Objective To study the relationship between the stenosis content of carotid artery and the degree of homocysteine (Hcy) by analyzing the level changes of Hcy in ischemic stroke patients.Methods One hundred and thirty patients of carotid artery stenosis with ischemic stroke were divided into four groups based on the degrees of carotid artery stenosis determined by ultrasound Doppler flow imaging:the slight stenosis group (32patients),the moderate group (35 patients),the severe group (33 patients) and the total occlusion group (30 patients).Meanwhile,30 healthy people were chosen as the control group.The levels of serum homocysteine (Hcy),total cholesterol (TC),triglyceride (TG) and low density lipoprotein (LDL) of each group were measured,and the results were compared with those of the control group.Results It was found that the serum Hcy level in ischemic stroke group were significantly higher than that in the control group (P＜0.05),whereas the variations ofTC,TG,HDL and LDL levels between the ischemic stroke group and the control group could be neglected (P＞0.05).An obviously increased order of serum Hcy levels was observed as the following,the control group,the slight stenosis group,the moderate group,the severe group as well as the total occlusion group.The difference between every two groups was statistically significant (P＜0.05).The level of serum Hcy was positive correlated to the content of carotid artery stenosis (r=0.835,P＜0.05).However,no correlation was found between the carotid artery stenosis and each level of TC,TG and LDL (r=0.031,0.012,0.018,respectively,P＞0.05).Conclusions The serum Hcy level is closely related to the degree of carotid artery stenosis,and high level Hcy is suggested to be one of major risk factors for ischemic stroke.

Objective: To investigate the benzo[a]pyrene ( B[a]P ) diolepoxide ( BPDE )-DNA adduct levels in offspring rats with intrauterine exposure to B[a]P, and examine the effects of BPDE-DNA adduct levels on pancreatic functional impairment and glucose metabolism in offspring rats. Methods: Forty pregnant rats were randomly divided into the blank control group, standard-dose group, low-dose group, medium-dose group and high-dose group (daily dose of 0, 2, 200, 800, 1 600 μg/kg B[a]P, respectively), of 8 animals in each group. Rats in the B[a]P treatment groups were administered by oral gavage with a mixture of B[a]P and corn oil at a dose of 0.2 mL/100 g body weight since day 1 of pregnancy until 21 days after delivery, while rats in the blank control group were given the same volume of coin oil by oral gavage. The BPDE-DNA adduct levels were measured and the pancreatic development was observed in the offspring rats 2 and 21 days and 12 weeks after birth, and the correlation between pancreas volume index and dose of exposure to B[a]P was examined using Spearman's rank correlation analysis. In addition, glucose metabolism was measured in offspring rats 12 months after birth using glucose tolerance test ( GTT ) and insulin tolerance test ( ITT ). Results: There was no abnormal appearance, death, abortion or preterm birth in pregnant or offspring rats in the five groups, and no significant differences were seen in activity, diet, drinking water or mental status in rats. The greatest level of BPDE-DNA adducts was measured in offspring rats 2 days after birth, with median levels ( interquartile range ) of 1 089.60 ( 586.10 ) to 1 405.49 ( 346.47 ) pg/mL, and no BPDE-DNA adducts were found in offspring rats 12 weeks after birth. The pancreas volume index correlated negatively with the dose of exposure to B[a]P in offspring rats 2 ( rs=-0.620, P=0.001 ) and 21 days after birth ( rs=-0.801, P=0.001 ). Hypoplasia of pancreas with loose tissues was seen in offspring rats 2 days after birth, while well pancreatic development was found in offspring rats 12 weeks after birth, with tight exocrine portion. GTT showed an increase in glucose levels in offspring rats in all five groups following abdominal injection of glucose and declined 30 min post-injection ( F=365.578, P<0.001 ), and ITT showed a tendency towards a decline in glucose levels in offspring rats in all five groups ( F=461.215, P<0.001 ). Conclusions The levels of BPDE-DNA adducts in offspring rats increase with the dose of intrauterine B[a]P exposure, and insulin resistance and impaired glucose tolerance occur 12 months post-exposure to B[a]P. Intrauterine B[a]P exposure affects pancreatic development in offspring rats and causes abnormal glucose metabolism in adult offspring rats.

Objective: To observe the effects of perinatal exposure to benzo[a]pyrene (B[a]P) on the expression of pancreatic duodenal homeobox-1 (PDX-1) and mitochondrial transcription factor A (TFAM) and mitochondrial DNA copy number in offspring mice, and to explore the role of maternal exposure to B[a]P in the pancreatic function damage of offspring mice. Methods: Forty pregnant rats were randomly divided into the control group, the lowest dose group (2 μg/kg), the low dose group (200 μg/kg), medium dose group (800 μg/kg) and high dose group (1 600 μg/kg), with 8 rats in each group. From day 1 of pregnancy, each exposed group was given 0.2 mL/100 g body weight of B[a]P and corn oil mixture by gavage once a day until 3 weeks after delivery, while the control group was given the same dose of corn oil. The pancreatic tissue of three-week-old mice were collected after abdominal anesthesia for insulin immunohistochemical detection. The protein and mRNA expression levels of PDX-1 and TFAM, as well as mitochondrial DNA copy number were detected. Spearman rank correlation analysis was used to analyze the correlation between B[a]P exposure dose and the above indicators. Results: The insulin-positive area ratio and average optical density of insulin in the medium and the high dose groups were significantly lower than those in the control group (all P<0.05). The insulin-positive area ratio and average optical density of insulin were negatively correlated with the B[a]P dose (rs=-0.862 and -0.858, both P<0.05). The protein expression levels of PDX-1 and TFAM in the high dose group were significantly lower than those in the control group (both P<0.05). The protein expression levels of PDX-1 and TFAM were negatively correlated with the B[a]P dose (rs=-0.756 and -0.799, both P<0.05). The mRNA expression levels of PDX-1 and mitochondrial DNA copy number in the medium and high dose groups were significantly lower than those in the control group, and the mRNA expression level of TFAM in the high dose group was significantly lower than that in the control group (all P<0.05). The mRNA expression levels of PDX-1, TFAM, and mitochondrial DNA copy number were negatively correlated with the B[a]P dose (rs=-0.722, -0.550 and -0.840, all P<0.05). Conclusion Perinatal exposure to B[a]P can induce the damage of islet β cells in offspring rats, which may be related to the decreased expression of PDX-1 and TFAM and the copy number of mitochondrial DNA.

The purpose of this study is to investigate the enantioselectivity of norgestrel (NG) transdermal permeation and the potential influence of linalool and lipids on the enantioselectivity. In vitro skin permeation studies of NG across the excised rat skins were performed with Valia-Chien diffusion cells, and the permeation samples were analyzed by enantioselective HPLC. The possible enantioselective permeation of NG across intact rat back skin and lipids extracted rat back skin and the influence of linalool were evaluated. The skin permeation rate of dl-NG was two times higher than that of l-NG when donor solutions (EtOH/H2O 2 : 8, v/v) containing l-NG or dl-NG. It may be mainly attributed to the solubility discrepancy between enantiomer and racemate. The enantioselective permeation of dl-NG across intact rat skin was observed when the donor solutions containing dl-linalool. The permeation flux of l-NG was 22% higher than that of d-NG. But interestingly, the enantioselective permeation of dl-NG disappeared under the same experimental condition except that the lipid extracted rat skin was used. Attenuated total reflection-fourier transform infrared spectroscopy analysis of stratum corneum showed that the wave number for asymmetric CH2 stretching vibrations of lipids treated with dl-linalool was greater than that of the control. The results indicated that the enantioselective permeation of NG may be contributed by the interaction between dl-linalool and lipids. More than half of lipids were composed of ceramides. The stereospecific interaction maybe existed among chiral enhancer (linalool), lipids (ceramides) and/or chiral drugs (NG).
Administration, Cutaneous ; Animals ; Lipids ; pharmacology ; Monoterpenes ; pharmacology ; Norgestrel ; pharmacokinetics ; Rats ; Skin Absorption ; drug effects ; Spectroscopy, Fourier Transform Infrared ; Stereoisomerism

OBJECTIVE To investigate the prevalence of DHA AmpC ?-lactamases mediated by plasmid in Klebsiella pneumoniae in China.METHODS Antimicrobial susceptibility test was conducted by the methods of double agar dilution and ESBLs confirmatory in K-B method according to the criteria of guidelines of CLSI.AmpC ?-lactamases were detected on the basis that AmpC ?-lactamases could be inhibited by 3-aminophenylboronic acid(APB).Gene chip technology and PCR were used to detect ESBLs and AmpC gene.RESULTS Among total 34 isolates of K.pneumoniae 32(94.1%) produced AmpC ?-lactamases and ESBLs.The most common(38.3%) were types DHA and TEM and SHV.MIC50 and MIC90 of all strains to all tested antimicrobial agents were lower than 34 strains tested 0.25?g/ml and 0.5?g/ml.Fourteen strains AmpC and ESBLs were conjugated successfully.CONCLUSIONS DHA AmpC ?-lactamases mediated by plasmid are the most common in K.pneumoniae in General Hospital of PLA of China.The most common(38.3%) are types DHA and TEM and SHV.Fourteen(41.2%) strains can be spreaded by plasmid.

Objective To analysis the effect of conbercept ophthalmic injection on peripheral blood vascular endothelial growth factor,intraocular pressure and visual acuity in patients with age related macular degeneration.Methods 60 patients who were diagnosed with age related macular degeneration were collected.All patients were randomly divided into experimental group and control group,30 cases in each group, lucentis intravitreal injection was given into the vitreous cavity in control group, experimental group give conbercept injection into the vitreous cavity,once a month.All patients were treated for 3 months,after the end of treatment,all patients serum VEGF and CRP,CMT,CNV and fluorescence leakage rate,intraocular pressure and visual acuity were detected.ResuIts After treatment,compared with control group,the content of serum VEGF was significantly lower in the experimental group(P<0.05).The content of serum CRP was significantly lower in experimental group(P<0.05).The CMT,CNV and fluorescence leakage rate were significantly lower in the experimental group (P<0.05).The level of IOP and visual acuity(logMAR)were significantly lower in the experimental group (P<0.05).ConcIusions Conbercept ophthalmic injection can reduce age-related macular degeneration patients week blood VEGF and CRP levels and decrease the CMT, CNV and fluorescence leakage rate, reduce intraocular pressure, visual acuity improvement and have guiding significance for clinic.

Objective To evaluate the value of ultrasound (US) in assessment knee joint inflammation in patients with juvenile rheumatoid arthritis(JRA).Methods US scans of the knees obtained in 30 children at clinically active stage; JRA was compared with those obtained in 30 healthy children and 10 JRA patients in clinical remission.Results Changes in synovial membrane thickness and presence of fluid in suprapatellar bursa showed statistically significant differences between JRA patients with active disease and the other subjects.Alterations in contour of the articular cartilage were demonstrated in 10 knees of patients with JRA.Conclusion US is a simple sensitive and reliable methods for the assessment and monitoring of knee joint involvement in JRA.

Objective To investigate the efficacy and safety of vasopressin receptor antagonist tolvaptan for treating dilutional hyponatremia casused by decompensated liver cirrhosis.Methods Ninety-six subjects with decompensated liver cirrhosis complicated by dilutional hyponatremia were divided into test group (n =56) and control group (n =40) by double blind method.Test group were treated with a single dose of tolvaptan 15 mg orally in addition to routine therapy,while control group were treated with routine therapy plus placebo.The changes in serum sodium concentration,liver functions,ascites,and edema of lower extremities between the two groups were observed.Measurement data were compared by t test,and categorical data were compared by chisquare test.Results On day 7,36 (64.3％) patients in test group and 9 (22.5％) patients in control group reached normal serum sodium concentrations (x2 =5.241,P＜0.01).All the patients in test group and only 3 patients in control group had 24 hours' total urine volume above 3500 mL.Difference of 24 hours' total urine volume during 7-day treatment between the two groups was statistically significant (t=20.899,P＜0.01).Thirty-nine patients in test group and 15 patients in control group had reduced ascites (x2 =4.260,P=0.039),but improvement of edema in lower extremities of both groups was comparable (12 cases in test group and 7 cases in control group; x2 =0.227,P=0.634).Serum alanine aminotransferase (ALT),total bilirubin (TBil),potassium and creatinine levels of test group were improved after treatment,but were not significantly different from either baseline levels (t=1.509,0.783,1.107,1.237; both P＞0.05) or those of the control group (t=1.712,1.635,1.121,0.873; both P＞0.05).Conclusions Single dose of tolvaptan (15 mg) exerts a strong diuretic effect and is able to greatly increase serum sodium concentration,thus has distinct therapeutic effect for patients with decompensated liver cirrhosis complicated by dilutional hyponatremia.

Background The molecular biological mechanisms of diabetic retinopathy(DR)is unclear up to now.Researches have proved that endoplasmic reticulum stress(ER Stress)-associated factors are elevated in peripheral blood in patients with diabetic retinopathy.and P58 IPK can inhibit those factors.So the relationship between P58 IPK and DR is worth to investigate. Objective The aim of this study was to detect the dynamic expression of P58 IPK in the retina of diabetic rats. Methods The diabetic animal models were established in 18 clean male SD rats by intraperitoneal injection of stilptozotiein(STZ)at a dose of 60 mg/kg.The rats were sacrificed in 1,3,6 months after injection.The expression change of P58 IPK mRNA in the rats retina was detected by quantitative real-time RT-PCR.Other 6 matched normal rats were used as control groups.This experiment followed the Regulations for the Administration of Affair Concerning experimental Animals by State Science and Technology Commission. Results The rats showed more drinking,more food and more urine after STZ injection with the blood glucose level≥ 16.5 mmol/L.The success rate of diabetic models was 100%.The A value of P58 IPK mRNA/β-actin in rat retina was 0.800±0.005 and 0.975±0.008 after injection of STZ.and that of control rats was 0.725±0.006,showing statistically significant difference between them(t=22.589,t=62.784,P＜0.05).In 6 months after injection of STZ,the expression of P58 IPK mRNA in experimental diabetic rat retina was evidently lower than the eontrol rats(0.671±0.004 versus 0.725±0.006,t=-17.984,P＜0.05).Conclusion P58 IPK has a close relation to the pathogenesis of DR,and it plays a retarding role for DR.

Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.