BACKGROUND:Stem cell transplantation provides a new approach to treat chronic renal disease.Specific marking and in vivo tracing of stern cells are the basis of studies in this field.However,the marking methods appropriate for all cells remain uncertain.OBJECTIVE:To observe the in vivo location and differentiation of 4',6-diamidino-2-phenylindole (DAPI) and green fluorescence protein (GFP)-Iabeled cells in adriamycin nephrosis rats so as to explore an efficient labeling and tracing method for metanephric mesenchymal cells (MMCs) derived from embryonic rats.DESIGN,TIME AND SETTING:Grouping comparative observation was performed at the Second Xiangya Hospital of Central South University from April to December 2007.MATERIALS:A total of 60 female SD rats,weighing 180-220 g,of dean grade,were used to establish models of adriamycin nephrosis.METHODS:DAPI and MMCs infected with GFP and DAPI were respectively injected into addamycin nephrosis via the tail vein.DAPI and GFP distribution in the frozen sections was detected at 1,3,and 5 weeks,postoperatively.In addition,GFP expression in renal tissues was detected by ABC immunoenzymatic staining method.MAIN OUTCOME MEASURES:DAPI and GFP-labeled Cell grafts in adriamycin nephrosis rats were compared.The changes of GFP-transfected MMCs at different time points were observed.RESULTS:DAPI positive cells were observed in tubular structures after 1 weeks of injection of DAPI-labeled cells and DAPI alone,and remained existing at 5 weeks,but the florescence was reduced with time.GFP-transfected MMCs were able to survive and integrate into tubular structures after 1 week,and remained existing at 5 weeks.Moreover,the fluorescence was not reduced.ABC immunoanzymatic staining showed that only a few GFP-positive MMCs appeared in glomerular tufts,and mainly distributed in cytoplasm.Semi-quantitative evaluation of GFP show that the positive cell rate in rats with early application was greater than that with advanced application,and the positive rate was increased with time.CONCLUSION:Liposome mediated GFP gene transfer was an efficient labeling in vitro and suitable tracing method for cell differentiation experiment in vivo,suitable for short-term tracing and observation of transplanted cells.

Objecfive To detect the functional repair of metanephric mesenchymal cells (MMCs) transplantation in adriamycin (ADR)-induced glomerulopathy rats. Methods A total of 90 Sprague-Dawley female rats were randomly divided into three groups:ADR group (n=40,rats were injected via the tail vein with O.25 mg ADR/100 g body weight on days 1 and 21),ADR- MMCs group(n=40,rats were injected via the tail vein with 5×10~6-7×10~6 MMCs 8 weeks after the second ADR administration),control(n=10).All the rats were scarified 8 weeks after MMCsinjection.Pathology and collagen IV expression in renal tissue were examined.Moreover,matrix metalloproteinases 2 (MMP-2) and matrix metallopmteinases 9 (MMP-9) expression in the renal tissue were also detected with immunohistochemistry,and quantity analysis of protein and gene was further demonstrated with Westem blot and RT-PCR analysis,respectively. Results There were no significant differences in tubulointerstitial injury score and glomerulosclerosis degree between ADR group and ADR-MMCs group(P>0.05).Compared with ADR group,collagen Ⅳ and MMP-2 expression decreased, MMP-9 expression incrased in renal tissue of ADR-MMCs group, and the difference was significant (P<0.05). Conclusion MMCs transplantation may have potentially therapeutic effect on renal tissue fibrosis of adriamyein-induced glomerulopathy in rats, and the signaling pathways of MMPs appear to be involved in these processes.

OBJECTIVETo explore the efficacy of inductible nitric oxide synthase (iNOS) inhibitor 1400W in vivo in blocking the death pathway of lipopolysaccharide (LPS)-induced activated-microglia to preoligodendrocytes (preOLs) in neonatal rats with infective-type periventricular leukomalacia (PVL) induced by LPS.

METHODSTwo-day-old neonatal rats were randomly divided into: a sham-operated group, an untreated PVL group, and four 1400W-treated PVL groups that were subcutaneously administrated with 20 mg/kg of 1400W at 0 h, 8 hrs, 16 hrs, and 24 hrs after LPS induction, respectively. The brain specimens were obtained 5 days after LPS induction. The pathological assessment of cerebral white matter was performed under a light microscope. Concentrations of nitric oxide (NO) were measured by nitric acid-deoxidize colorimetry. Synthesis of iNOS was determined by Western blot analysis. Peroxynitrite (ONOO(-)) level and the amount of preOLs were determined by immunocytochemistry. RETHODS: The obvious injuries of periventricular white matter, massive loss of positive O4-labelled preOLs, and increased levels of NO, ONOO(-) and iNOS were observed in neonatal rats with PVL. Compared to the untreated PVL group, the use of 1400W at 0 h, 8 hrs and 16 hrs after LPS induction significantly improved white matter injuries, reduced the levels of NO, ONOO(-) and iNOS, and increased the amount of O4-labelled preOLs. However, the use of 1400W at 24 hrs after LPS induction did not result in the improvements.

CONCLUSIONSiNOS inhibitor 1400W can effectively block the toxicity of LPS-activated microglia to preOLs and protect cerebral white matter through inhibiting iNOS and reducing the production of NO and ONOO(-). The use of 1400W within 16 hrs after LPS induction may provide cerebral protections in neonatal rats with PVL.

Amidines ; pharmacology ; Animals ; Apoptosis ; drug effects ; Benzylamines ; pharmacology ; Brain ; drug effects ; pathology ; Enzyme Inhibitors ; pharmacology ; Lipopolysaccharides ; toxicity ; Microglia ; cytology ; drug effects ; Nitric Oxide ; biosynthesis ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; Oligodendroglia ; cytology ; Peroxynitrous Acid ; biosynthesis ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology

OBJECTIVETo explore the toxicity of LPS-induced activated microglia to preoligodendrocytes (preOLs) and the effect of 1400W, a selective inhibitor of inducible nitric oxide synthetase (iNOS), on the blockage of the toxicity.

METHODSCo-cultured microglia and preOLs obtained from two-day-old Sprague-Dawley (SD) rats were divided into three groups: co-culture control group, co-culture LPS group and co-culture LPS plus 1400W group. After cultured cells were induced by LPS (100 ng/ml) for 48 hours, the concentration of nitric oxide (NO) was measured by nitric acid-oeoxidize-colorimetry, the level of peroxynitrite (ONOO(-)) was determined by immunocytochemistry, and the synthetic level of iNOS was detected by Western blotting, respectively. The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously. Data were analyzed with SPSS 11.0 software.

RESULTSCompared to co-culture control group, there was significant increase in levels of NO [(82.27+/-3.41) micromol/L vs. (167.86+/-9.87) micromol/L, t=8.593, P<0.01], ONOO(-)[(6.14+/-1.27) x 10(7)/L vs. (34.38+/-7.75) x 10(7)/L, t=5.892, P<0.01], and iNOS [(0.18+/-0.027) vs. (0.79+/-0.068), t=9.26, P<0.01] induced by LPS in co-culture LPS group, and with a higher apoptotic rate of preOLs [(6.73+/-1.39)% vs. (24.77+/-2.05)%, t=12.619, P<0.01]. However, all levels of NO [(69.55+/-5.07) micromol/L, t=8.896, P<0.01], ONOO(-) [(10.33+/-3.47) x 10(7)/L, t=14.96, P<0.01] and iNOS (0.35+/-0.042, t=5.506, P<0.01) decreased significantly with the use of 1400W at a dose of 10 micromol/L in co-culture LPS plus 1400W group, and the apoptotic rate of preOLs [(11.8+/-2.06)%, t=7.715, P<0.01] was also reduced evidently.

CONCLUSIONSNO, ONOO(-) and iNOS, etc. play important roles in the death pathway of preOLs induced by LPS. 1400W can block effectively the toxicity of LPS-activated microglia toxicity to preOLs through inhibiting iNOS selectively and reducing the production of NO and ONOO(-), and improve the survival rate of preOLs.

Amidines ; pharmacology ; Animals ; Benzylamines ; pharmacology ; Cells, Cultured ; Lipopolysaccharides ; toxicity ; Microglia ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; antagonists & inhibitors ; metabolism ; Oligodendroglia ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo find an inhibitor to reduce the volatilization of formalin.

METHODThe saturated solution of sodium hydrosulphite (SHS) was sprayed on the surface of the anatomy specimens, then the concentration of formaldehyde in the air was tested.

RESULTSThe concentration of formaldehyde in the air of SHS sprayed group [(3.10 +/- 1.22) mg/m3] was significantly lower than that of the control group [(8.36 +/- 4.11) mg/m3, P < 0.01].

CONCLUSIONSHS may be a volatilization inhibitor for formalin, which could reduce the concentration of formaldehyde in the air.

Air Pollution, Indoor ; prevention & control ; Anatomy ; Formaldehyde ; analysis ; chemistry ; Sulfites ; chemistry ; Volatilization

OBJECTIVETo evaluate the effects of glial cell line-derived neurotrophic factor (GDNF) and memantine on the long-term prognosis in neonatal rats with ischemia-induced periventricular leukomalacia (PVL).

METHODSThirty-two 5-day-old neonatal rats were randomly divided into 4 groups: sham-operated, PVL, GDNF-treated and memantine-treated. PVL was induced by right carotid artery ligation and hypoxia in the PVL, GDNF-treated and memantine-treated groups. GDNF (100 μg/kg) or memantine (20 mg/kg) was injected in the two treatment groups immediately after PVL inducement. The weight of the rats was measured immediately before and after hypoxia ischemia (HI). Both of Morris water maze test and Rivlin inclined plane test were performed at 26 days old (21 days after HI). The values of the escape latency (EL) and swimming distance, and the maximum inclined plane degree which the rats could stand at least 5 seconds were compared among the four groups.

RESULTSThe lower weight, the prolonged mean values of EL and swimming distance and the reduced maximum inclined plane degree were observed in the PVL group compared to those in the sham-operated, GDNF-treated and memantine-treated groups. There were no significant differences in the weight, the values of EI and swimming distance and the maximum inclined plane degree between the two treatment groups and the sham-operated group.

CONCLUSIONSThe administration of either GDNF or memantine can markedly increase the abilities of spatial discrimination,learning and memory, and motor coordination, promote weight gain, and improve long-term prognosis in rats with PVL.

Animals ; Animals, Newborn ; Body Weight ; Excitatory Amino Acid Antagonists ; therapeutic use ; Glial Cell Line-Derived Neurotrophic Factor ; therapeutic use ; Humans ; Infant, Newborn ; Leukomalacia, Periventricular ; drug therapy ; psychology ; Maze Learning ; drug effects ; Memantine ; therapeutic use ; Motor Activity ; drug effects ; Rats

OBJECTIVETo establish a reliable neonatal rat model of periventricular leukomalacia (PVL) which is expected to be similar to PVL of human preterm infants pathologically, and to explore the concomitant eye lesions in the PVL model.

METHODSTwo-old-day neonatal rats were randomly divided into a PVL group and a sham-operated group (n=19 each). The PVL model was established by the ligation of bilateral common carotid arteries, followed by a 30-min exposure to 8% oxygen. The cerebral infarction area was assessed with TTC staining 1 day after operation. Cerebral pathology was examined under a light micsrocope 2 and 21 days after operation. The examinations of eyes under a slip lamp and the pathology of eyeballs under a light microscope were performed 21 days after operation.

RESULTSThe TTC staining cerebral slices showed there were extensive white areas of infarction in the brain of the PVL group, with an infarction area of 53.45 +/- 33.90 mm3 and a percentage of infarction of (24.98 +/- 15.44)% . Significant cystic necrosis and apoptosis around the periventricular and subcortical white matter and mild damage in cortical neurons were observed in the PVL group 2 days after operation. The more obvious cystic necrosis around the periventricular area was found in the PVL group 21 days after operation. There were no pathological changes in the brain of the sham-operated group. All of rats in the PVL group had bilateral cataracts, however, no pathological changes were observed in their postbulbar tissues. The sham-operated group did not show eye abnormal.

CONCLUSIONSThe PVL animal model that was similar to PVL of human preterm infants pathologically was successfully established by the ligation of bilateral common carotid arteries, followed by 30-min hypoxia exposure, with a positive effect and a good repeatability. Cataract can also be induced by the method.

Animals ; Animals, Newborn ; Brain ; pathology ; Cataract ; etiology ; pathology ; Disease Models, Animal ; Female ; Humans ; Hypoxia-Ischemia, Brain ; complications ; Infant, Newborn ; Leukomalacia, Periventricular ; etiology ; pathology ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the physiopathologic basis of Weikangfu Granule (WKFG) in treating precancerosis of gastric mucosa in patients of chronic gastritis with Pi-deficiency syndrome (CG-PDS).

METHODSOne hundred and fifteen patients of CG-PDS who suffered from intestinal metaplasia (IM) and atypical hyperplasia (ATHP) of gastric mucosa, were divided into two groups. The treated group (n = 61) was treated by WKFG with its ingredients modified according to the syndrome type of patients. The control group (n = 54) was treated with Weishu granule. The histopathological and subcellular ultrastructural changes were detected by optical microscope, screening electronic microscope, transmission electronic microscope and histochemical staining; the nuclear and mitochondrial ultrastructure of gastric mucosa were analyzed with energy dispersion X-ray analyser and image analysis system. And the changes of cAMP, lipid peroxide (LPO), superoxide dismutase (SOD) before and after treatment in the treated group were measured and compared with those of the health control group consisting of 15 volunteers.

RESULTSThe symptomatic and pathological therapeutic effect in the treated group were significantly superior to those in the control group (P < 0.05). The contents of Zn, Cu, cAMP, SOD and (3)H-TdR LCT in gastric mucosa of the treated group before treatment were all lower than those of the healthy control group, yet all these indexes markedly increased after treatment, while serum LPO level, which increased before treatment was lowered after treatment. All the changes showed statistical significance (P < 0.05 or P < 0.01).

CONCLUSIONWKFG can reverse IM and ATHP in patients of CG-PDS, and the effect may be realized by way of increasing the level of Zn, Cu, cAMP and SOD in gastric mucosa, promoting cell differentiation, enhancing cellular immunity and reducing oxygen free radicals and lipid peroxidation.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Chronic Disease ; Copper ; analysis ; Cyclic AMP ; analysis ; Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; chemistry ; pathology ; ultrastructure ; Gastritis, Atrophic ; pathology ; Humans ; Lipid Peroxides ; analysis ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Precancerous Conditions ; pathology ; Stomach Neoplasms ; pathology ; Superoxide Dismutase ; analysis ; Syndrome ; Yang Deficiency ; complications ; Zinc ; analysis

OBJECTIVETo explore the association of CCR5δ32, CCR2-64I and SDFl-3 A gene polymorphisms with HIV-1-infection in Chinese population.

METHODSA meta-analysis was performed to identify case-control studies of CCR5δ32, CCR2-64I and SDFl-3 A polymorphisms from the literatures.

RESULTSFourteen studies of CCR5δ32 were found, involving a total of 1607 cases and 1632 controls. Compared with the wild-type homozygote wt/wt, the pooled odds ratios (95%CI) of wt/mt, mt/mt, and wt/mt+mt/mt genotypes of CCR5δ32 gene polymorphisms were 1.156 (0.808, 1.654), 0.997 (0.198, 5.022), and 1.149 (0.808, 1.634), respectively. Twelve studies of CCR2-64I were identified, including 1415 cases and 1239 controls. Compared with the wild-type homozygote wt/wt, the pooled odds ratios (95%CI) of wt/mt, mt/mt, and wt/mt+mt/mt genotypes of CCR2-64I gene polymorphisms were 1.005 (0.844, 1.197), 1.191 (0.808, 1.754), and 1.028 (0.870, 1.214), respectively. Ten studies of SDFl-3 A were found, involving 1179 cases and 1003 controls. Compared with the wild-type homozygote wt/wt, the pooled odds ratios (95%CI) of wt/mt, mt/mt, and wt/mt + mt/mt genotypes of SDF1-3 A gene polymorphisms were 1.010 (0.830, 1.228), 1.188 (0.860, 1.643), and 1.038 (0.861, 1.250).

CONCLUSIONCCR5δ32, CCR2-64I and SDFl-3 A gene polymorphisms do not show strong correlations to HIV-1-infection in Chinese population. These 3 genes may not have protective effect against HIV-1 infection in Chinese population, suggesting the susceptibility of Chinese population to the infection.

Alleles ; Asian Continental Ancestry Group ; genetics ; Chemokine CXCL12 ; genetics ; Gene Frequency ; Genotype ; HIV Infections ; genetics ; HIV-1 ; Humans ; Polymorphism, Genetic ; Receptors, CCR2 ; genetics ; Receptors, CCR5 ; genetics