Objective To investigate pathological changes of lens epithelium cells (LECs) and expressions of matrix metalloproteinase-3 (MMP-3),alpha-smooth muscle actin (α-SMA),and fibronectin (Fn) in lens epithelial cells of experimental diabetic cataract rats,and to evaluate the roles of MMP-3,α-SMA,and Fn in the pathogenesis of diabetic cataract.Methods A total of 105 healthy Sprague-Dawley (SD) rats without lens diseases was randomly divided into normal control (n =45) and diabetic model (n =60) groups.Diabetic model rats were subjected to intraperitoneal injection of streptozotocin (STZ) (60 mg/kg),and those in the normal control group received injection with 0.1 mmol/L citric acid buffer solution of the same volume.The diabetic models were affirmed upon a fasting blood ≥ 16.65 mmol/L at the 3rd days after the injection.Once a week,the changes of blood glucose and body weight were monitored and the progression of cataract formation in both lenses of all rats was recorded with slit lamp observation.At end of 4 weeks,8 weeks,and 12 weeks after STZ injection,lenses were isolated and embedded in paraffin.The LECs histopathology was examined with HE staining.Immunohistochemical method was used to detect expressions of MMP-3,α-SMA,and Fn in normal and diabetic LECs.The related comparisons and statistic analysis were carried out.Results The lenses of control group were always completely transparent throughout the period of experiment with 31.48％,77.78％,and 100％ of lenses in diabetic model group swelling at 4th,8th,and 12th week,respectively.Under the light microscopic level,it has been showed that lens epithelium cells,which was occurred aggregate plaque and arranged in many layers,presented some morphologic characteristics of fibroblasts by HE staining.In control group LECs regularly,MMP-3,α-SMA,and Fn did not express equally;MMP-3,α-SMA,and Fn expressions increased obviously highly,difference had statistical significance compared to diabetic cataract group LECs (P ＜ 0.01).With the development of course of disease,the differences in expression of MMP-3,α-SMA,and Fn were significant between the diabetic cataract LECs and the control group (P ＜ 0.01).During the progression of diabetic cataract,the expression of Fn was positively correlated with that of α-SMA and MMP-3 (r =0.994,P ＜ 0.01;r =0.993,P ＜ 0.01).Conclusions The diabetic cataract liyingbody animal model was established successfully,which has laid down necessary basis to expound the morbidity mechanism of diabetic cataract.In the lens of diabetic cataract group,the expressions of α-SMA,MMP-3,and Fn were significantly increased.They participated in the occurrence and development of diabetic cataract.The expression of Fn was positively correlated with α-SMA and MMP-3.The lens epithelium cells which took on elongated aspect like fibroblast cells appeared epithelial-mesenchymal transformation (EMT),which mediated the occurrence and development of LECs fibrosis.

Objective To study bioactive metabolite produced by marine actinomyces M326.Methods Based on the index of antimicrobial activity,the relation between activity of metabolite and conditions of culture in different media and time were researched,and the stability of bioactive metabolite treated with different temperatures and pH were obseved,and the bioactive ingredients were extracted and separated by organic solvent and by macroporous resin.Results The GBP medium made by artificial seawater or distilled water were suitable for production of strong bioactive metabolites.The antimicrobial substances were stable under pH 2～11 and were heat-resistant under strong acidic condition.It could not be extracted by usual organic solvent,but could be absorbed by AB-8 macroporous resin under alkaline situation.Conclusion The metabolites have strong activity against gram-positive bacteria and have different antimicrobial activities against gram-negative bacteria and drug-resistant strains moreover,the polarity of antimicrobial substances is strong.

Objective To extract and purify a polysaccharide SEP from eggs of sea urchin Strongylocentrotus nudus. and to determine its purity, molecular weight and immunological activity in vitro. Methods The orthogonal design was employed to obtain the best possible combination of the critical parameters for polysaccharide extraction. By ultrafiltration, DE-AE Sepharose Fast Flow anion-exchange chromatography and Sephacryl S-400 gel filtration chromatography, the deproteinated crude polysaccharide was purified. The homogeneity of SEP was proved by HPLC, polyacrylamide gel electrophoresis and paper chromatography. Its molecular weight was determined by HPGPC in reference to standVd T-series Dextran. Lymphocyte proliferation assay was made to investigate the immuno-modulating activity of SEP. Results and Conclusion The results indicated SEP was a homogeneous polysaccharide. Its molecular weight was about 1950KD. SEP increased remarkably spleen lymphocyte proliferation. The homogeneous polysaccharide SEP showed significantly immunological activity in vitro.

Objective To evaluate the dosimetric differences of one RapidPlan Model on different Radiotherapy devices.Methods A RapidPlan Model was built based on 30 reoptimization IMRT plans of cervical cancer patients on typeA LA.Dosimetric differences of automatic optimized IMRT plans using this model on 4 different type LAs,named respectivelyA,B,C andD,were compared with 12 test cervical cancer cases.These four LAs were well commissioned in the treatment planning system (TPS).Student t test was applied for statistical analysis on dosimetric differences.Results Dosimetric differences between A vs.B,C and D were observed on Dmean,HI,CI of PTV50 and PTV45,as well as on V50,V40,V30 of rectum and bladder.Significant dosimetric differences were observed between A and D (P<0.05).Conclusions Automatic planning with RapidPlan model may result in dosimetric differences on different Radiotherapy devices.These differences should be aware of with caution in its clinical application.

Marine actinomyces LYG-1 was isolated from marine mud flats in Lianyungang,China.Strain LYG-1 was identified using the methods of morphology,physiological and ehemotaxonomic characterization and 16S rRNA gene sequence analysis.The results showed that strain LYG-1 was a marine variable species of Streptomyces roseosporus.The fermentation broth of strain LYG-1 exhibited conspicuous antitumor activity against HepG2,MCF-7,HCT116 and MDA-MB-231 cell lines,and the IC50 values were defined by MTT method respectively.

Aim: To construct a prokaryotic expression vector carrying NuBCP-9-tumstatin(74-98) (abbreviated as NT) gene and to obtain the fusion peptide with antitumor activity. Methods: Nucleotide sequences of antitumor peptides, NuBCP-9 and Tumstatin( 74-98), were connected via a linker(G_4S)_3 based on biased codons of E. coli the fused NT gene was reconstructed using SOE PCR, and inserted into pET32a(+) vector, and transformed in E. coli BL21(DE3). After expression, the novel fusion peptide was purified through nickel-affinity chromatogra-phy, Factor Xa digestion and ultrafiltration. Biological activity of the fusion peptide on ECV304 and A549 cells was evaluated by MTT assay. Results: A prokaryotic expression system with NT gene was successfully constructed. The soluble fusion peptide was accounted for approximately 25% when induced by 0. 5 mmol/L IPTG at 30 ℃ for 4 h. The purified fusion peptide could inhibit cell growth of ECV304 and A549 with inhibition rates of 60. 8% and 65. 2% at 20 μmol/L, respectively. Conclusion: A novel fusion peptide with antitumor activity was cloned, expressed and purified.

Background Phakic posterior chamber implantable contact lens (PPC-ICL) or phakic posterior chamber Toric implantable contact lens (PPC-TICL) implantation is an effective way for the correction of high myopia or high myopia with astigmia,but it often has residual myopic power.Excimer laser-assisted subepithelial keratectomy (LASEK) can correct the residual myopia following PPC-ICL or PPC-TICL,but its effectiveness and safety deserve attention.Objective This study was to analyze the clinical effectiveness and safety of LASEK for residual myopia after PPC-ICL implantation for extreme high myopia.Methods A prospective cases-observational study was performed,and written informed consent was obtained from each patient before any surgery.Fourteen eyes of 9 patients with residual myopia following PPC-ICL or PPC-TICL for the eyes with spherical equivalent refraction of ≥-20.00 D were collected in the Affiliated Hospital of Guizhou Medical University from July 2010 to March 2015,including PPCICL implantation in 8 eyes and PPC-TICL implantation in 6 eyes.LASEK were performed on the eyes to correct the residual myopic power.Uncorrected visual acuity (UCVA),best corrected visual acuity (BCVA),haze,the distance of intraocular lens to lens,corneal thickness,corneal topography,corneal endothelial cell counting,intraocular pressure (IOP) and fundus were examined and compared before and after surgery.The effectiveness and safety of the surgery were evaluated.Results The operation was smooth and no complication was found after surgery in all of the eyes.The UCVA and BCVA were significantly different in the eyes among before surgery,6 months after PPC-ICL implantation and 12 months after LASEK (F =31.360,1.778;both at P＜0.05),and the UCVA after LASEK was higher than BCVA before LASEK.The refractive powers were (-22.27-±4.29),(-3.75±2.25) and (-0.42±0.63) D before surgery,6 months after PPC-ICL implantation and 12 months after LASEK,showing a significant difference among them (F=46.370,P＜0.05),and the refractive power was considerably lower after LASEK than that before surgery and after PPC-ICL implantation (both at P＜0.05).No significant difference was found in IOP or corneal endothelial cell counting in operated eyes among before surgery,6 months after PPC-ICL implantation and 12 months after LASEK (F=1.663,1.055;both at P＞0.05).The distance of intraocular lens to lens was (0.69±0.26)mm in the eyes after LASEK and (0.71 ±0.29)mm in the eyes after PPC-ICL implantation,with no significant difference between them (t =0.192,P＞0.05).Conclusions PPC-ICL or PPC-TICL implantation for the correction extreme high myopia often remains a certain degree of myopia,and LASEK for the correction of residual refractive power is safe and effective.

The article discussed the necessity of strengthening continuous medical education in general hospitals,and puts forward that to establish high quality medical staff that can keep up with time,take the challenges and promote the innovation and the development of hospital,We must attach importance to strengthening and continuing medical education,and at the same time,improving and perfecting the education system is also very important.

Hepatitis B virus (HBV) DNA replication takes place in the viral capsid that consists of 180 or 240 copies of HBV capsid (HBc or core) protein. The monomeric core protein contains an apical loop region that forms the spikes on the surface of viral capsid upon core dimerization and capsid assembly. To investigate the impact on HBV DNA replication through gene engineering at the spike of HBV capsid. plasmids expressing engineered HBc with linker-fused enhanced green fluorescent protein (EGFP) or shortened EGFP insertion at the spike region were constructed by Restriction Digestion and Ligation-independent Cloning (RLIC). The wildtype or mutant HBc construct was cotransfected with HBV1.1c(-), a plasmid containing 1.1 unit-length HBV genome with deficiency in HBc expression, into HEK293 cells, respectively. GFP signal was observed through a fluorescence microscope and HBV DNA replicative intermediates were assayed by Southern blotting to determine the expression and functions of different recombinants. Our results demonstrated that the RLIC method was effective to generate deletion or insertion in the apical loop region of HBc. Both HBc-EGFP recombinants with different linkers produced green fluorescence but with different subcellular distribution pattern. However, HBV DNA replication was not detected with the trans-complementation of these two HBc recombinants. In addition, other recombinants including the one only with the deletion of aa79-80 failed to support HBV replication. Taken together, our results suggest that RLIC is a robust method which can be broadly applied in gene engineering; different peptide linkers may have different influences on the functions of an engineered fusion protein; and HBc aa79-80 play a critical role for HBc to support HBV DNA replication.
Capsid Proteins ; genetics ; Cloning, Molecular ; Genetic Engineering ; methods ; Green Fluorescent Proteins ; biosynthesis ; genetics ; HEK293 Cells ; Hepatitis B Core Antigens ; biosynthesis ; genetics ; Hepatitis B virus ; genetics ; physiology ; Humans ; Mutation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection ; Virus Replication

Objective To investigate the effect of different doses of ioversol on renal function,and to explore early renal injury biomarkers on contrast induced kidney injury and safe ioversol dosage.Methods A total of 158 cases (98 males and 60 females) undergoing cerebral vascular intervention (CVI) in our department was selected with age ranging from 23 to 81 years old (average age 59.70 ± 12.02).Based on ioversol dosage in surgery,patients were divided into three groups:low dose group (≤ 150 ml,n =49),middle dose group (151-200 ml,n =74),and high dose group (＞200 ml,n =35).U-κ,U-λ,urinary transferrin (UTRF),urine microalbumin (UMA),urinary immunoglobulin IgG (UIgG),urine beta2-microglobulin (Uβ2-MG),Uα1-MG,urinary N-acetyl-beta-D-glucosaminidase (UNAG),plasma cystatin C (CysC) and Scr were detected by scattering turbidimetry,immune turbidimetry and fully automatic biochemical analysis pre-surgery 24 h and post-surgery 72 h.Contrast-induced acute kidney injury (CI-AKI) was defined as laboratory increase of Scr value≥44.2 μmol/L or ≥25％ from baseline measurement at 48 hours after surgery.The relationship in ioversol dosage and various factors was assessed by Single and multiple factors binary logistic regression analysis.Results According to the criterion that Scr increase value were ≥44.2 μmol/L,of 158 cases,3 cases occurred CI-AKI,the AKI incidence was 1.90％.Based on the criterion that Scr increase value was ≥25％,33 cases occurred CI-AKI,the incidence was 20.89％.The concentration of U-κ,UTRF,Uα1-MG,UNAG and plasma CysC were significantly different in high dose group compared to low ioversol dose group (P ＜ 0.05),while the other biomarkers had no significant difference (P ＞ 0.05).Conclusions The contrast media-ioversol could lead to CI-AKI;when the dosage of ioversol was more than 200 ml one-time,the concentration of U-κ,UTRF,Uα1-MG,UNAG and plasma CysC increased significantly.U-κ,UTRF,Uα1-MG,UNAG and plasma CysC could predict the early renal injury in patients who undergoing CVI.The rise of U-κ,UTRF,Uα1-MG,UNAG and plasma CysC are related to the dosage of ioversol.Furthermore,possibility of kidney injury is significantly high when ioversol dosage is more than 200 ml one-time.