Objective To evaluate the effects of two different chronic glaucoma models on intraocular pressure elevation and retinal structure changes in rat.Methods Two different chronic ocular hypertension (COH) models were made by three episcleral vein cauterization or latex microspheres injection into anterior chamber,6 cases of each model.IOP measurements of right eyes (COH eye) and left eye (control eye) were taken weekly by TonoPen (an applanation tonometer).Retinal ganglion cells (RGCs) were retrogradely labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) seven weeks later.Retina structure was observed by immunofluorescence.Results IOP was elevated at postoperative 1-8 weeks,and the mean IOP in the episcleral vein cauterization group was (27.20 ± 1.83) mmHg (1 kPa =7.5 mmHg),whereas the control group was (19.80 ± 1.35) mmHg (P =0.001,n =6).The mean IOP in the latex microspheres injection group was (27.40 ± 1.88) mmHg,whereas the control group was (19.40 ± 1.00) mmHg (P =0.000,n =6).Compared with control rat at postoperative 8 weeks,RGCs loss in episcleral vein cauterization group were 37.9％,39.6 and 33.5％ (all P =0.000,n =6),latex microspheres injection group were 37.3％,39.4％ and 33.5％ (all P =0.000,n =6).There was no statistical difference between episcleral vein cauterization group and latex microspheres injection group (P =0.855,0.949,0.634,n =6).Compared with control rat at postoperative 8 weeks,GCL thickness in both COH models were also significant reduced,but there was no statistical difference in GCL thickness among control group (7.32 ± 0.39) μm,episcleral vein cauterization group (4.97 ±0.33) μm,latex microspheres injection group (5.00 ±0.31) μm.Misoperation or careless operation may lead to microspheres particle residual on flat-mounted rat retina.Conclusion The episcleral vein cauterization or latex microspheres injection into anterior chamber can all increase the IOP.However,there are some advantages in episcleral vein cauterization such as few costs than latex microspheres injection and no microsphere contamination.

Objective To investigate the relationship between the content of stromal cell-derived factor-1 (SDF-1) in myocardium in different period of myocardial infarction and left ventricular function.Methods Twenty three Chinese miniature pigs were randomly divided into the experimental group and the control group.The swines in experimental group were prepared as acute myocardial infarction model by ligating anterior descending coronary artery and were randomly divided into 6 subgroups according to the different time points after infarction.The left ventricular end-diastolic diameter (LVDd),left ventricular ejection fraction (LVEF) and left ventricular fraction shortening (LVFS) were measured respectively.Global circumferential strain (GCS) and global radial strain (GRS) of left ventricle were measured.The content of SDF-1 were also measured by real-time quantitative PCR.Results Compared with the control group,SDF-1 levels were significantly elevated,and LVEF,LVFS,GCS and GRS were reduced.However,LVDd were significantly increased.The content of SDF-1 and GCS has a negative correlation (r =-0.580,P =0.000).Conclusions The content of SDF-1 in myocardial tissue have a certain relationship with GCS of left ventricular myocardium.

Objective To study the best titer of recombinant adeno-associated virus serotype 2 (rAAV2) delivered to myocardium by targeted ultrasound microbubbles. Methods Twenty one adult SD rats were divided into seven groups. SonoVue attached with different titer [1.5 × 109~ 3.0 × 1011 vg/ml (virus genome/ml)] of rAAV2-EGFP was infused into the tail vein of rats, following ultrasound mediated microbubbles destruction,as experiment groups. Normal saline was infused into the tail vein of rats as the control group (without rAAV2). Rats were killed after 14 days and hearts were harvested. GFP protein expression which showed rAAV2 transfer was observed under fluorescence microscope in frozen section and integrate optical density(IOD) was measured by Image Pro Plus software. Results When the titer of rAAV2 was 1.5 × 1011 vg/ml infused into the tail vein of rats there was much more GFP expression in myocardium than lower titers (P <0.01). Conclusions The best titer of rAAV2 delivered to myocardium by targeted ultrasound microbubbles is 1.5 × 1011 vg/ml in the tail vein infusion of rats.

0.05).Conclusion The implanted radioactive 125I seeds can damage all kinds of tissues at different degrees,but this kind of damage is reversible,the dog may repair the damage through its own repair ability,its clinical application is safe.

Objective To investigate the safety and efficacy of protective measures(hydration treatment)during perioperative period of interventional therapy for renal function of gerontic patients with coronary artery disease(CAD)complicated by chronic renal insufficiency(CRI).Methods The relation between hydration therapies and the renal function of 168 CAD patients(≥60 years old)complicated by CRI were analyzed.These patients were divided into three groups:in the group A,patients underwent selective coronary angiography(SCA)with conventional treatment during interventional perioperative period(IPP);in the group B,patients underwent SCA and treated with the NaCl plus low dose NaHCO3 during IPP;in the group C,patients underwent percutaneous coronary intervention(PCI),and meanwhile treated with the same dose NaCl and NaHCO3 as the group B during IPP.Results Coronary interventions were successful in all 168 patients.Contrast medium associated nephropathy(CAN)was found in 30 patients,and incidences of the CAN were 17(27.4%)in the group A,3(7.5%)in the group B and 10(15.2%)in the group C,respectively.The accompanying diseases and characteristics of the patients also effected the incidence of CAN:the CAN incidence of patients with hyperlipidemia was significantly higher than that with normal cholesterol(P<0.001);patients with diabetes was significantly higher than that with normal serum glucose(P<0.001);patients with stenosis of renal artery was significantly higher than that with normal renal artery(P<0.001);patients with heart failure(grade Ⅲ～Ⅳ)was significantly higher than that with the grade Ⅰ～Ⅱ(P<0.001).Conclusion The hydration treatment with the NaCl plus low dose NaHCO3 during IPP can protect renal function and increase tolerance for gernotic CAD patients complicated by CRI during IPP,and improve prognosis.

Aim To investigate the influence of down-regulating adenosine A1 receptor and adenosine A2 A receptor gene expression on proliferation and activation of acetaldehyde-induced hepatic stellate cell-T6 cells through siRNA. Methods Alcoholic liver fibrosis in vitro model was constructed by inducing HSC-T6 cells with acetaldehyde. siRNA targeting A1R and A2AR were designed and synthesized according to its mRNA. The siRNA was transfected into rat HSC-T6 cells by li-posome LipofectamineTM 2000. HSC cell proliferation was measured by MTT. The mRNA levels of A1R, A2AR, α-SMA, Collagen I in the supernatant of the cell culture were measured by Quantitative Real-Time PCR. The protein levels of A1R, A2AR, α-SMA, Collagen I were measured by Western blot. Results A1 R and A2 AR siRNA effectively inhibited the cell proliferation, and they also significantly decreased the levels of A1R, A2AR,α-SMA, Collagen I, suggesting that A1 R and A2 AR might be potential target genes in the alcoholic liver fibrosis. Conclusions Silencing A1 R or A2 AR by RNAi can significantly inhibit the HSC proliferation, A1R and A2AR may be potential therapeutic target genes for alcoholic liver fibrosis.

Objective To explore the teaching effects of physiology lectures and students' attitude to lectures,their suggestions for contents,forms and other aspects of lectures.Methods Random sampling questionnaire survey was conducted among 500 students of 2008 grade and 2009 grade in Xiangya medical school of central south university.The percentage of the option in the questionnaire was calculated and the findings were directly described.Results All students thought that physiology lectures were necessary.Teachers should arrange lectures which students were interested in,pay attention to inspire thinking and enhance teacher-student interaction.67.6％ of students thought that the knowledge learned from lectures can be used in expanding scientific thinking.Conclusions The lectures are popular among most students,playing an important role in cultivating students' ability of strict scientific thinking.

Objective To express the anti-IL-1βscfv and soluble TNF receptor 1 (sTNFR1),and analyze their bio-activities.Methods sTNFR1 was obtained by RT-PCR from the total RNA of HeLa cells,and fused with IL-1βscfv by the hinge fragment of IgG molecule.The fusion gene IL-1scfv:TNFR1 was cloned into the expression vector pET27b(+).The fusion protein was expressed and purified from inclusion bodies.Results The ELISA analysis showed that the fusion protein could bind hIL-1β and hTNF-α respectively in a dose-dependent manner,indicating that scfv and sTNFR in the fusion protein can form the correct spatial configuration.The dolt-blot analysis showed that the fusion protein could concurrently bind with hIL-1β and hTNF-α,indicating that the combination of the two parts of the fusion protein does not influence each other for binding to their target molecules.The bioactivity assay showed that the fusion protein could inhibit both the cytotoxicity of hTNF-α on L929 cells and hIL-1β-induced proliferation of L929 cells,indicating that the fusion protein has the ability to neutralize hTNF-α and hIL-1β.Conclusion A bispecific fusion protein IL-1scfv:TNFR1 was successfully constructed.The fusion protein has the ability to inhibit the biological activity of hTNF-α and hIL-1β,and provides a drug candidate for the treatment of rheumatoid arthritis.

The aim of this project is to establish a fibroblast growth factor-21 (FGF-21) signaling pathway targeted cell model, for screening a class of FGF-21 receptor agonists as anti-diabetic candidates. FGF-21 requires beta klotho transmembrane proteins as co-receptor for the activation of tyrosine kinase FGF receptor (FGFR) signaling, thereby activating a series of intracellular signaling pathways and regulating gene transcription for glucose metabolism. Firstly a recombinant plasmid expressing co-receptor beta klotho and EGFP reporter genes was constructed. After introducing the recombinant plasmid into package cells, the cell culture supernatant was used to infect 3T3-L1 cells, which were then screened for stably expressing beta klotho gene. Administration of FGF-21 increased the expression of GLUT1 and stimulated GLUT1-mediated glucose uptake. This novel cell model can be conveniently used in high-throughput drug screening of FGF-21 or FGF-21 analogues.