Objective To investigate the effects of esculin and digitalisglycosides eye drops combined with 532 laser on the treatment of diabetic retinopathy.Methods 65 cases(130 eyes)with diabetic retinopathy in our hospital from November 2012 to November 2015 were selected and divided into observation group(33 cases)and control group(32 cases).The control group was treated with 532 laser treatment,and the observation group was combined with esculin and digitalisglycosides eye drops.Two groups of treatment were one months.Therapeutic effect of two groups were compared,improvement of visual acuity,retinal hemorrhage,exudation,edema absorption time,before and after macular retinal thickness changes.Results After treatment,the total efficiency of the observation group(96.97％)was higher than the control group(75.00％),the difference was statistically significant(P<0.05),the visual acuity of the observation group was significantly higher than that of the control group,and decreased significantly less than the control group,the difference was statistically significant(P<0.05),the retinal hemorrhage,exudation and edema were observed in the observation group faster than the control group,the difference was statistically significant(P<0.05),the macular retinal thickness decreased in the two groups after treatment,the difference was statistically significant(P<0.05),the retinal thickness of the macular area was lower in the observation group than in the control group,the difference was statistically significant(P<0.05).Conclusion Esculin and digitalisglycosides eye drops combined with 532 laser in the treatment of diabetic retinopathy has significant effect,and can reduce the macular retinal thickness,which has important clinical significance.

Objective:To establish the quantitative method for eucalyptol in Chimonanthus salicifolius S. Y. Hu by GC. Methods:The GC method was performed on a Zebron ZB-WAX column (60 m × 0. 32 mm,0. 5 μm) with programmed temperature of 60-200℃, an FID detector was used, the detector temperature was 250℃, the inlet temperature was 220℃, and the carrier gas was nitrogen with high purity. Results:A good linearity of eucalyptol was within the range of 0. 012 6-0. 503 4 mg·ml-1(r=0. 999 8), and the average recov-ery was 100. 68%(RSD=1. 51%,n=6). Conclusion:The method is simple, accurate and quick with good reproducibility, and suitable for the quality control of Chimonanthus salicifolius S. Y. Hu.

A quail-origin subtype of the influenza virus was isolated from a human-infecting H7N9 subtype of the avian influenza virus found in a live poultry market and was given the name A/Quail/Hangzhou/1/ 2013 (H9N2). We analyzed the whole genome of this virus and its biologic characteristics. Sequence analyses suggested that the: HA and NS genes belonged to a CK/BJ/1/94-like lineage; NA, NP, PA and PB1 genes belonged to a SH/F/98-like lineage; M and PB2 genes belonged to a G1-like lineage. Analyses of key amino acids showed that the cleavage site in HA protein was PSRSSR ↓ GL, and that the HA protein had a human receptor-binding site with Leu226. Deletion of amino acids 69 - 73 was detected in the stalk of NA protein, the M2 protein had an Asn31 mutation, and the NS1 protein had two mutations at Ser42, Ala149. The intravenous pathogenicity of this virus was 0.36. A study in chickens suggested that all inoculated birds shed the virus from the trachea and cloaca on the third day post-infection (p. i. ) until 11 days. All chickens that had direct contact shed the virus on the second day p. i. until 8 days. Results of virus reisolation suggested that lung and tracheal tissues could shed the virus in 5 days, whereas the other organs could shed the virus in 3 days. These results suggest that this virus strain is H9N2 subtype LPAIV, whose lineage is prevalent in mainland China. This research provides evidence on how to monitor and prevent the H9N2 subtype of the avian influenza virus.
Animals ; Chick Embryo ; Chickens ; China ; Genotype ; Influenza A Virus, H9N2 Subtype ; classification ; genetics ; isolation & purification ; Influenza in Birds ; virology ; Phylogeny ; Quail ; virology

Objective To investigate the effect of purified urinary IgG from patients with minimal change nephrotic syndrome and membranous nephropathy on the expression of macrophage migration inhibitory factor (MIF) in human proximal tubular epithelial cells (HK-2). Methods Proteins were precipitated from urine with (NH4)2SO4, and urinary IgG was purified by protein G column chromatography. SDS-PAGE and Western blotting were performed to analyze the urinary IgG. HK-2 cells were incubated with urinary IgG (0,0.5,1.0,2.5,5.0,10.0 mg/ml) from either minimal change nephrotic or membranous nephropathy patients. The expression of MIF mRNA was assessed by RT-PCR, and MIF protein was assessed by Western blotting. Results SDS-PAGE showed that there were four bands, which were all IgG fractions confirmed by Western blotting. The urinary IgG up-regulated the expression of MIF mRNA and protein in HK-2 cells. The expression increased in a dose-dependent manner. The expression of MIF mRNA and protein in HK-2 cells increased significantly when they were incubated with urinary IgG from membranous nephropathy patients at 1mg/ml (P

Purpose　To investigate the role of urokinase plasminogen activator (uPA),uPA receptor (uPAR),tissue type plasminogen activator (tPA) and plasminogen activator inhibitor 1 (PAI-1) in the invasiveness of human breast cancer cells.　Methods　Three human breast cancer cell lines with different invasive ability were taken as research targets.RT-PCR and milk plates methods were used to detect the expression of uPA system members and the PA activities,respectively.Modified Boyden's chamber model was employed to detect the invasive ability of cancer cell.　Results　MDA-MB-231 could express high level of uPA,uPAR,PAI-1 and low level of tPA.MDA-MB-435 could express lower level of uPA and hight level of tPA,but no PAI-1 and uPAR were detected.MCF-7 could express lower level of uPAR and high level of PAI-1,but no uPA and tPA were detected.MDA-MB-231 cells showed the highest total PA and uPA activity.MDA-MB-435 cells also showed high total PA activity,but almost all the activity owed to tPA.MCF-7 showed almost no PA activity.Correlated with their PA activities,MDA-MB-231 was found the most invasive in vitro,followed by MDA-MB-435,and MCF-7 almost had no invasive ability.The antibodies against uPA and uPAR were significantly effective in reducing the matrigel invasiveness of MDA-MB-231 by approximately 83.1% and 43.9% respectively (P<0.05).　Conclusions　Co-expression of uPA,uPAR and PAI-1 in human breast cancer highly correlates with the invasiveness in vitro.

Objective To identify a rapid and efficient fungal genomic DNA extraction method for PCR amplification.Methods Genomic DNA was extracted from Penicillum marneffei,Rhizopus microsporus,Cryptococcus neoformans and Candida albicans by heating pyrolysis,microwave,repeated freezing and thawing,lysozyme digestion,overnight snail enzymatic and Qiagen kit methods.DNA electrophoretogram was observed by gel imaging system.The concentration and purity of extracted DNA were determined with an ultramicro nucleic acid protein tester and the yields were calculated.PCR amplification and sequencing were also performed.ANOVA and SNK-q test were used for data analysis.Results There were statistical differences in concentrations and yields of the fungal DNA extracted from Penicillum marneffei (hyphal phase and yeast phase),Rhizopus microsporus,Coptococcus neoformans and Candida albicans by six methods (F=750.83,220.95,669.35,132.01,510.20 and 1658.35,287.10,963.64,1147.77,4521.22,all P ＜0.01).Of six methods,microwave method gained the highest DNA concentration and yield,followed by heating pyrolysis method,while Qiagen kit method obtained the lowest concentration and yield.All DNA extracted by 6 kinds of methods were positive in PCR amplification.Conclusion All of the six methods can be used for fungal DNA extraction which is sufficient for PCR amplification,but microwave and heating pyrolysis methods are more easy and simple to perform.

Objective: To establish an HPLC method for the simultaneous determination of three flavonoids in Shi Liang Cha. Methods:HPLC was applied with the chromatographic conditions as follows: the chromatographic column was Agilent Zorbax SB-C18 (250 mm × 4. 6 mm, 5μm) at 30℃;acetonitrile-0. 1% H3 PO4 solution was used as the mobile phase with gradient elution; the flow rate was 1. 0 ml·min-1, and the detection wavelength was 360nm. Results: Good linearity of rutin, quercetin and kaempferol was within the range of 0.0409-1.637 0mg·ml-1(r=0.999 2), 0.44-88.00μg·ml-1(r=0.999 8) and 0.41-77.63μg·ml-1(r=0. 999 2), respectively; the average recovery was 99.35%(RSD =1. 64%),101. 14% (RSD =1. 88%) and 99. 69% (RSD =1. 92%) , respectively. Conclusion:The method is simple, accurate and repeatable, and suitable for the simultaneous determination of rutin, quercetin and kaempferol in Shi Liang Cha.

Objective:To establish an HPLC method for the determination of oleanolic acid in She medicine radix of Aralia chinen-sis L. . Methods:The HPLC analysis was performed on a Waters XBridge-C18 (250 mm × 4. 6 mm, 5 μm) column. The mobile phase was methanol-0. 1 mol·L-1 ammonium acetate solution (83∶17) and the flow rate was 1. 0 ml·min-1 . The detection wavelength was 210 nm, the column temperature was 20 ℃, and the injection volume was 10 μl. Results:Oleanolic acid in radix of Aralia chinensis L. had a good separation from the other components, a good linearity was obtained within the range of 72. 52-725. 2 μg·ml-1 ( r=1. 000 0), and the average recovery was 98. 05%(RSD=1. 89%, n=6). Conclusion:The method is simple, accurate, reproducible and applicable in the assay of oleanolic acid in radix of Aralia chinensis L. .

Objective To study the changes of intestinal epithelial barrier function in rats with aging.Methods SD rats were divided into 3 groups:3-month-old group (group A),12-month-old group (group B) and 24-month-old group (group C,established by D-galactose injection with the dose of 0.125 g· kg-1 · d-1subcultaneously for 6 weeks) (n=10,each).The terminal ileum was obtained to make microtome section,and the morphology of small intestine mucous membrane,trophonema altitude and thickness were observed under light microscope.Occludin and ZO-1 protein expressions in terminal ileum mucous membrane were detected by immunohistochemistry.The expressions of Occludin and ZO 1 mRNA were determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).Results The small intestinal mucosa thickness and villus height were lower in group C and B than in group A [thickness:(87.6± 6.32) μm,(131.8± 5.22) μm vs.(162.9±7.28) μm; villus height:(56.4±5.38) μm,(76.7±5.40) μm vs.(108.1±6.42) μm;both P＜0.05].The small intestinal mucosa thickness and villus height was lower in group C than in group B (both P＜0.05).Occludin and ZO-1 protein expressions in small intestine tissue were reduced in group C and B as compared with group A [Occludin protein:(2.23±0.60)％,(4.21±0.61)％ vs.(12.31±0.94)％; ZO-1 protein:(2.03±0.54)％,(4.02±0.65) ％ vs.(12.21±0.81)％ ; both P＜0.05],and Occludin and ZO-1 protein expressions were less in group C than in group B (both P＜0.05).The levels of Occludin and ZO-1 mRNA in small intestine tissue were reduced in group C and B as compared with group A [Occludin:(0.20±0.03),(0.38±0.02) vs.(0.66±0.03) ; ZO-1:(0.18±0.03),(0.37±0.02) vs.(0.63±0.03); both P＜0.05],and Occludin and ZO-1 mRNA expressions were less in group C than in group B (both P ＜ 0.05).Conclusions The small intestinal mucosa thickness and villus height are reduced,the levels of Occludin and ZO-1 expressions are significantly decreased in small intestinal mucosa,and the intestinal barrier function is impaired with rat aging.

Objective To investigate the epidemiological characteristics of mycoplasma pneumoniae (MP),Epstein-Barr virus (EB virus) and cytomegalovirus (CMV) infection in children with acute upper respiratory tract infections in Hangzhou.Methods Throat swabs and sputum samples were collected from 5942 children with acute upper respiratory tract infections in Hangzhou First People's Hospital during January 2011 and December 2012.MP,EB virus and CMV were detected using quantitative PCR.The distribution and seasonal changes of the above pathogens in children of different ages were analyzed using Chi-square tests.Results MP,EB virus and CMV were positive in 29.91％ (1777/5942),22.92％ (1362/5942) and 8.55％ (508/5942) children,respectively.Mixed infections were found in 556 (9.36％) children.The positive rates of MP varied among different age groups (x2 =113,P =0.000),and the highest one was detected in children ＞ 6-year old (448/1012,44.36％).EB virus infection was rare in age group 0-1 year,and the positive rate was of statistical difference from those in other age groups (x2 =167,181 and 187,P =0.000).The highest positive rate of CMV (23.78％) was found in children aged 0-1 year old.The positive rates of MP varied in different months of the year (x2 =208 and 211,P =0.000),and the highest positive rate was found in July and August.Conclusion The predominant pathogen of acute upper respiratory tract infection in children is MP in Hangzhou,and MP plus EB virus infection is common,particularly in older children;while CMV infection more likely occures in 0-1 year old babies,and usually in summer.