Objective To explore the effect of bionic urine bags on dysuria in stroke patients and improve the effect of bladder function training. Methods Forty patients with urinary incontinence were selected and randomly divided into control group (n=46) and observation group (n=44) by random number table. The patients in the control group were treated with Kangwei anti-reflux urine bags and the urine was discharged every 3 hours at daytime, once every 4 hours at night. The patients in the experimental group were treated with OT-U bionic urine bags for drainage urine, the threshold of urine bag pressure set:when the bladder pressure reached more than 35cm H2O, the bladder urine flowed into the urine bag. The two groups were compared in view of catheter catheterization time, urination after urination time, self-urination, re-intubation rate, urine overflow, urinary tract infection. Results The duration of indwelling catheters in the observation group was significantly higher than that of the control group (P <0.05). The rates of re-intubation and urinary tract infection were significantly lower as well (P<0.05). Conclusion Biomimetic urine bag can effectively promote the training of bladder function, shorten the time of indwelling catheter and reduce the occurrence of urinary tract infection after extubation.

Objective To investigate the expression and distribution of aquaporin 3 (AQP3) in mouse early embryos at different stages.Methods Controlled ovarian hyperstimulation model of Kunming mouse was used to collect four-cell embryos,eight-cell embryos,morula stage,and early blastocysts.Immunofluorescence microscopy and laser confocal microscopy were used to detect expression and distribution of AQP3 channels in these stages.Results Fluorescence signal of AQP3 was found in four embryonic stages of mice.Distribution within embryo was different at different embryonic stages.AQP3 was mainly expressed on the karyotheca of blastomeres at four-cell and eight-cell stage.In morula stage,AQP3 was mainly expressed on cell membrane of each blastomere.In early blastocysts,AQP3 was predominantly expressed on the cell membrane and cytoplasm of trophoblastic cell.Conclusions AQP3 trans-membrane channel might have potential regulation function on mouse embryonic development.

Background Oxidative stress is a major mechanism underlying the pathogenesis of cardiovascular disease. It can trigger inflammatory cascades which are primarily mediated via nuclear factor-κB (NF-κB). The NF-κB transcription factor family includes several subunits (p50, p52, p65, c-Rel, and Rel B) that respond to myocardial ischemia. It has been proved that persistent myocyte NF-κB p65 activation in heart failure exacerbates cardiac remodeling. Mechods A recombinant adeno-associated virus serotype 9 carrying enhanced green fluorescent protein and anti-NF-κB p65 ribozyme (AAV9-R65-CMV-eGFP) was constructed. The cells were assessed by MTT assay, Annexin V–propidium iodide dual staining to study apoptosis. The expression of P65 and P50 were assessed by Western blot to investigate the under-lying molecular mechanisms. Results After stimulation with H2O2 for 6 h, H9c2 cells viability decreased significantly, a large fraction of cells underwent apoptosis. We observed a rescue of H9c2 cells from H2O2-induced apoptosis in pretreatment with AAV9-R65-CMV-eGFP. Moreover, AAV9-R65-CMV-eGFP decreased H2O2-induced P65 expression. Conclusions AAV9-R65-CMV-eGFP protects H9c2 cells from oxidative stress induced apoptosis through down-regulation of P65 expression. These observations indicate that AAV9-R65-CMV-eGFP has the potential to exert cardioprotective effects against oxidative stress, which might be of great importance to clinical efficacy for cardiovascular disease.

Objective To establish a single nucleotide polymorphisms genotyping (SNP) method for a convenient, accurate, and routine analysis of clinical samples. Methods Based on the design of oligonucleotide probe, the assay was performed through three steps:the conjunction of the detection probe, universal amplification, labeling and ELISA reaction. The genotype of each SNP was revealed by reading signals of each set of reaction tubes. This assay was applied to detect sixty-two plasma samples of lung cancer for circulating DNA for three SNPs of EGFR, c.2573T＞G(L858R), EGFR, c.2582T＞G＞T(G719C). Results were compared with those obtained by direct sequencing. Results The heterozygote mutation was identified for L858R by both methods, although no mutation was detected for L861Q and G719C. Six samples were identified as heterozygotes with the new method, and only two samples were unambiguously identified as heterozygotes by the direct sequencing. Two additional samples could not be identified as heterozygotes because the peak of mutant allele was very low compared with that of wild allele. Conclusion The developed method enabled accurate identification of SNP in a convenient manner, and which is adapted to routine analysis from heterogeneous samples unambiguously.

Objective To investigate the relation between homeostasis model assessment of insulin resistance(HOMA-IR) and the results of coronary angiography in coronary artery disease(CAD) patients complicated with hypertension.Methods One hundred and two CAD patients complicated with hypertension were enrolled into the investigation group and another 80 CAD patients without hypertension were considered as the control group.The HOMA-IR and the results of coronary angiography were compared between the 2 groups and their correlation was further analyzed.Results The HOMA-IR of the investigation group was higher than that of the control group(8.10?1.25 vs 4.70?2.13,P

BACKGROUND:A lot of work has been carried out on the development of the primary cultured rat myocardial cel s at home and abroad. The primary culture technology of rat myocardial cel s becomes more mature, but myocardial cel s from neonatal mice are not easy to be obtained under the same experimental conditions. The mouse genome has more similarities with the human genome, which has a higher research value. OBJECTIVE:To improve the primary culture method of neonatal mouse myocardial cel s, and to obtain myocardial cel s with high purity, vitality and original structure and function. METHODS:The mouse cardiac tissues were treated using an enzyme digestion method to isolate isolated single myocardial cel s:first, the cardiac tissues were digested using trypsin, and then col agenous fibers were treated with col agenase to isolate single myocardial cel s. The concentration and action time of trypsin and type II col agenase were adjusted, and the pH values of reagents and temperature of each step were strictly control ed. RESULTS AND CONCLUSION:At 24 hours after inoculation, the myocardial cel s began to be adherent;at 48 hours, independent pulsation of myocardial cel s could be observed;at 72 hours, myocardial cel s were cross-linked;and at 96 hours, myocardial cel s formed cel clusters and presented with consistent beating. The survival rate and purity of myocardial cel s were both over 95%. This modified method could successful y culture myocardial cel s with high purity and viablility from neonatal mice, and the structure and function of myocardial cel s could be retained. Therefore, it is a feasible culture method.

BACKGROUND:Platelet-derived growth factor-B (PDGF-B) is an effective pro-angiogenic growth factor, and adeno-associated virus type 9 (rAAV9) has a strong cardiomyocyte targeting affinity, which is an ideal vehicle for ischemic heart disease gene therapy.
OBJECTIVE:To explore the PDGF-B gene transfection of in vitro neonatal rat myocardial cells mediated by rAAV9 against ischemia and hypoxia-induced cardiomyocytes apoptosis.
METHODRat neonatal myocardial cells were isolated and cultured, and then transfected by rAAV9-PDGF-B and empty virus, rAAV9 with enhance green fluorescent protein (eGFP), under multiplicity of infection (MOI) of 105, 106 and 107, respectively. We observed the expression of eGFP under fluorescence microscopy every day, and used flow cytometry to measure transfection efficiency of vector rAAV9. Western blot and immunofluorescence were used to evaluate protein expression of PDGF-B. Myocardial ischemia and hypoxia injury model was established in vitro on the 5th day of transfection of rAAV9-eGFP and rAAV9-PDGF-B with 107 MOI. The number of myocardial apoptosis was measured by TUNEL assay. Western blot was employed to detect the protein expression of Bax and Caspase-3 which were related apoptosis, and the effect and its possible mechanism of PDGF-B gene overexpression against myocardial apoptosis were explored.
RESULTS AND CONCLUSION:rAAV9 vector can efficiently transfect neonatal rat myocardial cells. eGFP and PDGF-B protein expressed in myocardial cells correctly and efficiently, and the expression intensity increased gradual y with the increasing of time course and MOI. The expression became stable on the 5th day, and the transfection efficiency showed significant difference among these groups (P<0.01). Myocardial apoptosis rate was significantly reduced in the rAAV9-PDGF-B group than the rAAV9-eGFP group (P<0.05), and protein levels of Bax and Caspase-3 in the rAAV9-PDGF-B group were significantly lower than those of the rAAV9-eGFP group (P<0.05). These data indicate that overexpression of PDGF-B gene can effectively reduce ischemia and hypoxia-induced myocardial apoptosis, and the possible mechanism might be by inhibiting Bax and Caspase-3 protein expression, which can provide evidence of rAAV9-PDGF-B vector in the gene therapy of ischemic heart diseases.

BACKGROUND:Recombinant adeno-associated virus serotype 9 has a high affinity in myocardial tissue, and the expression of recombinant adeno-associated virus serotype 9-enhanced green fluorescent protein (rAAV9-eGFP) in the aorta of atherosclerosis mice is not clear. OBJECTIVE:To explore the optimal time point of rAAV9-eGFP expression in the aorta of atherosclerosis mice. METHODS:Atherosclerosis model was established with high-fat diet in 30 ApoE-/-mice for 16 weeks. Among them, 25 mice were injected with 5.0×1011 vg (virus genomes) rAAV9-eGFP through the tail vein, while the remaining 5 mice were injected with saline, serving as the control group. The virus-transfected mice were kil ed at 14, 21, 28, 35 and 60 days after transfection, and aortic tissue was harvested. The expression of enhanced green fluorescent protein was detected with laser scanning confocal microscope. Western blot assays were used to detect the expression of enhanced green fluorescent protein in aorta. The expression of enhanced green fluorescent protein in vivo was observed and the optimal expression time point was determined. RESULTS AND CONCLUSION:rAAV9-eGFP effectively transfected the aorta of atherosclerosis mice, enhanced green fluorescent protein was expressed in aortic tissue, and the expression intensity increased gradual y with the increasing transfection time. The highest expression level was found at 35 days after transfection and then maintained stable at 60 days. There were significant differences at different time points after transfection (P<0.001). These data indicate that rAAV9-eGFP can be effectively expressed in the aorta of atherosclerosis ApoE-/-mice and rAAV9-eGFP can be regarded as the optimal vector in the treatment of atherosclerosis.

BACKGROUND:Studies have shown that fibrin glue can promote the survival of myoblast grafts, reduce infarct size and induce neovascularization of infarct zone. OBJECTIVE:To understand the condition of revascularization of infarcted heart muscle using endothelial progenitor cells combined with degradable fibrin glue materials. METHODS:A total of 27 Sprague-Dawley rats were randomized into three groups, 9 rats in each group:non-myocardial infarction group, immediate transplantation group and 1-week post-infarction transplantation group. Then, these three groups were sub-grouped into two groups, respectively:endothelial progenitor cells+fibrin glue group (experimental group) and fibrin glue group (control group). At 3 and 8 weeks after transplantation, the rats were sacrificed in each group. The revascularization and function of infracted heart muscle were observed by microscope, immunohistochemistry and echocardiography. RESULTS AND CONCLUSION:Under the microscope, there were some lax connective tissues between the heart and chest in the experimental groups, but no difference existed between the experimental and control groups. The heart structure was normal relatively and difficult to be distinguished between the experimental and control groups histological y and immunological y, and there was no angeioma, vascular malformation and tumor. The number of revascularization of heart muscle showed no difference between experimental and control groups as wel as between different experimental groups. Additional y, there was no significant difference in cardiac function between experimental and control groups. Although there are no positive results of endothelial progenitor cells, we wil modify and improve the strategy and believe that the celldelivery system is of benefit and efficacy.

Objective To investigate the cell survival of the combination of fibrin glue and adiposederived stem cells (ADSCs) in rats when implanted into ischemic myocardium and the improvement of heart function.Methods The rat ADSCs were isolated from the subcutaneous adipose tissues.The surface phenotype of these cells was analyzed by flow cytometry.Myocardial infarction was induced in female rats using coronary artery ligation.One week after MI,surviving rats were randomized (random nuber) into 4 groups,control group (n =10),fibrin group (n =10),cell group (n =10) and combination group (n =10).100 μl of PBS was injected into the ischemic myocardium in control group.100 μl of Fibrin glue were injected into ischemic myocardium in fibrin group.100 μl of ADSCs labeled with DAPI were injected into the infract along the border zone in cell group.ADSCs in 100 μl of fibrin glue were injected into the infract in combination group.Four weeks after the injection the surviving rats underwent examination of heart functions by the Hemodynamics.The rats were killed and their hearts were taken out to undergo immunohistochemistry with 4,6-diamidino-2-phenylindole (DAPI) and actin and factor Ⅶ to measure the area of cardiac infarction and the capillary density.The heart infarcted size was calculated by masson trichrome staining.All data was analyzed by software SPSS 15.0,ANOVA comparison tests and the student t test were used,and P ＜ 0.05 was considered as statistically significant.Results Four weeks after the cells were transplanted,LVSP and + dp/dtmax of combination group were highest among all groups.The heart infarcted size of the combination group was (28.5 ± 3.6) ％,significantly less than those of the cell group (33.33 ± 2.3) ％ and fibrin group (35.96 ± 2.11) ％,both P ＜ 0.05.The capillary density of the combination group was (108.7 ± 11.38) /mm2,significantly greater than those of the cell group and that of the fibrin group,and greater than that of the control group.DAPI and actin double staining detected a varied increase in the number of surviving cardiomyoctyes at the heart infarcted area.Conclusions Transplantation of ADSCs with fibrin glue brings better improvement in cell survival and in restoration of heart function than either cellular or fibrin therapy alone.