AIM: To observe effect of ? radioactive [103Pd] stent on the proliferation and apoptosis of smooth muscle cells, the mechanism of radioactive stent preventing in-stent restenosis was explored. METHODS: Fifty male New Zealand rabbits were randomized into stent group and [103Pd] stent group. Control group was set up. The materials were harvested on 3, 7, 14, 28, 56 days after operation and the following investigation were carried out, including pathomorphology, immunohistochemistry, apoptosis (TUNEL) and in situ hybridization studies.RESULTS: ① The severity of the stenosis in [103Pd] stent group was less severe than that in stent group. It was most obvious on 56 th day (P

【Objective】To evaluate the impact of stent implantation on proliferation and apoptosis in media vascular smooth muscle cells and to explore the mechanism of restenosis after stent implantation.【Methods】Fifty male New Zealand rabbits were randomized into balloon group and stent group.Control group were set up.The materials were harvested on 3,7,14,28 and 56 day after operation and the following investigation were carried out.① Assessing the expression of proliferating cell nuclear antigen (PCNA) and Cyclin E of media smooth muscle cells with immunohistochemistry;② Analyzing apoptosis of media smooth muscle cells by TUNEL technique.【Results】The expressions of PCNA,Cyclin E and apoptosis in stent and balloon groups were markedly increased compared to control groups.① Stent group induced significant increased expression of PCNA and Cyclin E in the media smooth muscle cells compared to balloon group.On day 7,the positive rates of PCNA and Cyclin E were 24.36±0.55% vs 18.74±1.09% (P<0.01) and 22.65±1.00% vs 17.68±1.10% (P<0.01) respectively;② Stent group induced much more significant apoptosis than balloon group.The highest rate of apoptosis appeared on day 7:12.46±1.13% vs 5.54±0.53% (P<0.01);③By calculating the ratio of positive rates of PCNA to apoptosis and Cyclin E to apoptosis respectively,the ratio of balloongroup was higher than that of stent group.【Conclusion】Stent group induces augmented proliferation and much more significant apoptosis of media smooth muscle cells compared to balloon group.It shows that the severity of restenosis is relieved after stent implantation.

With a comparison between the Non-paper Test model and the traditional one in pharmacology test, an objective analysis was made about the strengths and weaknesses of two test models. This practice shows that the paperless examination can promote teaching and help to improve the teaching quality, but need to be further improved.

BACKGROUND: Brain ischemia-reperfusion (IR) injury is closely connected with the activity of Kinesin. Previous research believes that reduced activity of Kinesin, a mierotubule based motor protein, is an early mark for nerve cell death induced by brain ischemia. Erigeron breviscapus can prevent brain IR-induced proteinase C activation, reduce calcium overload, and reduce ischemic infarctional volume, thus attenuating brain IR injury. However, it still remains less reported at present whether the neuroprotective role of erigeron breviscapus is related to Kinesin activity.OBJECTIVE: To observe the effect of erigeron breviscapus on the activity of Kinesin, a microtubule based motor protein, in hippocampal pyramidal cells during brain IR.DESIGN: Randomized controlled study.SETTING: Anesthesia Department, Affiliated Hospital of Jining Medical College; Anesthesiology Key Laboratory, Affiliated Hospital of Xuzhou Medical College.MATERIALS: This experiment was carried out in the Anesthesiology Key Laboratory, the Affiliated Hospital of Xuzhou Medical College, between February and August 1999. Totally 35 male gerbils were included.METHODS: Gerbils were randomized into sham-operation group (n=5), ischemia-reperfusion control group (n=15) and erigeron breviscapus group (n=15), the latter two of which were further divided into three subgroups according to reperfusion time, namely reperfusion group Ⅰ (reperfusion of 6 hours), reperfusion group Ⅱ (reperfusion of 48 hours) and reperfusion group Ⅲ (reperfusion of 96 hours) with 5 in each subgroup. Gerbils in IR group and erigeron breviscapus group were subjected to IR model preparation before experiment by brain arterial occlusion for 10 minutes, while gerbils in sham-operation group had only bilateral common carotid artery isolated without occlusion. Gerbils in erigeron breviscapus group were pretreated 15 minutes before ischemic inducement with intraperitoneal injection of breviscapine (its effective component is erigeron breviscapus) at a dosage of 45 mg/kg, which was replaced with the same volume of isometric normal saline in sham-operation group and IR group. IHC staining was used to detect hippocampus microtubule based motor protein-Kinesin activity with the assistance of computer imaging analysis technology. MAIN OUTCOME MEASURES: Activity and changes of Kinesin of animals in each group.RESULTS: Totally 35 animals were enrolled in this experiment and all entered the result analysis with no one lost during the experiment. In hippocampal CA1 region, Kinesin activity in IR group was found to descend to 58%, 38% and 12% respectively of that in sham-operation group at IR 6 hours, 48 and 96 hours (P < 0.01). In erigeron breviscapus group at IR 6 hours, 48 hours and 96 hours it was 81%, 61% and 21% of that in shamoperation group, and was obviously higher than that in IR control group (P < 0.05). However, the changes of Kinesin activity were not obviously different in hippocampal CA2, CA3 and CA4 regions.CONCLUSION: Erigeron breviscapus can exert brain-protecting function by reducing hippocampal CA1 Kinesin activity during brain IR injury.

Objective:To set up an effective and simple purification method to obtain highly purified prokaryotic protein of PDCD5 and study its stability. Methods:Recombinant PDCD5 protein expressed in E. coli was accumulated as an inclusion body. After washing, the inclusion body was denatured, renatured, digested with thrombine and then purified by two steps of chromatography. The purity of the products was analyzed by capillary electrophoresis and the stability was identified by SDS-PAGE. Results:Capillary electrophoresis showed that the purity of protein was 100%, and molecular weight was 15 800 with pI 5.9. Further bioactivity assay indicated that the purified PDCD5 could enhance the apoptosis of HL 60 cells withdrawing cytokine, which was in a dose dependent manner. Stability analysis showed that the PDCD5 protein was sensitive to temperature and easy to degrade at 4 ℃ and 25 ℃. However, it was relatively stable at -20 ℃ or lyophilized. Conclusion:Highly purified and stable recombinant PDCD5 protein was obtained, which lays a foundation for the functional study and application investigation of PDCD5 .

We analyzed genetic evolution characteristics of avian influenza A (H7N9) virus isolated in Zhaoqing,China,2014-2016.Nucleic acid were extracted and sequenced from 17 samples of H7N9 positive cases in Zhaoqing.Genetic characteristics of homology and important amino acid sites were analyzed by using BioEdit5.0 and MEGA6.0.The evolutionary trees were constructed by Neighbor-Joining and the referenced sequences were downloaded from GenBank,Eight nucleic acid fragments from 7 strains of H7N9 viruses were successfully generated.The highest homology was found in HA gene with A/chicken/Dongguan/695/2014(H7N9),and NA gene with A/chicken/Dongguan/1075/2014(H7N9).The internal genes were high homology with avian H7N9 and H9N2 virus from Dongguan and Shenzhen in Guangdong,China.The HA and NA genes were directly evolved in the Pearl River Delta evolution branch with the H7N9 sequences from the cities of Dongguan,Guangzhou and Shenzhen,while the sequences from the provinces of Anhui,Zhejiang,and Jiangsu were in the Yangtze River Delta evolution branch.There were 2 alkaline amino acids in cleavage site of HA,2 mutations (G186V and Q226L) in the crucial sites related with the receptor of HA protein,1 mutation (E627K) in PB2 protein,and 1 drug resistance mutation (S31N) in M2 protein.And no evidence of neuraminidase resistance in NA protein was found.In conclusion,the H7N9 virus for human infection in Zhaoqing may originate from avian H7N9 and H9N2 viruses,which circulated in the Pearl River Delta region of Guangdong from 2013 to 2014.The mutations of G186V,Q226L and E627 K might be related with high susceptibility to human beings.

AIM: To investigate the changes of intrarenal endothelin system in the course of congestive heart failure. METHODS: A canine congestive heart failure model induced by rapid right ventricular pacing was used in the present study. Twenty-one mongrel dogs divided randomly into 3 groups: control, congestive heart failure 2 weeks(CHF2) and congestive heart failure 4 weeks (CHF4). The severity of heart failure was evaluated by means of hemodynamic measurement. The concentration of plasma endothelin was detected via RIA, and the expression of endothelin was detected by RT-PCR. RESULTS: The concentration of plasma endothelin in both of CHF2 and CHF4 elevated significantly. In CHF2, the expression of endothelin receptor B(ETB) in renal medulla increased significantly. And in CHF4, the expression of preproendothelin, endothelin receptor A(ETA) and ETB increased both in renal cortex and medulla. Furthermore, in cortex, the expression of ETA increased more significantly than ETB, while in medulla, ETB expressed much more than ETA. CONCLUSION: The changes of renal endothelin system expression plays a role in the regulation of water and electrolyte balance during the progress of congestive heart failure.

AIM: To evaluate expression of the components of endothelin system in pulmonary tissue in a canine pacing-induced congestive heart failure model. METHODS: Twenty-one dogs were divided into 3 groups received 2 (pacing 2 group) or 4 weeks (pacing 4 group) rapid right ventricular pacing, respectively, whereas the other group consisted of 7 dogs received sham-operation as control group. Haemodynamic parameters were detected via left and right heart catheterization. Plasma endothelin-1 was determined by means of RIA. In addition, RT-PCR was used to quantify expression of mRNA of components of endothelin system using ?-actin as internal control. One-way ANOVA and linear regression analysis were used for statistical study. RESULTS:Plasma endothelin-1 increased significantly in heart failure animals. The ratio of preproET-1 to ? actin mRNA was significantly increased from 0.14?0.06 in control group to 0.35?0.08 in pacing 2 group and 0.53?0.08 in pacing 4 group ( P

Objective To explore the investigation method of complicated and profound traditional Chinese herbal medicine,the potential action mechanisms of flavonoids from Scutellaria baicalensis were studied by docking calculation.Methods In total, eight flavonoids (aglycones and their glicosides) from S. baicalensis were selected as ligands.The crystalline structures of targets related to common diseases were used as the receptors for calculation. The calculations were conducted with Schr(o)dinger software package. The grading standard of selectivity was developed according to G-score between ligands and receptors. Results Twenty-six pharmacologic actions have been reported.Among all effects in literature, nine of them can be deduced from the docking calculation of aglycone. From glycosides with grade ++, 25 reported effects can be estimated by calculation. Apparently, the target selectivity of aglycones and their glycosides are different form the virtual evaluation. The virtual evaluation results of glycosides were closer to the reported effects. Conclusion Our proposed virtual evaluation method seems an effective way to investigate the complicated system of traditional Chinese herbal medicine. It suggests that aglycones may be effective as the form of glucoside in vivo, and metabolism is a very important factor for virtual evaluation.

ObjectiveTo investigate the association of the expression of the NK cell-activating receptor NKG2D, its ligand major histocompatibility complex class I chain-related gene A (MICA), and related cytokines ［interferon-γ (IFN-γ), interleukin-10 (IL-10), and interleukin-15 (IL-15)］ with intrahepatic inflammation in primary biliary cholangitis (PBC). MethodsLiver biopsy specimens were collected from 30 patients with PBC (PBC group), 15 patients with chronic hepatitis B (CHB group), and 10 patients with nonalcoholic fatty liver disease (NAFLD group), who were hospitalized in The Second Affiliated Hospital of Kunming Medical University from August 2014 to June 2015. The degree of liver inflammation (G) and fibrosis degree (S) of the liver specimens were determined, and immunohistochemistry was used to measure the expression of NKG2D, MICA, IFN-γ, IL-10, and IL-15 in liver tissue (the scores were determined based on the number of cells stained and the degree of staining to evaluate the expression of each marker). A one-way analysis of variance was used for comparison of continuous data between multiple groups, and the t-test was used for comparison between two groups; a Spearman correlation analysis was used to investigate correlation. ResultsIn the PBC group, the expression of NKG2D increased with the degree of inflammation, and the patients with G3-4 inflammation had significantly higher expression than those with G1-2 inflammation (G1 vs G2 vs G3 vs G4: 1.4±0.05 vs 1.56±0.05 vs 1.86±0.11 vs 2.60±0.17, F=150.8, P＜0.05); the expression of NKG2D decreased with fibrosis degree (S3 vs S4: 2.30±0.17 vs 1.56±0.05, t=-1.52, P＜0.05). In the PBC group, there was no significant difference in MICA between G3 and G4 (0.11±0.01 vs 0.20±0.03, t=-2.20, P＞0.05) and between S3 and S4 (0.12±0.02 vs 0.18±0.03, t=-2.64, P＞0.05). In the PBC group, there was a significant difference in the expression of IL-15 between the patients with different degrees of inflammation (G1 vs G2 vs G3 vs G4: 0.70±0.10 vs 1.50±0.10 vs 1.93±0.11 vs 2.60±0.17, F=251.3, P＜0.05), while there was no significant difference between the patients with different fibrosis degrees (S3 vs S4: 2.00±0.05 vs 2.40±0.30, t=-1.62, P＞0.05). In the CHB group, there was a significant difference in the expression of IL-15 between the patients with different degrees of inflammation (G1 vs G2 vs G3: 0.73±0.15 vs 1.96±0.15 vs 2.50±0.17, F=150, P＜0.05) and between the patients with different fibrosis degrees (S1 vs S2 vs S3: 0.70±0.10 vs 21.96±0.15 vs 2.50±0.17, F=158.7, P＜0.05). In the PBC group, the expression of IL-10 was only observed in the patients with G1 inflammation (0.16±0.01), and in the CHB group, the expression of IL-10 was observed in the patients with G1 and G2 inflammation, with no significant difference (G1 vs G2: 0.19±0.01 vs 0.13±0.01, t=-1.522, P＞0.05). In the patients with PBC, the expression of IL-15 in liver tissue was positively correlated with the levels of alkaline phosphatase (ALP) and gamma-glutamyl transpeptidase (GGT) (r=0.241 and 0.407, P=0.014 and 0.045). ConclusionThe NK cell-activating receptor NKG2D affects the degree of intrahepatic inflammation in PBC, and the NKG2D ligand MICA is expressed in the advanced stage of PBC and can downregulate NKG2D. The expression of IL-15 increases with the degree of inflammation in PBC and is positively correlated with the levels of ALP and GGT, suggesting that the activation of NK cells and abnormal secretion of cytokines are involved in the development and progression of PBC and IL-15 may be used as an auxiliary index for the diagnosis of PBC.