转染ANGPTL-1基因对人皮肤成纤维细胞胶原表达的影响

目的探讨重组ANGPTL-1基因转染人皮肤成纤维细胞的可行性,以及转染转染后对细胞增殖速率和Ⅰ、Ⅲ型胶原合成的影响。方法体外重组ANGPTL-1基因,借助真核表达载体系统,转入体外培养的人皮肤成纤维细胞,流式细胞仪检测基因表达及细胞转染效率;RT-PCR比较转染前后ANGPTL-1 mRNA及Ⅰ、Ⅲ型胶原mRNA的表达。结果基因转染后,(63±7.1)%的被转染细胞表达目的基因;转染组ANGPTL-1 mRNA相对表达量较未转染组和转染空载体组明显增高(P<0.01,n=5);转染组Ⅰ、Ⅲ型胶原mRNA相对表达量较未转染组和转染空载体组均明显增高(P<0.01,n=5)。结论人皮肤成纤维细胞可作为ANGPTL-1转染的靶细胞,ANGPTL-1基因可能是促进Ⅰ、Ⅲ型胶原合成的重要基因之一。

Skin care efficacy study of recombinant humanized collagen based on in vitro level

Studying the skin care efficacy of recombinant humanized collagen based on in vitro level.The stability of the recombinant humanized collagen was first analyzed by treating at different temperatures,then its skincare efficacy based on in vitro level was evaluated by detecting the inhibition rate of elastase,the inhibition rate of collagenase,the protein content of type I collagen in human fibroblasts,the inhibition of reactive oxygen species(ROS)with human keratinocytes,and the effects of the recombinant humanized collagen on the expression of hyaluronic acid(HA),filaggrin(FLG)and transglutaminase 1(TGM1)in keratinocytes.The results showed that recombinant humanized collagen was able to maintain stability at temperatures below 70℃.With regard to its skincare efficacy,recombinant humanized collagen could inhibit elastase and collagenase activities and promote the increase of type I collagen content in human fibroblasts.It also showed good inhibition of ROS in keratinocytes in vitro and could increase the expression of HA,FLG,and TGM1 in keratinocytes.In short,the recombinant humanized collagen exhibited a favourable skin care effect in vitro level.This study proved that it has potential firming,anti-wrinkle,moisturizing,and repairing efficacy,and is a valuable cosmetic raw material.

Real-Time Face Tracking and Recognition in Video Sequence

A framework of real time face tracking and recognition is presented, which integrates skin color based tracking and PCA/BPNN (principle component analysis/back propagation neural network) hybrid recognition techniques. The algorithm is able to track the human face against a complex background and also works well when temporary occlusion occurs. We also obtain a very high recognition rate by averaging a number of samples over a long image sequence. The proposed approach has been successfully tested by many experiments, and can operate at 20 frames/s on an 800 MHz PC.

Effects of Tremella Polysaccharide on Immune Function of Physically-Stressed Mice

An immunosuppressive animal model induced by physical stress that forced mice to swim in cold water(14±1℃)and the restorative effect of Tremella polysac charide(TP)on the suppressed immune function by stress were studied in mice.It was found that the spleen plaque forming cell(PFC)response to sheep red blood cells,delayed cuta- neous hypersensitivity(DCH)induced by dinitrochlorobenzene and the lymphocyte prolifer- ation stimulated by concanavalin A(Con A)were significantly decreased in stressed mice. In addition.the maximal decrease of PFC was reached in 9-12 days after stress.A- drenolectomy could not affect the decrease of PFC in stressed mice.TP(200.400mg/kg) ig for 8-14 days significantly restored the PFC.DCH and lymphocyte proliferation to nor- mal level in stressed mice.

Silk Fibroin-modified Poly(butadiene) urethane Films and Their Effects on Fibroblast Viability

Surface-modified poly(butadiene)urethane (PBTU) films with silk fibroin (SF) were prepared by simple chemical method under the normal temperature. The physical properties and biological behaviour of the SF-modified PBTU film were evaluated. The results showed that the SF-modified PBTU films kept the tenacity and pliability very well, and could overcome rigid and brittle weaks of silk fibroin films. The morphology of SF in the PBTU film was dendritic aggregations, and the water-contact angle measurement indicated that the surface hydrophilicity of modified films was apparently enhanced. The biocompatibility of PBTU films was improved due to the change of surface components. The degree of platelet adhesion and the cell viability of rat embryo dermal fibroblasts seeded on PBTU films, SF films, and SF-modified PBTU films were measured by counting platelets before and after they contacted the films and MTT assay, respectively. The results indicated that platelet adhesion resistance and cell viability on the modified film were greatly superior to those on the PBTU film and the compound interface had good stability in the air.

Incorporation of Bromelain into Dermatological Bases： Accelerated Stability Studies

This work aimed the incorporation of the enzyme bromelain from pineapple peel into dermatological bases for potential therapeutic application. The enzyme was incorporated into Lanette cream and lotion, as well as Carbopol gel and Chemyunion cream and lotion, at a concentration of 0.5% （w/w）. These bases were subjected to centrifugation test and accelerated stability test during 90 days at 25 ℃（with and without sunlight）, 37 ℃ and 4 ℃, in order to evaluate bromelain stability in dermatological bases. Total protein content and enzymatic activity were measured. The bases were stable as its organoleptic characteristics （appearance, color, smell and sensitivity to touch） only when kept at 4 ℃ with activity remaining 95.5%, 84,9%. 77.7%, 73.8% and 72.3%, alter 90 days of testing in Carbopol gel, Lanette and Chemyunion cream, and Lanette and Chemyunion lotion, respectively. Based on the results, it was possible to incorporate bromelain into dermatological bases, and the activity was well preserved when these bases were kept in refrigerator at 4 ℃.

Matrine inhibits proliferation of mouse skin fibroblasts induced by platelet-derived growth factor-BB

Objective:To studythe effectof matrineon proliferationof mouseskinfibroblastsinducedby platelet-derivedgrowthfactor-BB(PDGF-BB).Methods :MouseskinfibroblastswereobtainedfromnewbornI-CRmiceandpropagatedin vitro.Proliferationof cellwasanalyzedby mitochondrialreductionof tetrazoliumsalt MTT andactualcellcount.Re sults:Matrine(50to500μg/ml)causeddose-dependentreductionof serum-stimu-latedcellgrowth.Growthinhibitionwas totallyreversedafterremovalof the drug.Matrinealso inhibited PDGF-BBinducedcellgrowthdose-dependently.Conclusion:Matrineexhibitspotentanti-proliferationeffecton mouseskinfibroblast.Thiseffectappearsto be mediatedby decreaseof PDGF-inducedgrowth.Theseresultssug-gestthatmatrinemighthavepreventiveandtherapeuticimplicationin skinfibrosis.

Intracolonic multiple pebbles in young adults:Radiographic imaging and conventional approach to a case

Most of the foreign bodies detected in adult gastroin- testinal systems are accidentally swallowed pins. In this study, we presented a case with intracolonic multiple pebbles. A 20-year-old man was admitted to emergency surgery policlinic for abdominal pain for 2 d without any alleviation or aggravation. His upright plain abdominal radiographic imaging revealed about 30-40 overt dense opacities in lumen of colonic segments, with oval and well shaped contours, each approximately 1 cm × 1 cm in size. The multiplanar reconstructions and three- dimensional images combined with sectional screening showed that all pebbles had passed completely into the colon and no foreign bodies had remained in the ileal segments. On psychiatric assessment, he was found to have immature personality features, difficulty in over- coming stressors and adaptation disorder. He recovered by conservative management and radiographic monitoring applied during his follow-up. Thus, it can be concluded that, in differential diagnosis of abdominal pain in adult ages, though less frequently seen than in children, gastrointestinal system foreign bodies should always be kept in mind and it should be considered that ingestion of pebbles may be one of the factors contributing to abdominal pain particularly in young adults with psychiatric problems. In such cases suspected of having foreign bodies which cannot be detected by plain films, abdominal tomography can be an alternative for diagnostic imaging.

Mechanistic Insights of Cells in Porous Scaffolds via Integrated Culture Technologies

This research aimed to combine 3 cell and tissue culture technologies to obtain mechanistic insights of cells in porous scaffolds. When cultivated on 2D （2-dimensional） surfaces, HDFs （human dermal fibroblasts） behaved individually and had no strict requirement on seeding density for proliferation; while HaCat cells relied heavily on initial densities for proliferation and colony formation, which was facilitated when co-cultured with HDFs. Experiments using a 3D CCIS （3-dimensional cell culture and imaging system） indicated that HDFs colonised openpores of varying sizes （125-420 ~tm） on modular substrates via bridge structures; while HaCat cells formed aperture structures and only colonised small pores （125 txm）. When co-cultured, HDFs not only facilitated HaCat attachment on the substrates, but also coordinated with HaCat cells to colonise open pores of varying sizes via bridge and aperture structures. Based on these observations, a 2-stage strategy for the culture of HDFs and HaCat cells on porous scaffolds was proposed and applied successfully on a cellulosic scaffold. This research demonstrated that cell colonisation in scaffolds was dependent on multiple factors; while the integrated 2D＆3D culture technologies and the 3D CCIS was an effective and efficient approach to obtain mechanistic insights of their influences on tissue regeneration.

OVER-EXPRESSION OF A TRUNCATED TYPE I TGF-P RECEPTOR IN NORMAL DERMAL FIBROBLASTS DECREASES TGF-β1 AUTOPRODUCTION

Objective To determine over-expression of a truncated type ⅡTGF-β receptor in down-regulating TGF-β1 auto production in normal dermal fibroblasts. Methods In vitro cultured dermal fibroblasts were treated with rhTGF-β1 (5ng/ml) or recombinant adenovirus containing α truncated type Ⅱ TGF-β receptor gene (50 pfu/cell). Their effects on regulating gene expression of TGF-β1 were observed with Northern Blot. Results rh TGF-β1 up-regulated the gene expression of TGF-β1, (34 %-150%) and type Ⅰ pro-collagen( 13 %- 190%). Overexpression of a truncated receptor Ⅱ decreased the gene expression of TGF-β1 (53%-66%). Conclusion Over-expression of the truncated TGF-β receptor Ⅱdown-regulated TGF-β1 autoproduction via blocking signal transduction of TGF-β. This study may provide a new strategy for scar gene therapy.

Isolation and Characterization of Collagen from Squid(Ommastrephes bartrami) Skin

Collagen of squid （Ommastrephes bartrami） skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen （ASC） and pepsin-solubilized collagen （PSC） were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type 1 collagen, containing α1 and α2 chains. FTIR （fourier transform infrared spectroscopy） investigations confirmed the existence of helical arrangements in PSC of squid skin. The denaturation temperature （Td） and shrinkage temperature （T5） of PSC were 29.4℃ and 52.8℃, respectively.

Distribution and ultrastructure of pigment cells in the skins of normal and albino adult turbot, Scophthalmus Maximus

The distribution and ultrastructure of pigment cells in skins of normal and albino adult turbots were examined with transmission electron microscopy （TEM）. Three types of pigment cells of melanophore, iridophore and xanthophore have been recognized in adult turbot skins. The skin color depends mainly on the amount and distribution of melanophore and iridophore, as xanthophore is quite rare. No pigment cells can be found in the epidermis of the skins. In the pigmented ocular skin of the turbot, melanophore and iridophore are usually co-localized in the dermis. This is quite different from the distribution in larvae skin. In albino and white blind skins of adult turbots, however, only iridophore monolayer still exists, while the melanophore monolayer disappears. This cytological evidence explains why the albino adult turbot, unlike its larvae, could never resume its body color no matter what enviroumental and nutritional conditions were provided. Endocytosis is quite active in the cellular membrane of the iridophore. This might be related to the formation of reflective platelet and stability of the iridophore.

Under the Skin. Intangible Components as the Basic Realities of Contemporary Post-bureaucratic Organizations： A Sociological Perspective

This article attempts to analyze, through a re-reading of the present sociological literature on the organization （that explores the organizations themselves in terms of ＂tacit＂ rather than ＂explicit＂, ＂socio-cultural systems＂ rather than ＂socio-technical systems＂）, the main intangible components that may affect the organizational live and all the organizing in general for the good or the evil, with respect to the ＂success＂ rather than the ＂failure＂ of the ＂in and out＂ organizational acting. There are then some sorts of ＂basic assumptions＂, deemed indispensable, also because they are still there and live in organizations but they do not always have a careful importance. Therefore the objective of the article is to describe them highlighting their salience and attempt to define parameters for further in-depth studies.

Treatment of Cough and Dyspnea due to Acute Bronchitis by Plaster for Cough and Dyspnea——A Report of 735 Cases

In the light of the theory of treating the internal disease externally, an externally used plaster for treating cough and dyspnea due to acute bronchitis (Ke Chuan Yi Tie Kang [symbol: see text]-[symbol: see text]) was successfully applied to 735 cases of acute bronchitis (the treatment group), with the other 423 cases treated with routine western drugs as controls. The results showed that the cure rate in the treatment group was significantly higher than that in the control group (P

Application of GlycoProtein IIb/IIIa antagonist( Integrilin) in peri-percutaneous transluminal catheter angioplasty and stent implantation in Chinese

Objective: to report the application of GPIIb/IIIa antagonist (Integrilin) in peri percutaneous transluminal catheter angioplasty and stent implantation in Chinese. Method: Twenty one patients who underwent percutaneous transluminal catheter angioplasty(PTCA) and stent implantation were included. After arterial puncture, integrilin was injected at dose of 180ug/kg, and then maintained at 2μg/kg·min for 18 hours. Asprine, plavix(clopidogrel) and heparin were used at the same time. ACT, PT, KPTT and blood routine were routinely monitored. Results: All sheaths were drawn out 2 to 4 hours after the procedure.. There was no severe complication such as hematoma, acute and subacute thrombosis in coronary artery, or thrombocytopenia. ACT returned to less than 150 seconds in 2 hours in 15 patients; in 4 hours in 6 patients. There was no significant difference between the pre and post procedure value of PT and platelet count. KPTT was significantly higher than pre procedure value at 2 hours after the procedure .No recurrence of angina pectoris was observed in the first nine patients within one year follow up, and no restenosis occurred in stents in the five patients who had coronary angiography one year later. Conclusion: Application of GPIIb/IIIa receptor antagonist (Integrilin) in peri percutaneous transluminal catheter angioplasty and stent implantation in combination with aspirin and plavix could significantly reduce the dosage and duration of heparin with benefit of shortening the indwelling time of sheaths, but did not increase risk of bleeding or lead to thrombosis in stent.

Ex vivo differentiation of multipotent adult progenitor cells to skin epidermal cells

Objective： By establishing the indirect contact co-culture system, we studied the in vitro condition for MAPCs differentiating into epidermal cells and the transformation of MAPCs into epidermal cell phenotype. Methods： Cell culture insert membrane was used for substitute basal membrane and MAPCs, fibroblast cells （FCs） and mixture of MAPCs and epidermal cells and FCs were separately implanted into 2 sides of it. PKH26 was used to label cloned MAPCs; type IV collagen rapid adhering method was used to isolate and culture the skin epidermal cells from l-day-old SD rat. Results： Part of the MAPCs transformed into cells expressing keratin in the presence of peripheral epithelia and FCs. Type Ⅳ collagen rapid adhering method successfully selected rats＇ epidermal stem cells. The mixture of the 2 kinds of cells or indirect culture might promote the differentiation through mesenchymal factors secreted by dermis FC. Conclusion： We were the first to have established the in vitro model of MAPCs differentiation into epidermal cells, in which MAPCs were transformed into epithelium-like cells.

Cytotoxicity and non-specific cellular uptake of bare and surface-modified upconversion nanoparticles in human skin cells

The cytotoxicity and non-specific cellular uptake of the most popular composition of upconversion nanoparticle （UCNP）, NaYF4：Yb^3＋：Er^3＋, is reported using normal human skin cells, including dermal fibroblasts and immortalized human epidermal linear keratinocytes （HaCaT）. A new hydrophilization reaction of as-synthesized UCNPs based on tetramethylammonium hydroxide （TMAH） enabled evaluation of the intrinsic cytotoxicity of bare UCNPs. The cytotoxicity effects of the UCNP surface-coating and polystyrene host were investigated over the concentration range 62.5-125 μg/mL with 24-h incubation, using a MTT test and optical microscopy. The fibroblast viability was not compromised by UCNPs, whereas the viability of keratinocytes varied from 52% ± 4% to 100% ± 10% than the control group, depending on the surface modification. Bare UCNPs reduced the keratinocyte viability to 76% ± 3%, while exhibiting profound non-specific cellular uptake. Hydrophilic poly（D,L-lactide）- and poly（maleic anhydride-alt-l-octadecene）-coated UCNPs were found to be least cytotoxic among the polymer-coated UCNPs, and were readily internalized by human skin cells. Polystyrene microbeads impregnated with UCNPs remained nontoxic. Surprisingly, no correlation was found between UCNP cytotoxicity and the internalization level in cells, although the latter ranged broadly from 0.03% to 59%, benchmarked against 100% uptake level of TMAH-UCNPs.

Analysis of differentially expressed genes in fetal skin of scarless and scar-forming periods of gestational rats

Objective ：To study the differences of gene expression between earlier gestational skin and later gestational skin of rats with the aids of single primer amplification （SPA） and high-density oligonucleotide DNA array to understand the molecular mechanism of scarless healing. Methods： Total RNAs were isolated from fetal rat skin of the scarless（E15） and scar-forming （ E18 ） periods of gestation （term = 21.5 days）. The RNAs from earlier gestational skin （ EGS ） and later gestational skin （ LGS ） were both reversely transcribed to cDNAs, then labeled with the incorporation of fluorescent dCTP for preparing the hybridization probes by SPA method. The mixed probes were then hybridized to the oligonucleotide DNA arrays which contained 5 705 probes representing 5 705 rat genes. After highly stringent washing, these DNA arrays were scanned for fluorescent signals to display the differentially expressed genes between the 2 groups of skin. Results. Among 5 705 rat genes, there were 53 genes （0.93%） with differentially expressed levels between EGS and LGS groups, 27 genes, including fibroblast growth factor 2 （ FGF2 ） and follistatin were up-regulated （0.47%） and 26 genes were down-regulated （0.46%） in fetal skin during scarless period versus scar-forming period. Higher expressions of FGF2 and follistatin in EGS than those in LGS were also revealed by RT-PCR method. Conclusions： High-density oligonucleotide DNA array provided a powerful tool for investigating differential gene expression in earlier and later gestational fetal skins. This technology validates that the mechanism of fetal scarless healing is very complicate and the change of many gene expressions is associated with fetal scarless healing.

Construction of the subtractive cDNA library of injured adult and fetal rabbit skins

Objective: Early gestational mammalian fetuses possess the amazing ability to heal cutaneous wounds in a scarless fashion. Over the past years, scientists have been working to decipher the mechanisms underlying this regenerative repair. The remarkable phenotypic differences between fetal and adult healings behoves us to learn their characteristics in genetics, which represents potentially important mechanisms involved in wound repair observed in fetal versus adult tissues. In this sense, it is reasonable to construct subtractive cDNA library for future research. Methods: Middle laparotomy and hysterotomy were performed on pregnant rabbits at 20 day gestation to expose the fetal back, and a longitudinal incision through the skin was made on the back of the fetus. The traumatized fetal skin was harvested 12 hours post operation, the fetus control and traumatized adult skin specimens were taken at the same time. dscDNA was synthesized from total RNA of skin samples with SMART technology. Taking one of the three samples as Tester respectively and the other two as Drivers, we obtained 1 forward and 2 reverse hybridization products. After being amplified with selective polymerase chain reaction, the products were inserted into a vector, and then transferred into E.coli HB101. The colonies were screened afterwards. Results: The wounded fetuses were alive for a long time even after birth. Every determinant step, such as RNA isolation, cDNA synthesis, Rsa I digestion, adaptor ligation and hybridization, was well operated. Subtractive efficiency identification demonstrated that the suppression subtractive hybridization (SSH) was successful. Insertion into vector and transferring to E.coli were satisfactory. Conclusions: Instead of classic SSH, an improved SSH with 2 Drivers was applied for the experiment. Results confirmed that the improved program was reasonable and correct in both theory and practice. The subtractive cDNA library we have obtained is going to be used for future researches to reveal scarless healing related gene(s) and its (their) expression.

Intramedullary nailing of clavicular midshaft fractures in adults using titanium elastic nail

Objective： Studies showed elastic stable intramedullary nailing （ESIN） of displaced midclavicular fractures has excellent outcomes, as well as high complication rates and specific problems. The aim was to discuss ESIN of midshaft clavicular fractures.Methods： Totally 60 eligible patients （aged 18-63 years） were randomized to either ESIN group or non-operative group between January 2007 and May 2008. Clavicular shortening was measured after trauma and osseous consolidation.Radiographic union and complications were assessed. Function analysis including Constant shoulder scores and disabilities of the arm, shoulder and hand （DASH） scores were performed after a 15-month follow-up.Results： ESIN led to a signifcantly shorter time to union, especially for simple fractures. In ESIN group, all patients got fracture union, of which 5 cases had medial skin irritation and 1 patient needed revision surgery because of implant failure. In the nonoperative group, there were 3 nonunion cases and 2 symptomatic malunions developed requiring corrective osteotomy. At 15 months after intramedullary stabilization, patients in the ESIN group were more satisfied with the appearance of the shoulder and overall outcome, and they benefited a lot from the great improvement of post-traumatic clavicular shortening. Furthermore,DASH scores were lower and Constant scores were significantly higher in contrast to the non-operative group.Conclusion： ESIN is a safe minimally invasive surgical technique with lower complication rate, faster return to daily activities, excellent cosmetic and better functional results,restoration of clavicular length for treating mid-shaft clavicular fractures, resulting in high overall satisfaction, which can be regard as an alternative to plate fixation or nonoperative treatment of mid-shaft clavicular fractures.