Objective To evaluate the influence of a selective Rho-associated protein kinase inhibitor (fasudil hydrochloride) on outflow facility in enucleated porcine,rabbit and bovine eyes.Methods At the constant perfusion pressure of 15 mm-Hg (1 kPa =7.5 mmHg),the baseline coefficient of outflow facility (C0) of the isolated porcine,rabbit and bovine eyes was recorded respectively.The enucleated porcine eyes were divided into two groups randomly (n =6),and they were control group and experimental group.The same grouping method was also used-C0 the ribbit and bovine eyes.The control group was subjected to GPBS perfusion,while the experimental group was treated with 100 μmol · L-1 fasudil solution,followed by recording the experimental coefficient of outflow facility (C1),as well as calculating ΔC (ΔC =C1-C0) and ΔC％ (ΔC％ =ΔC/C0).Finally,the paired t test and one-way analysis of variance were performed using SPSS 17.0.Results As for porcine eyes,the ΔC％ of the control group was (17.83 ± 3.84) ％ while the experimental group was (44.00 ± 6.44) ％;as for rabbit eyes,the ΔC％ of the control group was (15.50 ± 2.93) ％,while the experimental group was (31.67 ±6.54)％;as for bovine eyes,the ΔC％ of the control group was (11.67 ± 1.17)％,while the experimental group was (37.17 ± 4.48)％.The ΔC％ in the experimental group was significantly increased when compared with the control group in three animals,with significant difference (all P ＜ 0.05).There was no statistical difference in ΔC％ of three experimental groups among different kinds of animals (all P ＜ 0.05).Conclusion Fasudil can improve outflow facility in enucleated eyes of animals,and it can redistribute aqueous humor drainage to a wider area through directly regulating the cytoskeleton of cells and matrix,resulting in increased coefficient of outflow facility.

Objective Compare and analyze the characteristics of the coronary angiography between Han nationality and Uygur nationality in Xinjiang autonomous region Methods 2?200 clinical data and angiographic data from 1991 to 1999 were reviewed There were divided into two groups according to different nationality The extent of the lesions and the proximal calibres of coronary artery were measured and compared Results The proximal calibres of coronary arteries of Uygurs are greater than those of Hans The lesions of Uygurs′ coronary heart disease (CHD) patients are more severe than Hans′: more three branch lesion, more C type lesion and total occlusion cases. There are statistical differences between two nationalities Conclusion Proximal calibres of coronary branches of Uygures are bigger than Hans, but Uygur patients suffer more severe lesions of coronary arteries as compared with Hans

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 observe the time characteristics of acute aortic dissection (AAD) in Urumqi and its peripheral areas in Xinjiang Uygur autonomous region, and to explore the effect of meteorological conditions on the onset of AAD.Methods Retrospective analysis of the related data of the adult AAD patients diagnosed by imaging examination, and admitted to the First Affiliated Hospital of Xinjiang Medical University from March 2010 to July 2016 was performed. The onset time, the contemporaneous meteorological data, including daily minimum, average, and the highest temperature, daily average air pressure, daily average relative humidity, daily precipitation, daily average wind speed, daily maximum wind speed, and sunshine duration per day were collected. Analysis of concentrated distribution trend of the AAD onset was done by circular distribution statistics. Meteorological conditions were compared between the AAD day and no AAD day. The changes of air temperature 5 days before onset were observed.Results A total of 379 patients were enrolled, with 307 male and 72 female. The ratio of male to female was 4.19:1, the average age was 52.27±12.18; and the underlying diseases was hypertension (about 58.05%); the incidence sites were Urumqi and Changji city. ① The incidence of AAD in Urumqi and its peripheral areas had clear concentrated trend, and the peak period of AAD corresponded to January 1st to 2nd (homogeneity testr = 0.104,r0.05 = 0.009,P < 0.05). ② The minimum, mean and maximal atmospheric temperatures on the AAD day were lower than no AAD day [℃: 4.10 (-9.55, 14.60) vs. 7.75 (-6.70, 16.20), 14.10 (-1.50, 25.00) vs. 17.50 (0.60, 26.78), 8.50 (-6.22, 19.45) vs. 12.10 (-3.60, 20.90), allP < 0.05], sunshine duration was significantly shorter than no AAD day [hours: 8.50 (4.60, 10.70) vs. 8.90 (5.50, 11.50), P< 0.01], and atmospheric pressure were higher than no AAD day [kPa: 91.24 (90.66, 91.75) vs. 91.12 (90.62, 91.61), 19.11 (18.99, 19.22) vs. 19.09 (18.98, 19.19), bothP < 0.05]; there were no significant differences in the incidence of daytime temperature difference, average relative humidity, precipitation, average wind speed, and maximum wind speed between the two groups. ③ the peak period of AAD occurrence was December (38 cases), which was followed by January (36 cases); and there were 26 cases of each month in May, June, July, and September, which were lower than the rest of months. It was shown by the temperature distribution that the minimum average temperature was in January (-12.46 ℃), followed by December (-9.03 ℃); the maximum average temperature was in July [(24.59±3.21)℃], followed by August [(23.14±3.64)℃]. ④ The patients was sorted according to the quartile of daily mean temperature, into < -5.2 ℃, -5.2-10.7 ℃, 10.7-20.4 ℃, and > 20.4 ℃ groups, and each group had 104, 99, 98, 78 cases of AAD, respectively. The number of AAD in lowest temperature group was 1.33 times of the highest temperature group. In the 45-59 and 60-74 years, the number of AAD in lowest temperature group was higher than that in the highest temperature group (50 vs. 36, 26 vs. 13). In patients with hypertension, the number of AAD in lowest temperature group was higher than that in the highest temperature group (60 vs. 44). ⑤ The maximum air temperature, mean air temperature of the day from 5 days before to the onset day of AAD, and the lowest temperature from 3 days before to the onset day of AAD display slowly decrease trends, but there was no significant difference in daily temperature.Conclusions The incidence of AAD in Urumqi and its peripheral areas was concentrated from January 1st to 2nd of each year. When the temperature was lower than -5.2 ℃, AAD were more likely to attack. The 5 days changes of temperature and daily temperature before the attack were relevant with onset of AAD. It was speculated that the weather conditions such as air temperature may affect patient who has a past history of cardiovascular disease to attack AAD.

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.

Objective To study the association between serum concentrations of total testosterone levels and coronary heart disease (CHD) in the postmenopausal women. Methods The study was designed as a case-control study.394 postmenopausal female patients were selected from Cardiology Center of the First Affiliated Hospital of Xinjiang Medical University.The case group included 183 women patients with CHD aged (62.7±8.0) years,the control group,211 women with normal coronary aged (60.0 ± 8.8) years. Blood samples were collected to determine total testosterone,fasting glucose and lipid profile. CHD severity was expressed as the numbers of coronary arteries that had a stenosis ≥50％.According to the level of testosteron,all cases were divided into 4 groups by interquartile range method:Q1＜3.5 nmol/L(n=190),3.5 nmol/L≤Q2 ＜10.4 nmol/L(n=64),10.4 nmol/L≤Q3 ＜26.0 nmol/L(n=120) and Q4≥26.0 nmol/L(n=20).The association between the serum total testosterone levels and severity of coronary atherosclerosis was analyzed. Results The average total testosterone was higher in case group than in control group[(10.4 ± 24.3 ) nmol/L vs. ( 6.9 ± 17.4 ) nmol/L,Z =0.79,P =0.555].In Quantitative adjusted models,higher levels of total testosterone had strong correlation with CHD,Q4 incidence of CHD (75.0％,15 cases) was significantly higher than Q1 (46.8％,89 cases),Q2 (40.6％,26 cases)and Q3 (44.2％,53 cases) (x2 =7.69,P=0.048).After adjustment for other risk factors,women in the top quartile of total testosterone levels had a more than 3-fold increase in odds of CHD(OR=3.47,95％CI:1.06-11.32,P＜0.05).In addition,the serum concentrations of total testosterone level were significantly associated with the severity of CHD (F=12.94,P＜0.05). Conclusions Higher levels of total testosterone may be associated with high prevalence and severity of CHD as an independent factor in postmenopausal women.

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:Macrophage migration inhibitory factor (MIF) is a multi-potent cytokine that makes considerable contribution to the regulation of inflammatory response and immune response in the body. MIF rs1007888 is associated with various inflammatory diseases, but the correlation between rs1007888 and coronary heart disease in the Kazakhs of China has been rarely explored. OBJECTIVE:To investigate the relationship between rs1007888 gene polymorphisms in MIF gene and coronary heart disease in the Kazakhs from Xinjiang Uygur Autonomous Region, China. METHODS:A total of 230 Kazakh patients with coronary heart disease evidenced by coronary arteriography between December 2012 and July 2014 were recruited, and another 478 Kazak controls were free from coronary artery disease with normal angiograms. Real-time fluorescence quantitative PCR assay was used to detect the rs1007888 polymorphisms of MIF gene. Alele and genotype distributions of the rs1007888 polymorphism were compared between patients and controls. RESULTS AND CONCLUSION:Distribution of genotypes in the two groups appeared to be in Hardy-Weinberg equilibrium (P> 0.05). The alele frequencies and genotypes of MIF-rs1007888 showed no significant difference between the two groups (P > 0.05). Therefore, the genetic variation of rs1007888 in MIF gene is not associated with coronary heart disease in the Kazakhs of China.

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.