Polymorphisms of human leukocyte antigen B*27 on clinical phenotype of spondyloarthritis in Chinese.

BACKGROUND: In recent years, an ever-increasing number of alleles of human leukocyte antigen B*27 (HLA-B*27) have been identified. This study aimed to establish an updated method for HLA-B*27 subtyping, and to investigate the impact of HLA-B*27 polymorphisms on the clinical phenotype of spondyloarthritis (SpA). METHODS: Overall, 184 SpA patients were recruited for analyzing diversity of HLA-B*27 via an updated high-resolution polymerase chain reaction amplification with sequence specific primers (PCR-SSP). RESULTS: The prevalence of HLA-B*27 was 94.0%, and four subtypes were identified including HLA-B*2704 (77.5%), B*2705 (20.2%), B*2707 (1.7%), and B*2724 (0.6%). There was an obvious male predominance (P=.05) and markedly elevated C-reaction protein (CRP) in B*27 positive SpA (P<.01). In multivariate linear regression analysis, the elevated CRP was positively associated with HLA-B*27 positivity (regression coefficient B=46.1, P=.0003), grade of sacroiliitis (B=47.5, P=.0032), and male gender (B=20.4, P=.0041). Notably, a male predilection was also found in B*2705 positive SpA while B*2707 was associated with older age, higher positive family history, and higher prevalence of extra-articular features (all P<.05). CONCLUSIONS: In this study, an updated PCR-SSP technique to identify increasing alleles of HLA-B*27 was developed and their different effects on clinical manifestations of SpA were demonstrated. Genotyping of HLA-B*27 would shed light on our understanding of the pathogenesis of SpA.

Overexpression of miR-34c inhibits high glucose-induced apoptosis in podocytes by targeting Notch signaling pathways.

Recent findings have shown that microRNAs play critical roles in the pathogenesis of diabetic nephropathy. miR-34c has been found to inhibit fibrosis and the epithelial-mesenchymal transition of kidney cells. However, the role of miR-34c in diabetic nephropathy has not been well studied. The current study was designed to investigate the role and potential underlying mechanism of miR-34c in regulating diabetic nephropathy. After treating podocytes with high glucose (HG) in vitro, we found that miR-34c was downregulated and that overexpression of miR-34c inhibited HG-induced podocyte apoptosis. The direct interaction between miR-34c and the 3'-untranslated region (UTR) of Notch1 and Jagged1 was validated by dual-luciferase reporter assay. Moreover, Notch1 and Jagged1 as putative targets of miR-34c were downregulated by miR-34c overexpression in HG-treated podocytes. Overexpression of miR-34c inhibited HG-induced Notch signaling pathway activation, as indicated by decreased expression of the Notch intracellular domain (NICD) and downstream genes including Hes1 and Hey1. Furthermore, miR-34c overexpression increased the expression of the anti-apoptotic gene Bcl-2, and decreased the expression of the pro-apoptotic protein Bax and cleaved Caspase-3. Additionally, the phosphorylation of p53 was also downregulated by miR-34c overexpression. Taken together, our findings suggest that miR-34c overexpression inhibits the Notch signaling pathway by targeting Notch1 and Jaggged1 in HG-treated podocytes, representing a novel and potential therapeutic target for the treatment of diabetic nephropathy.

Heat shock protein 70 protects rat peritoneal mesothelial cells from advanced glycation end-products-induced epithelial-to-mesenchymal transition through mitogen­activated protein kinases/extracellular signal-regulated kinases and transforming growth factor-ß/Smad pathways.

Epithelial­to­mesenchymal transition (EMT) may result in damage to the peritoneum and the development of fibrosis in peritoneal mesothelial cells (PMCs). However, the mechanism underlying EMT in peritoneal mesothelial cells is not well understood. Heat shock proteins (HSPs) were initially identified as proteins that are expressed following exposure of cells to environmental stress. However, their function in the development of EMT in PMCs remains to be fully elucidated. In the present study, the effect of HSP70 on advanced glycation end­products (AGEs)­induced EMT in peritoneal mesothelial cells was investigated by overexpression of this protein using a plasmid and knockdown of HSP70 using small interfering RNA. In addition, the underlying molecular mechanisms were explored. The results demonstrated that AGEs activated changes associated with EMT, including the loss of E­cadherin and the increase in α­smooth muscle actin. Furthermore, AGEs also induced the upregulation of HSP70, which led to the partial inhibition of EMT in PMCs. HSP70 inhibits EMT by modulating transforming growth factor­ß (TGF­ß)/Smad expression and the mitogen­activated protein kinases (MAPK)/extracellular signal­regulated kinases (ERK) signaling pathways. The findings suggested that HSP70 augments the cellular defense capacity through inhibition of TGF­ß/Smad and MAPK/ERK signaling pathways, thereby protecting PMCs from AGEs­induced EMT.

Hypoxia­inducible adrenomedullin ameliorates the epithelial-to-mesenchymal transition in human proximal tubular epithelial cells.

Renal tubular epithelial cells can enter the epithelial­to­mesenchymal transition (EMT) in response to chronic hypoxia. EMT is a process which involves the phenotypic conversion of epithelial cells, that is believed to have an important role in renal fibrosis. However, the underlying mechanisms of the involvement of EMT in renal fibrosis remain to be elucidated. Adrenomedullin (AMD) has been implicated in renal fibrosis and is induced by hypoxia. The aims of the present study were to determine whether ADM signaling was active in human proximal tubular epithelial cells cultured under hypoxic conditions, and to observe the activity of ADM during EMT. The expression levels of ADM were significantly increased, in a time­dependent manner, in HK­2 and HKC human proximal tubular epithelial cells, cultured under hypoxic conditions. Overexpression of exogenous ADM was accompanied by increased expression levels of the epithelial markers E­cadherin and tight junction protein­1, and decreased expression levels of the mesenchymal markers vimentin and α­smooth muscle actin, during hypoxia. Knock­down of ADM expression by small hairpin RNA, or co­administration of an ADM peptide inhibitor, in HK­2 cells significantly exacerbated hypoxia­induced EMT, as compared to the lack of effect observed in the untransfected controls. ADM was shown to suppress EMT by inhibiting the activation of extracellular signal­regulated kinase (ERK), and this effect was prevented by the ERK activator apigenin. The results of the present study suggest that ADM has an important role in promoting EMT in hypoxic human proximal tubular epithelial cells. ADM may therefore represent a novel therapeutic target in the treatment of injured kidneys.

Molecular cloning, sequence analysis and expression of a GHF 43 xylanase from Aspergillus niger in Escherichia coli.

A new xylanase gene (xyn43A) from Aspergillus niger XZ-3S was cloned and expressed in Escherichia coli BL21-CodonPlus (DE3)-RIL. The coding region of the gene was separated by only one intron 86 bp in length. It encoded 318 amino acid residues of a protein with a calculated molecular weight (MW) of 33.47 kDa plus a signal peptide of 19 amino acids. The amino acid sequence of the xyn43A gene showed 77.56% amino acid identity to A. nidulans xylanase, and the phylogenetic tree analysis revealed that xyn43A had close relationships with those of family 43 of glycosyl hydrolases reported from other microorganisms. Three-dimensional structure modeling showed that Xyn43A had a typical five-blade ß-propeller fold. The mature peptide encoding cDNA was subcloned into pET-28a (+) expression vector. The resultant recombinant plasmid pET-28a-xyn43A was transformed into Escherichia coli BL21-CodonPlus (DE3)-RIL, and xylanase activity was measured. A maximum activity of 61.43 U/mg was obtained from the cellular extract of E. coli BL21-CodonPlus (DE3)-RIL harboring pET-28a-xyn43A. The recombinant xylanase had optimal activity at pH5.0 and 45°C. Fe(3+), Cu(2+) and EDTA had an obvious active effect on the enzyme.

Study on in vitro anti-tumor activity of Bidens bipinnata L. extract.

We studied the in vitro anti-tumor activity of Bidens Bipinnata L. extract. MTT assay was used to investigate the inhibitory effect of different concentrations of the extracts on human hepatocellular carcinoma (HepG2) cell lines and human cervical carcinoma (Hela) cell lines, and the IC50 values were calculated. The Bidens Bipinnata L. extract had different degrees of inhibitory effects on these two cells, and when exposure time was 48 h, the inhibition rate reached its peak, with IC50 values of 14.80 µg/mL and 13.50 µg/mL respectively. The Bidens Bipinnata L. extract had a good inhibitory effect on human HepG2 cell lines and Hela cell lines, and thus has certain development prospects.

Comparison of optic nerve morphology in eyes with glaucoma and eyes with non-arteritic anterior ischemic optic neuropathy by Fourier domain optical coherence tomography.

The aim of this study was to compare the optic nerve head (ONH) and peripapillary retinal nerve fiber layer (RNFL) thickness in eyes with glaucoma and non-arteritic anterior ischemic neuropathy (NAION) by Fourier domain optical coherence tomography (FDOCT), and to evaluate the diagnostic capability of FDOCT in glaucoma and NAION. This study included 26 eyes with glaucoma (36.6%), 15 eyes with NAION (21.1%) and 30 eyes of normal subjects (42.3%). Those with the following conditions were excluded; a visual field defect greater than one hemifield, spherical equivalent (SE) more than ±6 D, or the onset of NAION within 6 months. FDOCT was used to analyze the characteristics of ONH and RNFL thickness. Among the three groups of subjects, glaucomatous eyes had the largest cup area and cup volume, and the smallest rim area, rim volume and disc volume (P<0.05). NAION eyes had the smallest cup area and cup volume (P<0.05), but their rim area, rim volume and disc volume were comparable to those of control eyes (P>0.05). The cup-to-disc (C/D) ratio was increased in glaucomatous eyes but reduced in NAION eyes compared with control eyes. Glaucomatous eyes had the greatest loss of RNFL thickness in the temporal upper (TU), superior temporal (ST) and temporal lower (TL) regions (P<0.05), whereas NAION eyes had the smallest RNFL thickness in the superior nasal (SN) and nasal upper (NU) regions (P<0.05). The areas under the receiver operator characteristic curve (AROCs) of the temporal, superior and inferior RNFL in glaucomatous eyes were greater compared with that of the disc area (P<0.05). In addition, the AROCs of the temporal, superior and inferior RNFL were higher compared with that of nasal RNFL (P<0.05). The AROCs of all parameters for NAION were not significantly different, with the exception of superior, nasal superior and inferior temporal RNFL (P<0.05). In conclusion, FDOCT is able to detect quantitative differences in the optic disc morphology and RNFL thickness between glaucomatous and NAION eyes. These differences may provide new insights into the clinical characteristics and diagnosis of the two diseases.

Type 1 insulin-like growth factor receptor monoclonal antibody (HX-1162) treatment for liver cancer.

One of the most important molecules mediating the proliferation, growth, and metastasis of cancer cells is insulin-like growth factor (IGF), with its receptor IGF-1R. Here, we describe the potential of an IGF-1R monoclonal antibody, HX-1162, on liver cancer apoptosis in vitro and in vivo. We found that HX-1162 could induce the apoptosis of cultured liver cancer cells. Additionally, HX-1162 treatment inhibited the tumor growth after cancer cell grafting and enhanced the cell apoptosis inside the tumor tissue. We conclude that IGF-1R targeting therapy provides a new avenue toward treating liver cancer.

Analysis of kidney biopsy data from a single center in the midland rural area of china, 1996-2010.

OBJECTIVE: To survey the clinical epidemiology and correlations between pathology and clinical features of major groups of kidney diseases in a rural area of China. METHODS: From January 1996 to December 2010, histologic diagnosis of renal disease was made on samples collected from 919 patients from a single center in the midland rural area of China. Demographic data were obtained from all patients, and clinical profiles were analyzed in 917 patients. RESULTS: The mean age of the whole group was 33.13 (14.13) years (range 16-72 years). Men accounted for 55.28% (n = 508) and women made up 45.72% (n = 408). Patients aged 16 to 50 years comprised 83.75% of the sample (n = 770). Lupus nephritis was the predominant diagnosis in women; renal diseases were predominant in men. In patients with nephrotic syndrome, mesangial proliferative glomerulonephritis was the most frequent pathologic pattern (39.46%), followed by IgA nephropathy (18.39%), whereas in patients with nephritic syndrome, IgA nephropathy (39.64%) was the most frequent pathologic pattern, followed by mesangial proliferative glomerulonephritis (32.38%). The most common pathologic pattern in patients with secondary glomerulonephritis was Henoch-Schoˇnlein purpura nephritis, followed by lupus nephritis. CONCLUSIONS: Mesangial proliferative glomerulonephritis was the most common renal pathologic pattern. Male adolescents were predominant in this group of patients. The most common clinical syndrome was nephrotic syndrome.