmission electron microscope. MTT test was used to assess the effect of PTEN on proliferation and anticancer effects for mitomycin and theraubicin. The change of bcl-2 expression in the cells was measured by Western blot. The empty plasmid was used as control.

RESULTSWestern blot analysis showed that EJ cells expressed high level of PTEN protein after transfection with WT-PTEN or mutant PTEN plasmid. Abnormal morphological changes of the cells were observed in WT-PTEN transfected groups. The growth of EJ cells treated with WT-PTEN was significantly inhibited by 40.1% and anticancer effects were enhanced by mitomycin and theraubicin, but the cells transfected with mutant PTEN plasmid did not show such similar biological behavior.

CONCLUSIONWT-PTEN gene transfection can suppress the in vitro growth and induce apoptosis of bladder transitional carcinoma cell line EJ cells. Mutant PTEN does not show similar biological behavior. Overexpression of WT-PTEN inhibits cancer cell proliferation by down-regulating bcl-2 expression in the cells.

Antibiotics, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Blotting, Western ; Carcinoma, Transitional Cell ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Doxorubicin ; analogs & derivatives ; pharmacology ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Electron, Transmission ; Mitomycin ; pharmacology ; Mutation ; PTEN Phosphohydrolase ; genetics ; metabolism ; physiology ; Plasmids ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism ; physiology ; Transfection ; Urinary Bladder Neoplasms ; genetics ; metabolism ; pathology

This study was aimed to investigate the quantity and subtypes of dendritic cells (DC) in patients with immune related pancytopenia (IRP) and to explore the role of DC in pathogenesis of IRP. The quantity of plasmacytoid dendritic cells (pDC, Lin(-)HLA-DR(+) CD123(+) cells) and myeloid dendritic cells (mDC, Lin(-)HLA-DR(+) CD11c(+)cells) in peripheral blood of 65 patients with IRP (37 new diagnosed and 28 remitted) and 17 healthy controls were analyzed by flow cytometry. The results indicated that the ratio of pDC in peripheral blood mononuclear cells (PBMNC) was (0.91 ± 064)% in new diagnosed group, which was significantly higher than that in remission group (0.39 ± 0.11)% and control group (0.29 ± 0.13)% (P < 0.01), while this ratio of pDC in remission group was higher than that in control group (P < 0.05). The ratio of mDC in PBMNC was (0.21 ± 0.20)% in new diagnosed group and (0.34 ± 0.21)% in remission group respectively, there was no statistical difference as compared with control group (0.29 ± 0.09)% (P > 0.05). The ratio of pDC to mDC in new diagnosed group was 6.75 ± 7.11, which was significantly higher than that in remission group (1.55 ± 0.93) and control group (1.07 ± 0.43, P < 0.01), there was no statistical difference between the ratio of remission group and control group (P > 0.05). The ratio of pDC in PBMNC of IRP group negatively correlated to ratio of Th1/Th2 (r = -0.347, P < 0.05), and positively correlated to the ratio of auto-antibody on membrane of BMMNC (r = 0.606, P < 0.05) and to the quantity of CD5(+)B cells (r = 0.709, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.381, P < 0.01) and platelets (r = -0.343, P < 0.01). The ratio of mDC in PBMNC positively correlated to the ratio of Th1/Th2 (r = 0.595, P < 0.05) and the level of hemoglobin (r = 0.292, P < 0.05). The ratio of pDC/mDC negatively correlated to ratio of Th1/Th2 (r = -0.395, P < 0.05), it positively correlated to the level of antibody on membrane of BMMNC (r = 0.421, P < 0.05) and the quantity of CD5(+)B cells (r = 0.423, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.304, P < 0.05) and platelets (r = -0.287, P < 0.05). It is concluded that the quantity of pDC in peripheral blood of IRP patients increases, which may be related to the immunopathogenesis of IRP.
Adolescent ; Adult ; Blood Cell Count ; Case-Control Studies ; Child ; Child, Preschool ; Dendritic Cells ; cytology ; immunology ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; blood ; immunology ; Young Adult

OBJECTIVETo investigate the function of dendritic cells (DC) of patients with immune related pancytopenia (IRP) and explore the role of DC in IRP.

METHODSThe expression of CD80 and CD86 on myeloid DC (mDC, Lin-HLA-DR(+) CD11c(+) cells) and plasmacytoid DC (pDC, Lin-HLA-DR(+) CD123(+) cells) of 65 IRP (37 untreated and 28 remitted) patients and 17 healthy controls were analyzed by flow cytometry.

RESULTSThe expression of CD86 on pDC was (82.47 ± 13.17)% in untreated group and (60.08 ± 14.29)% in remission group, which were significantly higher than that of controls (47.95 ± 18.59)% (P < 0.05), while the expression in untreated group was higher than that of remission group (P < 0.05). The expression of CD80 on pDC was (6.31 ± 4.49)% in untreated group, which was significantly higher than that of remitted patients (3.09 ± 2.93)% and controls (2.33 ± 2.25)% (P < 0.05). The expression of CD86 on mDC was (97.06 ± 4.82)% in untreated group and (91.35 ± 12.20)% in control group, while the expression in untreated group was higher than that of control group (P < 0.05). The expression of CD80 on mDC was (6.20 ± 5.44)% in untreated group and (3.97 ± 3.24)% in remission group, which were significantly higher than that of controls (1.86 ± 1.73)% (P < 0.05). The expression of CD86 on pDC was negatively correlated to Th1/Th2 (r = -0.733, P < 0.05), it was positively correlated to the antibody on membrane of BMMNC (r = 0.283, P < 0.05) and the quantity of CD5(+)B cells (r = 0.436, P < 0.05), while it was negatively correlated to the level of hemoglobin, platelets and white blood cells (r = -0.539, P < 0.05; r = -0.519, P < 0.05; r = -0.567, P < 0.05, respectively). The expression of CD80 on pDC was negatively correlated to the level of hemoglobin and platelets (r = -0.431, P < 0.05; r = -0.464, P < 0.05).

CONCLUSIONThe function of pDC in PB of IRP were strengthened, which was relevant to the immunopathogenesis of IRP.

Adolescent ; Adult ; Autoimmune Diseases ; complications ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Case-Control Studies ; Child ; Child, Preschool ; Dendritic Cells ; metabolism ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Pancytopenia ; blood ; etiology ; pathology ; Young Adult

BACKGROUND: We previously found that the intestinal epithelial chemokine (C-C motif) ligand 7 (CCL7) plays an important role in the development of toxin-induced acute liver damage. The detailed effects of intestinal epithelial CCL7 on chronic diseases; however, are still unclear. Here, we aimed to investigate the impact of intestinal epithelial CCL7 overexpression on high-fat diet (HFD)-induced obesity and steatohepatitis in mice. METHODS: Intestinal epithelial CCL7 overexpression (CCL7) mice and their wild-type (WT) littermates were fed with normal chow or HFD for 16 weeks to induce obesity and non-alcoholic fatty liver disease. Body weight gain, as well as adipose tissue index were assessed. Liver injury was monitored by histological analysis and real time polymerase chain reaction. Gut microbial composition was analyzed by 16S rRNA gene sequencing. RESULTS: We found that the CCL7 mice on a HFD had markedly decreased weight gain (8.9 vs. 17.0 g, P < 0.05) and a lower adipose tissue index that include mesenteric fat (1.0% vs. 1.76%, P < 0.05), gonadal fat (2.1% vs. 6.1%, P < 0.05), subcutaneous fat (1.0% vs. 2.8%, P < 0.05) compared to WT animals. HFD-induced glucose intolerance and insulin resistance were also significantly improved in CCL7 mice compared to WT. Furthermore, HFD-fed CCL7 mice displayed less hepatic lipid accumulation and lower expression of inflammatory factors than WT mice. 16S rRNA gene sequencing demonstrated that CCL7 overexpression in intestinal epithelial cells improved HFD-induced gut microbial dysbiosis. CONCLUSIONS Our study revealed that CCL7 overexpression in the intestinal epithelium protects mice against the progression of diet-induced obesity, hepatic steatosis, and enteric dysbiosis.