Objective. To investigate the effect of mechanical tensile load on the proliferation of cells in rat facial connective tissue. Methods. Nine male Sprague-Dawley rats were randomized into control group (A), acupuncture without rotation group (B) and tensile stimulation group (C). The rats in group C were subjected to mechanical tensile stimulation of the facial connective tissue in the groin area with an acupuncture needle, which was rotated after insertion to cause tissue stretching. And in group B, needle was inserted into the rats' tissues without rotation. 5-bromo-2' deoxyuridine incorporation assay was used to label the proliferative cells after the treatments. Results. Tensile stimulation caused a significant enhancement of the cell proliferation activity in the fascia, whereas acupuncture did not produce significant changes in the cell proliferation as compared with group A. Tensile stimulation also resulted in the loss of normal structure of the inguinal fat pad, where numerous blood vessels and hemorrhagic foci were observed. Conclusion. Mechanical tensile stimulation can significantly enhance cell proliferation in the fascia. The mechanism underlying the therapeutic effect of complement therapies may involve the reconstruction of the loose connective tissue and enhancement of the cell proliferative activity under tensile load.

The role of FEN1 genetic variants on gallstone and gallbladder cancer susceptibility is unknown. FEN1 SNPs were genotyped using the polymerase chain reaction-restriction fragment length polymorphism method in blood samples from 341 gallbladder cancer patients and 339 healthy controls. The distribution of FEN1-69G > A genotypes among controls (AA, 20.6%; GA, 47.2% and GG 32.2%) was significantly different from that among gallbladder cancer cases (AA, 11.1%; GA, 48.1% and GG, 40.8%), significantly increased association with gallbladder cancer was observed for subjects with both the FEN1-69G > A GA (OR = 1.73, 95% CI = 1.01–2.63) and the FEN1-69G > A GG (OR = 2.29, 95% CI = 1.31–3.9). The distribution of FEN1 -4150T genotypes among controls (TT, 21.8%;GT, 49.3% and GG 28.9%) was significantly different from that among gallbladder cancer cases (TT, 12.9%; GT, 48.4% and GG 38.7%), significantly increased association with gallbladder cancer was observed for subjects with both the FEN1-4150T GT(OR = 1.93, 95% CI = 1.04–2.91) and the FEN1-4150T GG(OR = 2.56, 95% CI = 1.37–5.39). A significant trend towards increased association with gallbladder cancer was observed with potentially higher-risk FEN1-69G > A genotypes (P < 0.001, χ2 trend test) and FEN14150G > T (P < 0.001, χ2 trend test) in gallstone presence but not in gallstone absence (P = 0.81, P = 0.89, respectively). In conclusion, this study revealed firstly that FEN1 polymorphisms and haplotypes are associated with gallbladder cancer risk.

Cells that morphologically and functionally resemble male germ cells can be derived from bone marrow stem cells. However, autologous bone marrow procurement has potential limitations. Thus, in the present study, germ cell-associated genes including Oct4, Dazl, Nobox, Piwil and Rnf17 were examined in differentiating ADSCs in the presence of 10(-5) RA for 14 days, 21 days and 10(-6) RA for 14 days, 21 days and 28 days. The effect of all-trans retinoic acid (RA) which is a known inducer of primordial germ cell (PGC) proliferation/survival in vitro was also determined. Oct4, Dazl and Nobox showed the highest expression in the presence of 10(-6) mol/L AR after 21 days RA treatment, the three genes showed the same expression pattern. While Piwil showed the highest expression in the presence of 10(-6) mol/L AR after 28 days RA treatment, Rnf17 showed no expression at any time point. Our findings establish a comprehensive profile of male germ cell gene expression during differentiation of rat ADSCs and describe the capacity of RA to stimulate the expression of these genes. Furthermore, these data represent an important first step in designing a plausible directed differentiation protocol for male germ cells.

In this study, we investigated structural changes in alpha-glucosidase during urea denaturation. Alpha-glucosidase was inactivated by urea in a dose-dependent manner. The inactivation was a first-order reaction with a monophase process. Urea inhibited alpha-glucosidase in a mixed-type reaction. We found that an increase in the hydrophobic surface of this enzyme induced by urea resulted in aggregation caused by unstable folding intermediates. We also simulated the docking between alpha-glucosidase and urea. The docking simulation suggested that several residues, namely THR9, TRP14, LYS15, THR287, ALA289, ASP338, SER339, and TRP340, interact with urea. Our study provides insights into the alpha-glucosidase unfolding pathway and 3D structure of alpha-glucosidase.

We studied the inhibitory effects of trifluoroethanol (TFE) on the activity and conformation of tyrosinase. TFE increased the degree of secondary structure of tyrosinase, which directly resulted in enzyme inactivation. A reciprocal study showed that TFE inhibited tyrosinase in a slope-parabolic mixed-type inhibition manner (K-I=0.5 +/- 0.096 M). Time-interval kinetic studies showed that the inhibition was best described as first order with biphasic processes. Intrinsic and 1-anilinonaphthalene-8-sulfonate-binding fluorescences were also measured to gain more insight into the supposed structural changes; these showed that TFE induced a conspicuous tertiary structural change in tyrosinase by exposing hydrophobic surfaces. We also predicted the tertiary structure of tyrosinase and simulated its docking with TFE. The docking simulation was successful with significant scores (binding energy for Autodock4=-4.75 kcal/mol; for Dock6=-23.07 kcal/mol) and suggested that the TRP173 residue was mainly responsible for the interaction with TFE. Our results provide insight into the structure of tyrosinase and allow us to describe a new inhibition strategy that works by inducing conformational changes rather than targeting the active site of the protein.