Clinical Efficacy for Treatment of Patients with Peripheral T Cell Lymphoma by HyperCVAD and CHOP/CHOP-like Protocols / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the clinical efficacy for treatment of patients with peripheral T cell lymphoma (PTCL) by HyperCVAD and CHOPCHOP-like protocols.</p><p><b>METHODS</b>A total of 97 patients with peripheral T cell lymphoma were divided into observation group (50 cases) and control group (47 cases). The patients in observation group were treated by HyperCVAD, and the patients in the control group were treated by CHOP/CHOP-like protocol. The clinical efficacy, accumulate survival rate and side effect of the 2 groups were compared.</p><p><b>RESULTS</b>The remission rate in the observation group were higher than that in control group (P < 0.05); The PFS rate (1 year) in observation group was higher than that in control group (P < 0.05); the PFS rate (2 years, 3 years) was not significantly different (P > 0.05). The OS rate (1 year, 2 years, 3 years) did not show difference (P > 0.05); the number of patients with neutropenia in observation group was higher than that in control group (P < 0.05); the levels of CD4(+) CD45RA(+), CD4(+) CD45RO(+) in observation group were lower than those in control group (P < 0.05); the levels of CD4(+) and CD4(+)/CD8(+) in observation group was lower than those in control group (P < 0.05); the level of CD8(+) in observation group was higher than that in control group (P < 0.05); the incidence of pneumonia, cardiotoxicity, severe anemia, and thrombopenia were not significantly different (P > 0.05).</p><p><b>CONCLUSION</b>HypeCVAD protocol shows good clinical effects on the patients with peripheral T cell lymphoma, displaying a high remission rate and PFS rate (1 year), but it has a high incidence of neutropenia, thus it needs more attention in clinic.</p>

Research Progress on Gene Alterations of Amelogenin Locus in Gender Identification / 法医学杂志

There are two kinds of amelogenin gene mutation, including mutation in primer-binding region of amelogenin gene and micro deletion of Y chromosome encompassing amelogenin gene, and the latter is more common. The mechanisms of mutation in primer-binding region of amelogenin gene is nucleotide point mutation and the mechanism of micro deletion of Y chromosome encompassing amelogenin gene maybe non-allelic homologous recombination or non-homologous end-joining. Among the population worldwide, there is a notably higher frequency of amelogenin gene mutations in Indian population, Sri Lanka population and Nepalese population which reside within the Indian subcontinent. Though amelogenin gene mutations have little impact on fertility and phenotype, they might cause incorrect result in gender identification. Using composite-amplification kit which including autosomal STR locus, amelogenin gene locus and multiple Y-STR locus, could avoid wrong gender identification caused by amelogenin gene mutation.

Species identification and absolute quantification of biological samples by droplet digital PCR / 法医学杂志

OBJECTIVE@#To test droplet digital PCR for species identification and absolute quantification of biological sample.@*METHODS@#Specific primers and probes for human mtDNA encoding gene ND4 and 16S rRNA were designed, and the species-specificity was assessed on DNA samples derived from human and common animals. To determine the sensitivity and stability of droplet digital PCR for species identification and absolute quantification, gradient dilution series of recombinant plasmid and 16 human DNA samples were analyzed.@*RESULTS@#Human recombinant plasmid FAM (ND4) could be used in detecting the samples of human. And the results of detecting were consistent with all levels of diluted concentrations. Droplet digital PCR was able to detect low and single copy of target DNA.@*CONCLUSION@#Droplet digital PCR, with high sensitivity and specificity, is fully amenable for species identification and absolute quantification of biological samples, also it can be applied on routine forensic examination.

The study of pedicle imaging and reformation with the multi-slice spiral CT in the surgical treatment of lumbar spondylolisthesis / 中国骨伤

<p><b>OBJECTIVE</b>To evaluate the value of pedicle parameter obtained by the reformation images on multi-slice spiral CT (MSCT) in the surgical treatment of lumbar spondylolisthesis.</p><p><b>METHODS</b>From January 2009 to March 2010, 60 patients with lumbar spondylolisthesis failing in conservative treatment were enrolled into the study and divided into experimental and control group randomly (each group with 30 patients). There were 26 males and 34 females ranging in age from 18 to 59 years with an average of (42.60 +/- 9.36) years. The experimental group was examined with volumetric scanning on MSCT before operation. Reformation such as multiplanar reconstruction (MPR) and volume rendering (VR) were carried out at the work station. Transverse section angle (TSA), sagittal section angle (SSA), pedicle length (PL), pedicle width (PW) and pedicle height (PH) were measured on different images and pedicle screws were implanted according pedicle parameter. In control group, the pedicle screws were implanted according to conventional anatomic landmark. Preparative time of screw canal and accuracy of screw were compared between two groups.</p><p><b>RESULTS</b>A hundred fifty-six screws were inserted in experiment group,143 screws were excellent, 11 good, and 2 poor. A hundred fifty screws were inserted in control group, 101 screws were excellent, 26 good, and 23 poor. There was significant difference in accuracy of screw between two groups (P < 0.001). The preparative time of screw canal in experiment group was (66.20 +/- 7.31) s, and was shorter than that of control group [(104.11 +/- 9.51) s, P < 0.001)].</p><p><b>CONCLUSION</b>Abundant information and parameter could be obtained with the MSCT reconstruction images. The images and parameters could make a perfect operative strategy before operation, adjust the direction of pedicle screws during operation, avoid and decrease operative complications effectively.</p>

Forensic analysis of LCN DNA using sample concentration methods followed by miniSTR genotyping / 法医学杂志

OBJECTIVE@#To optimize low copy number (LCN) DNA analysis methods for forensic STR genotyping.@*METHODS@#Two groups of DNA sample, extracted using either Magnetic bead method or Chelex-100 methods, were previously amplified with a Identifiler PCR Amplification kit, but no genotype was detected. The DNA samples were concentrated using either a drying method or the Microcon-100 method, then amplified using an miniFiler PCR Amplification kit and genotyped.@*RESULTS@#Among the 127 DNA samples, 47 samples, previously extracted using the Magnetic bead method, were genotyped with 36% success rate. Eighty samples, previously extracted using the Chelex-100 method, were genotyped with 30% success rate.@*CONCLUSION@#The application of sample concentration methods and miniFiler kit can improve the success rate of LCN STR analysis.

Polymerase region mutations and hepatitis B virus genotypes in chronic hepatitis B patients with poor response to lamivudine / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the mutations in Polymerase region and hepatitis B virus (HBV) genotypes in chronic hepatitis B patients with poor response to Lamivudine treatment.</p><p><b>METHODS</b>631 chronic hepatitis B patients with poor response to Lamivudine were recruited in this study. Real-time PCR and DNA sequencing were used to determine HBV genotypes; direct sequencing was performed to detect mutations, and real-time PCR was used to quantify HBV DNA load. Mutations in polymerase region were investigated in different HBV genotypes.</p><p><b>RESULTS</b>272 patients were infected with HBV of genotype B, and 359 patients were infected with HBV of genotype C. The mean age of patients infected with HBV of genotype C (39.1+/-11.4 years old) were significant higher than that of patients infected with HBV of genotype B (33.7+/-9.7 years old) (t = -6.55, P less than 0.01). The patients infected with HBV of genotype C had relatively higher HBV DNA load [(5.96+/-1.22) log10 copies/ml] than the patients infected with HBV of genotype B [(5.58+/-1.21) log10 copies/ml] (t = -2.01, P less than 0.05). The overall incidence rate of A181V/T mutation in genotype C (5.3%) was significantly higher than that in genotype B (0.4%) (x2=12.23, P less than 0.01), but the incidence rate of M204I/V, L180M, T184A/G/I/S, S202G/I and V173L mutations was not significantly different between genotype B and C (each P more than 0.05). M204I mutation in genotype B (20.6%) was more frequent than that in genotype C (13.9%) (x2=4.91, P less than 0.05). The Lamivudine resistance mutations were not significantly different between genotype B and genotype C (x2 = 0.00, P more than 0.05).</p><p><b>CONCLUSIONS</b>The incidence rate of lamivudine resistance mutation is not significantly different between genotype B and genotype C, but patients infected with HBV of genotype C have higher HBV DNA load than patients infected with HBV of genotype B.</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>

Clinical application on professor Yang Zhao-gang's elongated needle therapy / 中国针灸

Professor YANG Zhao-gang graduated from the Tianjin University of TCM in 1965. As a director doctor and a famous expert on the elongate needle therapy, he is engaged in medicine for more than 40 years. Through his untiring investigation on the acupuncture techniques, more than 10 specialized books have been published. He summarized some of basic acupuncture techniques and principle of acupoints combination on the elongated needle therapy, such as, 'selecting acupoints located on the higher place of the body, and combining the acupoints of Shangwan (CV 13), Zhongwan (CV 12) and Xiawan (CV 10)' 'dredging the pivot of meridian, restoring the Conception Vessel and regulating qi'. Its clinical effect is definite for treating the diseases of nervous, digestive and urinary systems. Basing on the traditional acupuncture techniques of the elongated needle, he explored and innovated some of new methods, such as pricking the auricular acupoints and shallow puncturing with the filiform needle. These efforts make the acupuncture treatment more secure, effective and reliable.

Resistin Binding Peptide Stimulates Basal Insulin Secretion of RINm5F Insulinoma Cells / 实用儿科临床杂志

Objective A resistin binding peptide (RBP) was selected by phage display in our previous work. Studies had shown that RBP could antagonize the role of resistin on the lipid metabolism and endocrine function of adipose tissue, but whether RBP affects the insulin secretion of pancreatic cells is still unknown. The aim of this study is to assess the effect of RBP on basal insulin secretion in RINm5F insulinoma cells. Methods The cell viability was measured by 3-[4,5-dimethyhhiazol-2-yl]-2,5-diphenyltetra-zolium bromide (MTT) cytotoxicity assay. The supernatants were assayed for insulin content by enzyme linked immunosorbent assay (ELISA). Reverse transcriptase-PCR assay and Western blotting were used to determine the expression of glucose transporter 2 (GLUT2) involved in insulin secretion. Cytosolic Ca2+, the trigger of insulin exocytosis, was analyzed with the fluorescent probe FURA-3/AM. Results RBP did no effect on the cell viability with a concentration of 10-8-10-12mol/L of 2 hours intervention. But it stimulated basal insulin secretion of RINm5F cells, accompanied by up-regulated increased expression of GLUT2 and elevated concentration of cytosolic Ca2+. Conclusion RBP could stimulate basal insulin secretion without affecting the cell viability.

Effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells / 中华儿科杂志

<p><b>OBJECTIVE</b>The prostate apoptosis response factor-4 (Par-4) gene was originally identified by differential screening for genes that are up-regulated when prostate cells are induced to undergo apoptosis. Par-4 was found to possess potent apoptotic activity in various cellular systems in response to numerous stimuli. The aim of this study was to explore the effects of small interfering RNA (siRNA) against Par-4 gene on the apoptosis of human bone marrow mesenchymal stem cells (hBMSCs) exposed to glutamate.</p><p><b>METHODS</b>Primary culture of hBMSCs was carried out and siRNAs targeted Par-4 gene (Par-4-SiRNA) were chemically synthesized. Eukaryocytic expression vector was built and were transfected into hBMSCs with liposome. After selecting with G418, the stable cell clones were treated with glutamate. The expression of Par-4 mRNA was determined by real-time PCR. The apoptosis of hBMSCs was quantified by flow cytometry. Western blotting was used to detect the protein levels of phosphorylated Akt1 (Thr308). Relative Caspase-3 activity was determined by colorimetric assay.</p><p><b>RESULTS</b>The Par-4-SiRNA-1 and Par-4-siRNA-2 could markedly down-regulate the mRNA levels of Par-4 gene in hBMSCs. With the transfections of Par-4-SiRNA-1 and Par-4-SiRNA-2, the levels of Par-4 mRNA were respectively decreased by 88% and 67%. Both Par-4-SiRNA-1 and Par-4-SiRNA-2 inhibited significantly the apoptosis of hBMSCs induced by glutamate, in which the percentages of apoptotic cells were respectively decreased to 38.80% +/- 3.97% (P < 0.01) and 45.49% +/- 4.32% (P < 0.01) from 60.30% +/- 6.82%. Western blot assays demonstrated that, glutamate down-regulated the expression of phosphorylated Akt1 proteins in hBMSCs (89.07 +/- 6.42 and 28.30 +/- 5.65, respectively, P < 0.01). However, Par-4-SiRNA-1 and Par-4-SiRNA-2 could markedly recover the down-regulation of Akt1 proteins induced by glutamate (63.56 +/- 6.75 and 45.59 +/- 4.88, respectively, P < 0.01). And the relative Caspase-3 activity which was enhanced by the treatment with glutamate (0.1428 +/- 0.0495 and 0.8616 +/- 0.1051, P < 0.01), was suppressed by Par-4-SiRNA-1 and Par-4-SiRNA-2 (0.8616 +/- 0.1051 and 0.6581 +/- 0.0555, respectively, P < 0.01).</p><p><b>CONCLUSION</b>SiRNA against Par-4 gene could inhibit the apoptosis of hBMSCs induced by glutamate, and its inhibitory effects may be mediated by the up-regulation of phosphorylated Akt1 and the suppression of the relative Caspase-3 activity.</p>

Association of RANTES gene promoter -28C/G polymorphism with respiratory syncytial virus bronchiolitis / 中华儿科杂志

<p><b>OBJECTIVE</b>Respiratory syncytial virus (RSV) infects nearly all children under two years of age. It is poorly understood why a few children who were infected with RSV develop bronchiolitis that require hospital admission while most have a relatively minor illness. Several recent studies have obtained some indications for the involvement of genetic heterogeneity in RSV bronchiolitis, implying that the clinical outcome of RSV infection perhaps is determined by genetic factors. Regulated on activation, normal T cell expressed and secreted RANTES plays a key role in the pathophysiology of RSV bronchiolitis. The purpose of this study was to explore the genetic association between the RANTES gene promoter -28C/G polymorphism and RSV bronchiolitis in Chinese Han ethnic group population.</p><p><b>METHODS</b>The study recruited 238 hospitalized patients (186 male and 52 female) under 12 months of age, with a clinical diagnosis of bronchiolitis due to RSV, the sex, age, hospital stay, SaO2 at the time of admission, personal and family history of atopy were recorded. The 288 healthy control subjects (206 male and 82 female), who had no evidence of personal or familial history of atopy and no history of wheezing, were chosen at the same time. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to identify the polymorphism at position -28C/G of the RANTES promoter. The total IgE concentrations in serum samples were measured by enzyme-linked immunosorbent assay (ELISA). The absolute peripheral blood eosinophil counts were measured by using an automated hematology analyzer.</p><p><b>RESULTS</b>The distribution of RANTES -28C/G gene polymorphism was in accordance with Hardy-Weinberg equilibrium. Compared to control subjects, significant difference was demonstrated for genotypes and allele frequencies of the RANTES -28C/G polymorphism in patients with RSV bronchiolitis (G = 10.22, P < 0.01; chi2 = 9.708, P < 0.01). Compared with the wild type CC, the -28G allele carriers demonstrated a 2.09-fold increased risk of RSV bronchiolitis (OR = 2.09, 95% CI = 1.32 - 3.30, P < 0.01). Interestingly, both the percentage of personal history of atopy and the percentage of family history of atopy for the -28G allele carriers were significantly higher (P < 0.05) than that for those CC homozygotes carriers in RSV bronchiolitis. Compared with the wild type CC, the -28G allele carriers demonstrated a 1.85-fold increased risk of the personal history of atopy (OR = 1.85, 95% CI = 1.01 - 3.38, P = 0.045) and a 1.91-fold increased risk of the family history of atopy (OR = 1.91, 95% CI = 1.03 - 3.54, P = 0.037), and the absolute peripheral blood eosinophil counts for the -28G allele carriers were significantly higher (P < 0.05).</p><p><b>CONCLUSION</b>The RANTES gene promoter -28C/G polymorphism is associated with the susceptibility to RSV bronchiolitis, and the -28G allele is an important predisposing factor for the personal history of atopy and the family history of atopy in RSV bronchiolitis.</p>

DNA genetyping of the trace bloodstains on the adsorbent object / 法医学杂志

OBJECTIVE@#To study the extraction and analysis of DNA from trace bloodstains on the adsorbent object.@*METHODS@#DNA was extracted by Chelex-100, QIAamp Mini Kit, and modified QIAamp Mini Kit, respectively. The extracted DNA was amplified and short tandem repeat (STR) loci were genotyped by multiplex PCR procedures, and then analyzed by ABI 3100 Genetic Analyzer.@*RESULTS@#The STR loci could be better genotyped with modified QIAamp Mini Kit, compared to poorer genotyping results obtained with Chelex-100 extraction method and QIAamp Mini Kit.@*CONCLUSION@#Our data indicate that DNA extracted from trace bloodstains on adsorbent object with modified QIAamp Mini Kit serves a better DNA template. for amplification and genotyping.

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>

Comparison of urinary proteomics between steroid-sensitive and steroid-resistant minimal change nephrotic syndrome in children / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the differentially expressed proteins in the urine of children with steroid-sensitive and steroid-resistant minimal change nephrotic syndrome (SRINS and SSINS, respectively).</p><p><b>METHODS</b>Urine samples were collected from 10 children with SRINS and 70 with SSINS as well as 30 healthy volunteers (control). Isoelectric focusing and two-dimensional electrophoresis in combination with matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry was performed for analysis of the urine proteins.</p><p><b>RESULTS AND CONCLUSION</b>In the urine samples, 30 protein spots were identified to have differential expression between SRINS and SSINS. Further analysis of 14 protein spots identified 12 proteins expressing in SRINS, namely kinesin family member 27, PITPNB, bullous pemphigoid antigen, alpha-1 protease inhibitor, Zn-alpha-2GP, alpha-1B-glycoprotein, serum albumin precursor, haptoglobin precursor, kinesin like motor protein, IRAK4, cytoplasmic dynein and cytokeratin 9. Nine of these 12 proteins were up-regulated (U1-U3, U5, U7-U9, U11-U12) and 3 down-regulated (D4, D6, D10) in SRINS, suggesting that these proteins may serve as the potential therapeutic targets and as new diagnostic markers for steroid-resistant nephrotic syndrome.</p>

Overexpression of resistin affect 3T3-L1 adipocyte lipid metabolism / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To investigate the effect of resistin overexpression on 3T3-L1 adipocyte lipid and glucose metabolism.</p><p><b>METHODS</b>Expression vector for rat resistin gene was constructed and transfected into 3T3-L1 adipocytes. Cell differentiation and lipid accumulation was determined by Oil Red O staining. Differentiation marker genes (pref-1, C/EBPalpha, FAS) and glucose transporter 4 (GLUT4) gene mRNA expressions were evaluated by reverse transcription-PCR (RT-PCR). Triglyceride (TG) and free fatty acids (FFAs) in adipocytes were measured by colorimetric kit.</p><p><b>RESULTS</b>(1) In resistin-overexpressed adipocytes, the lipid droplets presented at the second day which was earlier than the control cells. (2) The expression of C/EBPalpha and FAS genes in resistin-overexpressed adipocytes were up-regulated and the pref-1 was down-regulated compared with that of the control cells. (3) In resistin-overexpressed adipocytes, cellular TG and FFAs levels were significantly increased (P<0.05). (4) There was no difference in the expression of GLUT4 gene between 3T3-L1 adipocytes and resistin-overexpressed adipocytes (P> 0.05).</p><p><b>CONCLUSION</b>Overexpression of resistin can affect 3T3-L1 adipocyte lipid metabolism and thereby result in obesity and insulin resistance, but have no effect on GLUT4 gene expression.</p>

Association between interleukin-8 gene-251 locus polymorphism and respiratory syncytial virus bronchiolitis and post-bronchiolitis wheezing in infants / 中华儿科杂志

<p><b>OBJECTIVE</b>Respiratory syncytial virus (RSV) infects nearly all children under two years of age. It is not understood why some develop serious bronchiolitis. Whether there is a genetic component is not known. The nature of the association between RSV bronchiolitis and subsequent wheezing remains unknown. interleukin-8 (IL-8) is a potent neutrophil chemokine and activator, which plays a role in virus-induced wheezing diseases. The purpose of this study was to assess the genetic association between the IL-8 gene promoter -251A/T polymorphism and RSV bronchiolitis and post-bronchiolitis wheezing in children.</p><p><b>METHODS</b>Totally 320 children who were hospitalized for bronchiolitis together with positive immunofluorescence for RSV were recruited in this study from Jan. 2002 to Jan. 2004. A group of 272 healthy children were enrolled as controls. The age of these children ranged from 1 to 12 months. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to identify the polymorphism at position-251 of the IL-8 promoter in RSV bronchiolitis and control groups. The total IL-8 and IgE concentrations in serum samples were measured by enzyme-linked immunosorbent assay (ELISA). The patients with RSV bronchiolitis were followed up in order to analyze the occurrence of wheezing post-bronchiolitis.</p><p><b>RESULTS</b>(1) Both A allele and T allele were detected at -251 of the IL-8 promoter; the prevalence of the A allele in RSV bronchiolitis group was 45.6%, as compared with 37.7% in normal group. The prevalence of IL-8-251A allele was significantly different between the two groups (P < 0.05). (2) For genotypes T/T, A/T, A/A in RSV bronchiolitis, level of serum IL-8 were (17 +/- 6) ng/L, (21 +/- 7) ng/L, (24 +/- 9) ng/L, respectively, the difference was significant among the three genotypes (P < 0.01). (3) The prevalence of the A allele in the group who wheezed after the episode of RSV bronchiolitis was 54.6%, as compared with 35.8% in the group who had bronchiolitis but did not go on to wheeze. The prevalence of IL-8-251A allele was significantly different between the two groups (P < 0.05).</p><p><b>CONCLUSION</b>Polymorphism of IL-8 promoter-251A/T was associated with susceptibility to RSV bronchiolitis in children. The association of IL-8-251A with severe RSV bronchiolitis is most marked in the children who go on to wheeze.</p>

A resistin binding peptide selected by phage display inhibits 3T3-L1 preadipocyte differentiation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Resistin, a newly discovered cysteine-rich hormone secreted mainly by adipose tissues, has been proposed to form a biochemical link between obesity and type 2 diabetes. However, the resistin receptor has not yet been identified. This study aimed to identify resistin binding proteins/receptor.</p><p><b>METHODS</b>Three cDNA fragments with the same 11 bp 5' sequence were found by screening a cDNA phage display library of rat multiple tissues. As the reading frames of the same 11 bp 5' sequence were interrupted by a TGA stop codon, plaque lift assay was consequently used to prove the readthrough phenomenon. The stop codon in the same 11 bp 5' sequence was replaced by tryptophan, and the binding activity of the coded peptide [AWIL, which was designated as resistin binding peptide (RBP)] with resistin was identified by the confocal microscopy technique and the affinity chromatography experiment. pDual GC-resistin and pDual GC-resistin binding peptide were co-transfected into 3T3-L1 cells to confirm the function of resistin binding peptide.</p><p><b>RESULTS</b>Three cDNA fragments with the same 11 bp 5' sequence were found. The TGA stop codon in reading frames of the same 11 bp 5' sequence was proved to be readthroughed. The binding activity of RBP with resistin was consequently identified. The expression of the resistin binding peptide in 3T3-L1 preadipocytes expressing pDual GC-resistin significantly inhibited the adipogenic differentiation.</p><p><b>CONCLUSION</b>RBP could effectively rescue the promoted differentiation of resistin overexpressed 3T3-L1 preadipocyte.</p>

Transdifferentiation and collagen-synthesis effects of exogenous connective tissue growth factor on renal tubular epithelial cells in vitro / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the effects of recombinal human connective tissue growth factor (rhCTGF) stimulation on epithelial-myofibroblast transdifferentiation (EMT) and collagen-synthesis in human renal tubular epithelial cell line (HK2) in vitro.</p><p><b>METHODS</b>The cultured HK2 cells were stimulated with rhCTGF of 5 ng/mL. The morphological changes were observed under an inverted microscope. The cells were collected at 0, 3, 6, 12, 24 and 48 hrs after rhCTGF stimulation. The expression of E-cadherin,alpha-smooth muscle actin (alpha-SMA), collagen Ialpha1 (Col Ialpha1) and collagen IValpha1 (Col IValpha1) mRNAs were detected by RT-PCR.</p><p><b>RESULTS</b>rhCTGF stimulation changed the HK2 cell appearance from oval to fusiformdown-regulated the E-cadherin mRNA expression and up-regulated alpha-SMA mRNA expression, but had no effects the Col Ialpha1 and Col IValpha1 mRNA expression.</p><p><b>CONCLUSIONS</b>Exogenous CTGF can mediate the EMT but has no collagen-synthesis effects on HK2 cells.</p>

Thyroid autoantibodies in children with Turner's syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>Turner's syndrome (TS) is characterized by the absence of an X chromosome or the presence of a structurally abnormal X chromosome in a phenotypic female. It was recently reported that autoimmune thyroiditis (AIT) was found in 38% of white patients with TS, and few studies in this aspect have been conducted in China. The purpose of this study was to determine the frequency of AIT among TS patients and risk factors for development of thyroid dysfunction in Chinese children with TS.</p><p><b>METHODS</b>Serum antithyroglobulin antibody (TgAb), thyroperoxidase antibody (TPOAb) and thyroid function (T(3), T(4) and TSH) of 24 children with TS (mean age 12.9 +/- 2.4 years, range 4.8 - 16.8 years) were assessed. Their karyotype distribution was as follows: thirteen patients with 45, XO kayrotype, eight patients with structurally abnormal X chromosome, two with X mosaic kayrotype and one with 46, XX. Techniques including radioimmunoassy and elctro-chemiluminescence immunoassy were used in this study. All TS children were divided into two groups. Group one was thyroid autoantibodies (TAA)-positive group, the levels of TgAb and/or TPOAb in them were higher than the normal levels (TgAb < 30%, TPOAb < 20%), respectively, and the remaining patients were assigned into TAA-negative group.</p><p><b>RESULTS</b>Seven of the 24 (29%) patients had higher levels of TgAb and TPOAb than the normal values (< 30% and < 20%). The level of serum TSH [6.1 (3.6-100.0) mU/L] in TAA-positive group was significantly higher than that [3.9 (1.7-7.9) mU/L] in TAA-negative group (P < 0.05). The frequency of hypothyroidism or subclinical hypothyroidism in TAA-positive group (5/7) was higher than that in TAA-negative group (3/17) (P < 0.05).</p><p><b>CONCLUSION</b>The positive rate of serum TAA in children with TS was 29%. About 70% TS children with positive serum TAA developed hypothyroidism or subclinical hypothyroidism. The results have provided the basis for regular follow-up assessment of thyroid autoantibodies and thyroid function in children with TS, and these measures are of importance for timely diagnosis of thyroid dysfunction and application of appropriate treatment.</p>