The calcium antagonist action of arctigenin (ACT) was studied in order to verify the ef-fect of Fructus Arctii for the relieve of exterior syndrome. Muscular contraction of isolated rat trachea,colon, pulmonary artery and thoracic aorta induced by KC1, that of guinea pig trachea induced by CaCI2,before and after the addition of ACT were assessed and their contraction-response curves drawn and PD'2calculated according to Scott. The inhibition rate of two-phase contraction of guinea pig trachea induced byacetylcholine chloride (Ach) in comparison with verapamil (VER) was calculated. Results of the studyshowed that ACT could non-compatitively antagonize the muscular contraction of the test specimens withPD'2 of 4.01, 5.11, 5.98 and 6.05 respectively. Similar to VER, ACT could non-competitively antagonizethe isolated guinea pig trachea with PD'2 of 4.04 and 5.62 respectively. Both of them could inhibit thefirst phase contraction induced by Ach with inhibition rates of 66.14% and 81.42% respectively. It wasconcluded that ACT, as the active constituent of Fructus Arctii, relaxed smooth muscle contraction byblocking the potential dependant Ca2+ channel and the internal release of Ca2+.

Objective To explore the vitamin D nutritional status of urban residents in Jinan,in order to provide the basis for the prevention and treatment of chronic diseases associated with vitamin D insufficiency.Methods 1478 cases aged 30-90 years,including 602 men (59.8± 13.0) years on average and 876 women (57.4±12.9) years on average were selected by a stratified random sampling from 3 community medical care institutions in Jinan.Subjects were divided into six groups according to the 10-year interval.Serum 25-hydroxyvitamin D [25(OH) D] level was measured by enzyme-linked immunosorbent assay (ELISA).Serum [25(OH) D] level ≥ 75 nmol/L was defined as vitamin D sufficient,(50.0 ～ 74.9) nmol/L as vitamin D critical value,25.0 ～ 49.9 nmol/L as vitamin D insufficiency,＜ 25 nmol/L as vitamin D deficiency.All data processing and statistical analysis were finished by SPSS 13.0.Results The average level of [25 (OH) D] was 58.60 nmol/L in males [95％ CI:56.7-60.5 nmol/L] and 54.17 nmol/L in females [95％ CI:52.8-55.8 nmol/L],which were less than 75 nmol/L.The average level of [[25(OH) D] was 49.1nmol/L in females aged 70-79 years (95％CI:45.7～52.5 nmol/L),41.7 nmol/L in females aged 80-90 years (95％CI:38.2 ～46.7 nmol/L),which were less than 50 nmol/L.The percentage of cases with vitamin D deficiency,insufficiency,critical value,and sufficiency was 5.6％,33.4％,36.5％,24.4％ in males,and 6.5％,42.9％,32.6％,18.1％ in female respectively.The incidence of vitamin D deficiency and insufficiency was increased with ageing (male:x2 =33.68 P＜0.01,female:x2 =55.7,P＜0.001).The percentage of vitamin D deficiency and insufficiency was more in females than in males (60-69 years old:x2=9.387,P＜0.05,80-90 years old:x2=8.896,P＜0.05).The [25 (OH) D] level was lowest in winter (48.8 nmol/L,95％CI:46.8～50.9 nmol/L) and highest in autumn (60.2 nmol/L,95％ CI:58.0～62.3 nmol/L).The incidence of vitamin D deficiency and insufficiency was highest in winter (x2 =18.364,P＜0.05).Conclusions Vitamin D deficiency and insufficiency are widely prevalent in urban residents,which is more severe in the elderly and females.Serum [25(OH) D] level is related to season,and the vitamin D deficiency and insufficiency are severe in winter.

In order to research the biologic activity of osteoblast-stimulating factor 1 (OSF-1), the pPIC9K/osf-1 yeast expression vector was constructed to express and purify OSF-1. Firstly, the osf-1 gene sequence was obtained by artificial synthesis and cloned into Pichia pastoris expression vector pPIC9K to generate pPIC9K/osf-1. The recombinant plasmid was linearized by Sac I and transformed into P. pastoris GS115 by electroporation. Recombinant P. pastoris GS115/ pPIC9K/osf-1 was screened by MD and G418-YPD plates and further identified by PCR. The positive P. pastoris was induced with 1% methanol at 25 degrees C for 96 h. The target protein was analyzed by SDS-PAGE showing a special band about 18 kDa. The target protein was successfully purified from the supernatant of the broth using ion exchange chromatography of SP-Sephadex C-50. The purity of target protein was above 98%. Western blotting appeared a good antigenicity of the purified protein. Bioassay results show that the recombinant protein OSF-1 can promote the differentiation and proliferation of osteoblasts MC3T3-E1. We successfully expressed OSF-1 by recombinant P. pastoris for further development of anti-osteoporosis of research and industrial production of OSF-1.
Blotting, Western ; Carrier Proteins ; biosynthesis ; Chromatography, Affinity ; Cytokines ; biosynthesis ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Pichia ; metabolism ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis

(1.Department of Anesthesiology, Jinling Hospital, School of Medicine, Nanjing University, Nanjing 210002, Jiang-su, China;2.Department of Anesthesiology, General Hospital of Shenyang Military Region, PLA, Shenyang 110000, China)
le-blind, 60 patients scheduled for OPCABG were randomly divided into 4 groups: control group( group C) and 3 dexmedetomidine group(group D1、D2、D2).Patients in each group received saline or dexmedetomidine 0.2μg/kg, 0.4μg/kg, 0.6μg/kg respectively. Hemodynamic variables such as HR、MAP、CVP、MPAP、PCWP、SVRI and PVRI were recorded after entering into the operationroom ( T0 ) , before the anesthesia induction( T1 ) , at 5 minutes after tracheal intubation ( T2 ) , during the sternotomy( T3 ) , at the end of an-astomosis of left anterior descending(T4), at the end of anastomosis of right coronary artery or 1eft circum flex coronary artery(T5) and after operation ( T6 ) .Body tempreture and urine volume were recorded during the operation.Total quantity of analgesics consumption and heart adverse reaction( bradyarrhythmia, tachycardia, hypotension, hypertension etc) were also recorded. Results At T2 -T6 , there was a significant decline in HR, MAP, SVRI, PVRI and CI in group D2 and group D3 compared with group C (P<0.05).SVRI and PVRI in group C and group D2 significantly increased at T3 -T6 compared with those at at T0(P<0.05).At T2 and T6, HR and MAP in group group D2 signicantly increased compared with group D3([57 ±6]times/min vs [52 ±4] times/min, [72 ±7]mmHg vs [63 ±5]mmHg;[72 ±5]times/min vs [55 ±6]times/min, [68 ±5]mmHg vs [63 ±5]mmHg)].At T4 -T6, significant difference was found in CI, SVRI and PVRI between group D2and group D3(P<0.05).In comparison to group C, hypertension, tachycardia in-cidence, noradrenaline amount and isosorbide mononitrate amount declined significant in group D2and group D3(P<0.05);hypoten-sion and bradycardia incidence increased significantly while isosorbide mononitrate amount reduced significantly(P <0.05). Conclusion In OPCABG, dexmedetomidine infused at 0.4μg/(kg· h) after a loading dose of 0.5μg/(kg· 10 min) is useful in maintaining stable hemodynamics which contributes to oxygen balance and reduces myocardium injury in patients.

Objective To analyse the influencing factors of quality of life (QOL)among postoperative children with congenital anorectal malformation (ARM), so as to provide evidence for developing comprehensive interventions and improve QOL of children with ARM.Methods The QOL of 100 children aged at 2 and 5 years old were evaluated by using the Chinese version of PedsQLTM4.0 scale, and the total and 4 dimensions scores of QOL were calculated.Univariate and multiple stepwise regression model was applied to analyse the influencing factors of the QOL.Results The results of univariate analysis among ARM children aged at 2 years old showed that the clinical classification, bowel disorders and family residence have different influence on QOL scores of the 4 dimensions and total scale (P ＜ 0.05).The results of univariate analysis among ARM children aged at 5 years old showed that the clinical classification, bowel disorders and maternal education had different influence on QOL scores of the 4 dimensions and total scale (P ＜ 0.05).The results of multiple stepwise regression model among ARM children aged at 2 years old showed that the major influence factor of QOL scores in physical function was family residence and bowel disorders(P ＜ 0.05), the major influence factor of QOL scores in social functioning was bowel disorders(P ＜ 0.05), the major influence factor of the total scores was bowel disorders and family residence(P ＜0.05).The results of multiple stepwise regression model among ARM children aged at 5 years old showed that the major influence factor of QOL scores in physical function was maternal occupation and clinical classification(P ＜ 0.05), the major influence factor of QOL scores in emotional functioning was bowel disorders(P ＜ 0.05) ,the major influence factor of QOL scores in social functioning was clinical classification and bowel disorders (P ＜ 0.05), the major influence factor of QOL scores in school performance was maternal occupation(P ＜ 0.05), the major influence factor of total scores was bowel disorders and clinical classification(P ＜ 0.05).Conclusion Family residence and bowel disorders have effct on QOL of ARM children aged at 2 years old.Maternal occupation, clinical classification and bowel disorders have effct on QOL of ARM children aged at 5 years old.We should take positive interventions on these influencing factors to improve QOL of children with ARM.

OBJECTIVE:To investigate the effects of neoadjunctive chemotherapy(NAC)of docetaxel and epirubicin com-bined with cyclophosphamide on clinical efficacy and tumor markers of breast cancer patients with different molecular types. METH-ODS:A total of 88 female patients with locally advanced breast cancer collected from our hospital during Jan. 2014-Jan. 2016 were divided into Luminal A type(23 cases),Luminal B type(21 cases),basal-like type(11 cases),HER2-over expressing type(18 cases)and normal breast-like type(15 cases)according to molecular type. All patients were given Docetaxel injection+Epirubicin hydrochloride injection+Cyclophosphamide for injection for consecutive 6 cycles(21 d as a cycle). Total response rates and patho-logical complete remission(pCR)rates were compared among breast cancer patients with different molecular types. The expression of serum tumor markers [CEA,CA125,CA153] were compared before and after treatment,and the occurrence of ADR was record-ed. RESULTS:Total response rate of 88 patients was 63.64%,among which that of basal-like breast cancer patients was 72.73%, significantly higher than other molecular types,with statistical significance(P<0.05). There was no statistical significance in total response rates of pairwise molecular type comparison(P>0.05). The pCR rate of 88 patients was 27.27%,and that of basal-like breast cancer patients was the highest(45.45%). There was statistical significance in pCR rates of pairwise molecular type compari-son(P<0.05),except there was no significant difference in pCR rate between HER2-over expressing type and normal breast-like type(P>0.05). Before treatment,there was no statistical significance in the expression of CEA,CA125 and CA153 in breast can-cer patients with different molecular types(P>0.05). After treatment,the expression of CEA,CA125 and CA153 in different mo-lecular types were decreased significantly,with statistical significance(P<0.05). There was no statistical significance in the expres-sion of above markers among different molecular types(P>0.05). There was no statistical significance in the incidence of ADR among different molecular types(P>0.05). CONCLUSIONS:NAC plan of docetaxel and epirubicin combined with cyclophospha-mide can reduce the expression of tumor markers and shows certain therapeutic efficacy for breast cancer patients with different mo-lecular types. Total response rate and pCR rate of basal-like type are better than those of other molecular types,so NAC plan is the preferred treatment for basal-like type breast cancer.

AIM: To study the mechanism of diabetic cardiomyopathy and abnormality of oxygen free radicals. METHODS: The contents of myocardial cytosolic cytochrome C, mitochondria cytochrome C, mitochondrial calcium, NO, MDA and the activity of SOD and NOS were determined in diabetic rats induced by STZ. The pathological changes were observed under transmission electron microscope. RESULTS: Compared to the normal and ganoderma group, the levels of mitochondrial NO, iNOS, MDA, calcium and plasma Cyt-C in rat myocardium were higher (P<0.05), while mitochondrial Cyt-C and SOD were lowered in model group (P<0.05). The bouncary indistinct, disorganization, a focal loss of muscular fibril, myocardium mitochondria swelling, pulmonary vascular endothelial cellular swelling and obstructed lumen of the capillary were also observed under transmission electronic microscope. CONCLUSION: The findings indicate that oxyradical and lipid peroxidation might be associated with the damage of myocardial mitochondria in NIDDM rats. Cyt-C and mitochondrial calcium is also involved in the process.

Objective To systemically evaluate the efficacy of all kinds of therapies of cerebral venous sinus thrombosis (CVST) associated with intracranial hematoma (ICH) and provide reference for making treatment scheme.Methods We searched English databases (Pubmed,ISI Web of science and Cochrane library databases) for publications on CVST associated with ICH and the relevant references of those articles.Retrieval time deadline was up to January 2015,and the language was set to English.All publications were searched without restrictions of publication type or published fields.Documents were screened to extract data and to analyze systemically.Results In total,17 articles fulfilled the inclusion criteria and included 31 patients.Because of the severities of reported cases were greatly different and treatments were varied,these patients were divided into 3 levels to get a better understanding of the efficacy.In 6 patients of stage Ⅰ,3 accepted anticoagulation,1 operative treatment,1 endovascular intervention and the left one used more than one methods;the prognosis was good and modified Rankin scale (mRS) score was 0 no matter which kind of therapy was performed.In 11 patients of stage Ⅱ,2 accepted anticoagulation enjoyed mRS scores of 0 and 6,3 operative treatment enjoyed mRS scores of 0,1 and 6,1 endovascular intervention enjoyed mRS scores of 0,and the left 5 used more than one method enjoyed mRS scores of 0,1,1,2 and 2.In 8 patients of stage Ⅲ,1 accepted anticoagulation enjoyed mRS scores of 2,3 operative treatment enjoyed mRS scores of 1,2 and 4,and 4 endovascular intervention enjoyed mRS scores of 0,1,1 and 1.Conclusion It seems that anticoagulation therapy alone may not be effectively enough for those patients that with severe clinical or imaging features;individualized therapy strategies with different types would be more effective.

Objective To evaluate the probability of siblings identification in Identifiler system by using the software of automatic analysis.Methods Using the software of automatic analysis in siblings jdentification.STP genetic typing of 151 pairs of full siblings and 31224 pairs of unrelated individuals from manual simulation were analyzed in 15 STR loci of ldentifiler system.Results Kin probability(W_(FS))of 39.07% full siblings were more than 99.999% while W_(FS) of unrelated individual pairs were 0% .W_(FS) of 60.93% full siblings and 21.3% unrelated individual pairs were all at the range from 99.999% to 1% .W_(FS) of 78.7% unrelated individual pairs 0% full siblings individuals were less than 1% .Therefore,there were notability difference between full siblings and unrelated individual pairs.In addition,testing of 15 STR loci of Identifiler system,it suggested that the pair were siblings when the locus number of the entirely-same is not less than 5 or that of the entirely-different is not more than 1,and that the pair were unrelated individuals when the locus number of the entirely-different is not less than 6 or that of the entirely-8alne is not more than 1.Conclusion The software of automatic analysis and the Identifiler system call be used to siblings identification.

ObjectiveTo investigate the effects of dopamine receptor agonist pramipexole and levodopa on emotion and cognition, and mitochondrial membrane potential of rats after global cerebral ischemia-reperfusion injury. MethodsA total of 80 male Sprague-Dawley rats were divided into sham group (n = 20), model group (n = 20), pramipexole group (n = 20) and combined group (n = 20). The latter three groups were used to prepare the model of global cerebral ischemia-reperfusion injury with Pulsinelli's four-vessel occlusion. The pramipexole group was intraperitoneally injected pramipexole 0.5 mg/kg once a day, while the combined group was injected levodopa 50 mg/kg and pramipexole 0.5 mg/kg, for 14 days. Five rats in each group were tested with open field test three, seven and 14 days after modeling; five were tested with Y-maze test seven and 14 days after modeling; five were detected mitochondrial membrane potential three, seven and 14 days after modeling; and five were observed under Nissl's staining14 days after modeling. ResultsCompared with the model group, the number of entries into the central zone (P < 0.05), total distance travelled (P < 0.05) and average velocity (P < 0.05) in the open field test increased in the pramipexole and combined groups seven and 14 days after modeling, duration spent in the central zone increased in the pramipexole and combined groups seven days after modeling (P < 0.05); the rate of spontaneous alternation of Y-maze test increased in the pramipexole and combined groups 14 days after modeling (P < 0.05); mitochondrial membrane potential in hippocampus increased in the pramipexole and combined groups seven and 14 days after modeling (P < 0.05), and it was less in the pramipexole group than in the combined group 14 days after modeling (P < 0.05); and the number of surviving neurons in the hippocampal CA1 increased in the pramipexole and combined groups 14 days after modeling (P < 0.05). ConclusionPramipexole may improve emotion and cognition of rats after global cerebral ischemia-reperfusion injury, and it may be helpful for restoring mitochondrial membrane potential as combining with levodopa.