Twenty-five compounds of novel quinoxaline-based scaffold with antiplatelet activity were designed and synthesized on the basis of previous quinoxaline analogues, and the structures were confirmed by ¹H NMR, ¹³C NMR, and MS. The antiplatelet activity was evaluated, structure-activity relationship (SAR) study was summarized and the selectivity of PAR4 was confirmed by calcium mobilization assays. It was indicated that compound 14a, 14g, 13i, 13p showed moderate activity against PAR4, especially, the activity of compound 14g (IC₅₀ = 0.26 μmol·L^-1) was 6.7 times than the lead compound A (IC₅₀ = 1.73 μmol·L^-1). Therefore, 2,3-dihydro-[1,4]dioxino[2,3-g]quinoxaline and [1,3]dioxolo[4,5-g]quinoxaline derivatives are promising compounds for the discovery of novel antiplatelet agents. It is worthy of further research to develop highly effective and selective PAR4 antagonists.

Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

In order to prepare the recombinant vasoactive intestinal peptide (VIP) using intein mediated rapid purification system,the cDNA encoding the recombinant VIP was designed and synthesized according to the preference of E.coli,and then was cloned into the expression vector PTWIN. The recombinant plasmid PTWIN-VIP was transformed into expression host E.coli strain ER2566.The fusion protein consisting of the recombinant VIP,intein and chitin binding domain was expressed and purified by chitin affinity chromatography. The target peptide was released from the fusion protein by changing the temperature and the pH of the cleavage buffer. The molecular weight of the recombinant VIP was determined by the mass spectrometry and the results was conformity with the theoretical value. The preliminary bioactivity assay indicated that the recombinant VIP decreased the serum resistin levels significantly in LPS-induced acute inflammation. The preparation and the characterization of anti-inflammatory effects of the recombinant VIP layed the foundation for its further application.

BACKGROUND: Acidic peptide is the tripeptide composed of 3 glutamic acids, which cannot bring excitatory nerve signal transmission into playlike single glutamic acid through presynaptic release and integration withpostsynaptic NMDA receptor directly as excitable neurotransmitter. It is quite possible that acidic peptide plays its actions by integrating with multiple metabolic glutamic acidic receptors so as to promote neuron proliferation or release nerve growth factor (NGF). OBJECTIVE: To probe into whether acidic peptide induces changes in learning and memory of model rats with Alzheimer disease (AD).DESIGN: Randomized controlled single experiment was designed.SETTING: Teaching-Research Room of Biochemistry and Molecular Biology of Basic Medical College of Zhengzhou University.MATERIALS: The experiment was performed in 2nd Research Room and Experimental Animal Room of Teaching-Research Room of Biochemistry and Molecular Biology of Basic Medical College of Zhengzhou University.Totally 100 SD male rats were selected and some of them were excluded due to retarded response in step down test. Totally 84 rats were included in the experiment and randomized into 7 groups, named normal control,model group, physiological saline group (PS group), piracetam group, acidic peptide groups of 60, 30 and 15 mg/kg, 12 rats in each group. Acidic peptide is a new small molecular peptide separated from bovine brain in this research team and is tripeptide composed of three glutamic acids.METHODS: Except normal control, in the rest groups, after 1 week routine breeding, cerebral stereotactic microinjection was used to inject 5 μg ibotenic acid in hippocampus of rats to destroy bilateral Meynert's basal ganglia to establish AD model. In normal control and model group, no medication was applied. In PS group, physiological saline was used for gastric perfusion. In piracetam group, piracetam of 0.3 g/kg was used for gastric perfusion and in acidic peptide groups of 15, 30 and 60 mg/kg,acidic peptide of 60, 30 and 15 mg/kg was applied for gastric perfusion successively, continuously for 20 days, once per day, 2 mL/time. On the expiration of gastric perfusion, learning and memory of rats were examined with step down test in every group. The animal was placed on the safe table on step down platform to adapt to the environment for 3 minutes, afterwards, 36 V electric current was given. Error response was recorded if the animal jumped to the copper railings after electric shock and correct response was recorded if the animal jumped back the safe area. Step-up latent phase and frequency of correct response were recorded in 3 minutes.MAIN OUTCOME MEASURES: Comparison of learning and memory of rats in every group. RESULTS: Totally 84 rats were all included in the result analysis. ①Comparison of learning in every group: Compared with model group, stepup latent phase was shortened remarkably in every acidic peptide group[(102.03±5.33), (71.77±4.38), (68.28±9.53), (69.13±8.79) s, P ＜ 0.01] and the frequency of correct response was improved remarkably [(12.92±2.91),(16.17±2.79), (15.83±3.27), (16.33±2.53) times, P ＜ 0.01]. ② Comparison of memory in every group: Compared with model group, step-up latent phase was shortened remarkably in every acidic peptide group [(43.17±4.66),(29.78±4.48), (26.20±3.28), (22.09±4.43) s, P ＜ 0.01] and the frequency of correct response was improved remarkably [(15.67±2.15), (20.92±2.68),(20.83±2.29), (20.25±2.05) times, P ＜ 0.01].CONCLUSION: Acidic peptide can shorten remarkably the step-up latent phase of AD rats in step down test and improve the frequency of correct response. It is indicated that acidic peptide provides good intervention on learning and memory of rat model of Alzheimer disease.

Objective Using CT perfusion (CTP) technique,to investigate the graft perfusion changes in patients with hepatic artery stenosis (HAS) with or without ischemic-type biliary lesions (ITBL) after orthotopic liver transplantation (OLT).Methods Thirteen recipients with HAS received CTP scan of the liver,including 8 with ITBL and 5 without ITBL.For all patients,the diagnosis of HAS was made by CTA,and the diagnosis of ITBL by percutaneous transhepatic cholangiography.CT perfusion indices were obtained,including hepatic artery perfusion (HAP),portal vein perfusion (PVP),total liver perfusion (TLP) and hepatic perfusion index (HPI).Results Of the 13 patients with HAS,mean HAP in patients with and without ITBL was 59.8 and 35.1 ml·min-1 ·100 ml-1 (P =0.021,two-tailed paired Student t test) ; mean PVP was 125.4 and 166.2 ml·min-1·100 m1-1 (P =0.016) ; mean TLP was 185.2 and 201.3 ml· min-1 · 100 ml-1 (P =0.306) ; and mean HPI was 33.6 and 18.2 (P =0.005),respectively.Conclusion Using CTP technique,liver perfusion changes were reflected by measuring CTP indices noninvasively.Compared to those without ITBL in this study,HAP and HPI in patients with ITBL were higher and PVP was lower,which may be contributed to biliary inflammation.

Objective: To observe the clinical efficacy of spleen-invigorating and qi-benefiting pediatric massage (tuina) for treating recurrent respiratory tract infection in children with cerebral palsy due to qi deficiency of spleen and lung. Methods: A total of 70 children with cerebral palsy who suffered from recurrent respiratory tract infection due to qi deficiency of spleen and lung were randomized into an observation group and a control group by the random number table method, with 35 cases in each group. Both groups were treated with conventional rehabilitation training, while the observation group was given additional spleen-invigorating and qi-benefiting pediatric massage, and the control group additionally took oral Yu Ping Feng granule. The clinical efficacy was evaluated by the total effective rate, traditional Chinese medicine (TCM) symptom score, and serum levels of immunoglobulin (Ig) A, IgM and IgG. Results: The difference in total effective rate between the two groups after treatment was statistically significant (P<0.05). After treatment, the TCM symptom scores in both groups decreased to varying degrees than those before treatment, and the intra-group differences were statistically significant (all P<0.01); the differences in the scores of various TCM symptoms between the two groups were statistically significant (all P<0.05). After treatment, the levels of IgA, IgM and IgG of the children in both groups increased to varying degrees, and the intra-group differences were statistically significant (all P<0.05). The between-group differences in the IgA, IgM and IgG levels after treatment were statistically significant (all P<0.05). Conclusion: Spleen-invigorating and qi-benefiting pediatric massage can effectively treat recurrent respiratory tract infection due to qi deficiency of spleen and lung in children with cerebral palsy, relieve the clinical symptoms and improve immune function, and thus is worthy of clinical promotion and application.

Objective To construct a voxel mouse model combining with Monte Carlo method used for radiation dose calculation.Methods A set of slice images obtained from a nude male mouse (28 g) was utilized, all slice images were registered,some organs or tissues were identified, segmented and filled with specific color, and finally the physical property was defined by MCNP application.Results A mouse model with a voxel size of 0.2 mm×0.2 mm×0.2 mm, consisting of 14 organs or tissues,was constructed, which could satisfy the requirement of precision for radiation dose distribution calculation and moderate computing time.Conclusion The voxel mouse model can be used to calculate the quantity of ionization radiation dosimetry in related areas including radiological medicine, nuclear medicine and space radiation medicine.

It was hypothesized that the VPAC1 agonist may exert anti-obesity functions because VPAC1 is involved in the anorexigenic effects and the anti-inflammatory function of pituitary adenylate cyclase-activating polypeptide (PACAP)/vasoactive intestinal polypeptide (VIP). Furthermore, our in vitro test showed that the expression of VPAC1 increased significantly after the 3T3-L1 adipocytes were differentiated, and that incubation of adipocytes with VPAC1 agonist (10-1 000 nmol/L per 1x10(6) cells) resulted in stimulation of lipolysis. To test the effect of VPAC1 agonist [Lys15, Arg16, Leu27]-VIP (1-7) GRF (8-27) on diet-induced obesity (DIO), we further designed the following two in vivo experiments: (1) Mice were fed on high-fat diet (HFD) and intraperitoneally (i.p.) treated with VPAC1 agonist simultaneously for 28 d; (2) Mice were given HFD for 35 d, and subsequently fed on the same HFD and i.p. treated with VPAC1 agonist for the next 28 d. The physiological indices, including body weight, weight of white adipose tissue, plasma glucose and blood lipid, were collected. The results showed that treatment with VPAC1 agonist inhibited ingestion significantly and prevented the elevations in body weight and the weights of the white adipose tissues (epididymal and dorsal) induced by HFD. The increases in plasma glucose, cholesterol, triglycerides and LDL induced by HFD were also down-regulated in mice treated with VPAC1 agonist. VPAC1 agonist treatment also improved the glucose tolerance. Therefore, VPAC1 agonist treatment inhibits the development of the obesity induced by HFD and helps to improve the morbidities associated with DIO.
3T3-L1 Cells ; Adipocytes ; drug effects ; Animals ; Body Weight ; Diet, High-Fat ; Mice ; Obesity ; drug therapy ; Receptors, Vasoactive Intestinal Polypeptide, Type I ; agonists ; Vasoactive Intestinal Peptide ; pharmacology

Objective:To observe the effect of Governor Vessel-unblocking and mind-refreshing acupuncture plus functional training on neural development in infants with brain damage and seek an effective method for early intervention of infantile brain damage.Methods:Eighty infants with brain injury were recruited and allocated to a treatment group and a control group by their visiting sequence,with 40 cases in each group.The control group received exercise training,40 min each session and 6 sessions a week,and tuina treatment,30 min each time and 6 times a week.Based on the treatment protocol for the control group,the treatment group additionally received Governor Vessel-unblocking and mind-refreshing acupuncture,3 times a week and 10 sessions as a course at a 2-week interval.Before the treatment and after 14-week treatment,the gross motor function measure (GMFM) and developmental quotient (DQ) of Bejing Gesell developmental scale were used to evaluate the development of the infants.Results:After the treatment,the GMFM score and DQs of Gesell scale all increased by different levels in the two groups,and the intra-group differences were statistically significant (all P＜0.05);the scores of the treatment group were superior to those of the control group,and the between-group differences were statistically significant (all P＜0.05).Conclusion:Governor Vessel-unblocking and mind-refreshing acupuncture plus functional training can significantly promote the development of gross motor and cognitive functions in infants with brain damage,and it is an early and effective intervention for infantile brain damage.

Objective To exlore the influence of internal quality control and external quality control assessment(EQA) resulting from applicability of control samples in measurement of whole blood viscosity (WBV) through the analysis and comparison of applicability of 1 non-Newtonian fluid internal quality control sample in 3 viscometers. Methods Viscometer B, C and D were used to measure WBV of 30 blood samples in parallel under the shear rate(SR) of 1 s-1,30 s~(-1) and 200 s~(-1), then the blood SR-WBV curves of 3 viscometers were drawn according to the results. At the same time, viscometers B, C and D were used respectively to determine the WBV of control A 10 times in one day, then the control A SR-WBV curves were mapped. Three viscometers were used to measure the manufactory control samples and control A 5 times in one day for 4 days. Four groups of daily values of manufactory control samples and control A of each instrument were used to carry out F test to calculate whether 4 daily values are difference. Finally, the control A was dispensed in 49 laboratories nationwide chosen for measurement. On the basis of viscometer used, 20 laboratories were classified as group B, 20 laboratories were classified as group C and 9 laboratories were classified as group D. Then the data under SR of 1 s~(-1) were analyzed to calculate the coefficient of variation (CV) in the group. Results There was significant difference among the WBV of blood samples measured by the viscometers B, C and D. The results under SR of 1 s~(-1) declined in turn, and they were highest under SR of 30 s~(-1) followed by the values of viscometer D and B and they were (8.14±0.75), highest under SR of 30 s-1 followed by the values of viscometer B and D, and they were (7.35±0.07), daily values of manufactory control and control A of each instruments in four groups were compared. Under SR of 1 s~(-1), there was no difference between daily values of manufactory control and control A in viscometer B (F = 2.63, 1.37, P > 0.05), and there was no difference of daily values of manufactory control among viscometer C and D (F = 0.33,3. 14, P > 0.05), but significant daily difference existed when control A was tested by viscometer C and D (F = 5.76, 8.00, P < 0.05). Under SR of 30 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers(F =0.31, 0.18, 2.26, P >0.05), and there was no difference of daily values of control A among 3 viscometers' (F = 1.03, 1.83, 2.40, P > 0.05); Under SR of 200 s~(-1), there was no difference of daily values of manufactory control among 3 viscometers (F =2.59, 0.68, 2.96, P > 0.05), and there was no difference of daily values of control A among 3 viscometers (F=2.31, 3.01, 2.28, P>0.05). When control A was tested under SR of 1 s~(-1) in 49 laboratories nationwide, the WBV values in groups of viscometer B, C and D were (18.47±1.30), (11.17±2.38), viscometer D and C were 63.75% and 21.3%. Conclusions Control A could fully mimic the properties of whole blood steadily on viscometer B, but partially mimic viscometer C and D, so the control A is most appropriate for viscometer B. Because current non-Newtonian fluid internal quality control could mimic rheological properties of whole blood under specifically conditions, laboratories should evaluate the consistent degree between control and whole blood, only the candidates which can mimic the properties of whole blood approximately could be chosen as quality control of WBV. When third-party control is chosen to be samples of EQA, its applicability should be in consideration. Pretest should be performed adequately to define applicability of third-party control, so as to reduce the difference among laboratories due to applicability of control and reflect detection quality of laboratories exactly.