ObjectiveTo investigate the effect of puerarin on the expression of pigment epithelium-derived factor (PEDF) in the retina of diabetic rats.MethodsThe diabetic retinopathy rat model was established.Rats were randomly divided into three groups (normal,diabetic,and diabetic with puerarin injection).The pathological changes of the retinopathy were examined.PEDF was analyzed with PCR in different time point.Results Body weight of diabetic group [ ( 216.9 ± 8.37 ) g,and(179.1 ±7.56)g] was compared with normal group[ (298.2 -±6.78)g,and(323.5 -±6.45)g] in the 3 month and 5 month ( q =12.39,P ＜0.01 ; q =15.47,P ＜0.01 ).Blood glucose of diabetic group and diabetic with puerarin injection group was compared with normal group at each time point ( P ＜ 0.01,P ＜ 0.05 ).HE staining of the retina of diabetic rats indicated a diabetic retinopathy occurred in the early - stage in diabetic group,but changed slightly in the diabetic model with an injection of puerarin.The PEDF expression at the 5 month time point in normal group,diabetic group,and diabetic group with puerarin injection was 8.833 (1.200),1.650(3.300),and 8.350( 1.100),respectively.Compared with normal group and diabetic group with puerarin injection,the level of PEDF was significantly decreased in diabetic group (x2 =48.57,P ＜ 0.01 ;x2=48.46,P ＜ 0.01 ).ConclusionsPuerarin can alleviate the retinal pathological damage of diabetic rats,early intervention with puerarin can prevent from the expression of PEDF and delay the diabetic retinopathy process.

Objective To study the relationship between Xsp Ⅰ restriction fregment length polymorphism (RFLP) in exon 4 of low-density lipoprotein receptor (LDLR) gene and hypercholesterolemia. Methods PCR-RFLP method was applied to determine the polymorphism of LDLR gene in 446 cases of hypercholesterolemia, 284 of borderline hypercholesterolemia and 187 normal control subjects. Results Three genotypes, X+X+, X+X- and X-X-, were found in the population. (1) The frequencies of the X+X+ genotype and X+ allele in the group with hypercholesterolemia were higher than those of the other 2 groups (P<0.05). (2) From X-X- to X+X+ genotype, cholesterol and low-density lipoprotein-cholesterol increased gradually, while high-density lipoprotein-cholesterol decreased (all P < 0.05). (3) With logistic regression analysis, LDLR genotype was associated with hypercholesterolemia. Conclusion Polymorphism in Xsp I restriction site at the exon 4 of LDLR (X+ X+ genotype and X+ allele) may be a risk factor of hypercholesterolemia in Chinese population.

Objective To investigate the characteristics of executive function in patients with brain injury. Methods From March 1st, to June 30th, 2015, 44 patients with brain injury were investigated with Wisconsin Card Sorting Test (WCST), the indexes including Responses Answer, Categories Completed, Correct Responses, Errors Responses, Trials to Complete First Category, Percent Conceptual Level Respons-es Percentage, Perseverative Responses Errors, Nonperseverative Responses Errors, Failure to Maintain Set, and Learning to Learn. Results The abnormal rates were the most in Nonperseverative Responses Errors and Percent Conceptual Level Responses Percentage (61.36%), and then in Responses Answer/Categories Completed/Correct Responses (59.09%), Correct Responses (43.18%), Trials to Complete First Category (38.64%), Perseverative Errors (29.51%), Learning to Learn (25.00%), and Failure to Maintain Set (9.09%). The patients with trau-matic brain injury were different from those with stroke in Responses Answer, Errors Responses, Perseverative Responses Errors, Catego-ries Completed, Percent Conceptual Level Responses Percentage, and Learning to Learn (Z>2.444, t>2.156, P<0.05). The patients injured in frontal lobe were different from those in other areas in Perseverative Responses Errors (t=2.595, P=0.015). Conclusion Executive function damaged generally in patients with brain injury, which related to concentration, abstract, shifting attention, working memory, etc. The frontal lobe damage may associate with the disorder of shifting attention.

The primary neural stem cells were isolated from SD rat and formed the neuropheres, the neuropheres were passaged and planted on the dish coated with 0.1% gelatin, the colony was picked up under the microscope, then dispersed and cultured, to obtain the clone proliferated from one cell, passaging and picking up the cells 5～6 times at least. The NSC and its differentiated cells were identified with the marker genes respectively. The results showed that the neural stem cells were isolated from the SD rat embryos and the real clone were obtained by picking up the cells again and again, and then cultured in the form of monolayer. The marker genes of the neural stem cells and its differentiated cells could be detected at last. It will provide the rat model the resource of the cells for the treatment and the basic research for the morphology standard.

Twenty healthy subjects in each of 3 groups were fed with monounsaturated fatty-acid diet,polyunsaturated fatty-acid diet, or saturated fatty-acid diet separately for 3 days. It suggested that monounsaturated fatty-acids may ameliorate the oxidative stress and improve insulin sensitivity.

A baseline survey on health knowledge was conducted in Yayuncun (Asian Games Village) Community in 2004 and 2005 respectively. A sample consisting of 1765 residents aged 15 and above from 600 households, who had lived in the community at least 5 years, was randomly taken; the "Beijing Typical Community Resident Questionnaire" was used for survey and the participants were followedup annually from 2006 to 2008. The results showed that the awareness rate of food nutrition and disease prevention increased year by year; however, the participants with educational level lower than junior middle school had the lowest rate of overall health knowledge. The main channels for local residents to acquire health knowledge were activities, lectures and other programs with the title of the Olympic Games; the educational level and the year of survey were independent affecting factors on overall health knowledge among residents.

BACKGROUND: At present, there is few reports about using middl ecerebral artery obstraction (MCAO) model to determine the repair course of cerebral infarction during functional training.OBJECTIVE: To determine the effect of electro-stimulating therapy on promoting the rehabilitation of cerebral infarction and its mechanism.DESIGN: Randomized controlled study.SETTING: Animal Center and Electron Microscope Laboratory of Zhongshan University.MATERIALS: The experiment was carried out in the Animal Center of Zhongshan Medical College and Neurological Laboratory of the First Affiliated Hospital of Zhongshang University from January 2002 to December2004. A total of 200 healthy males SD rats, aged 3 months and weighing 90-110 g, were selected. According to the following criteria: SBP＞180mmHg (1 mmHg=0.133 kPa), BWT score of MCAO models which were reproduced by RHRSP was 1, totally 180 RHRSP were admitted to the research and divided into electro-stimulating therapy group (n=90) and control group (n=90).METHODS: Electro-stimulating was given to four accupuncture points of the paralyzed limbs of rats. The electro-stimulating treatment was given about 30 minutes once a day. And a therapy course was 6 days, and between two therapy courses there was one-day break. At the end of 1st, 3rd,6th and 9th therapy courses, the brain of motor function and tissue in marginal zone of cerebral infarction were assayed as follow: [1] The beam walking test (BWT, 1 as severe disorder and 7 as normal). [2] Electron microscope. [3] Astrpcyte glial fibriliary acidic protein, neurofilament protein and microtubule-associated protein-2 were assayed with immunohistochemistry. Five fields of each slice in the two groups were randomly selected to add up the positive cell number. Totally 30 positive cells of glial fibriliary acidic protein was selected to assay average absorbency (A) of positive cellular plasm. [4] Apoptosis of neurons were observed with in situ end-labeling (ISEL). [5] Brain-micro vasodilatatio was observed according to the criteria of one complete microvessel account under the field.MAIN OUTCOME MEASURES: [1] Scores of motor function; [2] Ultramicrostructure of cranial neurons and astrocyte; [3] Cranial glial fibriliary acidic protein, neurofilament protein and microtubule-associated protein-2;[4] Apoptosis of neurons; [5] Diastole of cerebral microvessel.RESULTS: Totally 180 rats were eligible while 20 rats were excluded because of their BWT score＞1 after MCAO operation. [1] Results of beam walking test (BWT): Functional recovery of paralysis limbs in electric stimulation group was better than that in control group from the third to the ninth course. In the ninth course, 6 points of rats in electric stimulation group was more than that in control group (42, 46, χ2=15.4, P ＜ 0.01). [2]Positive absorbency of cerebral glial fibriliary acidic protein: That in electric stimulation group was higher than that in control group in the 3rd, 6th,and 9th [(52.97±0.59)% vs (46.40±0.56)%; (49.44±0.80)% vs (46.40±0.56)%;(43.25±0.48)% vs (34.20±0.50)%, P ＜ 0.05]. [3] Assay of neurofilament protein: That in electric stimulation group was higher than that in control group in the 6th and 9th course [(22.9±2.7)% vs (11.9±2.3)%; (26.5±1.7)%vs (11.7±1.5)%, P ＜ 0.05]. [4] Assay of microtubule-associated protein-2:That in electric stimulation group was higher than that in control group in the 6th and 9th course [(21.7±1.3)% vs (11.3±1.1)%; (24.4±2.1)% vs(11.9±2.3)%, P ＜ 0.05]. [5] Apoptosis of neurons: There was not significantly different between the two groups. [6] Results of open number of cerebral microvessel: That in electric stimulation group was higher than that in control group in the 1st, 3rd, 6th and 9th course (33 vs 19; 48 vs 31;45 vs 25; 46 vs 23, Z=-2.309, P ＜ 0.05).CONCLUSION: Electro-stimulating treatment can promote motor function of paralyzed limbs, which was due to that electro-stimulating treatment may promote extinction of the swollen feet of astrocytes, reinforce neurons activity and arouse the dilatation of cerebral capillary which promote the microvascular dilatation in order to improve cerebral blood circulation.

Objective To investigate the suppressive effect of exogenous carbon monoxide (CO) on abnormal platelet exocytosis and its possible molecular mechanism. Methods Venous blood was collected from healthy volunteers. Platelet-rich plasma (PRP) was isolated from the blood by differential centrifugation. The PRP was randomly divided into five groups by random number table, namely normal control group, lipopolysaccharide (LPS) group (challenged with 10 mg/L LPS), inactively exogenous carbon monoxide releasing molecule 2 (iCORM-2) group (given 10 mg/L LPS + 50 μmol/L iCORM-2 for intervention), exogenous carbon monoxide releasing molecule 2 (CORM-2) 10 μmol/L and 50 μmol/L groups (given 10 mg/L LPS + CORM-2 10 μmol/L or 50 μmol/L for intervention). After 30 minutes, enzyme linked immunosorbent assay (ELISA) was used to determine the platelet-derived growth factor BB (PDGF-BB) and matrix metalloproteinase 2 (MMP-2). Chemical fluorescein method was used to determine the platelet adenosine triphosphate (ATP). Flow cytometer was used to determine the expression of P-selectin. The expressions of Toll-like receptor 4 (TLR4), phosphorylation of protein kinase Cθ (PKCθ) and syntaxin binding protein 1 (STXBP-1) were determined by Western Bolt. The soluble N-ethylmaleimide-sensitive factor-attachment protein receptors (SNAREs) complex formation [syntaxin 2-synaptosomal-associated protein 23-vesicle associated membrane protein 8 (STX2-SNAP23-VAMP8)] mediated by STXBP-1 was determined by immunoprecipitation. Results ① Compared with normal control group, the platelet release of PDGF-BB, MMP-2 and ATP was significantly increased after LPS challenge, and the P-selectin expression of platelet was also obviously up-regulated [PDGF-BB (μg/L): 127.53±1.78 vs. 94.35±5.84, MMP-2 (ng/L): 51.87±9.20 vs. 35.83±3.17, ATP (μmol/L): 1.288±0.056 vs. 0.975±0.010, P-selectin: (3.93±0.19)% vs. (0.44±0.10)%, all P < 0.05]. The increases in platelet release of PDGF-BB, MMP-2 and ATP were suppressed by 10 μmol/L or 50 μmol/L CORM-2 administration, as well as high-expression of P-selectin in a dose-dependent manner [PDGF-BB (μg/L): 114.68±1.35, 97.08±6.14 vs. 127.53±1.78, MMP-2 (ng/L): 32.67±8.00, 24.63±1.63 vs. 51.87±9.20, ATP (μmol/L): 0.999±0.015, 0.965±0.008 vs. 1.288±0.056, P-selectin: (1.95±0.27)%, (0.94±0.11)% vs. (3.93±0.19)%, all P < 0.05]. ② Compared with normal control group, LPS challenge resulted in a significant increase in the expression of TLR4 and the phosphorylation of PKCθ and STXBP-1 [TLR4 (gray value): 1.21±0.38 vs. 0.67±0.06, p-PKCθ (gray value): 1.36±0.20 vs. 0.44±0.03, p-STXBP-1 (gray value): 1.13±0.06 vs. 0.59±0.04, all P < 0.05]. The increases in above parameters were suppressed by 10 μmol/L or 50 μmol/L CORM-2 administration in a dose-dependent manner [TLR4 (gray value): 0.76±0.05, 0.65±0.04 vs. 1.21±0.38; p-PKCθ (gray value): 0.71±0.07, 0.47±0.10 vs. 1.36±0.20; p-STXBP-1 (gray value): 0.56±0.02, 0.48±0.01 vs. 1.13±0.06, all P < 0.05]. ③ Compared with normal control group, the SNAREs proteins in platelet that combined with STXBP-1, including STX2, SNAP23 and VAMP8, were obviously increased after LPS challenge [STX2 (gray value): 1.35±0.06 vs. 0.57±0.04, SNAP23 (gray value): 0.97±0.04 vs. 0.30±0.12, VAMP8 (gray value): 1.37±0.12 vs. 0.77±0.10, all P < 0.05]. The increases in SNAREs complex formation were suppressed by 10 μmol/L or 50 μmol/L CORM-2 administration in a dose-dependent manner [STX2 (gray value): 0.77±0.02, 0.39±0.03 vs. 1.35±0.06, SNAP23 (gray value): 0.41±0.03, 0.22±0.08 vs. 0.97±0.04, VAMP8 (gray value): 0.85±0.07, 0.66±0.07 vs. 1.37±0.12, all P < 0.05]. There was no significant difference in the above mentioned parameters between iCORM-2 group and LPS group. Conclusions LPS-induced abnormal secretion of platelet was suppressed by CORM-2 administration. The mechanism may involve the TLR4/PKCθ/STXBP-1 signaling pathway activation and the SNAREs complex formation.

Objective To evaluate the relationship between placental expression of Gab1 and neonatal birth weight in mothers with gestational diabetes mellitus (GDM).Methods From the singleton and full-term cesarean delivered women in Shengjing Hospital Affiliated to China Medical University between October 2014 and May 2015,30 macrosomia babies with maternal GDM were selected as GDM macrosomia group,30 cases of GDM with normal neonatal birth weight as GDM normal group,30 cases without GDM but with macrosomia as normal macrosomia group,and 30 cases without GDM and with normal neonatal birth weight as the normal control group.Gab1 protein and mRNA expression in placentas were detected using immunohistochemistry,Western blot and real-time quantitative-polymerase chain reaction.Analysis of variance,LSD,Dunnett's T3,Chi-square test and Pearson's correlation analysis were used for statistical analysis.Results (1) Gab 1 protein location and positive expression rate:Gab 1 protein expression in human placenta tissue was located in the nucleus.The positive epression rate of Gab 1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group [93％(28/30),73％(22/30) vs 73％(22/30)] and those in the normal macrosomia group and GDM normal group were higher than in the normal control group[47％(14/30)](x2=4.320,4.320,4.444 and 4.444,all P＜0.05).(2) The expression levels of Gabl protein and mRNA:The expression level of Gab1 protein in the GDM macrosomia group was higher than in the GDM normal group and normal macrosomia group (1.43 ± 0.58 vs 1.05 ± 0.67 and 0.95± 0.59),and that in the normal macrosomia group and GDM normal group were higher than in the normal control group (0.64±0.38) (LSD test,all P＜0.05).The expression levels of Gab1 mRNA showed the same trend as the expression levels of Gab1 protein in the four groups.(3) Gab 1 protein expression level was positively associated with neonatal birth weight (r=0.320,P=0.320).Conclusions The expression of Gab1 in placenta is involved in the regulation of birth weight in GDM mothers.