Occurrence of Breast Mucinous Carcinoma After Autologous Fat Grating for Breast Augmentation.

UNLABELLED: Autologous fat grafting is commonly performed in reconstructive breast surgery as well as in aesthetic breast augmentation surgery. Nevertheless, little is known about the interaction between fat grafts and cancer. A 36-year-old patient had undergone bilateral breast augmentation with autologous fat grafting. Two months after surgery, she perceived two small palpable indurations in the right breast. Nine months after the procedure, the lumps grew bigger and lumpectomy was performed. Histologic examination of the specimens showed mucinous carcinoma of the breast. This case raises once again the question about the possible links between breast cancer and fat grafts. The level of evidence is level V. NO LEVEL ASSIGNED: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266.

Application of Botulinum Toxin at the Yonsei Point for the Treatment of Gummy Smile: A Randomized Controlled Trial.

BACKGROUND: Demand for less-invasive procedures for treating gummy smile, such as botulinum toxin A injections, has increased substantially over the years. Meanwhile, the optimal injection site for botulinum toxin A injection is debated. The authors aimed to investigate the efficacy of botulinum toxin A injection at the Yonsei point for treating gummy smile. METHODS: In this double-blind, single-site, randomized clinical trial, healthy participants with a gummy smile (anterior gingival exposure of ≥3.0 mm) were enrolled and randomized (1:1 ratio) into two groups. The experimental group was administered 6 U of botulinum toxin A at the Yonsei point (a single-site injection of 3 U to the right Yonsei point and 3 U to the left Yonsei point), and the control group received the same dose in the bilateral levator labii superioris alaeque nasi muscle sites. The patients were assessed at baseline and 4, 12, 24, and 48 weeks after the first injection using a digital vernier caliper. RESULTS: A total of 49 participants were enrolled. Anterior and bilateral posterior gingival exposure were reduced at 4, 12, and 24 weeks ( P ≤ 0.05) and returned to baseline at 48 weeks in both groups; there was no difference between the groups at these time points. The increase in satisfaction among patients was significant, and few adverse events were observed. CONCLUSION: Both the Yonsei point and the levator labii superioris alaeque nasi muscle site can be used as botulinum toxin A injection sites for treating gummy smile. CLINICAL QUESTION/LEVEL OF EVIDENCE: Therapeutic, I.

A p-toluenesulfonamide-modified benzo[h]chromene hydrazone: Fluorescent turn-on detection of hypochlorite and its application to imaging the endoplasmic reticula of living cells and zebrafishes.

Hypochlorite (ClO-) is a ROS that plays a crucial role in the immune system in the body. As the largest organelle in the cell, the endoplasmic reticulum (ER) manages various life activities. Thus, a simple hydrazone-based probe was designed, which provided a fast turn-on fluorescent response toward ClO-. With a terminal p-toluenesulfonamide group as the endoplasmic reticulum (ER)-specific site, probe 1 was mainly accumulated at ER of living cells, and could be used for imaging endogenous and exogenous HClO in cells and zebrafishes.

[Application of magnetic materials in analysis on Chinese herb medicines].

China is the cradle of Chinese herb medicines,with rich plant resources. However, traditional processing methods have many disadvantages, such as high comsumption of organic solvent, long extraction time and high loss of effective constituents. For the purpose of rational use of Chinese herb medicines and accurate analysis on their constituents,the sample pre-treatment method with magnetic nanoparticles as the carrier brought new opportunities in recent years. after consulting literatures,the essay summarizes traditional extraction methods of Chinese herb medicines, characteristics of magnetic materials and their application in the analysis on Chinese herb medicines.

A ratiometric fluorescent probe for hydrogen sulfide in neat aqueous solution and its application in lysosome-targetable cell imaging.

Hydrogen sulfide (H2S) has been recently regarded as one of the most important gasotransmitters in the metabolic system, while abnormal H2S concentration is associated with various diseases. Although numerous fluorescent probes have been developed for the detection of cellular H2S, only a few of them can monitor lysosomal H2S with ratiometric fluorescent output. Here, we developed a water-soluble probe 1 toward H2S by introducing 2,4-dinitrophenyl ether into a novel merocyanine-based dye. As expected, H2S induced an obvious red-shift of the probe from 520 nm to 580 nm in neat aqueous solution, and this fluorescent ratiometric response is highly selective and sensitive (with the detection limit of 0.81 nM), rapid (within 10 s), and effective in a wide pH range (2.0-10.0). In particular, the probe was successfully applied for tracing H2S in the lysosomes of living cells and in zebrafish.

Simultaneous detection of lysosomal SO2 and viscosity using a hemicyanine-based fluorescent probe.

The changes in sulfur dioxide and viscosity of lysosomes are significant indicators in physiological processes and the cell microenvironment. This study aimed to synthesize a hemicyanine-based probe for simultaneous detection of SO2 and viscosity. The probe could not only rationally detect sulfur dioxide in a semi-aqueous solution with high sensitivity (limit of detection = 0.78 µM) and fast response (within 30 s) but also monitor viscosity via fluorescence emission enhancement at 580 nm. Further, the dual-response probe was successfully used to image SO2 and viscosity in the lysosomes of living cells.

A quinoline-based probe for the ratiometric fluorescent detection of sulfite in lysosomes of living cells.

A lysosome-targeting ratiometric fluorescent probe was synthesized for detecting sulfite based on sulfite-triggered nucleophilic addition reaction. Due to the specific reaction, the fluorescence intensity ratio (I530/I390) of the probe in an almost aqueous solution (0.5% DMSO) changed significantly after the addition of HSO3-, corresponding to the change in the fluorescence color of the solution from green to blue. The recognition was conducted using high-resolution mass spectrometry, proton nuclear magnetic resonance, and density functional theory calculations. The fluorescent probe could be utilized to quantitatively monitor HSO3- in lysosomes of living C6 glioma cells and real-water samples.

An AIRE-active far-red ratiometric fluorescent chemosensor for specifically sensing Zn2+ and resultant Zn2+ complex for subsequent pyrophosphate detection in almost pure aqueous media.

A simple Schiff-base fluorescent chemosensor (1) was synthesized by the reaction of 3-amino-pyrazine-2-carbohydrazide and 7-diethylamino-3-formylcoumarin; the sensor 1 displayed a notable green emission at 524 nm in DMSO and an aggregation-induced ratiometric emission (AIRE) at 555 nm in an almost buffered aqueous media (0.5% DMSO content). The AIRE of 1 was quenched following binding to Zn2+ ions, while the fluorescence emission in the far-red region was evidently enhanced at 628 nm. Notably, the ratiometric signal output could be utilized to specifically distinguish Zn2+ among various metal ions. Moreover, the 1-Zn2+ complex was effectively employed as a fluorescent ratiometric chemosensor for pyrophosphate (PPi) detection. The detection limit was 3.52 µM and 2.45 µM for Zn2+ and PPi, respectively. The binding mechanism was evaluated by 1H NMR, ESI-MS, single-crystal X-ray diffraction, TEM, time-resolved fluorescence spectrophotometry, and density functional theory studies. Overall, owing to its sensitive fluorescence behavior, cell imaging studies demonstrated that this sensor is capable of sensing Zn2+ and PPi in living cells.

[Expression of rat annexin 5 and its effect on human sperm motility in vitro].

OBJECTIVE: Gonadotropin releasing hormones (GnRH) regulate the expression of annexin 5 in Leydig cells, and annexin 5 is supposed to be a signal molecule in regulating testosterone secretion. This study aimed to investigate the function of annexin 5 in male reproduction by observing its effect on human sperm motility in vitro. METHODS: The encoding sequence of rat annexin 5 was chemically synthesized and inserted into the HIS fusion expression vector pET28a. The expression of the fusion protein HIS-annexin 5 was induced by isopropyl-beta-D-thiogalactoside (IPTG) under the control of the T7 promoter, and the products were purified by affinity chromatography. The anticoagulant activity of annexin 5 was determined by the modified activated partial thromboplastin time (APTT) test. Semen samples from 15 donors were assigned to a control and an annexin 5 group, the latter treated with recombinant annexin 5 at the concentration of 10(-8) mol/L. Sperm motility and the percentage of grade a + b sperm were measured by computer-assisted semen analysis (CASA) after 20 and 60 min exposure, and the sperm ascending experiment was done after 20 min treatment. RESULTS: The product of the synthesized target gene was 947 bp in length, and the inserted sequence corresponded to the published encoding sequence of rat annexin 5. The plasmid pET28a-annexin 5 was transformed into E. coli BL21(DE3) and IPTG induced a fusion protein with a relative molecular weight of about 36,000, a purity of 95% and a high anticoagulant activity. Compared with the control group, sperm motility and the percentage of grade a + b sperm were increased by 40% (P < 0.01) and 21% (P < 0.01), respectively, after 20 min treatment with annexin 5, but neither showed any significant improvement after 60 min. The sperm ascending altitude was remarkably elevated after annexin 5 treatment, with extremely significant difference from the control group (37.84 +/- 6.35 vs. 49.5 +/- 12.27, P < 0.01). CONCLUSION: An annexin 5 recombinant expression vector was successfully constructed. The protein annexin 5 can be efficiently expressed in E. coli and effectively improve human sperm motility in vitro.

[Effects of GnRH analogues on MAPK pathway in rat Leydig cells].

OBJECTIVE: To investigate the effects of GnRH analogues GnRHa and GnRHant on the MAPK pathway in rat Leydig cells. METHODS: Rat Leydig cells were primarily cultured for 24 hours in vitro and serum-starved for 2 hours, followed by treatment with GnRHa (10(-7) mol/L) or GnRHant (10(-6) mol/L) for 0, 5, 15, 30, 60 and 90 minutes, with the 0 min group as the control. Then the protein levels of phosphorylated ERK (p-ERK) and phosphorylated p38 (p-p38) were detected by Western blot, and that of p-ERK determined by the same means after co-incubation of GnRHa or GnRHant with the PKC inhibitor GF109203X at 1, 5, 10 and 20 micromol/L. RESULTS: After stimulation of the Leydig cells with GnRHa or GnRHant for different times, the protein level of p-p38 showed no significant difference from that of the control group (P > 0.05). Then the Leydig cells were treated with GF109203X at different concentrations for 20 minutes and with addition of GnRHa for another 10 minutes. The level of p-ERK was significantly decreased (P < 0.05) by GF109203X at 10 and 20 micromol/L. Compared with the control, the p-ERK expression was increased by 65% at 15 minutes (P < 0.05) in the GnRHant stimulation group, by 81% (to the peak) at 30 minutes (P < 0.05), began to fall at 60 minutes, and returned to the base level at 90 minutes. The p-ERK level exhibited no significant difference from that of the control (P > 0.05) after treatment of the Leydig cells with different concentrations of GF109203X for 20 minutes and then with GnRHant for 30 minutes. CONCLUSION: The ERK MAPK activation induced by GnRHa depends on the PKC pathway, but not that induced by GnRHant. The p-38 MAPK pathway may not be involved in the effect of GnRH analogues on rat Leydig cells.

Cloning, sequence and functional analysis of goat ATP-binding cassette transporter G2 (ABCG2).

ATP-binding cassette transporter G2 (ABCG2) gene encodes a protein that has a wide variety of substrates and is responsible for the active secretion of clinically and toxicologically important molecules into milk. Although known in many species, this marks the first time this gene product has been reported in goats. In this study, we cloned and sequenced goat ABCG2 gene complete coding sequence and predicted its putative translated protein structure with implicative functional domains. One six-transmembrane span on C-terminal region and at least one coiled-coil domain on N-terminal were predicted and compared primarily with those of other closely related species. In addition, three conserved cysteines (in positions 595, 606, and 611) were determined toward the C-terminal of goat's ABCG2. Two known functional motifs were identified in goat's protein through comparative studies with other species. The goat ABCG2 relative expression profile revealed that the gene expression was a function of lactation stage and parallel to goat lactation curve.

Construction and characterization of a goat mammary gland cDNA library.

A lactating goat mammary gland cDNA library was constructed by using a modified commercially available cDNA library construction kit protocol. The resulting clones were sequenced and functionally analyzed through cross-species genomic comparison to assess (1) the capacity and functional quality of the constructed library for subsequent research and (2) the efficiency of the procedural modifications. The study resulted in the construction of a high-quality mammary gland cDNA library, which was characterized by (1) the total recombinants number of 1.4 x 10(7) colony-forming units (cfus) that was at least 10 times greater than the number expected from the application of the standard kit protocol, (2) the recombinants rate of 96%, and (3) the average insert size of 1,082 bp. BLAST analysis of sequenced clones against GenBank databases determined 55.7% of clone redundancy, 22 known function gene clusters, and 29 novel gene clusters. The analysis of the primary gene expression profile showed that 59% of the tested clones were genes that coded for milk proteins while 16% of the clones coded for ribosomal, metabolism, immune response, and translation proteins. The remaining 25% of the tested clones were described as novel genes. Cross-species comparison showed that 77% of characterized gene clusters were successfully identified by using resources from other ruminants and unrelated species. This outcome is in consonance with the common belief that the genomic resources that have been generated across species are potentially powerful tools that could be used for enhancing the molecular understanding of less genomically studied species, such as goat.

Decellularized aorta of fetal pigs as a potential scaffold for small diameter tissue engineered vascular graft.

BACKGROUND: For cardiovascular tissue engineering, acellularized biomaterials from pig have been widely investigated. Our purpose was to study mechanical properties and biocompatibility of decellularized aorta of fetal pigs (DAFP) to determine its potential as scaffold for small diameter tissue engineered vascular graft. METHODS: Descending aorta of fetal pigs was removed cells using trypsin, ribonuclease and desoxyribonuclease. Mechanical properties of DAFP were evaluated by tensile stress-strain and burst pressure analysis. Assessment of cell adhesion and compatibility was conducted by seeding porcine aortic endothelial cells. To evaluate biocompatibility in vivo, DAFP was implanted subcutaneously into adult male Sprague Dawley rats for 2, 4 and 8 weeks. RESULTS: Histochemistry and scanning electron microscopy examination of DAFP revealed well-preserved extracellular matrix proteins and porous three-dimensional structures. Compared with fresh aorta, DAFP had similar ultimate tensile strength, axial compliance and burst pressure. Cell culture studies in vitro showed that porcine aortic endothelial cells adhered and proliferated on the surfaces of DAFP with excellent cell viability. Subdermal implantation demonstrated that the DAFP did not show almost any immunological reaction and exhibited minimal calcification during the whole follow-up period. CONCLUSION: The DAFP has the potential to serve as scaffolds for small diameter tissue engineered vascular graft.

Subtraction CT angiography for the detection of intracranial aneurysms: A meta-analysis.

The aim of this meta-analysis was to investigate the accuracy of subtraction computed tomography angiography (CTA) for diagnosing intracranial aneurysms. A systematic literature search up to January 1, 2013 was performed in PubMed. Two independent reviewers selected 8 studies that compared subtraction CTA with digital subtraction angiography. Data from the studies were used to construct a 2×2 contingency table on a per-patient basis in ≥5 diseased and 5 non-diseased patients, with additional data on a per-aneurysm basis. Overall, subtraction CTA had a pooled sensitivity of 99% [95% confidence interval (CI), 95-100%] and specificity of 94% (95% CI, 86-97%) for detecting and ruling out cerebral aneurysms, respectively, on a per-patient basis. On a per-aneurysm basis, the pooled sensitivity was 96% (95% CI, 90-99%), and the specificity was 91% (95% CI, 85-95%). In conclusion, subtraction CTA is a highly sensitive, specific and non-invasive method for the diagnosis and evaluation of intracranial aneurysms.

Inferring the Skeletal Muscle Developmental Changes of Grazing and Barn-Fed Goats from Gene Expression Data.

Thirty-six Xiangdong black goats were used to investigate age-related mRNA and protein expression levels of some genes related to skeletal muscle structural proteins, MRFs and MEF2 family, and skeletal muscle fiber type and composition during skeletal muscle growth under grazing (G) and barn-fed (BF) feeding systems. Goats were slaughtered at six time points selected to reflect developmental changes of skeletal muscle during nonrumination (days 0, 7, and 14), transition (day 42), and rumination phases (days 56 and 70). It was observed that the number of type IIx in the longissimus dorsi was increased quickly while numbers of type IIa and IIb decreased slightly, indicating that these genes were coordinated during the rapid growth and development stages of skeletal muscle. No gene expression was affected (P > 0.05) by feeding system except Myf5 and Myf6. Protein expressions of MYOZ3 and MEF2C were affected (P < 0.05) by age, whereas PGC-1α was linearly decreased in the G group, and only MYOZ3 protein was affected (P < 0.001) by feeding system. Moreover, it was found that PGC-1α and MEF2C proteins may interact with each other in promoting muscle growth. The current results indicate that (1) skeletal muscle growth during days 0-70 after birth is mainly myofiber hypertrophy and differentiation, (2) weaning affects the expression of relevant genes of skeletal muscle structural proteins, skeletal muscle growth, and skeletal muscle fiber type and composition, and (3) nutrition or feeding regimen mainly influences the expression of skeletal muscle growth genes.

[Experimental study of compatibility between chondrocytes and allogenic cartilage microparticle acellular tissue matris in vitro].

OBJECTIVE: To construct tissue engineered cartilage using cartilage microparticle acellular tissue matrix (CMACTM) as scaffold. METHODS: To determine the content of hydroxyproline, glycosaminoglycan and DNA of CMACTM prepared from sheep's articular cartilage with multistep enzymic method, and to analyze CMACTM with gross observation, histology and scanning electron microscopy. Allogenic chondrocytes were mixed with CMACTM and cultured in vitro from 0 to 35 days. Observations through inverted microscope, scanning and transmission electron microscope, quantifications of hydroxyproline, glycosaminoglycan and DNA in the composite, cells adhesion rate were applied to analyze the results. RESULTS: The diameter of CMACTM was 0.100-0.154 mm, which contain extracellular matrix only. Hydroxyproline, glycosaminoglycan and DNA quantifications in CMACTM were 204.374 +/- 3.120 microg/mg, 18.302 +/- 2.037 microg/mg and 0.042 +/- 0.013 microg/mg respectively. Allogenic chondrocytes enclosed CMACTM tightly, hydroxyproline, glycosaminoglycan and DNA quantifications in the composite of the two formers increased with difference on 7th day compared with that on 0 day, reached to the peaks on 14th day (hydroxyproline, DNA) and on 21st day (glycosaminoglycan), and retained at a high level on the following days. Cells adhesion rate was 92%. CONCLUSION: Allogenic CMACTM possessed satisfactory biocompatibility for chondrocytes and provided a new scaffold for cartilage tissue engineering.

[Management after expanded flap ulceration during auricular reconstruction].

OBJECTIVE: To investigate the feasibility of Brent I methods for total auricular reconstruction when expanded flap ulceration happened. METHODS: The expanded flaps in the retroauricular region ulcerated during total auricular reconstruction in 8 patients with microtia. Then the expanders were removed and autologous rib cartilage frameworks were implanted. Brent I techniques for the total auricular reconstruction were employed. RESULTS: All the wounds in 8 patients with microtia healed primarily. The expanded flaps survived completely. The reconstructed ears had good shape and appearance with little hair. The size, shape and position of reconstructed ears were coordinated with the face. CONCLUSIONS: Brent I technique is an alternative method for total auricular reconstruction when the expanded flap ulceration occurs during total ear reconstruction.

Bilateral congenital choanal atresia and osteoma of ethmoid sinus with supernumerary nostril: a case report and review of the literature.

INTRODUCTION: Congenital choanal atresia is a relatively rare deformity, especially bilateral congenital choanal atresia. We report a case of bilateral congenital choanal atresia in a 22-year-old Chinese man, who was also diagnosed with congenital right accessory nasal deformity, osteoma of his left ethmoid sinus and congenital keratoleukoma of his right eye. CASE PRESENTATION: A 22-year-old Chinese man presented with mouth breathing, sleep snoring and difficult feeding after birth, with no olfactory sensation. Three-dimensional computed tomography revealed bilateral choanal atresia and a high density bony shadow in his left ethmoid sinus that extended to his left frontal sinus. CONCLUSIONS: Choanal atresia is often accompanied by other congenital abnormalities. To the best of our knowledge, this is the first report of choanal atresia accompanied by congenital accessory nasal deformity and congenital keratoleukoma.

Effect of tea catechins on regulation of antioxidant enzyme expression in H2O2-induced skeletal muscle cells of goat in vitro.

Skeletal muscle cells (SMCs) of goats were stress induced with 1 mM H(2)O(2) in the absence or presence of 0.5, 5, and 50 µg/mL tea catechins (TCs) incubation. Cells were harvested at 48 h postincubation with TCs to investigate the effects of TCs on cell proliferation, cell membrane integrity, antioxidant enzyme activities, and antioxidant enzyme genes and protein expression levels. Results showed that H(2)O(2) induction inhibited cell proliferation with or without TC incubation; moreover, the inhibition effect was enhanced in the presence of TCs (P < 0.001). H(2)O(2)-induced stress increased the lactate dehydrogenase (LDH) activity in the absence or presence of TC incubation, but concentrations of TCs, less than 5 µg/mL, showed protective functions against LDH leakage than in other H(2)O(2)-induced treatments. The catalase (CAT) activity increased when SMCs were stress induced with H(2)O(2) in the absence or presence of TC incubation (P < 0.001). H(2)O(2)-induced stress decreased CuZn superoxide dismutase (CuZn-SOD) and glutathione peroxidase (GPx) activities, whereas this effect was prevented by incubation with TCs in a concentration-dependent manner. H(2)O(2)-induced stress with or without TC incubation had significant effects on mRNA and protein expression levels of CAT, CuZn-SOD, and GPx (P < 0.001). CAT and CuZn-SOD mRNA expression levels were increased by different concentrations of TC incubation, and this tendency was basically consistent with corresponding protein expression levels. The GPx mRNA expression level increased with a low concentration of TCs but decreased with concentrations greater than 5 µg/mL of TCs, whereas GPx protein expression in all TC-incubated groups was lower than in the control treatment. The current findings imply that TCs had an inhibitory effect on cell proliferation and enhanced damage to the cell membrane integrity, but TCs affected antioxidant status in SMCs by modulating antioxidant enzyme activities at mRNA and protein expression levels.

Effects of rice straw particle size on digesta particle size distribution, nitrogen metabolism, blood biochemical parameters, microbial amino acid composition and intestinal amino acid digestibility in goats.

Effects of rice straw particle size and physically effective neutral detergent fiber (peNDF) on particle size distribution of different digestive tract, nitrogen (N) metabolism, blood biochemical parameters, microbial amino acid (AA) composition and intestinal AA digestibility in goats were investigated. A 4 × 4 Latin square design was employed using four mature Liuyang black goats fitted with permanent ruminal, duodenal, and terminal ileal fistulae. During each of the four periods, goats were offered one of four diets that were similar in chemical composition, but varied in particle sizes and peNDF through alteration of the theoretical cut length of rice straw (10, 20, 40 and 80 mm, respectively). Dietary peNDF contents of four diets were 17.4, 20.9, 22.5 and 25.4%, respectively. Results showed that increasing particle size of rice straw and dietary peNDF significantly affected the particle size distributions of digesta in rumen, duodenum and ileum, except feces. However, increasing particle size of rice straw and peNDF did not affected N metabolism in goats, except the increased apparent N digestibility in rumen and large intestine, and the decreased apparent N digestibility in small intestine. Furthermore, increasing particle size of rice straw and peNDF showed little influence on the profile of blood biochemical parameters, microbial AA composition and intestinal AA digestibility in goats.