Environmental Pollutants ; toxicity ; Humans ; Hypoxia-Inducible Factor 1 ; metabolism

Objective To explore the predictive value of serum soluble fms-like tyrosine kinase-1 (sFlt-1) and vascular endothelia growth factor (VEGF) levels in preeclampsia at second trimester. Methods Serum sFlt-1,VEGF concentrations were determined in 172 initial normal pregnant women at 26-28 gestation week.The outcomes of pregnancies were followed.In a cohort of 172 pregnant women, 16 cases of preeclampsia were developed (preeclampsia group),and 156 cases were with no complication (control group).Results The serum levels of sFlt-1 in preeclampsia group (11.4?6.2)?g/L were significantly higher than that in control group(4.5?2.1)?g/L(P＜0.01).The serum levels of sFlt-1 in precelampsia women with the onset before 32 gestation week and fetal growth retardation,(14.0?6.8)?g/L,(14.4?6.7)?g/L were significantly higher than that in women with the onset after 32 gestation week and with no fetal growth retardation (9.0?4.1)?g/L,(8.9?4.0)?g/L,respectively (P＜0.05).There was no significant difference in the serum levels of VEGF between preeclampsia group and control group.A sFlt-1 cutoff value of 8.75?g/L at 26-28 gestation week yielded a sensitivity of 87.5%,specificity of 97.4%,positive predictive value of 80.0%,negative predictive value of 88.5%,respectively,for subsequent onset of preeclampsia.Conclusion Maternal serum sFlt-1 concentration at second trimester can be used as an early predictive marker of preeclampsia.

AIM: To investigate the expression of MAT1 protein in pancreatic cancers and the relationship between MAT1 and clinicopathological features of pancreatic cancers. METHODS: 94 surgical specimens, including 70 pancreatic cancers, 10 pancreatic benign tumors, 14 chronic pancreatitis and 10 autopsy normal pancreas tissues, were analyzed immunohistochemically, and then MAT1 expression and clinicopathological features were compared. RESULTS: MAT1 was expressed mainly in the cancer cells,and also in the fibroblasts, where it was localized within the cytoplasm and nuclear envelope. MAT1 expression was found in 75.7% (53/70) of the cancers, but not detected or weakly expressed in control tissues. There was a significant difference in expression of MAT1 among the above four tissues (P

High resolution melting (HRM), an important technology for genotyping and mutation scanning, has broad prospects in the authentification of traditional Chinese medicine. This paper selected universal trnH-psbA primers and used HRM to establish a new methods for identification of Akebia herbs. PCR was conduct at the annealing temperature of 58 degrees C and 35 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further analyzed. The results showed the Tm values of Caulis Akebiae was (81.84 ± 0.16), (85.28 ± 0.16) degrees C and Caulis Clematidis Armandii was (83.22 ± 0.19) degrees C and Caulis Aristolochiae manshuriensis was (81.67 ± 0.14) degrees C, (84.24 ± 0.10) degrees C with 5-125 mg - L-' DNA template, 0.4 μmol x L(-1) primer, 2.0 mmol x L(-1) Mg2+. This method can achieve the authentification of Akebia herbs and is simple, fast, high-throughput, visual.
Chemistry Techniques, Analytical ; methods ; DNA, Plant ; chemistry ; genetics ; Genotype ; Magnoliopsida ; chemistry ; classification ; genetics ; Phylogeny ; Transition Temperature

OBJECTIVETo detect the differentially expressed genes in oral lichen planus(OLP) by cDNA microarray.

METHODSThe PCR products of 4000 genes were spotted on chemical material coated glass plates in array. The total RNAs were isolated from OLP and normal oral mucosa tissue and were purified to mRNA. Both the mRNA were reversely transcribed to cDNA with the incorporations of fluorescent dUTP, for preparing the hybridization probes. The mixed probes were then hybridized to the cDNA microarray. Microarray was scanned for the fluorescent signals and showed the differences between the two samples.

RESULTSThe expressions of 122 genes were up regulated while the expressions of 91 genes were down regulated in OLP among the 4000 target genes. The up-regulated genes were mainly the ones of immunity related genes, metabolize related genes, oncogene, cytokine and cell signal transduction protein. The down-regulated genes were mainly the ones of DNA binding and transcription factors, cell signal transduction protein, immunity related genes, metabolize related genes, and cytokine.

CONCLUSIONTwo hundred and thirteen differentially expressed genes with different functions were revealed in OLP, which may play some roles in the progression of OLP.

Objective To explore the effect of pain intervention on limb function exercises in patients with peripheric fractures of knee joint.Methods 60 patients with peripheric fractures of knee joint were randomized in equal number into the observation group and control group.Both groups took functional exercises for affected limbs.Besides,the former and latter groups were administered with celecoxib at a dosage of 200mg twice a day from pre-operation to discharge and after operation,respectively.The two groups were compared in terms of pain degree at different time points as well as the functional recovery of affected limbs.Results The observation group was lower in pain scores than the control group at hours 6,8,12,24,36 and 48.The active and passive motions of the affected limbs in the observation group were significantly better than those in the control at days 1,2,3,4 and 5(all P<0.001).Conclusion Pain intervention with celecoxib before operation may help patients to take functional exercises as early as possible,promoting the rehabilitation of functions of affected limbs.

Objective:To differentiate human embryonic stem cells ( hESCs ) into keratinocytes ( K-hESCs) and analyse the expression characteristics of biomarkers of K-hESCs.Methods: The hESCs of line H9 were seeded on matrigel in mTeSR1 medium.The hESCs were directly differentiated into kerati-nocytes in epithelial differentiation medium with bone morphogenetic protein 4, retinoic acid and N2 sup-plement.The karyotype of K-hESCs was analyzed, comparing the gene expression differences of K-hESCs with human gingival epithelial cells (HGECs), human immortalized oral epithelial cells (HIOECs) and HaCaT by Real-time PCR.Molecular characteristics of the cell differentiation were observed throughout the process by immunocytochemical techniques.Results:H9-hESCs were successfully differentiated into the cells that exhibited characteristics of keratinocytes in epithelial differentiation medium.The karyotype of K-hESCs was 46, XX; and the keratinocyte gene p63 expression in K-hESCs was significantly lower than that in HaCaT ( P<0.05) , but there was no significant difference of p63 expression in K-hESCs, comparing with that in HGECs and HIOECs ( P >0.05 ) .Conclusion: H9-hESCs could be directly differentiated into K-hESCs.The gene expression of K-hESCs was similar to that of epithelial cells in the early stage of monolayer cells differentiation with high proliferative activity.

Objective To investigate the protective effects of atorvastatin on hyperphosphate-induced rat vascular smooth muscle ceils (RVSMCs) calcification and to discuss the mechanism. Methods RVSMCs were placed in various culture media, including normal phosphate medium, high phosphate medium, ZVAD-FMK medium and atorvastatin medium.Calcium content and cell protein content were quantified by the o-cresolphthalein complexone method and BCA protein assay respectively. Calcification was visualized by yon Kossa staining. And cell apoptosis was quantified by ELISA. Results (1)At day 3, 6, 9, RVSMCs calcification occurred more frequently in high phosphate medium than that in normal phosphate medium (P<0.05). (2)At day 6, RVSMCs calcification was significantly inhibited in 1.0 μmol/L and 2.0 μmol/LZVAD-FMK medium (P<0.05). And in 10 nmol/L and I00 nmol/L statin medium, RVSMCscalcium deposition significantly decreased (P<0.05). (3)RVSMCs apoptosis and calcification occurredfrequently in high phosphate medium. And atorvastatin significantly inhibited RVSMCs apoptosisboth in long-term and short-term (P<0.05). Conclusions Hyperphosphate can induce the calcium deposition of RVSMCs in vitro. Atorvastatin protects RVSMCs from phosphate-induced calcification by inhibiting apoptosis.

Objective To evaluate the effect of parathyroid hormone (PTH) on the expression of connective tissue growth factor (CTGF) in human renal tubular epithelial cells, and to explore the role of MAPK signaling pathway. Methods Real time RT-PCR, Western blot, and reporter gene assay were employed to detect PTH-induced CTGF expression in HK-2 cells. Inhibitors (PD98059 and U0126) of MAPK signaling pathway were used to confirm involved signal pathway. Results HK-2 cells had basic expression level of CTGF mRNA and protein, which were increased significantly after treatment with PTH. The luciferase activity was up-regulated to a higher level as compared with control group after treatment with 10-10 mol/L PTH for 12 h [(1.8884±0.0780) vs (0.9891±0.0300) A, P<0.01]. Moreover, a small amount of p-ERK1/2 was detected in normal HK-2 cells, but it was increased significantly in response to PTH activation, most remarkably when treated with 10-10 mol/L PTH for 30 min. Inhibitors of MAPK signaling pathway, PD98059 and U0126, noticeably inhibited the expression of CTGF mRNA and protein as well as gene promoters in HK-2 cells. Conclusion PTH can induce higher expression of CTGF in HK-2 cells probably via MAPK signaling pathway.

Objective To investigate the effect of parathyroid hormone (PTH) on the transition and connective tissue growth factor (CTGF) expression of human renal proximal tubular epithelial cell line HK-2 . Methods The expression of CTGF mRNA and protein of HK-2 cells were measured by real time RT-PCR and Western blot respectively . The effect of PTH on the phenotypic transformation of HK-2 cells was examined by light microscopy . The expression of α-smooth muscle actin (α-SMA) in HK-2 cells was detected by immunofluorescence . Results Basal level of CTGF mRNA and the protein expression were detected in HK-2 ceils . PTH upregulated the expression of CTGF mRNA and protein with the maximal response at the concentration of 10-10 mol/L and the best stimulating time was at 72 h . After exposure to PTH (10-10tool/L) for 12 hours, the highest level of luciferase activity was 1 .96 fold as compared to control (1 .888±0 .078 vs 0 .989±0 .030, P＜0 .01 ) . Untreated cells showed negligible expression of ±-SMA,whereas ±-SMA expression was significantly increased in cells treated with PTH . Conclusion PTH up-regulates CTGF expression and induces transition of HK-2 cells .