Clinical and immunological features of lupus nephritis in children: retrospective analysis of 40 cases / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To analyze the clinical and immunological features of children with lupus nephritis (LN).</p><p><b>METHODS</b>Chart records of 40 (4 male and 36 female) LN children who were admitted consecutively between January, 2005 and December, 2010 were reviewed. The baseline demographic, pathological and immunological data were analyzed.</p><p><b>RESULTS</b>In the 40 LN patients analyzed, the mean age of the disease onset was 10.6 ± 2.6 (range from 2.6 to 14.3) years, and 35 cases (88%) were school-age children. Proteinuria was detected in all 40 cases, including nephrotic-range proteinuria in 12 (30%) cases, and isolated proteinuria in 9 (22%) cases. Twenty-six (65%) patients had varying degrees of hematuria. Acute nephritis was the most common sub-type, accounting for 47% of the total cases. Among the 39 cases undergoing renal biopsy, 3 were unclassified and the remaining 36 were classified, respectively, as type IV LN (50%, 18 cases), type II LN (22%, 8 cases). In the histopathologcally classified case, 100% were antinuclear antibody-positive, 61% were anti-dsDNA-positive, and 89% showed varying degrees of decrease in serum C3 and C4 concentrations. Following treatment for 6 months, a high LN remission rate (95%) was achieved; the acute renal activity index remained higher in IV, V+III and V+IV subtypes than in other subtypes, while the chronic index and the degree of tubulointerstitial damage were not different between histopathological subtypes.</p><p><b>CONCLUSIONS</b>The clinical manifestations of LN children are diverse. Clinically, acute nephritis is the most common form of LN in children. Histopathologically, type IV is the most frequent subtype of LN. Early treatment may result in significant disease remission.</p>

Clinical and pathological features of Alport syndrome in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical and pathological features of Alport syndrome in children.</p><p><b>METHODS</b>The clinical and histopathological data of 10 hospitalized children with Alport syndrome from February 2007 to February 2009 were retrospectively reviewed.</p><p><b>RESULTS</b>There were 7 males and 3 females, with the age ranging from 2 years to 6 years and 7 months (mean 3 years and 2 months). Five of 10 cases had positive family history. X-linked dominant inheritance Alport syndrome was diagnosed in 8 cases, and autosomal recessive inheritance Alport syndrome in 2 cases. Recurrent gross hematuria was found in 5 cases, hematuria and proteinuria in 3 cases, massive proteinuria in 1 case, and nephritic syndrome in 1 case. Under the light microscope, 8 cases presented with mesangial proliferation glomerulonephritis, and 2 cases with focal segmental glomerulosclerosis. Immunofluorescence assay showed that all cases had IgM deposition in glomerulus. Only 1 case showed typical glomerular basement membrane (GBM) pathological changes. All cases showed abnormal alpha-chain distribution in renal collagen IV.</p><p><b>CONCLUSIONS</b>The children with Alport syndrome have diverse clinical manifestations. Characteristic histopathological presentations could not be found under a light microscope, mesangial proliferation glomerulonephritis is the dominant pathological change, and IgM deposition in glomerulus is common. The GBM pathological change in children is not common. Immunofluorescence assay of alpha-chain in collagen IV is needed for the diagnosis of Alport syndrome.</p>

Mechanisms of decorin inhibiting epithelial-to-mesenchymal transition induced by transforming growth factor beta1 in renal tubular epithelial cells / 中华儿科杂志

<p><b>OBJECTIVE</b>To investigate the mechanisms of decorin inhibiting epithelial-to-mesenchymal transition (EMT) induced by transforming growth factor beta1 (TGF-beta1) in renal tubular epithelial cells.</p><p><b>METHOD</b>HK-2 cells in vitro were divided into 4 groups: (1) negative control group; (2) decorin group, added with decorin 100 ng/ml ; (3) TGF-beta1 group, added with TGF-beta1 10 ng/ml; (4) decorin and TGF-beta1 group, added with decorin 100 ng/ml and TGF-beta1 10 ng/ml. The protein level of phosphor-ERK, phosphor-PI3K, phosphor-Smad(3) and beta-catenin was detected by Western blotting method. The snail mRNA level was tested by real time-PCR, while the lymphoid enhancer factor-1 (LEF-1) mRNA level was measured by RT-PCR.</p><p><b>RESULTS</b>The snail (2.59 +/- 0.70:1.02 +/- 0.13) and LEF-1 mRNA (1.85 +/- 0.08:0.30 +/- 0.11) were significantly up-regulated, meanwhile the protein level of phosphor-ERK (1.11 +/- 0.09:0.47 +/- 0.07), phosphor-PI3K (14.79 +/- 1.02:2.48 +/- 0.06), phosphor-Smad(3) (0.95 +/- 0.02:0.08 +/- 0.01) and beta-catenin (1.46 +/- 0.20:0.49 +/- 0.05) were significantly increased in TGF-beta1 group compared to control group, while there were no statistically significant difference in all figures between control group and decorin group. The phosphor-ERK protein level (0.58 +/- 0.08) and the snail mRNA level (1.24 +/- 0.03) were significantly down-regulated in TGF-beta1 and decorin group compared to TGF-beta1 group, however there were no statistically significant differences in the level of phosphor-PI3K (15.84 +/- 1.64), phosphor-Smad(3) (0.90 +/- 0.04) and beta-catenin (1.42 +/- 0.09) between these two groups.</p><p><b>CONCLUSION</b>Decorin inhibited EMT induced by TGF-beta1 which may be through blocking the ERK signal transduction pathway.</p>

NADPH oxidase-derived reactive oxygen species involved in angiotensin II-induced monocyte chemoattractant protein-1 expression in mesangial cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the origin of oxidative stress induced by angiotensin II (AngII) in human mesangial cells and the role of reactive oxygen species (ROS) in AngII-induced monocyte chemoattractant protein-1 (MCP-1) expression.</p><p><b>METHODS</b>MCP-1 expression was determined by real time RT-PCR. ROS production was measured by DCFDA fluorescence. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidase activity was examined by lucigenin chemiluminescence. p47phox and p67phox translocation was assayed by Western blot. Twenty-four male mice were randomly divided into three groups: the control, the AngIIinfusion [AngII 400 ng/(kg.min)], and the apocynin treatment. AngII was infused by subcutaneously osmotic minipump for 14 days. Urinary albumin and 8-isoprostane excretion were measured by ELISA.</p><p><b>RESULTS</b>In cultured human mesangial cells, AngII induced the MCP-1 expression in a dose-dependent manner with 3.56 fold increase as compared with the control. AngII increased intracellular ROS production as early as 3 min with the peak at 60 min and was in a time and dose-dependent. Incubation with different dosages of AngII (1 nmol/L, 10 nmol/L, and 100 nmol/L AngII) for 60 min, ROS production increased at 1.82, 2.92, and 4.08 folds respectively. AngII-induced ROS generation was sensitive to diphenyleneiodonium sulfate (DPI, 10 micromol/L) and apocynin (500 micromol/L), two structurally distinct NADPH oxidase inhibitors. In contrast, inhibitors of other oxidant-producing enzymes, including the mitochondrial complex Iinhibitor rotenone, the xanthine oxidase inhibitor allopurinol, the cyclooxygenase inhibitor indomethacin, the lipoxygenase inhibitor nordihydroguiaretic acid, the cytochrome P450 oxygenase inhibitor ketoconazole and the nitric oxide synthase inhibitor G-nitro-L-arginine methyl ester were without an effect. AngII-induced ROS generation was inhibited by the AT1 antagonist losartan (10 micromol/L) but not the AT2 antagonist PD123319 (10 micromol/L). AngII treatment induced translocation of cytosolic of p47phox and p67phox to the membrane. The antioxidants almost abolished AngII-induced MCP-1 expression. AngII infusion increased urinary and p67 translocation by 2.69-, 2.97-, and 2.67-fold, respectively.</p><p><b>CONCLUSIONS</b>NADPH oxidase-derived ROS is involved in AngII-induced MCP-1 expression. Inhibition of NADPH oxidase alleviates AngII-induced renal injury.</p>

Effects of core proteoglycan on the transdifferentiation of human renal tubular epithelial cell induced by transforming growth factor-beta1 in vitro / 中华儿科杂志

<p><b>OBJECTIVE</b>To study the effects of core proteoglycan on the transdifferentiation of human renal tubular epithelial cell induced by transforming growth factor beta1 (TGF-beta1) in vitro.</p><p><b>METHOD</b>The cultured HK-2 cells were divided into six groups: A. negative control group; B. 10 ng/ml TGF-beta1 group; C. 10 ng/ml core proteoglycan treated group; D. 100 ng/ml core proteoglycan treated group; E. 10 ng/ml TGF-beta1 + 10 ng/ml core proteoglycan group; F. 10 ng/ml TGF-beta1 + 100 ng/ml core proteoglycan group. The changes in configuration of HK-2 cells were inspected 48 hours after adding the stimulating factor. At the same time, changes in mRNA of keratin, alpha-smooth muscle actin, vimentin were analyzed.</p><p><b>RESULTS</b>Compared with group A, group B showed great changes in the morphology of cells, most cells converted into spindle shape, like fibroblast; groups E and F, especially group F showed significantly reduced spindle shape cells. Compared with group A, groups C and D had no significant changes in morphology of cells Compared with 10 ng/ml TGF-beta1 group and negative control, the mRNA expression of alpha-smooth muscle actin and vimentin had significant increase, but that of keratin reduced (P < 0.05). However, after combined treatment with TGF-beta1 and core proteoglycan, alpha-smooth muscle actin and vimentin expression were reduced significantly, while expression of keratin was up-regulated. Single core proteoglycan treated group and negative control group had no dramatic differences (P > 0.05).</p><p><b>CONCLUSION</b>TGF-beta1 can induce the transdifferentiation of human renal tubular epithelial cell and core proteoglycan has some inhibitory effect on transdifferentiation of human renal tubular epithelial cell induced by TGF-beta1 in vitro.</p>

Characterization of a bioflocculant from a newly isolated Vagococcus sp. W31 / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Screening of microorganisms producing flocculating substances was carried out. A strain secreting a large amount of bioflocculant was isolated from wastewater samples collected from the Little Moon River in Beijing. Based on the morphological properties and 16S rDNA sequence analysis, the isolate (designated W31) was classified as Vagococcus sp. A bioflocculant (named MBFW31) produced by W31 was extracted from the culture broth by ethanol precipitation and purified by gel chromatography. MBFW31 was heat-stable and had strong flocculating activity in a wide range of pH with relatively low dosage requirement. MBFW31 was identified as a polysaccharide with molecular weight over 2 x 10(6). It contained neutral sugar and uronic acid as its major and minor components, respectively. Infrared spectra showed the presence of hydroxyl, carboxyl and methoxyl group in its molecules. The present results suggested that MBFW31 had potential application in wastewater treatment.

Effects of colchicine on synthesis and excretion of cytokines and extracellular matrix by human renal fibroblasts / 中华儿科杂志

<p><b>OBJECTIVE</b>Renal interstitial fibrosis is the final common pathway leading to end-stage renal failure for progressive renal disease of various types. The present study was undertaken to add to the knowledge on colchicine's antiinflammatory and antifibrotic properties confirmed by both human and experimental studies. As the main effector cells, fibroblasts have a central role in the pathogenesis of renal fibrosis. This study aimed to investigate the effects of colchicine on the synthesis and excretion of cytokines transforming growth factor-beta1 (TGF-beta1), interleukin-1beta (IL-1beta) and extracellular matrix (collagen III, collagen IV) in human renal fibroblast.</p><p><b>METHODS</b>Various concentrations of colchicine (5.0 micromol/L, 10.0 micromol/L, 20.0 micromol/L, 40.0 micromol/L) were used to pretreat human embryo renal fibroblasts for 1 hour which were cultured in vitro and then stimulated by 10.0 microg/ml of lipopolysaccharide (LPS). After 18 hours, these fibroblasts and their supernatant were collected. The expression of TGF-beta1 mRNA, IL-1beta mRNA in the cells was studied by using RT-PCR, and the excretion of TGF-beta1, IL-1beta, collagen III and collagen IV by the fibroblasts was assessed by ELISA respectively.</p><p><b>RESULTS</b>(1) By pure stimulation with 10.0 micro g/ml LPS, the expression of TGF-beta1 mRNA and IL-1beta mRNA of fibroblasts was up-regulated 3 times (66.1 +/- 1.6 vs. 22.3 +/- 2.0, q = 590.5, P = 0.002) and 4.7 times (22.0 +/- 2.2 vs. 4.7 +/- 0.8, q = 106.8, P = 0.009), respectively. The protein excretion of TGF-beta1 and IL-1beta was remarkably increased as well compared with the control group [TGF-beta1: (516 +/- 14) pg/ml vs. (420 +/- 5) pg/ml (q = 80.3, P = 0.012), IL-1beta: (3.4 +/- 0.3) pg/ml vs. (0.3 +/- 0.1) pg/ml (q = 297.9, P = 0.003)]. (2) Colchicine could inhibit the expression of TGF-beta1 mRNA and protein in a dose-dependent manner. IL-1beta mRNA and protein were both up-regulated by colchicine. (3) LPS could not stimulate the excretion of extracellular matrix by fibroblasts, but the excretion of collagen III and collagen IV was down regulated by colchicine in a dose-dependent manner.</p><p><b>CONCLUSION</b>(1) The expression of TGF-beta1 mRNA and the excretion of TGF-beta1 protein in the fibroblasts was significantly suppressed by colchicine, while the expression of IL-1beta mRNA and the excretion of IL-1beta protein were enhanced. (2) Colchicine has significant inhibitory effect on the excretion of extracellular matrix such as collagen III and collagen IV in fibroblasts.</p>

Curcumin inhibited the proliferation and extracellular matrix production of human mesangial cells / 中华儿科杂志

<p><b>OBJECTIVE</b>Glomerulosclerosis is characterized by extracellular matrix accumulative and is often associated with mesangial cell proliferation. Curcumin showed a protective effect on anti-glomerular basement membrane (anti-GBM) nephritis in vivo, although their cellular localization and mechanism of action is still unclear. In this study, a glomerular mesangial cell line derived from fetus was used to determine whether curcumin could inhibit the cell proliferation and alter the extracellular matrix turnover.</p><p><b>METHODS</b>The cell activity was determined with MTT method. Mesangial cells were cultured in vitro and incubated with 0, 3.125, 6.25, 12.5, 25, 50, 100 and 200 micromol/L curcumin. In addition,human mesangial cells were cultured with or without LPS (10 microg/ml) in presence or absence of various concentrations of curcumin (4, 16 and 200 micromol/L), respectively. The supernatant and cells were collected. Then, the levels of the collagen type IV and III protein in the supernatant were determined by using enzyme-linked immunosorbent assay and the IL-1 beta and MCP-1 mRNA in the cells was measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) after subconfluent quiescent mesangial cells were incubated with various concentrations of curcumin for 24 h in vitro.</p><p><b>RESULTS</b>Curcumin at the concentration equal to or over 6.25 micro mol/L was able to inhibit the proliferation of mesangial cells in a dose-dependent manner, the optical density according to the sequential concentrations of curcumin was 0.65 +/- 0.02, 0.62 +/- 0.04, 0.56 +/- 0.01, 0.53 +/- 0.02, 0.51 +/- 0.03, 0.44 +/- 0.05, 0.41 +/- 0.07 and 0.38 +/- 0.06. Without any stimulation, human mesangial cells secreted some collagen type IV and III (10 +/- 9.13 ng/ml and 29.5 +/- 0.58 ng/ml, respectively) and expressed some MCP-1 mRNA, but did not express IL-1 beta mRNA. LPS increased the expression of collagen type IV and III in the culture medium of mesangial cells in vitro [(138.75 +/- 23.23) ng/ml and (38.25 +/- 5.38) ng/ml] and up-regulated the IL-1 beta and MCP-1 mRNA expression [(16.91 +/- 1.68)% and (76.6 +/- 6.59)%]. Yet curcumin could significantly decrease collagen type IV and III in the supernatant of cultured mesangial cells induced by LPS (20.5 +/- 1.00, P < 0.05 and 20.5 +/- 4.12 ng/ml, P < 0.05) and down-regulated the mRNA expression of IL-1 beta and MCP-1 in mesangial cells induced by LPS (P < 0.01).</p><p><b>CONCLUSION</b>Curcumin could inhibit the human mesangial cell proliferation and alter the extracellular matrix turnover, meanwhile it could down-regulate the IL-1 beta and MCP-1 mRNA expression induced by LPS, which may be valuable in decreasing the progression of glomerulosclerosis.</p>

Study on effect of Salvia injection in treating primary nephrotic syndrome and on endothelin and serum interleukin-2 receptor in children / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the effect and mechanism of Salvia Injection (SI) in treating primary nephrotic syndrome (PNS) in children.</p><p><b>METHODS</b>Forty-four PNS children were randomly divided into conventional steroid treated group (20 cases) and conventional plus SI intervention treated group (24 cases), the levels of serum endothelin (ET), soluable interleukin-2 receptor (sIL-2R) were observed.</p><p><b>RESULTS</b>Before treatment, plasma ET and sIL-2R in PNS children were higher than those in healthy children significantly (P < 0.01). After treatment, the ET and sIL-2R levels were all obviously improved in both treated groups (P < 0.05) and the improvement in the conventional plus SI intervention treated group was more obvious, the difference between the two treated groups after treatment was significant (P < 0.05).</p><p><b>CONCLUSION</b>Conventional treatment supplemented with SI could more effectively improve the levels of plasma ET and SIL-2R in treating PNS children, and hence alleviate the damage of renal tissue.</p>