BACKGROUND: Pillbugs have been proved to exert an effect on the proliferation and migration of vascular smooth muscle cells (VSMCs) through MTT assay and scratch assay, but the mechanisms are poorly understood. OBJECTIVE: To investigate the molecular mechanisms underlying pillbugs influencing osteopontin-induced VSMC migration and proliferation. METHODS: VSMCs from Sprague-Dawley rat thoraco-abdominal aorta were cultured in a common medium, and 3-5 generations of cells were selected. There were eight groups: control, osteopontin, pillbug water extract (0.5, 1.0, and 2.0 g/L) with osteopontin, and pillbug ethyl acetate extract (0.25, 0.5, and 1.0 g/L) with osteopontin groups. After 24 hours of culture, the expression levels of proliferating cell nuclear antigen (PCNA) and integrin kinase (ILK) were detected by western blot assay. RESULTS AND CONCLUSION: Pillbug water and ethyl acetate extracts both inhibited the expression of PCNA, especially when cultured in 0.5 g/L pillbug water extract (77.8% of the osteopontin group) and 1.0 g/L ethyl acetate extract (74.1% of the osteopontin group). 0.5 g/L pillbug water extract significantly downregulated ILK level induced by osteopontin, which was 81.4% of the osteopontin group, while pillbug ethyl acetate extract exposed no influence on ILK expression. These results reveal that pillbug water and ethyl acetate extracts both are able to inhibit osteopontin-induced VSMC proliferation, and the role of the pillbug water extract maybe associated with ILK protein pathway.

Objective To explore the value of dual energy CT imaging in the diagnosis of gouty tophus deposition.Methods Sixty-five cases of patients with gout treated in this hospital between December 2012 to December 2015 were selected.Another 65 cases of patients(rheumatoid arthritis,degenerative osteoarthritis and unexplained leg pain) were selected as control group.The re sults of dual energy CT imaging between two group were compared.Three experienced clinicians who didn't understand the imaging results were selected.They had a medical history and physical examination in patients with gout.The differences between clinical evaluation and imaging results were compared.Results Dual energy CT imaging showed that there were no tophi deposition in con trol group;the topi deposition in wrist joint,knee joint and ankle joint were 6.06％oo (33/34),100.00％ (22/22) and 96.49％ (55/57) in gout group.Dual energy CT imaging detected a total of 545 parts of tophus deposition.It was 4.36 times of the result of clinical evaluation(125 parts).Conclusion Dual energy CT imaging shows the advantages of simple noninvasive,high accuracy and low rate of misdiagnosis in diagnosing gout nodules.

Objective To observe the effects ofXiangsha Liujunzi Decoction (XSLJZ) on levels and mRNA of IL-6, IL-10 and protein expression of HSP70 of gastric mucosa in chronic atrophic gastritis (CAG) rats with spleen-stomach deficiency; To discuss its mechanism.Methods Rats were divided into 2 groups through random number table: normal group and model group. The model of CAG rat with spleen-stomach deficiency type was induced by synthetic methods. After successful modeling, rats were divided into model group, positive control group, XSLJZ high-, medium-, low-dose group. Rats in normal and model group received distilled water 10 mL/(kg?d) for gavage; XSLJZ high-, medium-, low-dose group received XSLJZ 24, 12, 6 g/(kg?d), respectively; positive control group received mycin 0.30 g/(kg?d) for gavage for 120 consecutive days. Generally living conditions, levels and mRNA of IL-6, IL-10 and protein expression of HSP70 in gastric mucosa tissue were detected by protein immunoblotting.Results Compared with the normal group, generally living conditions of rats in the model group were poor; mRNA and the content of IL-6 increased significantly, and mRNA and the content of IL-10 decreased significantly (P<0.05,P<0.01); mRNA and protein expression of HSP70 in gastric tissues was much lower than that of normal group (P<0.05,P<0.01). Compared with model group, generally living conditions of rats in the XSLJZ high-dose groups were improved significantly; mRNA and the content of IL-6 decreased significantly, and mRNA and the content of IL-10 increased significantly (P<0.05); XSLJZ high-, medium-dose groups mRNA and protein expression of HSP70 in gastric tissues increased significantly(P<0.05).Conclusion XSLJZ has protective effects on gastric mucosa of CAG rats with spleen-stomach deficiency.

Aim To observe the differences of tonify-ing blood effect for the combination of Danggui and Honghua ( GH) with different proportions on blood de-ficiency mice, and choose the proportion of GH which has the optimal tonifying blood effect. Methods The blood deficient mice model was induced by injection of phenylhydrazine and cyclophosphamide. On the basis of multi-attribute comprehensive index method, the op-timal dose was determined through three doses of GH 1: 1 , which was the highest frequency proportion of GH used in the “Dictionary of Chinese Medicine Prescrip-tions”. According to the optimal dose, the change reg-ulation of tonifying blood effect of GH with different proportions was observed. Results Among three do-
ses (1, 3 and 5 times of clinical dose), the tonifying blood effect of GH was best when the dosing concentra-tion of GH was clinical dose. Among nine proportions (1 : 0, 4 : 1, 2 : 1, 3 : 2, 1 : 1, 2 : 3, 1 : 2, 1 : 4 and 0 : 1), GH 1 : 1 had the best effect. Conclusion The results are consistent with regulations of medi-cine usage that GH 1 : 1 has the highest frequency used in Herbal Formulae, which could provide scientif-ic basis for more effective application of Danggui and Honghua in modern clinic medicine.

AIM:To explore the intracellular signaling transduction pathways in mouse neurons stimulated by octapeptide cholecystokinin(CCK8)on the base of phosphoproteome analysis and to evaluate the types of receptor by the use of CCK8 receptor specific antagonists.METHODS:Cultured neurons were divided into CCK stimulation group,CCK receptor antagonist group and control group at random.All of them were labeled with [32P] orthophosphate(2.03?108 Bq/L)for 2 h in 5% CO2 at 37 ℃.Control and stimulation groups were stimulated with DMEM and CCK8(10-7 mol/L)respectively for 60 min.The neurons in the antagonist groups were pre-incubated for 10 min with different type of receptor antagonists(10-8 mol/L),and then stimulated with CCK8(10-7 mol/L)for 60 min.Reactions were terminated by freezing neurons in liquid nitrogen,and then neuron proteins were collected.The [32P]-labeled lysates isoelectrically focused on IPG Drystrip pH4-7 linear gels and subsequently separated by second-dimensional SDS-PAGE.The 2-DE patterns of phosphoproteins were imaged with autoradiography for 7 days at-70 ℃.Then the phosphoprotein spots were inquired in Swiss-Prot and PPDB protein databases,and analyzed by PDQuest 2D software.RESULTS:Autoradiograph of the 2-DE maps showed that phosphorylation modifications of 46 proteins were involved in CCK signaling transduction in neuron,including protein kinase such as MK07,PI3K,PKC(?,?,?),PKA?,PKG?,AKT3 etc.Intracellular signal molecular such as RB17,GBO?,transcription factor,receptor for growth factor and nuclear protein were also observed.When the neurons were treated with L364 718,the phosphorylation of PKC? and P55G declined obviously,while the phosphorylation of PKC?,PKG?,OGFR and EGFR reduced when the CCK_B specific antagonist L365 260 was used.CONCLUSION:Both CCKA and CCK_B lead to the signaling transduction of CCK8 in neuron,but CCK_B probably play a more important role.The results suggest that cAMP-PKA,MAPK,JNK,PI3K-PKB and cGMP-PKG pathways are involved in CCK_B mediated signaling transduction.

The combination of Danggui and Honghua (GH) is a popular herb pair commonly used in clinic for the treatment of blood stasis syndrome in China. To evaluate the activating blood circulation and dissipating blood stasis effects of the combination of different proportions of Danggui and Honghua on acute blood stasis rats, and optimize the proportion of GH to have the best activating blood circulation and dissipating blood stasis effect. Acute blood stasis rat model was induced by subcutaneous injection of adrenaline and ice water bath. The blood stasis rats were administrated intragastrically with GH (1 : 0, 4 : 1, 2 : 1, 3 : 2, 1 : 1, 2 : 3, 1: 2, 1 : 4 and 0 : 1) extracts. The whole blood viscosity (WBV), plasma viscosity (PV), and high shear whole blood relative index (HSWBRI), low shear whole blood relative index (LSWBRI), and erythrocyte aggregation index (EAI) were tested to observe the effects of GH on hemorheology of blood stasis rats. And the maximum aggregation induced by adenosine diphosphate (ADP) was tested to observe the effect of GH on platelet aggregation index of blood stasis rats. In addition, the prothrombin time (PT), thrombin time (TT), activated partial thromboplastin time (APTT) and plasma fibrinogen (FIB) were tested to observe the effects of GH on blood coagulation function of blood stasis rats. Then principal component analysis and multi-attribute comprehensive index methods were both used to comprehensively evaluate the total activating blood circulation and dissipating blood stasis effects of GH. The results showed that the hemorheological indexes and coagulation parameters of model group both had significant differences with normal group. Compared with model group, GH (1 : 0, 4 : 1, 2: 1, 3 : 2, 1 : 1, 2 : 3, 1 : 2, 1 : 4 and 0 : 1) could improve all the blood hemorheology indexes and regulate part indexes of blood coagulation function and platelet aggregation in acute blood stasis rats. Based on principal component analysis and multi-attribute comprehensive index methods, GH 1 : 1 and GH 3 : 2 both had the best effect of blood circulation and dissipating blood stasis, and the effect of GH 1 : 1 was slightly better than GH 3 : 2. These results suggest that GH could obviously ameliorate the abnormality of hemorheology and blood coagulation function in acute blood stasis rats. The optimized proportion of GH was consistent with regulations of medicine usage that GH 1 : 1 had the highest frequency used in traditional Chinese formulae. It could provide scientific basis for more effective application of the compatibility between Danggui and Honghua in modern clinic medicine.

Objective To investigate the change of plasma vitamin D, A and E levels in infant with infantile hepatitis syndrome (IHS) and to provide reliable basis for clinical treatment. Methods Seventy-three infant with IHS (IHS group) from January 2015 to May 2016 and 82 cases of physical examination infant (control group) were enrolled in this study.The levels of plasma vitamin D, A and E were tested and compared between two groups. Results The levels of vitamin D, A and E in IHS group were significantly lower than those in control group:(37.871 ± 20.111) nmol/L vs. (97.708 ± 28.827) nmol/L, (1.082 ± 0.657)μmol/L vs. (1.810 ± 0.517)μmol/L, (21.252 ± 7.596)μmol/L vs. (26.647 ± 6.495)μmol/L, P<0.01 or <0.05. The nutritional status of vitamin D, A and E in two groups had significant differences (P<0.01). Conclusions There have obvious deficiency in fat-soluble vitamin D, A and E in infant with HIS , and vitamin D deficiency is the most obvious. The levels of vitamin D, A and E are the dynamic monitoring indicators of treatment, nutritional status evaluation and long-term complications development.

Objective To observe effects of Kangshuai Yizhi Capsule on ATP in brain tissue and IL-6, TNF-α in serum of aging model rats, and explore the protective effects of the capsule on brain tissue.Methods Totally 72 rats were randomly divided into a normal group and a model group. The subacutely aging model rats were made by injectingD-gal, then aging rats were numbered and grouped by random number table into the model group, Kangshuai Yizhi high-dose group, Kangshuai Yizhi Capsule low-dose group and Naofukang group. All dose groups were received gavage by giving corresponding doses, while normal group and model group were given the same amount of saline everyday. After treated for 60 days, the activity of Na+-K+-ATP and Ca2+-Mg2+-ATP in brain tissue, and IL-6, TNF-α in serum were detected.Results Compared with normal group, Na+-K+-ATP and Ca2+-Mg2+-ATP were less active (P<0.05), but levels of IL-6 and TNF-α in model group were significantly higher, with statistical significance (P<0.05). Compared with model group, after treated with Kangshuai Yizhi Capsule, Na+-K+-ATP and Ca2+-Mg2+-ATP were more active, and IL-6 and TNF-α levels were down-regulated significantly in dose groups, with statistical significance (P<0.05). Meanwhile, Kangshuai Yizhi Capsule high-dose group showed the most obvious effect among dose groups.ConclusionKangshuai Yizhi Capsule has effects of enhancing activity of ATP in brain tissue and reducing level of proinflammatory factors.

Objective To investigate the levels of serum AFP and CEA in healthy physical examination people and their sexual difference.Methods (1 )Healthy physical examination people were divided into the group A(510 males)and group B(477 fe-males);(2)these two groups were subdivided into the group A1 (<40 years old),A2 (40 - <50 years old),A3 (50 - <60 years old),A4(60-<70 years old)and A5 (70 -87 years old),and group B1 (<40 years old),B2(40 -<50 years old),B3 (50 -<60 years old),B4(60-<70 years old)and B5(70-87 years old).The serum AFP and CEA in 987 healthy physical examination peo-ple were measured by the electrochemiluminesence immunoassay,and then the detection results were compared among various groups.Results (1)The detection results of serum AFP and CEA had statistically significant differences between the group A and B (P <0.01);(2)there was no statistical differences among the group A and among the group B.Conclusion The serum AFP and CEA levels in healthy physical examination people in the local area have difference between males and females,which in males are significantly higher than those in females,indicating that the reference range of AFP and CEA should be repectively established ac-cording to different sexes;while the serum AFP and CEA levels have no differences among various age groups in males and fe-males.

Objective: To explore the clinical value of plasma copeptin level on major adverse cardiovascular event (MACE) occurrence in patients with acute ST-segment elevation myocardial infarction (STEMI) during hospitalization.
Methods: Our research included 2 groups:STEMI group, n=80 and Control group, n=80 patients with stable coronary artery disease (CAD). All patients were treated in our hospital from 2012-06 to 2014-06. Plasma level of copeptin was detected by ELISA, other relevant examinations were conducted to study the MACE occurrence in STEMI patients.
Results: Plasma copeptin level in STEMI group (523.26 ± 142.69) pg/ml was higher than that in Control group (345.25 ± 89.36) pg/ml, P<0.05. In STEMI group, there were 28/80 (35%) patients suffered from MACE, compared with non-MACE patients, they had increased plasma copeptin, cardiac muscle protein I (cTnI), kinase isoenzyme (CK-MB) and left ventricular ejection fraction (LVEF), P<0.05. Multivariate regression analysis indicated that plasma copeptin, cTnI and LVEF were the independent risk factors for MACE occurrence. According to occurred area under the curve, compared with cTnI and CK-MB, plasma copeptin level had the higher predictive value to judge the ROC, positive/negative possibility, sensitivity and speciifcity for MACE occurrence in STEAMI patients, P<0.05.
Conclusion: Plasma copeptin level could effectively predict MACE occurrence in patients with acute STEMI during
hospitalization, it may predict their prognosis at certain point.