Objective:To introduce a method combining thread-burying in eyebrow and construction of double eyelid for treatment of upper eyelid cutis laxa in the middle-aged and young.Methods: We used 3-0 non-invasive thread for intradermal suture and fixed the eyebrow to the superciliary periost,then double eyelid construction was performed to remove the superfluous skin of upper eyelid in 23 patients with upper eyelid cutis laxa.Results: All the 23 cases obtained satisfactory clinical outcomes, with the upper eyelid cutis laxa obviously improved.Conclusion: Thread-burying in eyebrow combined with double eyelid construction is a simple and effective strategy for treatment of middle-aged and young patients with upper eyelid cutis laxa.

Objective To evaluate the use of the kite flap (als o known as subcutaneous pedicled V-Y advancement flap)for the repair of moderate -sized anterior lamellar skin defects of eyelid. Methods Since 1994, kite flaps have been used to repair anterior lamellar skin defects of eyelids in 14 patients. Six patients were male and eight female, with a mean age of 43 years (range 15-64 years). The diameter of the largest defect reache d 1/3 length of the eyelid. The flaps were designed adjacent to the defects and the incision line corresponding to natural winkle lines on the eyelid. We underm ined the flap under the orbicular muscle, and advanced the flap to cover the def ect. The incisions were closed with 7-0 suture. Results After operation, all flaps survived with primary healing. Follow-up for 3 to 12 months showed that all cases had achieved satisfactory cosmetic effects without secondary deformity. Conclusions It is a simple, safe, an d reliable method to use kite flap for the repair of moderate-sized anterior la mellar eyelid skin defects that are too large to achieve primary approximation. Because kite-flap covers the defect through a direct advancement way without ro tation and twist, there are no dog-ear deformitys present at the pedical area a nd the repaired area looks smoothly. Because of its widely based muscle pedicle which incorporates venous and lymphatic drainage over most of its length, the un desirable pigment deposition is avoided.

OBJECTIVEThis study aims to investigate the expression of connexin 43 (cx43) gene during early development in zebra fish and provide a foundation for further research of cx43 gene in tooth development.

METHODSTotal RNA was extracted within 72 h after fertilization of zebra fish embryos and then reversed transcribed to generate the cDNA library. The specific fragments of the cx43 gene were then cloned and connected to the PGEMT vector. After confirming the constructed plasmid, the corresponding RNA polymerase was chosen, and the digoxin-labeled anti-sense mRNA probe of cx43 was synthesized in vitro. The cx43 gene expression of zebra fish indifferent stages was carried out by in situ hybridization. The relationship of the cx43 gene expression and anatomy of the pharyngeal teeth were compared by alizarin red staining.

RESULTSThe mRNA antisense probe of cx43 was acquired. The positive signal of sepia was observed in the different stages of zebra fish pharyngeal teeth after fertilization. After fertilization for 9 days, the expression site of cx43 in situ hybridization was overlapped in accordance with the anatomical site of the pharyngeal teeth.

CONCLUSIONcx43 gene participates in tooth development and mineralization process and plays a crucial role in later mineralization.

Animals ; Connexin 43 ; Gene Expression ; Genetic Vectors ; In Situ Hybridization ; Odontogenesis ; Plasmids ; RNA, Messenger ; Tooth ; Zebrafish

Objective To investigate the relationship between truncating mutations of APC gene and sporadic colorectal cancer,and analyze the feasibility of non-radioactive protein truncation test (PTT) in the detection of mutations of APC gene.Methods Ninety-six samples of sporadic colorectal cancer tissues ( including 44 patients with colonic cancer and 52 patients with rectal cancer) were obtained from the General Hospital of Ningxia Medical University from September 2008 to September 2010.The mutation cluster region of the APC gene was screened using digital PTT labeled with fluorescent Lys-t-RNA,with a polymerase chain reaction fragment amplified from genomic DNA serving as a tenplate for in vitro translation.The occurrence of gene mutation was determined according to the emergence of truncated peptides.The mutation cluster region of the APC gene in 46 samples of colorectal cancer tissues was analyzed by direct sequencing.The detection rates of the 2 methods were compared by chi-square test.Results Thirteen (26％) truncated peptides were detected in the 50 samples of colorectal cancer tissues.The mutation type of 4 samples is nonsense mutation,which resulted in emergence of truncated gene products.Eleven (24％) truncated peptides were detected in the 46 samples of colorectal cancer tissues.There was no significant difference in the detection rates between PTT and direct sequencing ( x2 =0.033,P ＞ 0.05 ).Conclusions Truncating mutations of APC gene are common alterations in sporadic colorectal cancer in the Chinese.Digital PTT labeled with fluorescent Lys-t-RNA is rapid and high-sensitive in screening gene mutations.

Objective To study the mechanism of tear trough deformity and palabromalar groove deformity.Methods Four old cadavers (2 male,2 female,an average age of 67.2 years) with obvious tear trough deformity and palpbromalar groove deformity and 4 young cadavers (2 male,2 female,an average age of 23.5 years) without tear trough deformity and palpbromalar groove deformity were selected and dissected and histological observation were performed on lower eyelid and periorbital region.Results Compared to young specimens,the skin and orbicularis oculi muscle of old specimens were atrophy and relaxed.Tear trough deformity and palabromalar groove deformity overlaid the junction of thinner eyelid skin and thicker cheek skin.The superior border of the malar fat pad covered the junction of the palpebral and orbital portions of the orbicularis muscle,and correlated with the tear trough and palpbromalar groove,but the superior border of the malar fat pad in young cadavers was found above the tear trough and palpbromalar groove line.The orbicularis retaining ligament arose from the orbital rim and caudal to the junction of the palpebral and orbital portions of the orbicularis muscle,and it was relaxed in old group than that in young group.Conclusions Tear trough deformity and palabromalar groove deformity result from combination of age-related relaxation,atrophy and descent of layers of tissues.The orbital septal and the orbicularis retaining ligament prevent tissues from descending,which makes tear trough deformity and palabromalar groove deformity more visible.

OBJECTIVETo investigate the mechanism of retinoic acid (RA) signal in dental evolution, RA is used to explore the influence of the mechanism on neural crest's migration during the early stage of zebra fish embryos.

METHODSWe divided embryos of wild type and transgenic line zebra fish into three groups. 1 x 10(-7) to 6 x 10(-7) mol x L(-1) RA and 1 x 10(-7) mo x L(-1) 4-diethylaminobenzaldehyde (DEAB) were added into egg water at 24 hpf for 9 h. Dimethyl sulfoxid (DMSO) with the concentration was used as control group. Then, antisense probes of dlx2a, dlx2b, and barxl were formulated to perform whole-mount in situ hybridization to check the expressions of the genes in 48 hpf to 72 hpf embryos. We observed fluorescence of transgenic line in 4 dpf embryos.

RESULTSWe obtained three mRNA probes successfully. Compared with DMSO control group, a low concentration (1 x 10(-7) mol x L(-1)) of RA could up-regulate the expression of mRNA (barx1, dlx2a) in neural crest. Obvious migration trend was observed toward the pharyngeal arch in which teeth adhered. Transgenic fish had spreading fluorescence tendency in pharyngeal arch. However, a high concentration (4 x 10(-7) mol x L(-1)) of RA malformed the embryos and killed them after treatment. One third of the embryos of middle concentration (3 x 10(-7) mo x L(-1)) exhibited delayed development. DEAB resulted in neural crest dysplasia. The expression of barxl and dlx2a were suppressed, and the appearance of dlx2b in tooth was delayed.

CONCLUSIONRA signal pathway can regulate the progenitors of tooth by controlling the growth of the neural crest and manipulating tooth development

Animals ; Branchial Region ; Cell Differentiation ; drug effects ; Embryo, Nonmammalian ; drug effects ; embryology ; metabolism ; In Situ Hybridization ; Neural Crest ; drug effects ; Odontogenesis ; Signal Transduction ; Tooth ; drug effects ; embryology ; metabolism ; Tretinoin ; pharmacology ; Zebrafish ; embryology ; genetics ; metabolism

ZnO nanoparticle-containing carbon composite nanofiber ( ZnO-CNF ) was prepared by the electrospinning of the ZnCl2-PAN precursor, followed by preoxidation and carbonization. The ZnO nanoparticles were uniformly distributed on the surface of the carbon nanofiber with the size of 20-30 nm, confirmed by scanning electron microscopy ( SEM ) . The wettability of the ZnO-CNF was studied by water contact angle test. With Nafion as an additive, the ZnO-CNF modified electrode was successfully constructed by dip-coating. The surface morphology and electrochemical properties of the modified electrode were investigated by SEM and cyclic voltammetry. There was a sensitive response of the ZnO-CNF modified electrode on Pb ions in solution, demonstrated by square wave stripping voltammetry. Under the optimized conditions, a good linear relationship between peak current and Pb2+concentration was obtained in the range of 2. 4×10-10-2. 4×10-7 mol/L (R=0. 998) by 10 min preconcentration at -1. 0 V in 0. 1 mol/L NaAc buffer solution (pH=4. 6). The detection limit was 4. 8×10-11 mol/L. The practical analytical application of the ZnO-CNF modified electrode was assessed by the measurement of the actual water sample and the result was consistent with that obtained by ICP-MS.

Objective To study the mechanism of p55 inducing cell apoptosis, the 180 bp fragment of Puma promoter was cloned into the pGL3-basic luciferase reporter vector. The biological activity of Pumareporter plasmid was verified by cell transfection. Methods The target fragments of Puma were amplified by RT-PCR method and the fragments were inserted into the pGL3-basic luciferase reporter vector. The acquired Puma-Luc plasmid was transfected into H1299 cell line and detected its activity. Results Sequencing indicated that the amplified Puma promoter is correct. Dual-luciferase Reporter Assay showed the Puma-Luc constructs have promoter activity. Conclusion The cloning of human Puma gene promoter and the construction of its reporter vector were successful. This study will lay the foundation for further research on the function of p53 inducing apoptosis through mitochondrial pathway.

Objective To observe morphological characteristics and activity distribution of T. asahii biofilm. Methods The morphological characteristics of T. asahii biofilm were observed under an inverted microscope and scanning electron microscope, and activity was measured and quantitatively analyzed by 2,3-bis (2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazo-lium hydroxide (XTT) assay and viable count, respectively. Spatial distribution of dead/vital cells, activity and thickness of biofilm at different layers were assessed under a confocal laser scanning microscope (CLSM) following double staining with FDA/PI. Results T. asahii formed a biofilm in vitro on the surface of polystyrene materials. Under a scanning microscope, the biofilm displayed a complex three-dimensional structure which composed of spores, pseudohy-pha and true hypha. As time prolonged, the activity and quantity of biofilm increased. The results of XTT assay were correlated with those of viable count (r = 0.94, P < 0.01). The activity was of no obvious difference between different layers of the biofilm. The thickness of biofilm varied from 14.3 μm to 31 μm. Conclusions The structure of T. asahii biofilm in vitro is more complex than that of planktonic T. asahii. The activity is of no significant difference between different layers of T. asahii biofilm.

Objective To analyze neuraminidase(NA) inhibitor resistance of seasonal H1N1 influenza A viruses isolated in Shenzhen during 2008 to 2009. Methods The NA gene of these viruses were sequenced. Phylogenetic analysis of the sequences was performed with Mega3. 1 software. Results In 2008, most isolates of the seasonal H1 N1 virus were susceptible to neuraminidase inhibitors, but the H275Y mutation in the neuraminidase gene region associated with high-level oseltamivir resistance had been detected in 92.6% of the strains isolated in 2009. Furthermore, a strain with Q136K was found, which showed the resistance to Zanamivir. Conclusion In the light of emerging resistance, close monitoring and understanding of the nature and dynamics of resistance mutations in influenza virus should be a priority.