Objective To investigate the relationship between the numbers of deep medullary veins (DMV) beside the ipsilateral lateral ventricle and the stenotic degree of large artery of ipsilateral internal carotid artery (ICA) system and cerebrovascular reactivity (CVR) in patients with acute cerebral infarction showed on susceptibility-weighted imaging (SWI) in middle cerebral artery (MCA) territory.Methods The patients with acute cerebral infarction in MCA territory who underwent SWI,cerebral angiography and the transcranlal Doppler carbon dioxide challenge test during the hospital stay were enrolled retrospectively.According to the numbers of DMV beside the lateral ventricle on SWI,they were classified into 3 grades:Grade 1:DMV ＜5; Grade 2:DMV 5-10; Grade 3:DMV ＞ 10.The correlations of the DMV grades and the degree of ipsilateral ICA/MCA and CVR were analyzed.Results A total of 44 patients with acute cerebral infarction in MCA territory were enrolled.Twenty-four patients were DMV 1,15 were DMV2,and 5 were DMV 3.The mean stenosis degree of the main arteries of the ipsilateral ICA system was 40.52％ ±30.62％.The mean CVR of MCA was 0.41 ± 0.16.The stenosis degrees in the DMA grade 1,2,and 3groups were 31.31％ ± 26.02％,44.93％ ± 31.03％,and 71.76％ ± 26.68％,respectively.There were significant differences among the 3 groups (F=4.589,P =0.018),and the stenosis degree in the DMV grade 3 group was significantly higher than that in the DMV grade 1 group (P=0.014).The CVRs in the grade 1,2,and 3 groups were 0.45 ± 0.12,0.41 ± 0.18,and 0.22 ± 0.12,respectively.There were significant differences among the 3 groups (F=6.179,P=0.005),and the CVR in the DMV grade 3 group was significantly lower than that in the DMV grade 1 group (P =0.006) and in the DMV grade 2 group (P =0.038).Spearman correlation analysis showed that DMV grade was significantly positively correlated with the stenosis degree (rs =0.381,P =0.013),and was significantly negatively correlated with CVR (rs =-0.375,P=0.012).Conclusions The DMV grade in patients with cerebral infarction was positively correlated with the stenosis degree of responsible large artery and was negatively correlated with the MCA CVR in the responsible side.

AIM: To determine whether chronic hypercholesterolemia affects ionic currents on cardiac ventricular myocytes of rats. METHODS: Whole - cell patch - clamp technique was used to record the ionic currents in single cardiac myocytes isolated from normal cholesterolemia and hypercholesterolemia rats. RESULTS: In the hypercholesterol group (group Ⅱ ), serum total - cholesterol level was significantly higher than that of normal group (group Ⅰ) [ (3. 10 ±tricular myocytes of rats, 50% repolarization of action potential duration (APD50) prolonged from (70. 86 ± 8.12) ms (group Ⅰ) to (116.16±6.90)ms (group Ⅱ) (n=10 in each group, P＜0.01); APD90 prolonged from (95.10±7. 27)ms (group Ⅰ ) to (144. 04 ± 7.39)ms (group Ⅱ ) (n = 10 in each group, P ＜ 0. 01 ); at the test potential of - 120 mV, Ik1 increased from ( - 16. 98 ±4. 54) pA/pF(group Ⅰ ) to ( - 19.92 ±4.08) pA/pF (group Ⅱ ) (n = 12 in each group, P ＜ 0. 05 ); at the test potential of 0 mV, ICa- L decreased from ( - 8.56 ± 1.29) pA/pF ( group Ⅰ ) to ( -5. 24 ± 0. 90) pA/pF ( group Ⅱ ) ( n = 10 in each group, P ＜ 0. 01 ); at the test potential of + 60 mV, Ito decreased from (13.20±1.97) pA/pF (group Ⅰ) to (10.30±1.97) pA/pF (group Ⅱ) (n=8 in each group, P＜0. 05). CON-CLUSION: Hypercholesterolemia affects the ionic currents on cardiomyocytes of rats greatly, which may be the ionic mechanism of cardiac toxicity induced by hypercholesterolemia.

AIM To elucidate the possible antiarrhythmic mechanism of matrine. METHODS Whole-cell patch-clamp technique was used to record ionic currents in ventricular myocytes. RESULTS In guinea pig ventricular myocytes, matrine 100 μmol·L-1 prolonged 90% action potential duration (APD90) by 40% at a stimulation of 0.1 Hz in a frequency-independent manner, inhibited IK1 by 47% at the test potential of -120 mV, reduced IKr,tail by 50% and had no effect on IKs,tail. CONCLUSION Matrine prolonged APD through blockade of multiple potassium currents, which may relate to its antiarrhythmic efficacy.

Aim To examine the role and uderlying mechanisms of Lin28 /let-7d axis in the proliferation of lung fibrobalsts and fibroblasts-into-myfibroblasts tran-sition,and provide novel strategy for the treatment of idiopathic pulmonary fibrosis (IPF).Methods We induced experimental lung fibrosis in mice by intratra-cheally injection of bleomycin (BLM).Ang Ⅱ and TGF-β1 were used to induce fibrogenesis in cultured MRC-5 cells;qRT-PCR and Western blot were applied to determine the changes of Lin28B,collagen 1 α1 and collagen 3α1 ;MTT assay,Edu satining and immun-ofluoresence were used to examine the cell viability, proliferation and fibroblasts-into-myofibroblasts transi-tion in MRC-5 cells.Results Lin28B was increased in the lung of mice with experimental lung fibrosis and in MRC-5 cells treated with AngⅡ or TGF-β1 .Moreo-ver,Lin28B enhanced collagen deposition via inhibi-ting expression of let-7d,which maybe contribute to the progression of IPF.In addition,further studies showed that Lin28B promoted proliferation and fibro-blasts-into-myofibroblasts in MRC-5 cells.Conclusion Lin28B /let-7d axis contributes to fibrogenesis via promotes fibroblasts-into-myofibroblasts transition, which may provide novel approaches for lung fibrosis treatment.

Aim To evaluate the effect of 15-hydroxy eicosatetraenoic acid(15-HETE) on the tension of isolated internal carotid artery(ICA) rings in normoxic or hypoxic rats in order to find out the pathophysiological effect of 15-HETE in ischemic/hypoxic brain tissues.Methods Sixteen healthy Wistar rats were randomly divided into two groups(n=8): group A with normal oxygen supply(FiO_2 21%) and group B with hypoxia(FiO_2 10%).9 days later,the rats were killed and the ICA rings were prepared.15-HETE in different concentration were added accordingly,and the after washing up,blockers of different kinds of K~+ channels such as 2 mmol?L~(-1) 4-AP,10~(-2) mol?L~(-1) TEA and 10~(-6) mol?L~(-1) GLYB were added to the KH solution separately,followed by the administration of 10~(-6) mol?L~(-1) 15-HETE.The isometric tension was recorded using a four-channel force-displacement transducer.Results The ICA tension was significantly increased both in groups A and B by 15-HETE in concentration-dependent manner(P0.05,n=8).Furthermore,10~(-2) mol?L~(-1) TEA,10~(-6) mol?L~(-1) GLYB had little effects on the tension of rings.Application of 10~(-6) mol?L~(1) 15-HETE after administration of TEA or GLYB induced nearly the same increase in ICA tone as it happened in the group administered with 10~(-6)mol?L~(-1) 15-HETE alone.Conclusion 15-HETE can induce brain vasoconstriction especially under the hypoxic condition.Voltage-dependent potassium channels play an important role in 15-HETE induced constriction.

Objective To study the antitumor effect of new photodynamic therapy (PDT) applying TMPyP4 combined with nucleolin silence on cervical cancer SiHa cells in vitro, and to explore an available combination treatment project for cervical cancer.Methods The SiHa Cells were divided into blank control group, RNAi group,PDT group and PDT-RNAi group.The proliferation activities of SiHa cells were assessed by Cell Counting Kit-8 (CCK8)assay.The apoptotic rates were measured by flow cytometry (FCM)with Annexin Ⅴ/PI staining. The invasiveness abilities were assessed by Transwell assay. Results Compared with blank control group, the inhibitory rates of growth of SiHa cells in RNAi group, PDT group and PDT-RNAi group were increased significantly (P<0.01);the inhibitory rates of growth of SiHa cells in PDT-RNAi group were higher than those in PDT group and RNAi group (P<0.05 ). The Q value was 1.27. Compared with blank control group, the apoptotic rates of SiHa cells in experiment groups were increased (P<0.05 ), and the late apoptotic rate in PDT-RNAi group was also increased;there were significant differences of the apoptotic rates between PDT-RNAi group and RNAi group, PDT group (P<0.05 ). Compared with blank control group, the cell invasiveness abilities of SiHa cells in experiment groups were decreased;there were significant differences of the invasiveness abilities of SiHa cells between PDT group and RNAi group (P<0.05).Conclusion New PDT shows a strong photodynamic effect on the SiHa cells,which can inhibit the proliferation and invasiveness and induce the apoptosis of SiHa cells invitro;nucleolin silence shows a good synergy effect to PDT.

Methods: (i) Animal experiments. This study conducted experiments using specific pathogen-free (SPF) grade male C57BL/6J wild-type (WT) mice and APP/PS1 double transgenic mice. The animals were divided into three groups: WT group (WT mice, n = 5, receiving distilled water daily), APP/PS1 group (APP/PS1 double transgenic mice, n = 5, receiving distilled water daily), and BSTSD group [APP/PS1 double transgenic mice, n = 5, treated with BSTSD suspension at a dosage of 27 g/(kg·d) for 90 d]. Cognitive function was assessed using the Morris water maze (MWM). Post-experiment, hippocampal tissues were collected for analysis of pyramidal cell and synaptic morphology through hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM). (ii) Cell experiments. The HT-22 cells were divided into control group (untreated), Aβ25-35 group (treated with 20 μmol/L Aβ25-35 for 24 h), icariin group (pre-treated with 20 μmol/L icariin for 60 min, followed by 20 μmol/L Aβ25-35 for an additional 24 h), and icariin + LY294002 group [treated with 20 μmol/L icariin and 20 μmol/L LY294002 (an inhibitor of the phosphoinostitide 3-kinases (PI3K) signaling pathway) for 60 min, then exposed to 20 μmol/L Aβ25-35 for 24 h], and cell viability was measured. Western blot was used to detect the expression levels of synapse-associated proteins [synaptophysin (SYP) and postsynaptic density-95 (PSD-95)] and PI3K signaling pathway associated proteins [phosphorylated (p)-PI3K/PI3K, p-protein kinase B (Akt)/Akt, and p-mechanistic target of rapamycin (mTOR)/mTOR]. Results: (i) Animal experiments. Compared with APP/PS1 group, BSTSD group showed that escape latency was significantly shortened (P < 0.01) and the frequency of crossing the original platform was significantly increased (P < 0.01). Morphological observation showed that pyramidal cells in the hippocampal CA1 region were arranged more regularly, nuclear staining was uniform, and vacuole-like changes were reduced after BSTSD treatment. TEM showed that the length of synaptic active zone in BSTSD treatment group was increased compared with APP/PS1 group (P < 0.01), and the width of synaptic gap was decreased (P < 0.01). (ii) Cell experiments. Icariin had no obvious toxicity to HT-22 cells when the concentration was not more than 20 μmol/L (P > 0.05), and alleviated the cell viability decline induced by Aβ25-35 (P < 0.01). Western blot results showed that compared with Aβ25-35 group, the ratios of p-PI3K/PI3K, p-Akt/Akt and p-mTOR/mTOR in icariin group were significantly increased (P < 0.01), while the protein expression levels of SYP and PSD-95 were increased (P < 0.01). These effects were blocked by LY294002 (P < 0.01). Conclusion BSTSD and icariin enhance cognitive function and synaptic integrity in AD models and provide potential therapeutic strategies through activation of the PI3K/Akt/mTOR pathway.

ObjectiveTo explore the clinical value of three serum tumor markers, Golgi protein 73 (GP73), alpha-fetoprotein (AFP), and AFP-L3, in the diagnosis of liver cancer and recurrence monitoring after radio frequency ablation. MethodsA total of 174 patients who visited our hospital from July 2012 to October 2013 were included in the study, consisting of 86 patients with newly diagnosed liver cancer, 39 with liver cirrhosis, 29 with hepatitis, and 20 healthy controls. Among the patients with newly diagnosed liver cancer, 37 were followed up for three months after the radiofrequency ablation. Serum levels of GP73, AFP, and AFP-L3 were measured by ELISA, electrochemiluminescence, and affinity adsorption chromatography, respectively. Nonparametric tests were performed on the results of serum samples from the four groups which showed skewed distribution and were represented by median (quartile interval) ［M(P25-P75)］. Overall comparison was made by Kruskal-Wallis H test, and comparison between groups was made by Mann-Whitney U test. Pair-matching rank-sum test was performed using Wilcoxon Signed Ranks, and categorical data were analyzed by χ2 test. ResultsThe levels of GP73, AFP, and AFP-L3 in the liver cancer group were significantly higher than those in other groups (all P＜0.05). The positive rates of GP73 and AFP-L3 in the liver cancer group were significantly higher than those in other groups (all P＜0.05), and the positive rates of the two markers were significantly higher than that of AFP among patients with liver cancer (P＜0.05). Thirty-seven patients with newly diagnosed liver cancer were reexamined three months after radiofrequency ablation, and the preoperational AFP-L3 level in the patients who had recurrence was significantly higher than that in the patients without recurrence (P＜0.05). ConclusionSerum GP73, AFP, and AFP-L3 show great values in the diagnosis of liver cancer. AFP-L3 can be used as an indicator for the identification of benign or malignant liver disease and risk of recurrence after radiofrequency ablation.

Objective To assess the anti-arrhythmic activity and cardioprotective effects of Wenxin Granula, a traditional Chinese formula (consisting of Salviae Miltiorrhizae Radix, Polygonati Rhizoma, Notoginseng Radix et Rhizoma, Nardostachyos Radix et Rhizoma, Angelicae Sinensis Radix, and Succinum), on heart in ischemic-induced myocardial infarction (MI) rats and compare with those of Amiodarone which have been demonstrated in clinic. Methods Rats were randomly divided into Sham-operated (control), Ml + Amiodarone [5 mg/(kg·d)] (MI), and MI + Wenxin Granula [10 mg/(kg·d)] groups and left anterior descending coronary artery was occluded in each group. After left anterior descending for 12 h, standard lead Ⅱ of administration electrocardiogram was recorded in order to analyze the occurrence of arrhythmia. After one month, the size of the infarct area of heart was evaluated by TTC staining method and haemodynamic function was assessed to detect the heart function. Laser scanning confocal microscope and the technique of patch clamp were used to detect the intracellular Ca2+ ([Ca2+]j) and L-type calcium current (ICa-L), respectively. Results Both Wenxin Granula [10 mg/(kg·d)] and Amiodarone [5 mg/(kg·d)] could markedly decrease the incidence of arrhythmia in heart of rats which were subjected to ischemic injury. After one month, Wenxin Granula could significantly decrease mortality to 22.22% and reduce the infarct area (P < 0.05), but Amiodarone did not. The mechanism may involve that Wenxin Granula attenuated [Ca2+]j decreasing in MI rats. Additionally, Wenxin Granula could obviously ameliorate the impaired heart function of MI rats by decreasing the elevated left ventricular end-diastolic pressure and increasing the attenuated maximum change velocity of left ventricular pressure in the isovolumic contraction or relaxation period. On the other hand, electrophysiological experiment results revealed that Wenxin Granula administration one month later also increased the reduced ICa-L density in rat ventricular myocytes in MI rats. The results of LSCM showed that Wenxin Granula could recover the amplitude of [Ca2+]j decreased by heart failure during long term. Conclusion Wenxin Granula could not only inhibit the incidence of arrhythmia but also decrease the mortality, which was accompanied by recovering the amplitude of [Ca2+]j. This protective effect of Wenxin Granula may partially be mediated through changing ICa-L.as well as increasing [Ca2+]j.

ObjectiveTo investigate the effect of FUT8-siRNA on transforming growth factor β (TGF-β)-Smad2/3 signalling pathway in renal tubular epithelial cells.MethodsHK-2 cells were divided into six groups:normal group,negative control group,TGF-β1 group,TGF-β1 with FUT8 interference group,TGF-β1 with negative control group,FUT8 interference group.RNAi was performed to silence the expression of FUT8 gene,then immunofluorescent analysis was used to detect the expression of core fucose in the HK-2,immunoprecipitation and lectin blotting were performed to detect the core fucosylation of TGF-βR Ⅱ and ALK-5,and detect the change of Smad2/3 and p-Smad2/3 in HK-2 cells after FUT8 gene was silenced.ResultsCompared with the normal and negative control group,incubation with 5 μg/L TGF-β1 for 48 h could significantly up-regulate the core fucosylation of HK-2 cells,enhance the protein expression of TGF-βR Ⅱ and ALK-5 (P＜0.05),markedly increase the expression level of p-Smad 2/3 (P＜0.05) and cause it to nuclear translocation in HK-2 cells.While FUT8siRNA could inhibit the above up-regulation of TGF-βR Ⅱ and ALK-5(P＜0.05),suppress the increase of p-Smad 2/3(P＜0.05) and its nuclear translocation without disturbing the protein expression of TGF-βR Ⅱ and ALK-5.Conclusion FUT8-catalized core fucosylation of TGF-βR Ⅱ and ALK-5 is needed to fulfill their functions,and blocking core fucosylation of TGF-βR Ⅱ and ALK-5 leads to the inhibition of TGF-β-Smad2/3signalling pathway in HK-2 cells.