Mechanism of Artesunate in Reversing Epithelial-mesenchymal Transition and Akt /Snail Signaling Pathway in Colorectal Carcinoma / 中国实验方剂学杂志

Objective:To investigate the effects of artesunate （ART） on epithelial-mesenchymal transformation （EMT） of colorectal cancer HCT-8 cells，and explore the effects of ART on cell migration，invasion，EMT ability， and protein kinase B （Akt）/Snail signaling pathway of colorectal cancer. Method:3-（4-5-Dimethylthiazol-2-yl）2，5-diphenyltetrazolium bromide （MTT） assay was used to detect the effects of ART at different concentrations on the proliferation of HCT-8 cells. Wound healing assay and Transwell assay were used respectively to detect the effects of ART on migration and invasion of colorectal cancer cells. The effects of different concentrations of ART on the distribution of EMT-related proteins vimentin and E-cadherin in HCT-8 cells were detected by double-immunofluorescent staining. The effects of ART on protein expression levels of EMT markers E-cadherin，vimentin and N-cadherin in HCT-8 cells and the expression of Akt1， p-Akt1， and Snail1 in the Akt/Snail signaling pathway were determined by Western blot. Result:The dose-dependent inhibitory effects of ART on the proliferation of HCT-8 cells were determined and the inhibition rate was calculated. A dose-response curve was plotted accordingly. The half-maximal inhibitory concentration （IC₅₀） of ART on HCT-8 cells was （16.67±1.95） μmol·L^-1. The following four groups were set up： a control group （0 μmol·L^-1），and low-， medium-， and high-dose ART groups（2， 10， 50 μmol·L^-1）. Compared with the results in the control group，ART inhibited the migration and invasion of HCT-8 cells（<italic>P</italic><0.05）. Specifically， the expression of E-cadherin in HCT-8 cells was significantly up-regulated，and that of vimentin and N-cadherin was significantly down-regulated （<italic>P</italic><0.05）. The expression levels of p-Akt1 and Snail1 were significantly decreased after ART treatment，thus inhibiting EMT（<italic>P</italic><0.05）. Conclusion:The findings of this study suggested that ART inhibited the EMT-triggered migration and invasion of HCT-8 cells presumedly by inhibiting the activation of the Akt/Snail pathway to reverse EMT.

Effect of matrine on Fas, VEGF, and activities of telomerase of MCF-7 cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of matrine on Fas, VEGF, and activities of telomerase of MCF-7 cells.</p><p><b>METHODS</b>In vitro cultured human breast cancer MCF-7 cells were randomly divided into the experimental group and the control group. The matrine solution was added in cells of the experimental group. Equal volume of culture medium was added in cells of the control group or the negative control group. Zedoary Turmeric Oil, the telomerase inhibitor was added in cells of the positive control group. Morphological changes were observed under an inverted microscope. The telomerase activity was detected by TRAP-ELISA. Expressions of Fas and VEGF protein were detected by immunocytochemical assay.</p><p><b>RESULTS</b>Matrine obviously inhibited the growth and induced apoptosis of breast cancer cells. MCF-7 cells were treated by matrine of different concentrations at 24, 48, and 72 h, the telomerase activity gradually decreased along with increased matrine concentration and prolonged action time, showing dose-effect and time-effect positive relations. Matrine could up-regulate Fas protein expression and downregulate VEGF protein expression of MCF-7 cells.</p><p><b>CONCLUSION</b>Matrine showed obvious effect in inhibiting the growth of MCF-7 cells and promoting the apoptosis, which might be achieved by up-regulating the expression of Fas protein, inhibiting telomerase activity induced apoptosis of breast cancer cells, down-regulating the expression of VEGF protein, and inhibiting the tumor vascular formation.</p>

Effects of deguelin on the proliferation and apoptosis of human esophageal cancer cell Ec-109: an experimental research / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effects of deguelin on the apoptosis and proliferation of human esophageal cancer cell Ec-109, and to explore its possible mechanisms.</p><p><b>METHODS</b>Human esophageal cancer cells Ec-109 were in vitro cultured. They were divided into the blank control group, and 5, 10, 20, and 40 nmol/L deguelin groups. The inhibition on the proliferation was detected at 24, 48, and 72 h using CCK-8 assay. The early apoptosis rate at 24 h was detected by flow cytometry. The expressions of apoptosis-related proteins Bcl-2 and Bax were detected at 24 and 48 h respectively.</p><p><b>RESULTS</b>Compared with the blank control group at the same point, the growth inhibition rate in all deguelin groups increased at 24, 48, and 72 h, showing statistical difference (P <0.05). The early apoptosis rate was 4.37% +/- 0.35%, 6.71% +/-0.14%, 15.62% +/- 0.21%, and 19.78% +/- 0.15% in 5, 10, 20, and 40 nmol/L deguelin groups, respectively, showing statistical difference when compared with that of the blank control group (1.10% +/- 0.08%, P < 0.05). Compared with the blank control group, Bcl-2 protein expression obviously decreased, and Bax protein expression obviously increased in 10, 20, and 40 nmol/L deguelin groups, showing statistical difference (P <0.05). The aforesaid indices were in time- and dose-dependent manners.</p><p><b>CONCLUSION</b>Deguelin showed obvious effects on inhibiting the proliferation of Ec-109 cells and promoting their apoptosis, which was correlated with up-regulating Bax protein expression and down-regulating Bcl-2 protein expression.</p>

Scaling behavior of nucleotide cluster in DNA sequences / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

In this paper we study the scaling behavior of nucleotide cluster in 11 chromosomes of Encephalitozoon cuniculi Genome. The statistical distribution of nucleotide clusters for 11 chromosomes is characterized by the scaling behavior of P(S) proportional, variant e(-alphaS), where S represents nucleotide cluster size. The cluster-size distribution P(S(1)+S(2)) with the total size of sequential C-G cluster and A-T cluster S(1)+S(2) were also studied. P(S(1)+S(2)) follows exponential decay. There does not exist the case of large C-G cluster following large A-T cluster or large A-T cluster following large C-G cluster. We also discuss the relatively random walk length function L(n) and the local compositional complexity of nucleotide sequences based on a new model. These investigations may provide some insight into nucleotide cluster of DNA sequence.

Activity changes of protein kinase C in effect of interferon-gamma on wound healing and cicatrisation / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the roles of protein kinase C (PKC) in effect of interferon-gamma (IFN-gamma) on wound healing and cicatrization.</p><p><b>METHODS</b>IFN-gamma was applied on the wound and into the scar tissues of rabbit ear before or after wound healing. PKC activities in the tissues from 0, 3, 6 d, 11-16 d post-wounding and from 14, 30 and 45d post-epithelization were measured by phosphorus (32p) incorporation. The time of wound epithelization and scar changes were also observed.</p><p><b>RESULTS</b>The PKC activity in granulation tissue, wound margin tissue and scar tissue elevated obviously in comparing with that of normal skin (P < 0.01). IFN-gamma did not change PKC activity (P > 0.05). But it delayed the wound healing (P < 0.01) and inhibited scar hyperplasia (P <0.05).</p><p><b>CONCLUSIONS</b>PKC might not mediate the signal of IFN-gamma inhibiting the wound healing and scar hyperplasia. But PKC might be related to the wound healing and scar hyperplasia.</p>

A hemifacial transplantation model in hares / 中华整形外科杂志

<p><b>OBJECTIVE</b>To design an animal model to study the facial transplantation of allografts in rabbits.</p><p><b>METHODS</b>Livid blue rabbits and New Zealand white rabbits was applied as experiment animal, to harvest hemifacial composite-tissue flap based in the common external carotid artery with the branch of the external mandibular artery and auricularis magna artery, then allotransplantation was performed with the livid blue rabbits as recipient while new Zealand rabbits as donor, the immunosuppressive agent comprised ciclosporin, azamun and prednisone. 25 couples of rabbits were divided three groups. Group A, 5 couples of rabbits, no administered immunosuppressive agent and the artery anastomosis with end-to-end. Group B, 10 couples of rabbits, administered immunosuppressive agent and the artery anastomosis with end-to-end. Group C, 10 couples of rabbits, administered immunosuppressive agent and the artery anastomosis with end-to-side. Postoperative, to observe the survive ratio of animal and composite-tissue flap, verified the practicability of model further.</p><p><b>RESULTS</b>The blood supply of hemifacial composite-tissue flap is rich after allotransplantation. The survive ratio wasn't different with different procedure of the external carotid artery anastomosis.</p><p><b>CONCLUSIONS</b>This is a successful model of composite face flap transplantation in the rabbits.</p>

Study of Screening Differential Genes of Malignant Melanoma Using Gene Chip Expression Profiles / 中国康复理论与实践

@#ObjectiveTo identify differential genes of malignant melanoma using gene chip expression profiles.MethodsAgilent Human 1A OligoDNA was employed to find out difference in gene expression between malignant melanoma and nevus. The total RNA was isolated from two type tissues, labled the fluorescent to the probe, hybridized, washed and analyzed.ResultsAmong the 21073 target genes, 1596 genes were differentially expressed in malignant melanoma, including 733 genes up-regulated, and 863 down-regulated.ConclusionThe gene chip technique can screen genes that may be specifically expressed in malignant melanoma.

Experimental study of transplanted endothelial progenitor cells transfected with VEGF165 gene augment the survival volume of transplanted fat tissue / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of transplanting endothelial progenitor cells (EPCs) transfected with VEGF165 gene to free transplanted fat tissue for increasing neovascularization and the survival.</p><p><b>METHODS</b>EPCs isolated from human cord blood were cultured in vitro and identified by immunocytochemistry. After transfection by VEGF165 gene, the expression of VEGF was assessed using ELISA. Then EPCs with (VEGF gene transfection group) and without VEGF165 gene transfection (EPCs group) were transplanted to free transplanted fat tissue at 18 nude mice's back, and nine nude mice transplanted with free fat tissue were injected with M199 (control group). CM-DiI was used to trace the transplanted cells. The capillary density of transplanted fat tissue was detected by CD34 immunohistochemistry.</p><p><b>RESULTS</b>EPCs expressed cell markers CD34, KDR and CD133. After transfection, the expression of VEGF was positive. Transplanted EPCs survived and proliferated, and transplanted EPCs were incorporated into the capillary networks in the transplanted fat tissue. The percent of survival volume of transplanted fat tissue of VEGF gene transfection group was (96.2 +/- 8.6)%, significantly higher than that of the EPCs group [(75.3 +/- 6.8)%, P < 0.05) and M199 group [(40.2 +/- 2.5)%, P < 0.05). The capillary density of transplanted fat tissue of VEGF gene transfection group was significantly higher than those of the EPCs group and M199 group (P < 0.05).</p><p><b>CONCLUSIONS</b>EPCs from human cord blood can increase free transplanted fat tissue neovascularization and the survival volume, and the ability of promoting neovascularization of EPCs transfected with VEGF165 gene is more potent than EPCs alone.</p>

Statistical properties of nucleotide clusters in DNA sequences / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Using the complete genome of Plasmodium falciparum 3D7 which has 14 chromosomes as an example, we have examined the distribution functions for the amount of C or G and A or T consecutively and non-overlapping blocks of m bases in this system. The function P(S) about the number of the consecutive C-G or A-T content cluster conforms to the relation P(S) proportional, variante(-alphas); values of the scaling exponent alpha(CG) are much larger than alpha(AT); and alpha(AT) of 14 chromosomes are hardly changed, whereas alpha(CG) of 14 chromosomes have a number of fluctuations. We found maximum value of A-T cluster size is much larger than C-G, which implies the existence of large A-T cluster. Our study of the width function xi(m) of cluster C-G content showed that follows good power law xi(m) proportional, variantm(-gamma). The average gamma for 14 chromosomes is 0.931. These investigations provide some insight into the nucleotide clusters of DNA sequences, and help us understand other properties of DNA sequences.

Experiment of augmenting the survival areas of ischemic flap by transplanting endothelial progenitor cells / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the feasibility of transplanted endothelial progenitor cells to ischemic flap with increased neovascularization and augmented the survival areas.</p><p><b>METHODS</b>EPCs were isolated from human cord blood, cultured in vitro, identified by immunohistochemistry. Then EPCs were transplanted to ischemic flaps of 9 nude mice's back (experimental group), and 9 nude mice's back flaps was injected with M199(control group). And pedicle division time was 4 days after operation. CM-DiI was used to trace the transplanted cells. The blood perfusion of flaps was monitored by the laser Doppler flowetry, and the capillary density of flaps was detected by CD34 immunohistochemistry.</p><p><b>RESULTS</b>EPCs expressed cell markers CD34, KDR and CD133. Transplanted EPCs survived and was incorporated into the capillary networks in the ischemic flaps of nude mice. The percent of experimental group's flap survival area was (60.3 +/- 2.1)%, significantly higher than the control group[ (34.2 +/- 1.8)%, P < 0.05 ]. The blood perfusion, capillary density of flaps of experimental group was significantly higher than the control group (P < 0.05).</p><p><b>CONCLUSIONS</b>EPCs from human cord blood can increase ischemic flaps neovascularization and augment the survival areas.</p>

Signal roles of protein tyrosine kinase in transforming growth factor-beta1, or interferon-gamma regulated proliferation and collagen synthesis by fibroblasts from hypertrophic scar and normal dermis / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the signal roles of protein tyrosine kinase (PTK) on proliferation and collagen synthesis of fibroblasts derived from hypertrophic scar(HS-FB) and normal skin (NS-FB) by interferon-gamma (IFN-gamma) or transforming growth factor beta1 (TGF-beta1).</p><p><b>METHODS</b>HS-FB and NS-FB were cultured and passaged in Dulbecco's modified Eagle's medium(DMEM). The PTK activity in unstimulated or IFN-gamma or TGF-beta1-stimulated HS-FB and NS-FB (10,30,60 and 120 min) were assayed by phosphorus (32P) incorporation. Cell proliferation was determined with MTT stain. The type III procollagen was measured by radioimmunoassay.</p><p><b>RESULTS</b>TGF-beta1 did not change PTK activity but it increased predominately proliferation and collagen synthesis of HS-FB and NS-FB in time-dependent fashion. Genistein, an inhibitor of PTK, inhibited HS-FB and NS-FB to proliferate and synthesize collagen but it could not change the roles on proliferation and collagen synthesis by TGF-beta1. IFN-gamma activated transiently PTK (P < 0.05) and increased proliferation and collagen synthesis of both fibroblast (P < 0.05, at 30 min, 60 min). As the recovery of PTK activity, the proliferation and collagen synthesis were inhibited by IFN-gamma at 120 min. Furthermore, Genistein abrogated the transient increased roles and partly reversed the longterm inhibitory functions by IFN-gamma (P < 0.05) . There were no difference on PTK activity, proliferation and collagen synthesis between HS-FB and NS-FB.</p><p><b>CONCLUSIONS</b>PTK did not mediate the signal of TGF-beta1 but transduced the signal of transient increased roles of IFN-gamma. Inhibited or activated PTK might mediate the signal of decreasing or increasing proliferation and collagen synthesis of fibroblast.</p>

The establishment of the two dimensional gel electrophoresis (2-DE) map of hypertrophic scar epidermal cells and the display of the differentially expressed proteins / 中华整形外科杂志

<p><b>OBJECTIVE</b>To establish and optimize the two dimensional gel electrophoresis (2-DE) map of epidermal cells separated from the hypertrophic scar tissues so as to explore the function of the non cultured epidermal cells during the course of the formation of hypertrophic scar.</p><p><b>METHODS</b>To separate the epidermal cells from the scar tissues, the scar epidermis was digested with Dispase II and trypsin. The total protein of the cells was then extracted and separated with 2-DE and visualized with silver stain. Spots detection and matching were performed with Melaine 3.0 gels analyzing software. The results were then compared with the normal epidermal cells' 2-DE map coming from the Danish Center for Human Genome Research' s 2-DE PAGE Databases.</p><p><b>RESULTS</b>Nearly more than 600 protein spots were identified in the final optimized 2-DE map. We found out 24 differentially expressed proteins by comparing the difference in composition, shape or density of all the spots. In the 24 proteins, there are 8 up-regulated ones, 9 down-regulated ones, 4 disappeared ones and 3 newly founded ones.</p><p><b>CONCLUSIONS</b>The method of digesting the epidermis with Dispase II and trypsin to separate the epidermal cells to establish the 2-DE map is feasible and it made the further study on hypertrophic scar proteomics possible. The 24 differentially expressed proteins revealed that epidermal cells might play a role in the formation of hypertrophic scar.</p>

The design of the expanded flap for the repair of the temporal and cheek area / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the design of an expanded flap at the temporal and cheek area.</p><p><b>METHODS</b>The expanded flap was used for the repair of 619 temporal and cheek defects secondary to scar, nevus or hemangioma excision. In the frontal area, the rotational flap was usually used. For the repair of the cheek, the applied flap included the rotational, advanced, and transposition flap from the neck, as well as the pedicle flap from the thoracic area.</p><p><b>RESULTS</b>Eight thoracic-deltoid flaps had distal necrosis of 1 approximately 5 cm. Of them, 5 flaps were repositioned with subsequent good result; the other 3 flaps underwent skin grafting. The five facial expanded flaps showed distal necrosis of 0.5 approximately 1 cm. Of them, 4 flaps occurred delayed healing, 1 flap underwent skin grafting. Expander extrusion happened in 41 cases (6.62%), which resulted in deficiency of the expanded area. Satisfactory results were achieved in all the other cases.</p><p><b>CONCLUSIONS</b>According to our experience, careful design of the flap is very important for obtainingbetter surgical results and decreasing complications.</p>

Fabrication of a blood vessel scaffold with a combined polymer for tissue engineering / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the possibility to fabricate a blood vessel scaffold with a combined polymer for tissue engineering.</p><p><b>METHODS</b>A blood vessel scaffold was designed with a combined polymer composed of rabbit vascular smooth muscle cells(VSMCs), collagen and a non-spinning fabric mesh of polyglycolic acid (PGA). VSMCs were implanted into collagen gel and their growth was observed. The mixed solution of VSMCs and collagen was dropped into the tubular scaffold, followed by 7-day culturing.</p><p><b>RESULTS</b>VSMCs formed many prominences after culturing in gelatinous collagen for 3-4 hours. With cells extending, some cells became shuttle- or spindle-shaped. After VSMCs-collagen complex was implanted into the PGA mesh, most of VSMCs remained in the pore of PGA mesh with the formation of gelation. VSMCs could adhere to and grow on the PGA fiber.</p><p><b>CONCLUSION</b>The non-spinning PGA porous biodegradable material coated with collagen is a good carrier for VSMCs to adhere and grow.</p>