Background Many studies showed that attenuation of retinal nerve fiber layer(RNFL)in early glaucoma is one of the important signs.How to accurately and quantitatively measure RNFL thickness is very important for the early diagnosis and monitoring of glaucoma.0bjective This study was to evaluate the clinical value of Optovue RTVue OCT and Heidelberg Retina Tomograph Ⅲ(HRT-Ⅲ)confocal scanning laser ophthalmoscopy in glaucomatous eyes. Methods This cross-sectional study included 40 eyes of 26 patients with suspected open-angle glaucoma(SOAG),48 eyes of 29 patients with open-angle glaucoma and 48 eyes of 27 healthy subjects.Optical nerve head(ONH) parameters and peripapillary RNFL thickness were measured in all the subjects with Optovue RTVue OCT and HRT-Ⅲ,meanwhile all the eyes received perimetry with Humphrey 750-I.Glaucoma variables obtained from Optovue RTVue OCT and HRT-Ⅲ were analyzed among the groups.Topographic Optovue RTVue OCT and HRT-Ⅲ parameters,including disc area(DA),cup area(CA),rim area(RA),rim volume(RV),cup volume(CV),cup/disc area ratio(C/DAR)as well as superior,temporal,inferior and nasal average RNFL thickness,were analyzed.The relationship of ONH parameters and RNFL thickness was analyzed using a linear correlation.The correlation between the mean defect(MD)of the visual field and the tomography parameters in glaucomatous eyes was described by bivariate Pearson correlation coefficients.Resuits The ONH parameters and RNFL thickness obtained by HRT-Ⅲ and OCT showed significant difference(P<0.05).ONH parameters such as RA,C/DAR,CA and RV were statistically changed in SOAG and the POAG patients compared with the normal subjects(q=6.47,q=7.67,P<0.05).The superior and inferior RNFL thickness parameters in three groups were positively correlated between HRT-Ⅲand OCT(r=0.362,r=0.441,r=0.395,P<0.05),Topographic Optovue RTVue OCT and HRT-Ⅲ parameters including CV,CA,RA and C/DAR fitted Pearson analysis(all P<0.05).In POAG group。The RA,RV,CV,C/DAR from Optovue RTVue OCT were correlated with MD with the significant coefficient 0.284,0.286,0.340,0.371 respectively(P<0.05),and evidently correlations also were found between RA,RV,C/DAR with MD respectively with the coefficient 0.339,0.859,0.422(P<0.05)by HRT-Ⅲ. Conclusion Both Optovue RTVue OCT and HRT-Ⅲ can difierentiate ONH analysis with a similar outcome in glaucomatous eye.The C/D value.RA,superior and inferior RNFL thickness based on Optovue RTVue OCT and HRT-Ⅲare distinguishing indexes in the diagnosis of early glaucomatous damage.

In order to investigate the genetic basis of morphological variation of tetraploid plantlets of Atractylodes macrocephala, diploid plantlets were taken as experimental material, sterile filtration colchicine was used to soak 0.5-1.0 cm long buds. The difference between morphology and stomatal of diploid and tetraploid of A. macrocephala was compared, and genome polymorphism was explored by AFLP. The results showed that the buds dipped in 0.1% colchicine solution for 36 h was optimal conditions to induce tetraploid of A. macrocephala with induction rate of 32.0%. Morphological indexes such as leaf area index, leaf length and width, the density of stomas and the number of chloroplast of tetraploid were distinctly different from diploid. Four hundred and fifty-one bands ranging with 80-500 bp were amplified with 24 pairs of primers, the rate of polymorphism was 32.59%. These amplification sites of diploid were different from tetraploid of A. macrocephala, and the differences in morphology of them were reflected in the DNA polymorphism.
Amplified Fragment Length Polymorphism Analysis ; methods ; Atractylodes ; genetics ; Sequence Analysis, DNA ; Tetraploidy

Objective To explore the feasibility of imaging and treatment of cervical cancer xenograft model using 131I mediated by hNIS gene transfection. Methods The cervical cancer xenograft models were established with Hela-NIS( +) cells and Hela cells, respectively. Five Hela-NIS( +) xenograft models and five Hela xenograft models were dynamically imaged at 0.5, 1, 2, 4, 8, 16 and 20 h postinjection of 131I(7.4 MBq). Five Hela-NIS( +) xenograft models were imaged at 0. 5,1,2,4,8,16, 20 and 25 h postinjection of 99TcmO4-(11.1 MBq). Twenty Hela-NIS( +) cervical cancer xenograft models were randomly divided into four groups: Three 131I treating groups and one control group. The therapeutic effects of 131I at threelevels (74,111,148 MBq) were investigated following intraperitoneal injection. Results Hela-NIS( +)human cervical cancer xenografts were established successfully in nude mice. The Hela-NIS( +) xenografts significantly accumulated radioactivity after intraperitoneal injection of 131I, and the radioactivity was persistently present until 20 h postinjection, but Hela xenografts had no radioactive accumulation. The T/B value of the Hela-NIS( +) xenografts reached 17.34 at 8 h postinjection. The imaging with 99TcmO4- showed that the radioactivity was persistently present in Hela-NIS( +) xenografts for almost 25 h. The Hela-NIS( +)xenografts shrinked after 131I treatment. The inhibition ratios of tumor growth in 111 MBq and 148 MBq groups were both significantly higher than that of 74 MBq group (t: 2.74-5.75, P ＜0.05). Conclusions Hela-NIS( +) cervical cancer xenografts in nude mice could persistently accumulate 131I and 99TcmO4- and could be treated successfully with 131 I. 131 I treatment mediated by hNIS gene transfection could be a promising cancer treatment method.

OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity

CTB protein possessed mucosal adjuvant immunoactivity. The CTB gene was amplified by PCR method from a strain V. cholerae. The nucleotide sequence of CTB gene was 375 bp and shared 96.0%~99.2% homology with other 6 CTB genes. The recombinant plasmid pTWIN1-CTB transformed E. coli strain BL21(DE3) expressed with 0.8 mmol/L IPTG. The molecular weight of expression products was identical with expectative weight by SDS-PAGE electrophoresis. The CTB fusion proteins mainly assembled inclusion bodies and the outputs of proteins were approximately 20% of the total bacterial proteins. The CTB proteins possessed mucosal immunoactivity by GM1-ELISA assay.

Objective We sought to establish the standard operation procedures in department of cardiology.Methods According to the method of constructing standard operation procedures,the standard operation procedures for the department of cardiology was constructed through induction and consulting literature materials.Results 16 standard operation procedures for the department of cardiology were developed,including 8 SOP of common diseases to rescue,2 SOP of common operation in perioperative period,4 SOP for common instruments,2 SOP for common drugs usage.Conclusions Construction of standard operation procedures in department of cardiology can provide the basis for clinical nursing work,it can also supply methodological reference to build the standard operation procedures in other departments.

Objective　To explore the relationship between angiogenesis and tumor metastasis and prognosis by studying the angiogen esis in colorectal carcinoma tissues and metastasized tissues. Me thods　The microvessel density (MVD) and the expression of vascular endo thelial growth factor (VEGF) were studied with immunohistochemical assays in 15 samples of colorectal carcinoma with lymph node and liver metastasis, 20 specime ns of colorectal carcinoma without metastasis. Normal rectal mocosal tissues wer e taken from 10 cases of colorectal carcinoma with metastasis and 10 without.Re sults　The MVD was obviously higher in the cases of colorectal carcinoma with or without metastasis than in normal rectal tissues, and that in those wit h metastasis was higher than that of those without. The MVD was significantly hi gher in those positive to VEGF staining than those negative in colorectal carcin oma tissues and metastasized tumors. Conclusion　The MVD and VEG F expression can be regarded as indexes for tumor metastasis and prognosis in co lorectal carcinoma.

This study is to investigate the anti-tumor effect in vitro of methotrexate modified by LH-RH peptide (LH-RH-MTX). LH-RH receptors highly expressing MCF-7 human breast carcinoma cell line and lowly expressing K562 human erythroleukemia cell line were served as the tested cells. The cell proliferation inhibition rates of LH-RH-MTX were detected by MTT colorimetric assay. The effects of LH-RH-MTX on the cell cycle and apoptosis rates were detected by flow cytometry. The inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that on K562 cells, and the inhibition rate of LH-RH-MTX on MCF-7 cells was much higher than that of free MTX at the same concentration. The inhibition rate of LH-RH-MTX on rat bone marrow mononuclear cells was less than that of free MTX. The number of MCF-7 cells in S phase increased after administration of LH-RH-MTX. The apoptosis rate of LH-RH-MTX group significantly increased compared with that of the control group and MTX group. The relative expression of LHRHR mRNA of LH-RH-MTX group markedly decreased compared with that of the control group and MTX group. LH-RH-MTX is realizable to reduce drug side effects, increase the therapeutic index and achieve tumor-targeted therapy.
Animals ; Antimetabolites, Antineoplastic ; chemical synthesis ; pharmacology ; Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Drug Delivery Systems ; Gonadotropin-Releasing Hormone ; chemistry ; pharmacology ; Humans ; K562 Cells ; Leukocytes, Mononuclear ; MCF-7 Cells ; Methotrexate ; chemical synthesis ; pharmacology ; RNA, Messenger ; metabolism ; Rats ; Receptors, LHRH ; biosynthesis ; genetics

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism

Objective: To clarify the association between occupational exposure to wood dust and chronic obstructive pulmonary disease risk by a meta-analysis. Methods: A systematic search of the studies was conducted using 3 English databases (Pubmed, Embase, and Cochrane library) and 3 Chinese databases (CNKI, WanFang, and VIP) before March 2019. The following key words was used: 1) wood, 2) hardwood, 3) softwood, 4) saw, 5) dust, 6) chronic obstructive pulmonary disease, 7) chronic obstructive airway disease, 8) lung function. A quality score was evaluated by Newcastle-Ottawa Scale, NOS (Wells, 2012). Pooled effect value with 95% confidence interval (CI) was calculated using fixed-effect model (Heterogeneity test I²<25%) or random-effect model (Heterogeneity test I²≥25%). Meta-regression was used to explore heterogeneous source. Sensitivity analysis was used to verify the stability of the results. Publication bias was assessed by Egger＇s test. Fill and trim method was used to correct the pool effect value with 95%CI for studies which wit publication bias. The TSA threshold was calculated by the O＇Brien-Fleming loss function in the TSA data. The studies were evaluated based on the accrued information size (AIS) . Results: A total of 9 studies were included in the analysis. The occupational exposure to wood dust was not significantly associated with increased chronic obstructive pulmonary disease risk (ES=1.01, 95%CI: 0.856-1.194). TSA showed that the cumulative Z-value curve neither gone beyond the traditional (Z=1.96) threshold line, nor exceed the TSA threshold, but has reached the expected amount of information. This result was consistent with the meta-analysis. Conclusion This study does not yet consider that COPD is associated with occupational wood dust exposure.