Background Dacryocystitis is one of the most common infectious eye diseases.The gold standard for the identification of bacteria causing dacryocystitis is bacterial culture.The combination of regular culture method with molecular biology techeniques will generate more reliable results.However,very few research data are available in ophthalmological studies in this area.Objective This study was to identify the genera and species of the dacryocystitis-causing bacteria by PCR amplification of the 16S rRNA sequences.Methods Ten cases of qualified standardized bacteria samples were taken,and the nucleic acids were released in the heating process of the PCR procedure.The 16S rRNA genes were amplified and sequenced,and the genera and species were identified using BLAST from GenBank,and the results were used to compare with the results from biochemical identification to test the reliability of this method.The cultured bacterial species from the lacrimal sac secretions from 30 cases of dacryocystitis patients were identified with the above method.Results The outcome of the PCR identification for the 10 cases of quality control standard bacterial specimens was consistent with the results from the biochemical identification.The identification of the 30 cases of dacryocystitis through sequencing the 16S rRNA revealed there were 13 cases of Staphylococcus epidermidis infection,2 cases of Staphylococcus warneri infection,1 case of Staphylococcus hominis infection,5 cases of Corynebacterium macginleyi infection,3 cases of Streptococcus pneumonia infection,2 cases of Bacillus cereus infection,1 case of Micrococcus luteus infection,1 case of Moraxella catarrhalis infection,1 case of Moraxella osloensis infection and 1 case of Pseudomonas aeruginosa infection.Conclusions Sequencing the 16S rRNA is an accurate and specific way for the identification of the genera and species of bacteria that cause dacryocystitis in patients.This sequencing method is feasible in monitoring a variety of dacryocystitis-causing pathogens.More information and epidemiological statistics about dacryocystitis can be obtained from 16S rRNA sequencing.

Gene Library ; Humans ; Nucleic Acid Hybridization ; methods ; Pulmonary Fibrosis ; diagnosis

Objective: To study the chemical constituents from the stems of Acanthopanax gracilistylus. Methods: The chemical constituents of the plant were isolated and purified by column chromatography and their structures were elucidated on the basis of physico-chemical properties and spectral data. Results: A new ent-kaurane glycoside, named kaurane acid glycoside A (16α, 17-dihydroxy-ent-kauran-19-oic 19-[β-D-glucopyranosyl-(1→2)-β-D- glucopyranosyl] ester) (1), was isolated from the n-butanol part. Conclusion: Compound 1 is a new one.

Aged ; Back ; Carcinoma, Medullary ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Melanoma ; metabolism ; pathology ; secondary ; surgery ; Melanoma-Specific Antigens ; metabolism ; S100 Proteins ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Thyroid Neoplasms ; metabolism ; pathology ; secondary ; surgery

Background Zero spherical aberration intraocular lenses(IOL)is designed to prevent the addition of positive spherical aberration after surgery.Research indicated that some positive spherical aberration can provide better depth distance of focus and pseudoaccommodation.Objective The present study was to compare the visual function and wavefront aberrations in pseudophakic eyes with zero spherical aberration IOL and spherical IOL.Methods A prespective case-controlled study was designed.Eighty eyes of 52 patients with age-related cataract were enrolled and divided into two matched groups based on random number table method.The regular phacoemulsification was performed on the eyes,and a zero spherical aberration IOL(Akreos AO)was implanted in the test group and a spherical IOL was used in the control group(Akreos Adapt IOL).The corrected distance visual acuity(CDVA),contrast sensitivity,depth of focus and wavefront aberrations were recorded and compared at 3 months after cataract surgery between these two groups.The trail was approved by the Ethic Committee of Eye Hospital of Wenzhou Medical College,and written informed consent was obtained from each patient prior to the program.Results The clinical demography from the two groups was matched(P ＞ 0.05).There were no significant difference in the CDVA (LogM AR)(-0.03 ±0.08 versus-0.02+0.10)(t =-0.50,P =0.61)and in depth of focus(3.48± 1.07 DS versus 3.20±0.77 DS)(t =1.15,P=0.25)between the zero spherical aberration IOL group and the spherical IOL group.The contrast sensitivities under the mesopic condition at 12.0 c/d and mesopic with glare at 3.0,6.0,18.0 c/d were 12.42 ± 13.16,42.58 ±24.96,30.19± 25.64 and 3.03 ± 5.49 in the zero spherical aberration IOL group,and those in the spherical IOL group were 5.59 ± 8.11,28.74 ± 18.69,17.07 ± 19.35 and 0.22 ± 1.15 without significant differences between these two groups(P＜0.05).Under the 5.0 mm pupil analyzing zone,the spherical aberration in zero spherical aberration IOL group was(0.13 ±0.07)μm,showing a significant reduction in comparison with spherical IOL group(0.21 + 0.07 μm)(P ＜ 0.05).No evidently differences were found in total high-order aberration,coma aberration and trefoil aberration(P＞0.05),but the sphere aberration was considerably lower in the zero spherical aberration IOL group compared with spherical IOL group(t=-4.19,P＝0.00).Conclusions The visual quality of the eyes implanted zero spherical aberration IOL is significantly better than ones implanted with spherical IOL.

【Objective】 To establish a new method for the determination of fibrinogen content in cryoprecipitated antihemophilic factor. 【Methods】 Fibrinogen (Fib) could bind with sheep anti-human fibrinogen (anti-Fib) specifically and further form antigen-antibody complex. When the Fib was present in the solution, the fluorescence of fluorescein isothiocyanate (FITC) labeled on the anti-Fib (FITC-anti-Fib) was quenched due to the formation of immune complex. The fluorescence quenching degree of FITC-anti-Fib was positively correlated with Fib concentration (cFib) in a certain concentration range. 【Results】 The linear relationship between fluorescence quenching degree [(I0-I)/I0] of FITC-anti-Fib and ln(cFib) was (I0-I)/I0=15.53ln(cFib)+ 80.79 (R²=0.99) when the cFib was in the range of (0.007 8-0.560 0) g/L. The recovery of Fib was (96.77-102.43) %. When the method was applied to determine Fib at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.31%, 0.56%, and 0.49%, respectively, and the inter-day variation coefficients were 3.81%, 3.06%, and 4.13%, respectively. There was no significant difference between the results measured by fluorescence quenching method and coagulation method (t=-0.075, P>0.05). 【Conclusion】 In this work, a new fluorescence method for the determination of Fib in cryoprecipitated antihemophilic factor was successfully established based on the specific combination of fib and FITC-anti-Fib. The method is simple and rapid. The obtained results were accurate and reliable by using this method to determine Fib.

【Objective】 To determine the volume range of suspended erythrocyte and establish its internal control standard. 【Methods】 The theoretical value of suspended erythrocyte volume was calculated according to the screening criteria of healthy blood donors and Quality Requirements for Whole Blood and Blood Components. A total of 2 410 bags of 1 U and 2 U suspended erythrocyte were randomly selected and weighed, and the volume range were formulated by ±2S and ±10% respectively and then compared to determine the volume range in line with the actual situation of our center. 【Results】 The theoretical volume range of 1 U and 2 U suspended erythrocyte were 117-160 mL vs 234-320 mL, and the actual volume range were 142-180 mL vs 276-393 mL. The volume range of 1 U and 2 U suspended erythrocyte formulated by ±2S were 145-181 mL vs 298-358 mL, and by ±10% were 147-179 mL vs 295-361 mL. The hematocrit and hemoglobin content of suspended erythrocyte within the actual volume range met the quality requirements. There were fluctuations in the volume of suspended erythrocyte from different regions. 【Conclusion】 Based on the actual situation of our center and the sampling results of suspended erythrocytes in recent two years, 163 mL±10% and 328 mL±10% were determined as the internal control standards of 1 U and 2 U suspended erythrocyte, respectively. Blood centers should establish accurate and feasible standard of suspended erythrocyte according to the actual situation.

【Objective】 To establish a simple, economical and rapid method for the determination of methylene blue (MB) release in virus inactivation bag. 【Methods】 Based on the fluorescence energy transfer between MB and BSA-stabilized gold nanoclusters (BSA-AuNCs), the standard curve of MB determination was established by measuring the fluorescence quenching degree of MB to BSA-AuNCs in different concentrations to conduct the determination of MB release in virus inactivation bag. 【Results】 There was a good linear relationship between the MB concentration (cMB) and the fluorescence quenching degree of BSA-AuNCs[ (I0-I)/I0=0.018cMB+ 0.021(r=0.996)] when the fluorescence emission wavelength was about 620 nm and the cMB was in the range of (0.9-36) μmoL/L. The recovery of MB was 98.00% -101.95 % when applied to determine MB at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.73%, 0.81% and 0.77% respectively, and the obtained inter-day variation coefficients were 3.92%, 3.81%, and 4.73% respectively. There was no significant difference between the results measured by this method and those measured by combination of solid-phase extraction and spectrophotometry(P>0.05). 【Conclusion】 The fluorescence energy transfer method could achieve simple and rapid determination of MB release in virus inactivation bag with accurate and reliable results.

Objective To investigate the correlation of the expressions of advanced glycation end products(AGE) and the receptor for advanced glycation end products(RAGE) in serum and placenta with the pathogenesis of preeclampsia. Methods From December 2013 to June 2014, 32 women with severe preeclampsia who received cesarean section in the Affiliated Hospital of Qingdao University were recruited in the study, defined as the severe preeclampsia group. 30 healthy pregnant women who received cesarean section in the same hospital were recruited as the control group. ELISA was used to measure the maternal serum AGE, soluble receptor for advanced glycation end products (sRAGE) and tumor necrosis factor-α(TNF-α) in these women. Furthermore, ELISA was also used to measure AGE and TNF-α in the placenta. The localizations of AGE and RAGE protein in placentas were detected by immunohistochemical SP method. RAGE and TNF-α mRNA expression in placentas were measured by real-time quantitative PCR. AGE, RAGE and TNF-αprotein expression in placentas were measured by western blot, respectively. Results (1) The serum levels of AGE,sRAGE and TNF-αin the severe preeclampsia group were (538 ± 75),(367 ± 86) and (322 ± 40) ng/L,respectively. They were significantly higher than those in the control group[(454 ± 50), (286 ± 35) and (270 ± 35) ng/L, respectively](P<0.05). The levels of AGE showed positive correlation with the levels of TNF-α(r=0.588,P<0.05),while the levels of sRAGE showed no correlation with TNF-α(r=-0.041, P>0.05). (2) In the severe preeclampsia group, the levels of AGE and TNF-αin placentas were (500 ± 82) and (334 ± 57) ng/L, which were higher than those in the control group [(431 ± 74) and (263 ± 46) ng/L, respectively](P<0.05). The levels of AGE showed positive correlation with the levels of TNF-ɑ(r=0.406,P<0.05). (3)AGE and RAGE protein mainly located in the syncytiotrophoblasts, macrophages and vascular endothelial cells in the placentas of the two groups. AGE expressed mainly in the cytoplasm, and RAGE expressed in the cytoplasm and cell membranes.(4)RAGE and TNF-αmRNA expression in the placentas of the severe preeclampsia group were 12.6 ± 4.6 and 10.4 ± 2.4, which were significantly higher than those in the control group (0.9 ± 0.4 and 3.5 ± 0.9,P<0.01). (5) The expressions of AGE、RAGE and TNF-αprotein in placentas of the severe preeclampsia group were 0.68 ± 0.06, 0.82 ± 0.08 and 0.76 ± 0.08. All were significantly higher than those of the control group (0.46 ± 0.05,0.42 ± 0.09 and 0.52 ± 0.07;P<0.01). Conclusions The levels of AGE and RAGE in serum and placentas elevated in the severe preeclampsia group, and the expression of TNF-αalso elevated. These indicated that AGE and RAGE might be involved in the systemic inflammatory response and local inflammatory response in placentas, and then caused the preeclampsia.

AIM:To investigate whether Notch1 changes stemness and chemotherapeutic sensitivity in human glioma U251 cells.METHODS: The lentiviral vectors, which expressed Notch1-shRNA or Notch1 intracellular domain ( NICD) , were transfected into U251 cells .Western blot and immunofluorescence staining were applied to monitor the va-lidity of the cells, down-regulation of Notch1 expression or over-expression of NICD.The proportion of CD133 +cells was analyzed by flow cytometry.The expression of nestin and GFAP was identified by immunofluorescence staining.The forma-tion rate of tumor cell spheres and the implanted tumor growth in SCID mice were observed.MTT assay was performed to e-valuate the chemotherapeutic sensitivity to VM-26 and BCNU of the cells with different treatments.RESULTS:Stemness was significantly enhanced in the cells over-expressing NICD.For example, the proportion of CD133 +cells was increased, the expression of nestin was up-regulated, the expression of GFAP was down-regulated, and the formation rate of tumor cell spheres and implanted tumor growth were increased.The chemotherapeutic sensitivity to VM-26 and BCNU of the cells was decreased.In the cells with Notch1 gene down-regulation by RNAi, the stemness was inhibited and chemotherapeutic sensi-tivity was increased.CONCLUSION:Notch1, which leads to the change of stemness and chemotherapeutic sensitivity in human glioma U251 cells, is likely to be a potential molecular target for treatment of glioma.