Long non-coding RNA CHCHD4P4 promotes epithelial-mesenchymal transition and inhibits cell proliferation in calcium oxalate-induced kidney damage

Kidney stone disease is a major cause of chronic renal insufficiency. The role of long non-coding RNAs (lncRNAs) in calcium oxalate-induced kidney damage is unclear. Therefore, we aimed to explore the roles of lncRNAs in glyoxylate-exposed and healthy mouse kidneys using microarray technology and bioinformatics analyses. A total 376 mouse lncRNAs were differentially expressed between the two groups. Using BLAST, 15 lncRNA homologs, including AU015836 and CHCHD4P4, were identified in mice and humans. The AU015836 expression in mice exposed to glyoxylate and the CHCHD4P4 expression in human proximal tubular epithelial (HK-2) cells exposed to calcium oxalate monohydrate were analyzed, and both lncRNAs were found to be upregulated in response to calcium oxalate. To further evaluate the effects of CHCHD4P4 on the cell behavior, we constructed stable CHCHD4P4-overexpressing and CHCHD4P4-knockdown HK-2 cells. The results showed that CHCHD4P4 inhibited cell proliferation and promoted the epithelial-mesenchymal transition in kidney damage and fibrosis caused by calcium oxalate crystallization and deposition. The silencing of CHCHD4P4 reduced the kidney damage and fibrosis and may thus be a potential molecular target for the treatment of kidney stones.

Diabetes mellitus affects the biomechanical function of the callus and the expression of TGF-beta1 and BMP2 in an early stage of fracture healing

Transforming growth factor beta 1 (TGF-β1) and bone morphogenetic protein-2 (BMP-2) are important regulators of bone repair and regeneration. In this study, we examined whether TGF-β1 and BMP-2 expressions were delayed during bone healing in type 1 diabetes mellitus. Tibial fractures were created in 95 diabetic and 95 control adult male Wistar rats of 10 weeks of age. At 1, 2, 3, 4, and 5 weeks after fracture induction, five rats were sacrificed from each group. The expressions of TGF-β1 and BMP2 in the fractured tibias were measured by immunohistochemistry and quantitative reverse-transcription polymerase chain reaction, weekly for the first 5 weeks post-fracture. Mechanical parameters (bending rigidity, torsional rigidity, destruction torque) of the healing bones were also assessed at 3, 4, and 5 weeks post-fracture, after the rats were sacrificed. The bending rigidity, torsional rigidity and destruction torque of the two groups increased continuously during the healing process. The diabetes group had lower mean values for bending rigidity, torsional rigidity and destruction torque compared with the control group (P<0.05). TGF-β1 and BMP-2 expression were significantly lower (P<0.05) in the control group than in the diabetes group at postoperative weeks 1, 2, and 3. Peak levels of TGF-β1 and BMP-2 expression were delayed by 1 week in the diabetes group compared with the control group. Our results demonstrate that there was a delayed recovery in the biomechanical function of the fractured bones in diabetic rats. This delay may be associated with a delayed expression of the growth factors TGF-β1 and BMP-2.

Seroepidemiology of Toxoplasma gondii and Neospora caninum infections in goats in Hubei province, China

Scarce information is available about the seroprevalence of Toxoplasma gondii (T. gondii) and Neospora caninum (N. caninum) infections in goats in Hubei province, China. In the present study, the prevalence of T. gondii and N. caninum infections in goats were investigated in Hubei province, China between 2014 and 2015. A total 2007 serum samples were collected from 6 counties of Hubei province, China and were tested for antibodies to N. caninum and T. gondii by an enzyme-linked immunosorbent assay (ELISA) and an indirect agglutination test (IAT), respectively. Antibodies against T. gondii and N. caninum were detected in 13.4% and 3.9%, respectively in goats. 2% (41) serum samples were positive to both parasites. There was no apparent association of T. gondii and N. caninum infection with gender of the animals. There were significant differences of T. gondii (p < 0.01), N. caninum (p < 0.05) and both parasites (p < 0.01) infection with season. This is the first time that antibodies to T. gondii and N. caninum have been detected in goats in Hubei province, China.

Toxicity and adverse effects of everolimus in the treatment of advanced nonsmall cell lung cancer pretreated with chemotherapy‑Chinese experiences.

BACKGROUND: There is not more treatment selection for advanced nonsmall‑cell lung cancer (NSCLC) patients who had disease progression after two previous treatments. Everolimus is an oral inhibitor of the mammalian target of rapamycin pathway, which is aberrantly activated in NSCLC. PATIENTS AND METHODS: Stage IV NSCLC patients, with one or multiple prior chemotherapy regimens, received everolimus 5–10 mg/day with or without chemotherapy until progression or unacceptable toxicity. The primary objective were toxicity of everolimus and overall disease control rate (DCR). RESULTS: 22 patients were enrolled. Common ≥grade3 events were stomatitis, dyspnea, vomiting, thrombocytopenia. Overall disease control rate was 54.5% among 22 patients, 1 had a partial response, and 11 had disease stabilization. Common ≥Grade 3 events were stomatitis, dyspnea, vomiting, and thrombocytopenia. CONCLUSION: Everolimus was well tolerated, showing the modest clinical activity in heavily pretreated advanced NSCLC.

BMP-2 and titanium particles synergistically activate osteoclast formation

A previous study showed that BMP-2 (bone morphogenetic protein-2) and wear debris can separately support osteoclast formation induced by the receptor activator of NF-κB ligand (RANKL). However, the effect of BMP-2 on wear debris-induced osteoclast formation is unclear. In this study, we show that neither titanium particles nor BMP-2 can induce osteoclast formation in RAW 264.7 mouse leukemic monocyte macrophage cells but that BMP-2 synergizes with titanium particles to enhance osteoclast formation in the presence of RANKL, and that at a low concentration, BMP-2 has an optimal effect to stimulate the size and number of multinuclear osteoclasts, expression of osteoclast genes, and resorption area. Our data also clarify that the effects caused by the increase in BMP-2 on phosphorylated SMAD levels such as c-Fos expression increased throughout the early stages of osteoclastogenesis. BMP-2 and titanium particles stimulate the expression of p-JNK, p-P38, p-IkB, and P50 compared with the titanium group. These data suggested that BMP-2 may be a crucial factor in titanium particle-mediated osteoclast formation.

Adenovirus-mediated siRNA targeting TNF-α and overexpression of bone morphogenetic protein-2 promotes early osteoblast differentiation on a cell model of Ti particle-induced inflammatory response in vitro

Wear particles are phagocytosed by macrophages and other inflammatory cells, resulting in cellular activation and release of proinflammatory factors, which cause periprosthetic osteolysis and subsequent aseptic loosening, the most common causes of total joint arthroplasty failure. During this pathological process, tumor necrosis factor-alpha (TNF-α) plays an important role in wear-particle-induced osteolysis. In this study, recombination adenovirus (Ad) vectors carrying both target genes [TNF-α small interfering RNA (TNF-α-siRNA) and bone morphogenetic protein 2 (BMP-2)] were synthesized and transfected into RAW264.7 macrophages and pro-osteoblastic MC3T3-E1 cells, respectively. The target gene BMP-2, expressed on pro-osteoblastic MC3T3-E1 cells and silenced by the TNF-α gene on cells, was treated with titanium (Ti) particles that were assessed by real-time PCR and Western blot. We showed that recombinant adenovirus (Ad-siTNFα-BMP-2) can induce osteoblast differentiation when treated with conditioned medium (CM) containing RAW264.7 macrophages challenged with a combination of Ti particles and Ad-siTNFα-BMP-2 (Ti-ad CM) assessed by alkaline phosphatase activity. The receptor activator of nuclear factor-κB ligand was downregulated in pro-osteoblastic MC3T3-E1 cells treated with Ti-ad CM in comparison with conditioned medium of RAW264.7 macrophages challenged with Ti particles (Ti CM). We suggest that Ad-siTNFα-BMP-2 induced osteoblast differentiation and inhibited osteoclastogenesis on a cell model of a Ti particle-induced inflammatory response, which may provide a novel approach for the treatment of periprosthetic osteolysis.

Optimization of Xhhh strain biodegradation with metal ions for pharmaceautical wastewater treatment.

Effects of three ions, Mn2+, Cu2+ and Zn2+ on biological treatment of pharmaceutical wastewater by a functional strain Xhhh were investigated. Through orthogonal tests, Cu2+ was determined to be the most important factor influencing Xhhh biodegradation performance. Biodegradation kinetic experiments demonstrated that with Cu2+concentration at about 2.00 mg l-1, the maximum of specific growth rate and specific degradation rate were obtained to be 0.033 and 0.075 d-1, respectively. The optimal levels of Mn2+ (5.00 mg l-1), Cu2+ (2.00 mg l-1) and Zn2+ (5.00 mgl -1) were achieved based on experimental results of their effects on the activities of manganese peroxidase and lignin peroxidase, and biodegradation kinetic parameters. Among three types of biodegradation kinetic models (Monod, Tessier and Contois), Tessier model was found most reasonable for kinetics description of Xhhh growth (R2 = 0.995) and pollutants degradation (R2 = 0.970) in the case of metals optimization. Both kinetics evaluation and experimental results demonstrated that optimization with the three metals made a great contribution to Xhhh growth and COD removal for pharmaceutical wastewater treatment.

Incidência da mola hidatiforme na Escola Paulista de Medicina / Incidence of hydatidiform mole at the Escola Paulista de Medicina

No período de janeiro de 1980 a dezembro de 1989 foram atendidas na Escola Paulista de Medicina, um centro de referência, 75 pacientes com mola hidatiforme, das quais 65 se submeteram a esvaziamento em nosso serviço, e dez em outros. Para o cálculo da freqüência da mola hidatiforme utilizamos somente as 65 pacientes cujo esvaziamento molar se procedeu na Escola Paulista de Medicina em relaçäo a 13.986 gravidezes (partos, abortos e gravidezes sectópicas) ocorridas no período estudado. Deste modo, a freqüência da mola hidatiforme foi de 1:215 gravidezes, valor comparável aos países asiáticos e africanos