Cornea is the major refractive components of the eye. As a viscoelastic tissue, cornea exhibits complicated biomechanical properties: non - linear elasticity, anisotropy and viscoelasticity. The biomechanical properties play an important role in keeping the normal structureand function. Changes in biomechanical properties are always earlier than the clinical symptoms. So quantitative measurement of the biomechanical properties benefits the early diagnosis and treatment of diseases. Different methods to measure the biomechanical properties of cornea were reviewed in detail, including classic ex vivo destructive tests, commercially available in vivo measuring methods and other emerging methods with the potential for clinical application but not validated for in vivo measurement. The operating principles, advantages as well as limitations of these methods were also described.

Objective · To observe the clinical outcomes of a modified laterally moved and coronally advanced flap combined with a connective tissue graft (CTG) for the treatment of severe Miller class Ⅱ or class Ⅲ isolated recession defects. Methods · Three patients with initial defect depths of more than 5 mm and malposition in some teeth were enrolled and underwent a modified laterally moved and coronally advanced flap combined with CTG. Recession depth (RD), keratinized tissue height (KTH) of both donor and adopted site, pocket depth (PD), and clinical attachment loss (CAL) at baseline and follow-up one-year after treatment were documented. Root coverage rate (RC) was calculated and visual analogue scale (VAS) was used to evaluate patient's satisfaction degree. Results · The mean RDs at baseline and followup were (5.3±0.5) mm and (0.3±0.5) mm. The mean RC at follow-up was (93.3±9.4)% and two cases had complete root coverage. The KTHs at adopted and donor sites were (0.3±0.5) mm and (6.0±0.8) mm at baseline and (4.3±0.5) mm and (5.7±1.3) mm at follow-up, respectively. PD and CAL were decreased from (1.7±0.5) mm and (7.0±0.8) mm at baseline to (1.3±0.5) mm and (1.3±1.2) mm at follow-up, respectively. The VAS value was 9.0±0.8 and subjective evaluation of patients was improved significantly at one-year follow-up, including root sensitivity and aesthetics. Conclusion · The modified laterally moved and coronally advanced flap with CTG has ideal clinical outcomes and satisfaction degree for the treatment of patients with severe recession defects that lack keratinized tissue and combine with buccal malposition.

Objective The presence of alloreactive memory T cells in recipient is a critical handicap to achieve transplantation tolerance.To make a mouse model which mimics the present transplant patient is important for research at this subject.Thus,we developed a novel re-transplant model and compared the alloresponse in this model with that in the conventional memory T cellstransfer model (transfer control).Methods The re-transplant model was established via microsurgery and vessel cannula techniques,and the experiment was composed of three groups:the re- transplant group,memory T cell-transfer group (transfer control) and the conventional blank group (blank control).The research indexes included survival time of donor heart,rejection score of allograft,and detection of proliferation and differentiation of the alloreactive memory/effector T cells by by flow cytometry (FCM) and in vitro mixed lymphocyte reaction (MLR).Results The median survival time of allograft in re-transplant recipients was significantly shortened compared to that of transfer control,but there was no significant difference in rejection score of graft between them (the score in retransplant group was the most intense of the three groups). Moreover, proliferation and differentiation of the alloreactive effector T cells were more intensive in re- transplant recipients than in the transfer control,which was confirmed by in vitro MLR and by FCM of the splenocytes for detecting CD44highCD62L-memory/effector phenotype cells.Conclusion The recall alloresponse in retransplantation is more intensive than that in memory-transfer setting and this re-transplant model is more close to the clinic situation than the memory-transfer model in rodents.

Objective To identify the genotypes of ESBLs-producing Klebsiella pneumoniae isolates from the First Affiliated Hospital,Shantou University Medical College.Methods The MICs of 10 antibiotics were determined by agar-dilution against the clinical isolates of ESBLs-producing K.pneumoniae.PCR were performed with specific primers for blaTEM,blaSHV, blaCTX-M and blaOXA respectively.PCR products were cloned and sequenced.Results The results of PCR showed that a- mong the 83 strains of ESBLs-producing K.pneumoniae,75 were positive for blaTEM,41 positive for blaSHV,25 poitive for blaCTX-M,9 positive for hlaOXA.Three genotypes were found in 13 strains(15.7%),2 genotypes in 59 strains (71.1%) and single genotype in only 11 strains(13.2%).The genes of CTX-M-3,TEM-1 and SHV were found co-existent in 9 strains. The strains carrying 2 or 3 ESBL genes were more resistant to antibiotics than those carrying only 1 ESBL gene.Conclusions The genotypes of ESBLs-producing Klebsiella pneumoniae in this hospital are blaTEM,blaSHV,blaCTX-M and blaOXA. Most strains carry 2 or 3 ESBL genes.

OBJECTIVETo establish a microemulsion liquid chromatography system with direct sample loading for determining the serum level of emodin in rats.

METHODSThe separation was performed on C₁₈ column (Hypersil BDS, 5 µm,150 mm×4.6 mm) with the microemulsion mobile phase consisting of 3.3% (w/V) SDS, 6.6% (V/V) n-butyl alcohol, and 1.0% (V/V) octane and water. The flow rate was 1.0 ml/min and the detection wavelength was 254 nm.

RESULTSThe linear range of emodin detection was 0.333-5.32 µg/ml. The average recovery was 99.65% with a RSD of 3.60%. The limit of quantification was 0.1386 µg/mL.

CONCLUSIONMicroemulsion liquid chromatography system with direct sample loading allows simple, accurate and rapid determination of emodin in rat serum.

Animals ; Chromatography, Liquid ; methods ; Emodin ; blood ; Male ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry

GL-PP-3A, an active polysaccharide peptide, was isolated and purified from Ganoderma lucidum, and then its structure was analyzed. Crude polysaccharide peptides were extracted from Ganoderma lucidum with hot water, precipitated with ethanol and then dialyzed from Ganoderma lucidum. Subsequently GL-PP-3A was isolated and purified from the crude polysaccharide peptides by fractional precipitation and chromatography of Bio-Gel P-10 column. The repetitive unit of GL-PP-3A was analyzed by high performance gel permeation chromatography (HPGPC), monosaccharide composition and methylation analysis, 1H NMR and 13C NMR. GL-PP-3A is a heteropolysaccharide which is composed mainly of glucose (Glc), and also contains saccharide residues such as rhamnose (Rha), xylose (Xyl), mannose (Man) and galactose (Gal) and 17 kinds of amino acids. Its weight-average molecular weight (Mw) and number-average molecular weight (Mn) were 1.7 x 10(4) and 1.1 x 10(4), respectively, with the ratio of Mw/Mn ( molecular weight distribution) being of 1.49. Its backbone chain is composed of 1,6-linked beta-D-Glcp and 1,3-liked beta-D-Glcp at a ratio of 2:1. Some of 1,6-linked glucose residuals of the backbone chain are substituted at 2-0 or 3-0, and there are 1 to 3 1,6-linked beta-D-Galp or 1,3-linked alpha-D-Manp in the branched chains, the nonreducing ends of which consist mainly of beta-D-Glcp and a few Rha.
Amino Acids ; analysis ; Chromatography, High Pressure Liquid ; Glucose ; analysis ; Magnetic Resonance Spectroscopy ; Molecular Weight ; Plants, Medicinal ; chemistry ; Polysaccharides ; chemistry ; isolation & purification ; Proteoglycans ; chemistry ; isolation & purification ; Reishi ; chemistry ; Rhamnose ; analysis ; Spectrophotometry, Ultraviolet

OBJECTIVETo investigate the protective effects of quercetin on cadmium-induced cytotoxicity in primary cultures of rat proximal tubular (rPT) cells.

METHODSPrimary cultures of rPT cells undergoing exponential growth were incubated with 1.0 μg/mL quercetin and/or cadmium (2.5, 5.0 μmol/L), in a serum-free medium at 37 °C at different time intervals. Commercial kits were used and flow cytometric analyses were performed on rPT cell cultures to assay apoptosis and oxidative stress.

RESULTSExposure of rPT cells to cadmium acetate (2.5, 5.0 µmol/L) induced a decrease in cell viability, caused an increase in apoptotic rate and apoptotic morphological changes. Simultaneously, elevation of intracellular reactive oxygen species, malondialdehyde and calcium levels, depletion of mitochondrial membrane potential and intracellular glutathione, and inhibition of Na+, K+-ATPase, Ca2+-ATPase, glutathione peroxidase (GSH-Px), catalase (CAT), and superoxide dismutase (SOD) activities were revealed during the cadmium exposure of rPT cells. However, simultaneous supplementation with 1 µg/mL quercetin protected rPT cells against cadmium-induced cytotoxicity through inhibiting apoptosis, attenuating lipid peroxidation, renewing mitochondrial function and elevating the intracellular antioxidants (non-enzymatic and enzymic) levels.

CONCLUSIONThe present study has suggested that quercetin, as a widely distributed dietary antioxidant, contributes potentially to prevent cadmium-induced cytotoxicity in rPT cells.

Animals ; Antioxidants ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Cadmium ; toxicity ; Cadmium Poisoning ; prevention & control ; Calcium ; metabolism ; Calcium-Transporting ATPases ; metabolism ; Cells, Cultured ; Kidney Tubules, Proximal ; drug effects ; metabolism ; Malondialdehyde ; metabolism ; Membrane Potential, Mitochondrial ; drug effects ; Quercetin ; pharmacology ; therapeutic use ; Rats ; Reactive Oxygen Species ; metabolism ; Sodium-Potassium-Exchanging ATPase ; metabolism

Sinisan is a widely used traditional Chinese medicine (TCM) in treating various diseases; however, the in vivo metabolic profile of its multiple components remains unknown. In this paper, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to identify the metabolites of Sinisan extract in rat plasma, urine, feces and bile after intragastric administration. Using MS(E) and mass defect filter techniques, 41 metabolites of 10 parent compounds (naringin, naringenin, hesperidin, neohesperidin, liquiritin, liquiritigenin, glycyrrhizic acid, glycyrrhetinic acid, saikosaponin a and saikosaponin d) were detected and tentatively identified. It was shown by our results that these compounds was metabolized to the forms of hydroxylation, glucuronidation, sulfation, glucuronidation with sulfation and glucuronidation with hydroxylation in vivo.
Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Flavanones ; analysis ; metabolism ; pharmacokinetics ; Glucosides ; analysis ; metabolism ; pharmacokinetics ; Glycyrrhizic Acid ; analysis ; metabolism ; pharmacokinetics ; Hesperidin ; analogs & derivatives ; analysis ; metabolism ; pharmacokinetics ; Hydroxylation ; Male ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVE Non-alcoholic fatty liver disease(NAFLD) encompasses a series of patho-logic changes ranging from steatosis to steatohepatitis,which may progress to cirrhosis and hepatocel-lular carcinoma.The purpose of this study was to determine whether Ganoderma lucidum polysaccha-ride peptide (GLPP) has therapeutic effect on NAFLD. METHODS ob/ob mouse model and ApoC3 transgenic mouse model were used for exploring the effect of GLPP on NAFLD. Key metabolic path-ways and enzymes were identified by metabolomics combining with KEGG and PIUmet analyses and key enzymes were detected by Western blotting. Hepatosteatosis models of HepG2 cells and primary hepatocytes were used to further confirm the therapeutic effect of GLPP on NAFLD. RESULTS GLPP administrated for a month alleviated hepatosteatosis, dyslipidemia, liver dysfunction and liver insulin resistance. Pathways of glycerophospholipid metabolism, fatty acid metabolism and primary bile acid biosynthesis were involved in the therapeutic effect of GLPP on NAFLD. Detection of key enzymes revealed that GLPP reversed low expression of CYP7A1,CYP8B1,FXR,SHP and high expression of FGFR4 in ob/ob mice and ApoC3 mice. Besides, GLPP inhibited fatty acid synthesis by reducing the expression of SREBP1c, FAS and ACC via a FXR-SHP dependent mechanism. Additionally, GLPP reduced the accumulation of lipid droplets and the content of TG in HepG2 cells and primary hepato-cytes induced by oleic acid and palmitic acid. CONCLUSION GLPP significantly improves NAFLD via regulating bile acid synthesis dependent on FXR-SHP/FGF pathway, which finally inhibits fatty acid synthesis,indicating that GLPP might be developed as a therapeutic drug for NAFLD.

BACKGROUNDFolic acid is very important for embryonic development and dihydrofolate reductase is one of the key enzymes in the process of folic acid performing its biological function. Therefore, the dysfunction of dihydrofolate reductase can inhibit the function of folic acid and finally cause the developmental malformations. In this study, we observed the abnormal cardiac phenotypes in dihydrofolate reductase (DHFR) gene knock-down zebrafish embryos, investigated the effect of DHFR on the expression of heart and neural crest derivatives expressed transcript 2 (HAND2) and explored the possible mechanism of DHFR knock-down inducing zebrafish cardiac malformations.

METHODSMorpholino oligonucleotides were microinjected into fertilized eggs to knock down the functions of DHFR or HAND2. Full length of HAND2 mRNA which was transcribed in vitro was microinjected into fertilized eggs to overexpress HAND2. The cardiac morphologies, the heart rates and the ventricular shortening fraction were observed and recorded under the microscope at 48 hours post fertilization. Whole-mount in situ hybridization and real-time PCR were performed to detect HAND2 expression.

RESULTSDHFR or HAND2 knock-down caused the cardiac malformation in zebrafish. The expression of HAND2 was obviously reduced in DHFR knock-down embryos (P < 0.05). Microinjecting HAND2 mRNA into fertilized eggs can induce HAND2 overexpression. HAND2 overexpression rescued the cardiac malformation phenotypes of DHFR knock-down embryos.

CONCLUSIONSDHFR plays a crucial role in cardiac development. The down-regulation of HAND2 caused by DHFR knock-down is the possible mechanism of DHFR knock-down inducing the cardiac malformation.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; physiology ; Female ; Heart ; embryology ; Heart Defects, Congenital ; etiology ; Tetrahydrofolate Dehydrogenase ; genetics ; physiology ; Zebrafish ; Zebrafish Proteins ; genetics ; physiology