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Objective:To explore the protective effect of rapamycin and the role of autophagy on podocyte injury induced by pu-romycin amino nucleoside.Methods:The mice podocytes cell 5(MPC5)were randomly divided into four groups:Control group,PAN group(50 μg/ml PAN);rapamycin group(RAP group,100 ng/ml,200 ng/ml,300 ng/ml rapamycin)and PAN+rapamycin group (PAN+RAP group,treated with rapamycin before PAN).The apoptosis of podocytes were detected by Annexin V/PI.Transmission electronic microscopy was used to observe the formation of autophagosome.The expression of autophagic markers,LC3,p62,and 4EBP1,P70S6K,mTOR were detected by Western blot.Results: The podocyte apoptotic rate was significantly decreased when the podocytes were treated with rapamycin compared with PAN group.Further,more autophagosomes appeared in cytoplasm of podocytes in RAP and PAN+RAP group compared with PAN group.And the expression of LC3Ⅱ was significantly increased in rapamycin treated podocytes,whereas the expression of p62 was decreased markedly.Phosphorylation of mTOR,4EBP1 and P70S6K was decreased rapamycin treated groups.Conclusion: Podocyte autophagy was inhibited by PAN, and the apoptosis of podocytes was promoted.Rapamycin protect puromycin amino nucleoside induced podocyte injury by upregulating autophagy and it may be concerned with mTOR 4EBP1 and P70S6K signaling pathway.
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Objective To investigate the prevalence of iron deficiency(ID)and iron deficiency anemia (IDA) in pregnant women in urban areas of China. Methods The study was a national cross-sectional survey conducted from September 19th, 2016 to November 20th, 2016. According to the classification of the National Bureau of Statistics, all survey sites were set up in 6 regions of the country. Pregnant women were continuously selected using multistage stratified sampling. A total of 12 403 pregnant women were collected and examined for serum ferritin and hemoglobin levels. Results The median serum ferritin level during pregnancy was 20.60 μg/L(11.78-36.98 μg/L), the hemoglobin level was(118±12)g/L. With the progress of pregnancy, the levels of serum ferritin and hemoglobin decreased gradually. The median serum ferritin levels in the first, second trimester and third trimester were 54.30 μg/L(34.48-94.01 μg/L), 28.60 μg/L(16.40-50.52 μg/L), and 16.70 μg/L(10.20-27.00 μg/L)respectively(P<0.01). The mean hemoglobin levels were(127 ± 10)g/L,(119 ± 11)g/L and(117 ± 11)g/L respectively(P<0.01). The prevalence of ID in urban pregnant women was 48.16%(5 973/12 403), and IDA prevalence was 13.87% (1 720/12 403). The prevalence of IDA in the first, second trimester and third trimester were 1.96% (20/1 019), 8.40%(293/3 487)and 17.82%(1 407/7 897), respectively(P<0.01). The prevalence of standardized ID and IDA were significantly different in various regions of China(P<0.01). The standardized prevalence of ID were relatively higher in East China and Northeast China, 57.37% and 53.41% respectively, while it was the lowest in Southwest China, 30.51%. The standardized prevalence of IDA in South Central, Northwest, and East China were relatively high, 21.30%, 16.97% and 17.53% respectively, and the standardized prevalence of IDA in Southwest China was the lowest, 5.44%,the differents in various regions were significant(all P<0.01). Conclusion The current phenomenon of ID and IDA in pregnant women is still very common,and nutrition and health care during pregnancy should be strengthened.
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OBJECTIVE: To investigate the changes of platelet mitochondrial adenosine triphosphate synthase 6( ATPase-6)gene in patients with occupational chronic myeloid leukemia( CML) caused by benzene. METHODS: Five occupational benzene induced CML patients were selected as the case group and 21 healthy workers without benzene exposure were selected as the control group,venous blood were drew from these study subjects. The number of platelet was counted. The level of mitochondrial ATPase-6 gene was examined by quantitative polymerase chain reaction. RESULTS: The platelet count in case group was lower than that of control group [( 135. 2 ± 32. 6) ×10~9/ L vs( 239. 7 ± 77. 5) ×10~9/ L,P < 0. 05]. The ratio of ATPase-6 gene / platelet was higher than that of control group( median: 3. 43 ×10~(-4)copies vs 1. 87 ×10~(-4)copies,P < 0. 05). The ATPase-6 gene level in case group showed no significant difference when compared with that of control group( median: 4. 83 ×10~7 copies / L vs 3. 24 ×10~7 copies / L,P > 0. 05). CONCLUSION: The benzene induced CML patients shows more sensitive change in ratio of ATPase-6 gene / platelet than in ATPase-6 gene level.
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To investigate the protective effects of eplerenone on adriamycin-induced renal injury and the possible mechanisms involved,36 male Sprague-Dawley rats were randomly divided into control group,adriamycin nephropathy (AN) group and eplerenone-treated group (100 mg·kg-1·d-1 eplerenone).Blood pressure,24-h urinary protein,serum potassium,sodium and creatinine were measured 28 days after adriamycin injection (a single tail intravenous injection of 6.5 mg/kg adriamycin).The morphological changes of renal tissues were observed by light and electron microscopy.lmmunohistochemistry and Western blotting were performed to examine the expression of TGF-β1 and desmin in renal cortex.The results showed that 28 days after adriamycin injection,there were no significant changes in the level of serum potassium,sodium,creatinine concentrations and blood pressure values in the rats of the three groups.Meanwhile,the 24-h proteinuria excretion in the AN group was significantly higher than that in the control group (P＜0.01),but that in the eplerenone-treated group was substantially reduced when compared with that in the AN group (P＜0.05).Mild mesangial cell proliferation and matrix expansion,diffuse deformation and confluence of foot processes in podocytes were found in the AN group.By contrast,rats in the eplerenone-treated group exhibited obvious attenuation of these morphological lesions.The protein expression of TGF-β1 and desmin in the AN group was markedly up-regulated in contrast to that in the control group (P＜0.01),whereas that in the eplerenone-treated group was much lower than in the AN group (P＜0.05).It was concluded that eplerenone may ameliorate the proteinuria and the development of pathological alteration in adriamycin-induced nephropathy presumably via the inhibition of cytokine release,and restore the morphology of podocytes independent of its blood pressure-lowing effects.
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<p><b>OBJECTIVE</b>To investigate the effects of hypertonic sodium chloride hydroxyethyl starch 40 (HSH) on brain edema and morphological changes during whole body hyperthermia (WBH) in rats.</p><p><b>METHODS</b>Sixty adult male SD rats were randomized into control group, WBH group without fluid infusion (group HT), WBH group with Ringer's infusion (group RL), WBH group with HAES + Ringer's infusion (group HRL) and WBH group with HSH infusion (group HSH). WBH was induced by exposure to 36 degrees celsius; for 3 h to achieve a rectal temperature of 41-42 degrees celsius;, and the corresponding fluids were administered intravenously within 30 min at the beginning of WBH. The control rats were housed at a controlled room temperature (22∓1) degrees celsius; for 4 h. After cooling at room temperature for 1 h, the rats were sacrificed and brain water content and morphological changes were evaluated.</p><p><b>RESULTS</b>Compared with the control group, all the WBH groups had significantly increased brain water content (P<0.05 or 0.01), but group HSH showed a significantly lower brain water content than group HT (P<0.05). The rats in groups HT, RL and HRL showed serious to moderate structural changes of the brain tissue and nerve cells, but HSH group had only mild pathologies.</p><p><b>CONCLUSION</b>HSH can reduce brain edema and ameliorate the damages to brain cells in rats exposed to WBH.</p>
Subject(s)
Animals , Male , Rats , Brain , Pathology , Brain Edema , Pathology , Hydroxyethyl Starch Derivatives , Therapeutic Uses , Hyperthermia, Induced , Rats, Sprague-Dawley , Saline Solution, Hypertonic , Therapeutic UsesABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of radiosensitivity and X-ray dose on the expression of miR-7 in nasopharyngeal carcinoma (NPC) cells.</p><p><b>METHODS</b>Low radiosensitive NPC cells CNE-1 and high radiosensitive NPC cells CNE-2 were exposed to 0, 2 and 8 Gy X-ray. The total RNAs of the cell lines were extracted 10 h after radiation for reverse transcription of miR-7 and 18S rRNA by stem-loop primer and random hexamers, respectively. The non-irradiated CNE-1 cells served as the control sample and the relative quantity of the expression level was calculated after real-time PCR using SyBR green.</p><p><b>RESULTS</b>miR-7 expression differed significantly between CNE-1 and CNE-2 cells (4.49-/+3.62 vs 1.29-/+1.10, F=135.483, P<0.001). The radiation dose also significantly affected the expression of miR-7 in NPC cells (F=39.565, P<0.001). CNE-1 cells with a 2 Gy exposure had the highest expression level of miR-7, while the non-irradiated CNE-1 cells had the lowest expression. CNE-2 cells exposed to 2 Gy X-ray had the lowest expression level of miR-7 and the non-irradiated CNE-2 cells had the highest.</p><p><b>CONCLUSION</b>Radiosensitivity and radiation dose of X-ray have significant effect on the expression of miR-7 in NPC cells, indicating that miR-7 plays an important role in radioresistance of NPC cells to X-ray, and suppressed miR-7 expression may elevate the radiosensitivity of NPC cells.</p>
Subject(s)
Humans , Apoptosis , Radiation Effects , Carcinoma , Cell Line, Tumor , Dose-Response Relationship, Radiation , Gene Expression Regulation, Neoplastic , Radiation Effects , MicroRNAs , Genetics , Nasopharyngeal Neoplasms , Genetics , Radiation Tolerance , Genetics , X-RaysABSTRACT
poptosis induced by PAN.
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To investigate the effects of albumin on the production of matrix metalloproteinases-2 (MMP-2) and matrix metalioproteinases-9 (MMP-9) in podocytes. Podocytes were treated with bovine serum albumin (BSA) at the concentration of 0.1,0.5,1,2 g/L,respectively. Conditioned media were harvested 12,24,48 and 72 h after the treatment. The expression of MMP-2 and MMP-9 was assayed by gelatin zymography,RT-PCR and Western blotting analysis. Our results showed that in comparison with the control group,BSA increased the expression of MMP-2 and MMP-9 mRNA and protein in a dose-and time-dependent manner (P＜0.05). Meanwhile,the enzymatic activities of MMP-2 and MMP-9 in the culture supernatants of podocytes were also increased (P＜0.05). It is concluded that albumin up-regulated the expression of MMP-2 and MMP-9 at gene and protein levels in a time-and dose-dependent manner.
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This study examined the effect of sulodexide on podocyte injury in rats with adriamycin nephropathy (AN). A total of 36 healthy male SD rats were randomly assigned to three groups:control group,AN group and sulodexide treatment group. Rat models of AN were established by a single tail intravenous injection of adriamycin (6.5 mg/kg) in both AN group and sulodexide treatment group. Sulodexide (10 mg/kg) was administered the rats in the treatment group once daily by garage from the fast day of model establishment until the 14th day or the 28th day. Samples of 24-h urine and renal cortex tissues were harvested at day 14,28 after the model establishment. Excretion of 24-h urinary protein was measured by Coomassie brilliant blue method. The pathological changes in renal tissues were observed by light microscopy and electron microscopy respectively. Heparanase mRNA was detected by RT-PCR. Expressions of desmin,CD2AP and heparanase were determined by immunohistological staining. The results showed that the expressions of heparanase mRNA and proteinwere increased in the glomeruli of AN rats at day 14 and 28 after the model establishment,which was accompanied by the increased expression of desmin and CD2AP. The mRNA and protein expression of heparanase was decreased in the sulodexide-treated rats as compared with AN rats at day 14 and 28. And,the protein expression of desmin and CD2AP was reduced as with heparanase in the sulodexide-treated rats. Proteinuria and podocyte foot process effacement were alleviated in the AN rats after sulodexide treatment. There was a positive correlation between the expression of heparanase and the expression of desmin and CD2AP (as well as 24-h urinary protein excretion). It was concluded that increased heparanase is involved in podocyte injury. Sulodexide can maintain and restore podocyte morphology by inhibiting the expression of heparanase in AN.