Background Choroidal neovascularization (CNV) is one of the causes of blindness in multiple eye diseases.Researches showed that complement system participates in the pathogenesis of CNV.Objective This study was to construct the recombinant of complement factor B-small interference RNA (CFB-siRNA) expression vector and to observe its inhibitory effect on human umbilical vein endothelial cells (ECV-304).Methods CFB gene primers were designed based on human CFB gene,and an expression vector of CFB-siRNA was constructed by inserting CFB-siRNA into pRNAT-U6.1/Neo plasmid.Recombinant plasmids were confirmed by the digestion analysis of restriction endonuclease,and all inserted sequences were verified by DNA sequencing.The recombinant pRNAT-U6.1/CFB-siRNA plasmid and the blank plasmid were transfected into ECV-304 cells in the CFB-siRNA group and blank plasmid group by electroblot,respectively,and non-transfected cells served as the normal control group.The cells were observed under the fluorescence microscope 48 hours after transfection,and the transfective efficiency was calculated.The relative expression of CFB mRNA in the cells of different groups was detected by semi-quantitative reverse transcription PCR (RT-PCR).MTT was employed to calculated the growth inhibitory rates of the cells 24,48 and 72 hours after transfection.The percentages of the cells in different cell cycles were detected by flow cytometry.Results The sequence of the target vector was identical to the designed sequence.The green fluorescence protein (GFP) was seen in both the CFB-siRNA group and the blank plasmid group.The relative expression levels of CFB mRNA were 0.07 ±0.04,0.14 ±0.02 and 0.14 ±0.03 in the CFB-siRNA group,the blank plasmid group and the normal control group,respectively,a significant difference was obtained among the three groups (F=233.05,P =0.00);the expression level of CFB mRNA in the CFB-siRNA group was significantly declined in comparison with the blank plasmid group and the normal control group (both at P＜0.05).The growth inhibitory rates of the cells were (23.45 ±0.01) ％,(33.48 ±0.02) ％ and (45.49±0.01) ％ at 24,48 and 72 hours after transfection,respectively,a significant difference was obtained among the three groups (Fgroup =212.99,P =0.00);the growth inhibitory rates in CFB-siRNA group were significantly higher than that in the blank plasmid group and normal control group (all at P＜ 0.05).The percentages of G1 phase cells were (44.4 ±0.5) ％,(25.8 ±0.4) ％ and (27.9 ± 0.6) ％ in the CFB-siRNA group,the blank plasmid group and the normal control group respectively,a significant difference was obtained among the three groups (F=58.98,P=0.00).The percentages of G1 phase and G2 phase cells in the CFB-siRNA group were significantly higher than those in the blank plasmid group and the normal control group (all at P＜0.05).Conclusions Recombinant pRNAT-U6.1/CFB siRNA inhibits the proliferation of ECV-304 cells effectively by arresting the cells in G1 intermediate phase of the growth cycle.

OBJECTIVETo observe the inhibitory effect of gefitineb on the proliferation and its inducing effect on the apoptosis of mouse I-10 Leydig testicular cancer cells in vitro.

METHODSWe treated I-10 Leydig testicular cancer cells of mice with gefitineb at 0, 1.25, 2.5, 5, 10, 20, and 40 µmol/L. Then we determined the inhibitory effect of gefitineb on the growth of the cells by MTT, detected their early and late apoptosis by Annexin V-FITC/propidium iodide double staining and Hoechst 33258 nuclear staining, respectively, and observed the expressions of apoptosis-related proteins Bcl-2, Bax and caspase 3/9 by Western blot.

RESULTSCompared with the blank control group, gefitineb significantly inhibited the proliferation of the I-10 cells at 10 and 20 µmol/L (P < 0.05). The survival rate of the cells was (32.4 ± 2.8)% (P < 0.01) and their early and late apoptosis rates were (26.7 ± 4.2)% and (59.33 ± 10.2)% in the 40 µmol/L group, significantly different from those in the control (P < 0.05 and P <0.01). In comparison with the blank control group, gefitineb at 10, 20, and 40 µmol/L increased the expression of pro-apoptotic protein Bax by (41.9 ± 7.1), (60.1 ± 9.8), and (69.0 ± 11.3)% (all P < 0.05), decreased that of apoptosis-inhibitory protein Bcl-2 by (50.3 ± 8.9), (63.9 ± 6.9), and (88.7 ± 13.9)% (all P < 0.05), and elevated that of the cleft proteins caspase-3 by (69.0 ± 6.9)% (P < 0.05), (71.5 ± 8.1)% (P < 0.05), and (110.9 ± 14.2)% (P < 0.01) and caspase-9 by (51.8 ± 4.9), (54.7 ± 6.7), and (43.8 ± 11.8)% (all P < 0.05).

CONCLUSIONGefitineb can increase the cytotoxicity of I-10 Leydig testicular cancer cells of mice and induce their apoptosis via the mitochondria-mediated apoptosis signaling pathway.

Animals ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Apoptosis Regulatory Proteins ; metabolism ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Proliferation ; drug effects ; Cell Survival ; Leydig Cell Tumor ; drug therapy ; metabolism ; pathology ; Male ; Mice ; Neoplasm Proteins ; metabolism ; Neoplasms, Germ Cell and Embryonal ; drug therapy ; metabolism ; pathology ; Quinazolines ; pharmacology ; Testicular Neoplasms ; drug therapy ; metabolism ; pathology ; bcl-2-Associated X Protein ; metabolism

AIM: To establish rat chronic obstructive pulmonary disease(COPD) models by passive cigarette smoking plus intratracheal instillation of lipopolysacchride(LPS) or passive cigarette smoking only, which would be similar to the pathogenesis of human COPD. METHODS: 48 Wistar rats were randomly divided into 4 groups.(1) Healthy control I group( n =12), rats were bred 4 weeks;healthy control II group( n =12), rats were bred for 3 months. (2) Model group I ( n =12), 200 ?g lipopolysaccharide(LPS) was instilled intratracheally once for every two weeks and the rats were exposured to 5% of cigarette smoke, 0.5 h/d for 4 weeks.(3) Model group II(n=12),rats were exposed to 5% of cigarette smoke, 0.5 h/d for 3 months. The pathologic changes of airways and lung tissues, pulmonary function and blood gas analysis were determined. The airway wall lymphocytes and alveolar macrophages were counted. The cross areas of epithelial layer, smooth muscle layer and lamina propria of bronchi were measured. The hydroxyproline of lung tissue homogenates was determined by biochemistry method. RESULTS: The pathologic changes of airways and lung tissue of two models were similar to but milder than those of COPD patients(biopsy data). The collagen deposition and the cross areas of epithelial layer and smooth muscle layer in airway walls of two model groups were significantly increased than those of control groups( P

Objective To explore the anatomical characteristic of the third palmar interosseous mus- cle as well its dominate nerve,and to investigate the anatomical basis of difficult recovery of digitus minimus adduction.Methods Twenty aduh fresh hands without deformity and trauma were obtained.Dissect and observe the third palmar interosseous muscle and its dominate nerve and adjacent structure under surgical mi- croseope,measure the size of the third pahnar interosseous muscle and its dominate nerve,the data were pro- cessed by stastistics method.Results Among palmar interosseous muscles and its dominate nerves,the third palmar interosseous muscle and its dominate nerve is the smallest.There are conspicuous tendon bundle on the surface of the third palmar interosseous muscle partly,which have a potential compression on the third palmar interosseous muscle dominting nerve.Conclusion The third palmar interosseous muscle is the smal- lest among palmar interusseous muscles and it is the only digitus minimus adduction muscle.The sominating nerve of the third palmar interosseous muscle is small anti the tendon bundle of the third palmar interosseous muscle have a potential compression.All these can cast light on diffcult recovery of digitus minimus adduction.

Objective:To make an assessment on the genotoxicity caused by black carbon ( BC ) and ozonized black carbon (O3-BC).Methods: In this study, 74 healthy male ICR mice [weighed (28 ± 1.5) g] were randomly divided into 7 groups, including one phosphate buffer solution ( PBS) control group and six particles exposed groups by intratracheal instillation with either BC or O 3-BC at the doses of 50, 100, 200 μg/mouse, respectively.There were 12 mice in the groups of 200μg/mouse and 10 mice in others.The mice were sacrificed 24 h after four intratrachealinstillations .The activities of catalase ( CAT) in serum and the levels of malondialdehyde ( MDA) in lung tissue homogenate were measured . As the DNA damage mark , 8-hydroxyguanosine ( 8-OHdG ) in urine and serum were quantified with ELISA method.Micronucleus test was used for potential genotoxicity of BC and O 3-BC.Hematoxylin and eosin staining was used to stain lung paraffin section .Results:The mice were in good condition during instillation , and the liver coefficient of the test groups was significantly lower than that of the control group (P<0.05).The activities of CAT in serum significantly increased in the 100 μg/mouse and 200μg/mouse groups after being exposed to these two kinds of particles .The micronucleus rate in allthe BC and O3-BC exposed groups increased ( P <0.05), but there was no statistically significant difference among the groups in the levels of 8-OHdG in serum and urine and MDA in lung tissue homogenate .In-flammatory response was found in the lung tissue under the microscope after exposure to BC and O 3-BC. Conclusion:Intratracheal instillation of BC and O 3-BC induced increasing of oxidative stress and genetic damage in mice .But there was no significant difference between these two particles in toxicity .Whether the genotoxicity of O 3-BC is higher than that of BC or not is uncertain .Further research is needed .

ObjectiveTo evaluate the effect and safety of different thrombolytic therapies for acute cerebral infarction due to occlusion of middle cerebral artery(MCA).MethodsOne hundred and thirty-two cases of acute cerebral infarction in territory of MCA were randomly divided into 3 groups,all of which were treated with alteplase.Group A (48 cases) was treated by intra-venous therapy with alteplase,group B (43 cases) was treated by infusing alteplase at the site of the internal carotid artery,and group C(41 cases) was treated by infusing alteplase into the thrombus.The improvement of neurological function,complications and mortality rate were recorded and statistically compared,with analysis of variance for counting data of normal distribution,x2 test for quantitative data,and the mean difference was significant at the 0.05level.ResultsThe effective rates of group A,B and C at 2 h,24 h,2 w were 18.8％ (9/48),39.6％ ( 19/48),45.8％ (22/48) ;39.5％ (17/43),53.5％ (23/43),58.1％ (25/43) ;78.0％ (32/41),85.4％ (35/41 ),87.8％ (36/41)respectively.The effective rate of group C was obviously better than group A( x2 =12.809,9.979,9.289,P ＜ 0.01 ) and B (x2 =31.295,19.425,17.161,P ＜ 0.01 ) with statistical significance.The effective rate of group B was better than group A at 2 h after thrombolytic therapy with statistical significance (x2 =4.801,P ＜ 0.05 ).The effective rate of group A and B did not have significant difference at 24 h,2 w after therapy ( x2 =1.765,1.375,P ＞ 0.05 ).The hemorrhage rates of group A,B and C were 14.6％ (7/48),14.0％ (6/43),7.3％ (3/41 ),the mortality rates of group A,B and C were 6.2％ (3/48),4.6％ (2/43),2.4％ (1/41),and there was no significant difference among the 3 groups ( x2 =1.328,0.786,P ＞ 0.05 ).ConclusionIt is suggested that the thrombus-imbeded thrombolytic therapy is a better way in treating acute cerebral infraction due to occlusion of MCA for its rapid and better therapeutic effect.

Objective To discuss the key points of endovascular therapy for complex subclavian artery occlusive diseases. Methods During the period from January 2012 to December 2013, a total of 92 patients with complex subclavian artery occlusive disease were admitted to Xuanwu Hospital of Capital Medical University, Beijing, China. The clinical data were retrospectively analyzed. The features of the lesions, the success rate of endovascular therapy, the use of combined approaches, the relief of symptoms after treatment, etc. were evaluated. Results The complex subclavian artery occlusive diseases could be divided into three types. Type Ⅰ: long segment of the left subclavian artery was occluded; type Ⅱ: ostial stenosis or occlusion of the right subclavian artery; and type Ⅲ: subclavian artery stenosis or occlusion was associated with the ostial disorder of the vertebral artery, or the opening of vertebral artery was affected by the subclavian artery stenosis or occlusion. The technical success rate was 82.6%. Combination use of femoral artery and brachial artery approach was employed in 27.2% of patients, which had improved the technical success rate. After the treatment the symptom improvement rate was 81.6%. Conclusion Upper limb artery approach can improve the re-canalization rate of left subclavian artery with long segment occlusion, and can ensure the accurate positioning of stent at the site of right subclavian artery opening. During the procedure of endovascular intervention for subclavian artery occlusion disease, attention should be paid to the protection of the vertebral artery.

With the prevalence of obesity and metabolic syndrome(MetS), nonalcoholic fatty liver disease(NAFLD) instead of chronic hepatitis B has been the most common cause of chronic liver disease in China. In order to standardize the diagnosis and treatment of NAFLD, the Fatty Liver Expert Committee of Chinese Medical Association issued “The guideline of prevention and treatment for nonalcoholic fatty liver disease” in March 2018, which was similar to the latest NAFLD Guidelines published by the AASLD, EASL and Asia-Pacific working party in the recent two years, but was different in details owing to the distinct condition of different countries. The purpose of the article is to help clinicians understand the guidelines more comprehensively and profoundly and guide their clinical work according to the interpretation of Chinese and overseas NAFLD guidelines.

PURPOSETo evaluate midazolam sequential with dexmedetomidine for agitated patients undergoing weaning to implement light sedation in ICU.

METHODSThis randomized, prospective study was conducted in Tianjin Third Central Hospital, China. Using a sealed-envelope method, the patients were randomly divided into 2 groups (40 patients per group). Each patient of group A received an initial loading dose of midazolam at 0.3-3mg/kg·h 24 h before extubation, followed by an infusion of dexmedetomidine at a rate of 0.2-1 μg/kg·h until extubation. Each patient of group B received midazolam at a dose of 0.3-3 mg/kg·h until extubation. The dose of sedation was regulated according to RASS sedative scores maintaining in the range of -2-1. All patients were continuously monitored for 60 min after extubation. During the course, heart rate (HR), mean artery pressure (MAP), extubation time, adverse reactions, ICU stay, and hospital stay were observed and recorded continuously at the following time points: 24 h before extubation (T1), 12 h before extubation (T2), extubation (T3), 30 min after extubation (T4), 60 min after extubation (T5).

RESULTSBoth groups reached the goal of sedation needed for ICU patients. Dexmedetomidine was associated with a significant increase in extubation quality compared with midazolam, reflected in the prevalence of delirium after extubation (20% (8/40) vs 45% (18/40)), respectively (p= 0.017). There were no clinically significant decreases in HR and MAP after infusing dexmedetomidine or midazolam. In the group A, HR was not significantly increased after extubation; however, in the group B, HR was significantly increased compared with the preextubation values (p < 0.05). HR was significantly higher in the group B compared with the group A at 30 and 60 min after extubation (both, p <0.05). Compared with preextubation values, MAP was significantly increased at extubation in the group B (p < 0.05) and MAP was significantly higher at T3, T4, T5 in the group B than group A (p < 0.05). There was a significant difference in extubation time ((3.0 ± 1.5) d vs (4.3 ± 2.2) d, p < 0.05), ICU stay ((5.4 ± 2.1) d vs (8.0 ± 1.4) d, p < 0.05), hospital stay ((10.1 ± 3.0) d vs (15.3 ± 2.6) d, p <0.05) between group A and B.

CONCLUSIONMidazolam sequential with dexmedetomidine can reach the goal of sedation for ICU agitated patients, meanwhile it can maintain the respiratory and circulation parameters and reduce adverse reactions.

Adult ; Aged ; Critical Care ; methods ; Delirium ; drug therapy ; etiology ; Dexmedetomidine ; administration & dosage ; Female ; Humans ; Hypnotics and Sedatives ; administration & dosage ; Intensive Care Units ; Length of Stay ; Male ; Midazolam ; administration & dosage ; Middle Aged ; Prognosis ; Prospective Studies ; Respiration, Artificial ; adverse effects ; methods ; Risk Assessment ; Statistics, Nonparametric ; Treatment Outcome ; Ventilator Weaning ; adverse effects ; psychology

OBJECTIVETo study the effects of prenatal exposure to Di-(2-ethylhexyl)-phthalate (DEHP) on genome-wide epigenetic alterations in ovary of adult offspring rat.

METHODSPregnant Wistar rats were randomly treated with DEHP (1000 mg/kg) or con oil at 12 - 17 days upon pregnance. DNA methylation changes in the ovary for the adult offsprings which were 70 days old were detected by Rat DNA methylation promoter plus CpG island arrays CpG island chip. Gene ontology (GO) method was performed to analyze the function of genes which were significantly different between exposed group and control group. Gene Igfbp1 (insulin-like growth factor binding protein 1) and Itga3 (integrin alpha 3) were randomly selected and the methylation status were verified by bisulfite genomic sequencing (BSP).

RESULTSThe methylation status were significantly different between exposed and control group in 406 genes (71 genes as hypermethylation and 335 genes as hypomethylation) (P < 0.05). GO analysis revealed that molecular transducer activity, cell part, cell, cellular process, multicellular organismal process, response to stimulus, biological regulation, regulation of biological process, reproduction, reproductive process, and rhythmic process were involved. The sequencing results were consistent with the data obtained by chips.

CONCLUSIONThis study provides evidence that prenatal exposure of DEHP may be associated with methylation changes on the genes in the rat ovary. Genes related to reproductive process have highly significant methylation changes, which may shed new light on mechanisms of reproductive and developmental toxicity after prenatal exposure to DEHP.

Animals ; CpG Islands ; genetics ; DNA Methylation ; Diethylhexyl Phthalate ; toxicity ; Female ; Genome ; Maternal Exposure ; Ovary ; pathology ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Wistar