An ultra high performance liquid chromatography-photodiode array detection-tandem quadrupole mass spectrometry ( UPLC-PAD-MS/MS) method was developed for the determination of total 13 flavonols and flavonol glycosides in red onion which including 6 quercetin and its glycosides, 4 isorhamnetin and its glycosides, 3 Kaempferol and its glycosides. The chromatographic separation was carried out by used a UPLC HSS T3 column and eluted under gradient with mobile phases of acetonitrile and water both contained 0 . 1%formic acid at a flow rate of 0. 3 mL/min. The results showed that the major flavonols and flavonol glycosides in red onion were quercetin-4’-glucoside, quercetin-3, 4’-diglucoside, quercetin and Isorhamnetin-4’-glucoside. The amounts and distributions of flavonols and flavonol glycosides among different parts of red onion were different. For the same amount of dry materials, the content ratio of total flavonols and flavonol glycosides in the outer two layers, the third layer and the inner layer was 60. 3:33. 0:6. 7, the amount of quercitin and its glycosides accounts above 92. 1% of total flavonols and flavonol glycosides for each part. In the outer two layers, the amount of flavonol monoglycosides are the highest, in the third layer, the amount of flavonol aglycones were the highest, but in the inner layer, the amount of flavonol diglycosides were the highest. Small amounts of Kaempferol and its glycosides were found in red onion, and mostly were found in outer layers. This method is simple, fast, accurate and convenient, and can be used to analyze flavonols and flavonol glycosides in onion product.

Objective To observe the clinical effect of eye drops of 0.1%olopatadine hydrochloride(patanol) and 0.05%FKS06 (tacrolimus) for treating severe vernal keratoconjunctivitis in children.Design Prospective case siries.Participants 62 cases(104 eyes) of severe vernal keratoconjunctivitis aged 3-14 years old.Methods We observed the symptoms and signs of severe vernal keratoconjunctivitis before and after the treatment of 0.1%olopatadine hydrochloride and 0.05%FKS06.We examined regularly the blood,liver and kidney functions for all patients during treatment.Main Outcome Measures Symptoms and signs.Results Following by 3- 34months,all patients were much better in different degree at 1 week than that before.The papillae were decreased and almost disappeared gradually at 1 month.The corneal lesions were cured rapidly in less than 2 weeks.2 cases(3.2%) complained a little disconfort just at the beginning.38 cases(61.3%) recurred from 2 weeks to 8 months after stopping the eye drops,and were controlled rapidly after reusing the eye drops.The blood routine and the biochemical functions of all patients were normal during treatment.Conclusions 0.1% olopatadine hydrochloride and 0.05%FKS06 have remarkable effects for alleviating symptoms and signs of severe vernal keratoconjunctivitis. The patients are comfortable and content with the treatment,and side effects are very few.

Chikungunya virus ; genetics ; isolation & purification ; China ; Genome, Viral ; RNA, Viral ; genetics ; Sequence Analysis, RNA

Objective To explore the best treatment for hepatitis B by observing the efficacy d interferon and adefovir dipivoxil (ADV) according to different combination of various quantification of HBV DNA and HBeAg. Methods One hundred and sixty-five hepatitis B antigen-positive patients were divided into four groups (A, B, C, D group) according to different quantification of HBV DNA and HBeAg. Each group was randomly divided into two sub-groups with interferon and the others with ADV. Items were observed in 48 weeks and statistic analysis was made to observe if there was significant difference between two methods in each group. Results There was no difference in the rate of HBV DNA inhibitory activity, HBV DNA clearance and ALT normalization as well as HBV DNA decreasing (level≥2lg copy/ml). But there was significant difference in the rate of HBeAg clearance, HBeAg seroconversion and HBeAg decreasing level ≥500.00 COL/ml (P<0.05). The efficacy decreased following the increasing of HBV DNA and HBeAg quantification, especially in ADV groups. The efficacy of interferon was better in seroconversion rate. Conclusion Quantification of HBV DNA and HBeAg can be the guidance to select different anti-virus methods.

Objective To sequence and analyze the envelope (E) gene of type Ⅰ dengue virus isolated from Guangzhou in 2009 for tracing the infection source. Methods The serum samples were collected from patients diagnosed with dengue fever in Guangzhou area during 2009. Dengue virus was isolated and cultured in C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced. The phylogenetic tree was drawn by neighbor-joining method. The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiology data. Results Four strains of type Ⅰ dengue virus were isolated from 19 samples. E gene of these strains was amplified and sequenced. The phylogenetic analysis showed that 09/GZ/9104 strain and 09/GZ/9236 strain had identical nucleotide sequence and fell within the American/African group, 09/GZ/11534 stain and 09/GZ/11562 strain had similar sequence homology and fell within the Asian group. Conclusion The typeⅠdengue viruses in Guangzhou area in 2009 are imported, which belong to two genotypes and may come from two independent origins respectively.

ObjectiveTo analyze the Envelope (E) gene of type 1,2,3 dengue virus isolated fromGuangzhouin2010, andtoinvestigatetheinfectionsourceandvirusgenotypes.MethodsEighty-five serum samples were collected from 85 patients in acute phase of dengue fever.Dengue virus was cultured and isolated by C6/36 cells.The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and then sequenced.The phylogenetic tree was drawn by neighbor-joining method.The bioinformatics analysis was performed by combining the phylogenetic information and the epidemiologic data.ResultsSix strains of type 1 dengue virus,two strains of type 2 dengue virus and six strains of type 3 dengue virus were isolated from 85 samples.The E gene sequence of these strains was obtained by sequencing.The phylogenetic analysis showed that type 1 and 3 dengue virus belonged to two genotypes (Asian and South Pacific ocean,India subcontinent and Southeast Asia/South Pacific ocean,respectively),and type 2 dengue virus belonged to one genotype (Malaysia/India subcontinent).ConclusionIt's presumed that all strains of type 2 dengue virus are imported,four strains of type 1 dengue virus are imported and four strains of type 3 dengue virus arc imported,the remaining two stains of type 1 and two stains of type 3 dengue virus need mosquito intermediary research further to prove their origins.

ObjectiveTo evaluate the therapeutical effect and toxicity of home-made paclitaxel and cisplatin combination in metastatic breast cancer.Methods Home-made paclitaxel 175 mg/m2,d1 intravenous drip,cisplatin 80 mg/m2,d1-3 intravenous drip,21-28 d was one cycle,all patients continuous course treatment for 2-6 cycles,then the effect was evaluated.ResultsAll the patients received meso-cycle were 4 (2-6) cycles chemotherapy.The total effective rate was 56.60％ (30/53),disease control rate was 81.13％(43/53 ).The rates of internal organ metastasis and non-internal organ metastasis,before menopause and after menopause had no significant difference [56.76％ (21/37) vs.56.25％ (9/16),59.09％ (13/22) vs.54.84％ ( 17/31 ),P ＞ 0.05 ].The median existence time for all the patients was 18.8 ( 12-27 ) months.The survival rate was 56.60％ (30/53) and 30.19％ (16/53) for 1 and 2 years respectively.The major toxieities were gastrointestinal reaction,alopeoia and myelosuppression,the rates of nerve and heart toxicities were less,15.09％ (8/53) and 7.55％ (4/53) respectively.ConclusionThe therapeutic effect of home-made paclitaxel and cisplatin combination in metastatic breast cancer is good,it is to be worth to spread in clinic.

Objective To dynamically observe the developing process and characteristics of the chronic intra-hepatic hyperplastic nodule to small hepatoma with ultrasound,and to evaluate the value of ultrasound in the diagnosis of small hepatoma.Methods One hundred twenty-two chrome hepatitis cases with HBsAg(+),HBcAb(+)and HBeAg(+)were prospectively examined by two-dimensional ultrasound and three-dimensional reconstruction.Characteristics and diagnostic accuracy of two-dimensional ultrasound and three-dimensional reconstruction were compared with the results of hepatic biopsy.Results Thirty-four of 122 patients developed small hepatoma.The duration between hepatic fibrosis and liver cirrhosis was (6.30±2.31)years,and time of hepatoma changed from liver cirrhosis was (8.01±2.10) years.The intrahepatic hyperplastic nodule,shape of small hepatoma and relationship between tumor and ambient tissue were displayed clearly with three-dimensional ultrasound,and the number of detection with threedimensional ultrasound was much higher than that of two-dimensional ultrasound significantly.Conclusions Dynamic ultrasound examination of the echogenicity change of chronic hepatic lesion caused by HBV is useful for early detection of small intra-hepatic nodule,three-dimensional ultrasound reconstruction may enhance the accuracy in diagnosing small hepatoma.

AIM: To study the inhibitory effects of nm23-H1 gene on proliferation and invasion of human lung adenocarcinoma A549 cell line. METHODS: Recombinant eukaryotic expression vector pcDNA3.1-nm23-H1 containing full length of human nm23-H1 cDNA was constructed and transfected into a human lung adenocarcinoma A549 cell line by lipofectamine. Cell strain that expressed nm23-H1 stably was screened out by G418 and named pcDNA-nm23-A549. Expression of nm23-H1 was identified by RT-PCR and immunohistochemistry. Growth curves were drawn to detect the inhibitory effects on cell proliferation. Cell cycle of pcDNA-nm23-A549 was examined by flow cytometry. Atomic force microscopy was used to observe the filopodia on the surface of the cells. RESULTS: Introduction of nm23-H1 obviously inhibited the proliferation of A549. Expression of nm23-H1 did not induce apotosis in A549 cells but increased the percentage of phase G_1 cells and decreased phase S cells. Meanwhile, phase G_1 to phase S transition was restrained. Filopodia in the cell surface was much fewer and its structure changed in cells transformed. CONCLUSION: nm23-H1 is capable of inhibiting A549 proliferation and decreasing its metastatic ability, probably by interfering with cell cycle and cell surface structure.

Objective To evaluate the efficacy of a sustained releasing mosquito larvicide package against larval breeding and its impact on water and plant,in order to provide a scientific evidence for its application in control and prevention of Dengue.Methods Guangzhou Center for Disease Control and Prevention was chosen as the test place.Twenty test sites were set up,2 bags of sustained releasing larvicides package,1 bag of sustained releasing larvicides package,3 g 1％ temephos granules and nothing were put into 4 glass bottles for each test site from July to December in 2014,respectively.The 4 glass bottles were called high dose (H) group,low dose (L)group,positive control (P) group and blank control (B) group,respectively.The 4 groups were observed at intervals of 10 days for 19 times.Environmental air temperature,turbidity of water,number of larvae and damage of plant were recorded.And 5 test sites were selected to collect water specimen.The chemical oxygen demand,ammonia nitrogen concentration and temephos concentration of water specimen were detected.Results The larval breeding rates were 0 (0/380),1.1％ (4/380),0.8％ (3/380) and 63.4％ (241/380),damage rates of plant were 5.0％ (19/380),5.5％ (21/380),4.7％ (18/380),4.7％ (18/380) and turbidty rates of water were 24.5％ (93/380),19.7％ (75/380),33.4％ (127/380) and 20.3％ (77/380) in H,L,P and B groups,respectively.Statistically significant differences were seen in larval breeding rate and turbidity rate of water between different groups (x2 =823.565,24.715,all P ＜ 0.05),but they were not seen in damage rate of plant (x2 =0.332,P ＞ 0.05).The temephos concentrations were 1.24,0.78 and 2.33 mg/L in H,L and P groups,respectively.Statistically significant differences were seen in temephos concentration between different groups (H =35.426,P ＜ 0.01),but they were not seen in chemical oxygen demand and ammonia nitrogen concentration (H =0.239,0.013,all P ＞ 0.05).Conclusions The sustained releasing package of mosquito larvicide makes less pollution to water and has no impact on water turbidity.Moreover,it doesn't damage the aquatic plant.The efficacy of the sustained releasing package of mosquito larvicide could effectively prevent mosquito larval breeding in Dengue epidemic period.