Objective:To investigate the role of integrin-linked kinase (ILK)-small interfering RNA (siRNA)-adeno-associated virus (AAV) in scar formation after glaucoma filtering surgery in rat eyes.Methods:Forty-eight Sprague Dawley rats of SPF grade, aged 8 to 9 weeks old, were selected and divided into blank control group, ILK-siRNA-AAV group, NC-siRNA-AAV group and mitomycin C (MMC) group by random number table method, with 12 rats in each group.Left eyes of the rats were taken as experimental eyes, and no intervention was administered to fellow eyes.The bulbar conjunctival filtering bleb after glaucoma filtration surgery in rats was established by anterior chamber drainage tube implantation.One day after operation, phosphate buffer solution, ILK-siRNA-AAV, and NC-siRNA-AAV were injected into the filtering bleb of blank control group, ILK-siRNA-AAV group and NC-siRNA-AAV group, 5 μl each group, respectively.Cotton tablets containing 0.4 mg/ml MMC were placed under conjunctival flap for 5 minutes during operation in MMC group.Intraocular pressure (IOP) was measured with a handheld tonometer before surgery and at 1, 2, 3, 7, 14, 21, 28 days after surgery.Formation of filtering blebs in rats was observed with a surgical microscope at 1, 2, 3, 7, 14, 21, 28 days after operation, and the bleb survival time was calculated by Kaplan-Meier survival analysis.The mRNA and protein expression levels of ILK in conjunctival and subconjunctival tissues at the surgical sites were detected by reverse transcription PCR and Western blot, respectively, on the 28th day after operation.Silencing of ILK gene was identified.Effect of ILK gene silencing on the morphology of drainage pathway was observed by hematoxylin-eosin staining.Effect of ILK gene silencing on collagen fiber deposition in the bulbar conjunctiva at filtration area was examined by Masson staining, and the percentage of positive area of collagen fiber staining in the total tissue visual field was calculated.The use and care of the animals complied with the ARVO Statement.This research protocol was approved by an Ethics Committee of Xi'an Jiaotong University (No.2013-772). Results:There were statistically significant differences in IOP at different time points between before and after surgery among four groups ( Fgroup=76.84, P<0.001; Ftime=114.49, P<0.001). The IOP of ILK-siRNA-AAV group on the 1st, 7th, 14th and 28th day after operation and the IOP of MMC group on the 2nd, 3rd, 7th, 14th and 28th day after operation were lower than those of blank control group, and the differences were statistically significant (all at P<0.05). The IOP of ILK-siRNA-AAV group was lower on 7th, 14th, 21st and 28th day after operation than those of NC-siRNA-AAV group, with statistically significant differences (all at P<0.05). The bleb survival time of blank control group, NC-siRNA-AAV group, ILK-siRNA-AAV group and MMC group was (3.50±1.51), (5.00±3.41), (31.50±3.15) and (31.33±2.46) days, respectively, with a significant difference among them ( F=395.83, P<0.05). The bleb survival time of ILK-siRNA-AAV group and MMC group was higher than that of blank control group and NC-siRNA-AAV group, and the differences were statistically significant (all at P<0.05). There were statistically significant differences in the relative expression levels of ILK mRNA and protein among four groups ( F=222.32, 752.69; both at P<0.05), and the relative expression levels of ILK mRNA and protein were significantly lower in ILK-siRNA-AAV group than blank control group and NC-siRNA-AAV group, and the differences were statistically significant (all at P<0.05). Proliferative fibrous connective tissue and a large number of cells at surgical sites were found in blank control group and NC-siRNA-AAV group, and the fibroblasts were of a high density and grew in clumps.In ILK-siRNA-AAV group, the bulbar conjunctiva was thin, and the arrangement of fibrous connective tissue was loose, and a few proliferative fibroblasts were found.In MMC group, the conjunctival fibrous layer was loose and thin, forming cavities, and scarce cells were found.There was statistically significant difference in the percentage of collagen fiber positive staining area among four groups ( F=741.66, P<0.05). The positive staining percentage of ILK-siRNA-AAV group and MMC group was significantly lower than that of blank control group, among which there was lower positive staining percentage in ILK-siRNA-AAV group than NC-siRNA-AAV group, and the differences were statistically significant (all at P<0.05). Conclusions:Silencing of ILK can inhibit the scar formation after glaucoma filtering surgery and maintain low IOP in rats.

Objective: To explore influence of bisoprolol on neuroendocrine factor levels in patients with chronic heart failure (CHF), and its therapeutic effects.Methods: A total of 116 CHF patients, who were treated in our department from Jan 2014 to Nov 2015, were selected.According to random number table, they were randomly and equally divided into routine treatment group and bisoprolol group.Left ventricular ejection fraction (LVEF), left ventricular end-diastolic dimension (LVEDd), 6min walking distance (6MWD), serum levels of brain natriuretic peptide (BNP), endothelin (ET) and atrial natriuretic peptide (ANP) before and three months after treatment, and therapeutic effect were compared between two groups.Results: There were no significant difference in above indexes between two groups before treatment, P>0.05 all.Compared with routine treatment group after three-month treatment, there were significant rise in LVEF [(28.13±4.16)% vs.(36.02±5.14)%] and 6MWD [(245.74±44.62)m vs.(315.23±53.12)m], significant reductions in LVEDd [(62.73±10.14)mm vs.(52.82±9.47)mm], serum levels of BNP [(424.51±62.26) ng/ml vs.(324.35±50.46) ng/ml], ET [(80.14±13.25) ng/ml vs.(57.91±10.32) ng/ml] and ANP [(233.24±42.35)ng/ml vs.(164.83±31.26) ng/ml] in bisoprolol group, P<0.05 all.Compared with routine treatment group, there was significant rise in total effective rate (72.4% vs.86.2%) in bisoprolol group, P=0.04.Conclusion: Bisoprolol can significantly reduce BNP, ET and ANP levels, then delay ventricular remodeling and improve cardiac function in patients with chronic heart failure.The therapeutic effect is significant, which is worth extending.

Objectives To investigate the effects of ethanol on neural development and kainate receptor expression in young mice. Methods Fetal alcohol spectrum disorder model was established by administration of 20% ethanol solu?tion to 7-day-old Kunming mice and control animals received physiological saline (The number of treatment and control were 80 and 40, respectively ). Body weight and general biological features were observed every day. Morris water maze was used to test learning and memory ability. Fluoro-Jade B was used to examine neural cells 24 hours after treatment in additional thirty 7-day-old Kunming mice which were further divided into two groups:a treatment group receiving 20%ethanol solution (n=15) and a control group receiving physiological saline (n=15). The development of neural cells and expression levels of kainite receptors were examined by using immunofluorescence staining. Results The body weight was significantly lighter in treatment group than in control group(control:21.13 ± 1.72g,treatment:13.96 ± 2.98g,P<0.05). Morris test showed that model group had longer latency than control group to find hidden platform(control:21.05± 5.31s,treatment:34.15±3.26s,P<0.05). Spatial probe test revealed that the number of passing through the platform were significantly smaller in model group than in control group(control:2.70 ± 1.25 times,treatment:0.93 ± 0.80 times,P<0.05). Astrocyte development anomaly was evident after ethanol treatment for 7 days. The expression levels of kainite re?ceptor GluR-6 and KA2 were up-regulated in the CA region of the hippocampus after ethanol treatment for 7 days. Con?clusion Kainite receptor GluR-6 and KA2 in CA region of the hippocampus may contribute to ethanol-induced hippo?campal neural development anomaly.

OBJECTIVE: To build a physiological aging scale for healthy people.
 METHODS: We collected age-related physiologic items through literature screening and expert interview. Two rounds of Delphi were implemented. The importance, feasibility and the degree of authority for the physiological index system were graded. Using analytic hierarchy process, we determined the weight of dimensions and items.
 RESULTS: Using Delphy mothod, 17 physiological and other professional experts offered the results as follow: coefficient of expert authorities Cr was 0.86±0.03, coordination coefficients for the first and second round were 0.264(χ2=229.691, P<0.001) and 0.293(χ2=228.474，P<0.001), respectively. The consistency was good. The aging scale for healthy people included 3 dimensions, namely physical form, feeling movement and functional status. Each dimension had 8 items. The weight coefficients for the 3 dimensions were 0.54, 0.16, and 0.30, respectively. The Cronbach's α coefficient of the scale was 0.893, the reliability was 0.796, and the variance of the common factor was 58.17%.
 CONCLUSION The improved Delphi method or physiological aging scale is satisfied, which can provide reference for the evaluation of aging.
Aging ; physiology ; Delphi Technique ; Humans ; Reproducibility of Results

Objective To sum up the techniques of radiofrequency ablation assisted laparoscopic liver resection .Methods A ret-rospective analysis was done based on the clinical data of 16 patients undergone radiofrequency ablation assisted laparoscopic liver resection from July 1 ,2011 to July 30 ,2012 .Results Sixteen patients were all received successful total laparoscopic liver resection . Anatomical liver resection was carried out on 5 patients including 2 left hemihepatectomy ,2 left lateral segmentectomy ,1 right pos-terior lobectomy ,and 11 patients underwent nonanatomical hepatectomy .None was transected under regional hepatic blood inflow occlusion .The mean operation time was 125 minutes(range 35-335 minutes) ,mean blood loss 310 mL(range 20~1 100 mL) ,and mean hospital stay 9 days(range 5 to 16 days) .No operation death and postoperative complications occurred .The patients were fol-lowed up for 2 to 12 months ,1 recurrence was found in patients with Ⅶ segment hepatocellular carcinoma 60 day after operation . Conclusion The application of radiofrequency ablation assisted laparoscopic liver resection can effectively control the resection mar-gin hepatic blood inflow to ensure the success of operation and reduction of complications .

Objective To investigate the rehabilitative effect of low frequency repetitive transcranial magnetic stimulation (rTMS) on convalescing patients with Broca's aphasia. MethodsTwenty-eight patients with Broca's aphasia recovering from cerebral infarction were randomly divided into a stimulation group and a control group with 14 subjects in each.Patients in the control group accepted conventional drugs,speech rehabilitation and sham stimulation,while patients in the stimulation group were in given low frequency rTMS in place of the sham stimulation.Their speech performance was evaluated using the China Rehabilitation Research Center's aphasia examination (CRRCAE) pre-stimulation,post-stimulation and 90 days later. ResultsCompared with before treatment and with the controls,the speaking scores of the stimulation group increased significantly after treatment and also 90 days later. ConclusionLow frequency rTMS can not only improve the speech performance of Broca's aphasia sufferers in the short term,but it also plays a lasting role.It may thus have clinical application for patients with Broca's aphasia.

This study is to investigate the transportation of scutellarin in cell and live models and study on mechanism of absorption and transport of scutellarin in mouse liver. The concentration of scutellarin in plasma and liver from control and pretreated groups was determined by high performance liquid chromatography. The uptake of scutellarin was examined in control hepatocytes group, induced hepatocytes group and induced hepatocytes plus pravastatin group. Pravastatin can affect the pharmacokinetics of scutellarin in mouse: CL is decreased while AUC is increased. The scutellarin absorption of hepatocyte induced group was higher than that of control group, but was decreased in the group with pravastatin added. The research showed that there was potential drug interaction between pravastatin and scutellarin. The drugs may compete for oatp2 mediated transport pathway consisted in the uptake of scutellarin in liver.

BACKGROUND:An abroad study repoRed the distribution and expression of augmenter of liver regeneration(ALR)in the central nervous system.There are few literatures on how to prepare and evaluate ALR protein polyclonal antibody in recombinant rats,and how to construct prokaryotic expression vector.There are no repots concerning ALR in the central nervous system in China.OBJECTIVE:TO express ALR fusion protein in E coli BL21 and prepare and identify polyclonal antibody.METHODS:RNA was extracted from the hippocampus of Sprague Dawley rats.The prokaryotic expression plasmid pET28a-ALR was constructed and the positive recombinant plasmid was transformed into BL21.Protein ALR was expressed by inducing transformed BL21 with Isopropyl-β-D-thiogalactopyranoside(IPTG)and purified by Ni~(2+)affinity chromatography column after immune the rabbit for 4 times.the serum of rabbits was extracted from hear as polyclonal antibody.The titer and specificity of the rabbit's antiserum was respectively measured by ELISA and Western blotting The following parameters were measured:construction of prokaryotic expression plasmid pET26a-ALR;pET28a-ALR recombinant enzyme digestion evaluation;results of ELISA and Western-blotting.RESULTS AND CONCLUSION:Expecting bands were obtained by double enzyme digestion electrophoresis,respectively 5.3 kb and 0.4 kb.Nucleotide sequence analysis verified that prokaryotic expression vector pET28a-ALR was successfully constructed.The 19 ku fusion protein was successfuIly expressed.The titer of the antiserum measured by ELlSA could achieve 1:2 000 This indicated that antibody and purified recombinant ALR had a good reaction.and high titer.could meet the experimental require.Western blotting analysis proved that the antibody could identify the prokaryotic expression product of ALR.Prokaryotic expression system expressed ALR fusion protein,prepared and purified polyclonal antibody of ALR protein,and could meet the experimental require of ALR immunoblotting.

Objectlve To investigate the value of basic and contrast-enhanced ultrasound in the diagnosis of superficial cervical lymph nodes.Methods Five hundred and forty-five cases of superficial cervical lymph nodes were sacned by basic ultrasound,in which 52 cases were also scaned by contrast-enhanced ultrasound.All cases were performed ultrasound-guided biopsy.Lymph nodes were divided into benign group and malignant group according to pathology reports.The differences of the two groups were analysed,and statistical analysis was performed.Results Two hundred and thirty cases were benign,315 cases were malignant.S/L(P＜0.01),RI(P＜0.01),vascular pattern(P＜0.01)and contrast enhancement pattern(P＜0.01)between benign and malignant group showed statistical significant differences,while no statistical difference in coefficient correlation of the time-intensity curve between the two groups was found.Conclusions A combination of basic and contrastenhanced ultrasound can significantly enhance the ability to identify malignant lymph nodes from benign lymph nodes.

Scutellarin is the major effective ingredient of breviscapine, which is widely used in the treatment of cardiovascular diseases and has a remarkable achievement. In recent years, more and more reseaches had focused on the pharmacokinetics of scutellarin and scutellarein. The article summarized the studies on pharmacokinetics of scutellarin and scutellarein have been performed by domestic and oversea scholars.
Animals ; Apigenin ; pharmacokinetics ; Flavonoids ; chemistry ; Glucuronates ; pharmacokinetics ; Humans