OBJECTIVE:To establish a RP-HPLC method to determine the content of ofloxacin in compound ofloxacin eye drops METHODS:Using Shim-pack ODS as fixed phase,a mixture to citric acid(0 05mol/L)-acetonitrile-ammonium acetate(0 5mol/L)-0 3% phosphoric acid(78∶22∶1∶3)as mobile phase,and detecting wavelength:293nm,flow rate:1 0ml/min RESULTS:The linear range of ofloxacin was 8 12?g～40 6?g(r=0 9 996,n=5) The average recovery was 99 29% and RSD(n=6)was 0 57% CONCLUSION:The method is rapid,simple and accurate

Objective To construct clinical thinking competence index system for undergraduate nursing students.Methods The study established the Clinical Thinking Competence Evaluation Index System through Delphi method.Results The evaluation index system included 5 level-1 dimensions and 28 level-2 Index connotations.Conclusions Through quantitative research methods,Clinical Thinking Competence evaluation index system is established for undergraduate nursing students,which provides an objective standard for cultivation and evaluation of undergraduate nursing students.

Objective To evaluate the diagnostic value of adenosine stress ~(99m)Tc-MIBI myocardial perfusion single photon emission computed tomography (ECT)in elderly patients with coronary artery disease (CAD). Methods Fifty-three patients aged ≥60 years without the history of myocardial infarction were included in the research. The average age of the patients was(65.5±6.5) years (60-78 years old). All patients underwent adenosine stress ~(99m)Tc- MIBI myocardial perfusion single photon ECT and coronary angiography (CAG) within 2 months, and the results were compared. Results Compared with CAG, the sensitivity, specificity, positive predictive value, negative predictive value and accurate index of ECT were 73.7%, 90.9%, 94.3%, 80.0% and 62.3%, respectively. The specificity for left anterior descending artery, left circumflex artery and right coronary artery of ECT were 90.9%, 85.7% and 100%, respectively, and the positive predictive values were 73.5%, 77.3% and 72.0%, respectively. Conclusions Adenosine is a strong vasodilator and it has multiple effects on cardiovascular system. Combination of angiocardiography and myocardial perfusion imaging can be used in diagnosis of coronary artery disease. ECT is a noninvasive, convenient and low cost method and plays an important role in diagnosing, directing treatment and judging prognosis of CAD in the elderly patients.

Objective To observe the effects on rabbit corneas and retinas after single intravitreal injection of voriconazole at different doses.Methods According to the randomization table,25 healthy rabbits were randomly divided into control group,and voriconazole 50,100,200,and 400 μg groups.Therefore,there were 5 rabbits in each group.The eyes of control group received intravitreal injection of 0.1 ml balanced saline solution,and those treatment groups received 0.1 ml voriconazole injection of corresponding dose.Before the injection and 1,7,and 14 days after the injection,endothelial cell counts and corneal thicknesses were measured;full-field electroretinogram were performed and b-wave amplitudes in maximal combined reaction (Max-R) were recorded.On 14 days after the injection,histologic structures were observed by light microscope and transmission electron microscope.Results There was no significant difference in endothelial cell counts (F=0.320,0.291,0.467,0.649) and corneal thicknesses (F=0.214,0.284,0.360,0.225) with those of control group at any time points (P＞0.05).Before and 1 day after the injection,b-wave amplitudes of each voriconazole group had no significant difference compared with those of control group (F=0.220,0.106;P＞0.05).On 7 days after the injection,b-wave amplitudes decreased significantly at doses of 200 μg and 400 μg (P＜0.05).On 14 days after the injection,there was no significant difference between the the amplitude of 200 μg group and that of control group (P＞ 0.05).However,the amplitude of the 400 μg group decreased continuously and there was still significant difference (P＜0.05).Light microscopy did not reveal any corneal abnormality in both control group and voriconazole groups.The retinas were normal except that of the 400 μg group,which had a thinner and degenerated inner nuclear layer and disordered photoreceptor layer.Under transmission electron microscope,there were no ultrastructure damages of corneas in both control group and voriconazole groups,either.The rabbit retinas of the 50 μg and 200 μg group have normal inner nuclear layer and photoreceptor layer,but degrees of changes in both layers were observed in the eyes of 200 μg and 400 μg group.Conclusions There is no obvious effects on rabbit corneas and retinas after single intravitreal injection of voriconazole at he dose less than or equal 100 μg.There are no obvious effects on rabbit corneas at the dose of 200 μg and 400 μg,while there are damages to the retinas in both functions and histological structures.

Objective:To evaluate the changes of neurocognitive function in early stage in patients with first-episode schizophrenia.Methods:In this study,73 cases of patients with first-episode schizophrenia (research group) and 75 cases of health person (control group) were selected from January 2015 to January 2016 in our hospital.The neurocognitive function was evaluated by neuro-psychological testing tool and the data between two group were compared.Results:Scores of delayed recall,total recall,3 trial,2 trial and 1 trial of research group were lower than those of control group in BVMT-R test,and the difference was statistically significant (P＜0.05).In HVLT-R test,the scores of total delay,3 trial and 2 trial of research group were significantly lower than those of control group (P＜0.05).The consuming time of dominant hands and subdominant hands in pegboard tasks were significantly higher in research group than in control group (P＜0.05).Conpletion time of connection test A,color connection 1 and 2 in connection test of research group were significantly higher than those of control group (P＜0.05).Attempt number and correct number in research group in PASAT test were significantly lower than in control group (P＜0.05).Number of search errors in research group was higher than in control group,while number of search correct,search total score and digital sign score were significantly lower than in control group (P＜0.05).Total number of words,color and color / word count in research group were lower than in control group in Stroop color word test,and the difference was statistically significant (P＜0.05).WMS-Ⅲ test results between two group had no significant difference (P＞0.05).Conclusion:The neurocognitive function in early stage in patients with first-episode schizophrenia has been extensively damaged.

BACKGROUND:The microRNAs are involved in regulation of stem cel proliferation, differentiation and aging. To study the effect of Let-7c, a member of Let-7, on the neural differentiation of bone marrow mesenchymal stem cels provides new ideas for stem cel therapy. OBJECTIVE: To investigate the role of Let-7c in the neural differentiation of bone marrow mesenchymal stem cels. METHODS: The lentiviral vectors of Let-7c-up and Let-7c-inhibition were constructed and transfected into rat bone marrow mesenchymal stem cels. Optimal multiplicity of infection was screened. The cels were divided into non-transfected group, negative control group (transfected with empty virus), transfected enhancement group (transfected with LV-rno-Let-7c-up), transfected inhibition group (transfected with LV-rno-Let-7c-5p-inhibition). Bone marrow mesenchymal stem cels were treated with fasudil as an inducer for triggering the cels to differentiate into neurons. The fluorescence expressed by transfected cels was observed under inverted fluorescence microscope. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The cel viability was determined by MTT method. RESULTS AND CONCLUSION:Under the inverted fluorescence microscope, the cels were successfuly transfected with LV-rno-Let-7c-up and LV-rno-Let-7c-5p-inhibition. Fasudil induced bone marrow mesenchymal stem cels to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in the transfected enhancement group were significantly higher than those in the negative control group (P < 0.05), while in the transfected inhibition group, they were lower than those in the negative control group (P < 0.05). These findings indicate that the differentiation percentage of bone marrow mesenchymal stem cels is increased by fasudil after transfection with LV-rno-Let-7c-up, and Let-7c may promote the differentiation of bone marrow mesenchymal stem cels into neurons.

BACKGROUND:MicroRNA plays an important role in the process of growth and aging of living body. To know the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons can promote the stem celltransplantation. OBJECTIVE:To investigate the role of let-7d in inducing bone marrow mesenchymal stem celldifferentiation into neurons. METHODS:(1) The lentiviral vector of let-7d was constructed and transfected into rat bone marrow mesenchymal stem cells. The cells were divided into non-transfected group, negative control group (transfected with FU-RNAi-NC-LV), transfected enhancement group (transfected with let-7d-LV), transfected inhibition group ( transfected with let-7d-inhibition-LV). (2) Rat bone marrow mesenchymal stem cells were treated with fasudil as an inducer for triggering the cells to differentiate into neurons. The expression of neuron-specific markers, neuron-specific enolase and microtubule-associated protein 2, were measured by immunocytochemical method. The mRNA expression of microtubule-associated protein 2 was detected by RT-PCR. The viability of bone marrow mesenchymal stem cells was determined by MTT method. RESULTS AND CONCLUSION:Under inverted fluorescence microscope, the cells were successful y transfected with let-7d. Fasudil induced bone marrow mesenchymal stem cells to differentiate into neurons. The transfection efficiency and expression levels of neuron-specific enolase and microtubule-associated protein 2 in transfected enhancement group were higher than those in the negative control group (P<0.05);while in the inhibition group, they were lower than those in the negative control group (P<0.05). These findings indicate that let-7d can promote the differentiation of bone marrow mesenchymal stem cells into neurons induced by fasudil, and by control ing the expression of let-7d we can influence the differentiation efficiency from bone marrow mesenchymal stem cells to neurons.

BACKGROUND:There is no clear understanding about the effect of let-7f and interleukin-6 (IL-6) on the proliferation of bone marrow mesenchymal stem cels and their relationship.
OBJECTIVE: To explore the effects of expression levels of let-7f and IL-6 on the proliferation of bone marrow mesenchymal stem cels and their relationship.
METHODS:(1) LV-rno-let-7f-up and LV-rno-let-7f-down were constructed and transfected into bone marrow mesenchymal stem cels of Sprague-Dawley rats, respectively. Then, there were four groups in the study: transfection upregulation group transfected with LV-rno-let-7f-up), transfection inhibition group (transfected with LV-rno-let-7f-down), negative control group (transfected with FU-RNAi-NC-LV), and untransfected group. The expression level of let-7f in each group was detected by qRT-PCR. The proliferation ability of cels and expression levels of IL-6 when let-7f expression was at different levels were detected by MTT, flow cytometry and ELISA. The expression of Cyclin D1 at mRNA and protein levels was detected by qRT-PCR and western blot, respectively. (2) To predict the potential target gene of let-7f, the wild-type/mutant IL-6 3’UTR reporter gene vectors were constructed, and cotransfected with let-7f/let-7f inhibitor respectively into the 293T cels to measure the luciferase.
RESULTS AND CONCLUSION: Compared with the negative control group, the proliferative and cloning capacities of cels in the transfection upregulation group were higher; the number of cels was significantly decreased at G1 stage and increased at S stage, and the apoptotic cels were reduced in number (P < 0.05). However, the transfection inhibition group had opposite results. The expression level of IL-6 in the transfection upregulation group was lower than that in the untransfected group and negative control group (P < 0.05); while in the transfection inhibition group, the expression level of IL-6 was significantly increased (P < 0.05). The expression of Cyclin D1 at mRNA and protein levels was up-regulated in transfection upregulation group (P < 0.05) and down-regulated in the transfection inhibition group (P < 0.05), but there was no significant difference between the negative control group and untransfected group (P > 0.05). Luciferase activity of cels transfected with wide-type IL-6 3’UTR and let-7f was significantly reduced (P < 0.05). These findings indicate that up-regulation of let-7f can promote the proliferative and cloning capacities of bone marrow mesenchymal stem cels and reduce cel apoptosis, but downrelation of let-7f exhibits an inhibitory effect. Overexpression of IL-6 can suppress the proliferation of bone marrow mesenchymal stem cels, which is considered to be a target gene of let-7f, and let-7f may suppress the expression of IL-6 to promote the cel proliferation.

Objective To construct eukaryotic expressing vector of mouse soluble CD160 and stably transfect into CHO cells for eukaryotic expression.Methods Recombinant soluble CD160(rsCD160) was constructed by gene recombination.Total RNA was extracted from the spleen of C57BL/6 mice.cDNA was amplified for the soluble form of CD160.Then,the PCR product was cloned tO pcDNA3.1 and pEGFP-N1.The recombinant plasmid was identified by restriction map and sequence analy-sis.The soluble CDl 60 expression in CHO cells transfected with recombinant psCDl 60 was verified by RT-PCR and Westernblot.The binding ability of psCD160 tO its ligand was detected by FACS.Results 520 bp mouse soluble CD160 gene was obtained.Recombinant mouse psCD160 was successfully constructed.After transfection,soluble CD160 expression in the culture supernatant of CHO cells was successfully detected.FACS analysis indicated that soluble CD160 could bind tO its ligand.Conclusion Recombinant mouse psCD160 is successfully constructed,which will benefit our further study on soluble CD160 for immune therapy against tumor in the future experiments.

A male patient of acute onset is reported, whose main clinical manifestations were ataxia and dysarthria, with elevated carcinoembryonic antigen, non-small cell lung cancer antigen, carbohydrate antigen 72-4, positive anti-Yo antibody. The patient′s gastroscopy and biopsy result suggested gastric cancer, and his symptoms got better after radical surgery. Anti-Yo-associated paraneoplastic cerebellar degeneration complicated with gastric adenocarcinoma was diagnosed. If encountering cases of ataxia or dysarthria in clinical work, the possibility of paraneoplastic cerebellar degeneration should be considered and evidence for tumor should be searched, so as to avoid missed diagnosis or misdiagnosis.