OBJECTIVE:To optimize the extraction of effective ingredients and purification for crude glycyrrhizic acid and liquiritin from Radix et Rhizoma Glycyrrhizae with different methods. METHODS:The contents and extraction percentages of glycyrrhizic acid and liquiritin were used as indexes to optimize the extraction solvent. Then the extraction process was optimized with orthogonal experiment taking the concentration of ammonium ethanol,the amount of solvent and the extracting times as indexes. Crude glycyrrhizic acid was purified with the method of recrystallization and crude liquiritin was purified with organic solvent method.RESULTS:The optimal extracting solvent was ammonium ethanol. The optimum extraction conditions were as follows:extracting the solvent 4 times(2 h/time) by adding 5 times volume of 60% ethanol(containing 0.3% ammonia water). The purity of glycyrrhizic acid was over 85% with total extraction rate at 43%,and content of liquiritin was above 15% with its total extraction rate at 67%. CONCLUSION:The method is simple and feasible and from which glycyrrhizic acid and liquiritin was been obtained,Radix et Rhizoma Glycyrrhizae can broaden the scope of the application.

OBJECTIVE：Probe into the feasibility of preparation of diclofenac sodium with β-cyclodextrins in 1∶1 and 2∶1 inclusion compounds ,and find out the affection in solubility of diclofenac sodium. METHODS:The 1∶1 and 2∶1 melusion compounds were prepared by liquid-phase an alysis the content and determination them solubility.RESULT:The 1 ∶1 and 2∶1 inclusion compounds were prepared by different ratio of reactants and etermined them mp were 299～300.5℃ and 302～303℃ respectively. CONCLUSION：Of us is that solubilits of the 1∶1 and 2∶1 inclusion compounds increase 0.65 and 1 times respetively comparing with declofenac sodium.

Objective To evaluate the clinical value of multiplex ligation-dependent probe amplification (MLPA) technique used in karyotype analysis of chorionic villi from missed abortion. Methods Feb 2008 to Oct 2008, 91 patients with missed abortion diagnosed by hormonal measurement, type B ultrasound and physical exam matched with 20 normal pregnant women undergoing artificial abortion were enrolled in this study. Chorionic villi was obtained by suction dilation and curettage in aseptic condition, then those villi was cultured and analyzed by traditional cytogenetic karyotyping method, in the mean time, the DNA extracted from villi was detected by MLPA. The results of chromosomal G-banding of chorionic villi were compared between two methods. Results The diagnostic concordance of MLPA and traditional karyotyping was observed in 92% (84/91) cases, there were 84 cases in the case group with diagnostic concordance by traditional karyotyping and MLPA except 7 cases of euploidy could not be detected by MLPA. The 84 cases included 40 normal karyotype,29 trisomy of euchromosome, 1 double trisomy of euchromosome, 10 monosomy X , 1 monosomy X combined with trisomy of euchromosome, 2 chimaera of X chromosome, 1 structural abnormity of euchromosome. Among 7 cases with discordance diagnosis, 2 cases with trisomy and 5 cases with tetrasomy of euchromosome were identified in traditional karyotyping, however, they were all diagnosed with normal disomy by MLPA. Of 20 villi from normal pregnancy, two methods got the consistent results. Conclusion The MLPA was rapid and efficacy method used for analyzing aneuploids in chorionic villi.

AIM:To observe the effect of Wnt/β-catenin signaling pathway on diabetic ulcer.METHODS:Diabetic animal model was established in the female Wistar rats by intraperitoneal injection of low-dose streptozotocin fol-lowing high-fat diet feeding.A circular wound was made on the dorsum of the rats in both control group and diabetic group. The condition of wound healing was recorded and the structures of the wound tissues were observed by HE staining in the 2 groups at 3, 7 and 14 d after wounding.The expression ofβ-catenin, GSK-3βand Rspo-3 at mRNA and protein levels in the wound tissues was detected by RT-PCR and ELISA.RESULTS:In diabetic group, the wound healing rate was lower (P<0.05), and the inflammatory cells, fibroblast cells and new capillaries in the wound tissues were fewer than those in control group.The expression of β-catenin and Rspo-3 at mRNA and protein levels in the wound tissues in control group was significantly higher than those in diabetic group, and the expression of GSK-3βwas exactly the opposite (P<0.05). CONCLUSION:The down-regulation of Wnt/β-catenin probably resultes from the decreased level of Rspo-3, which may be one of the reasons for delaying the diabetic ulcer healing.

Objective:Discussion MKK4 protein expression in nasopharyngeal carcinoma and -1044 A/T polymorphism correlation.Methods:90 patients with nasopharyngeal carcinoma , MKK4 protein expression was determined by immunohistochemical staining,polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP ) to detect the gene -1044A/T sites monocytes nucleotide polymorphism.Results:MKK4 protein expression in nasopharyngeal carcinoma (-) was 24.4%(22/90),(+) was 15.6%(14/90),(++) was 34.4%(31/90),(+++) was 25.6% (23/90).Low expression (-/+) patients with a total of 36 cases,-1044AA genotype accounted for 22 cases (61.11%),AT genotype accounted for 12 cases (33.33%),TT genotype accounted for two cases (5.56%),AT+TT gene type accounted for 14 cases (38.89%).The patients with high MKK4 expression of 54 cases,of which accounted for 38 cases of AA genotype (70.37%),AT genotype accounted with 15 cases (27.78%),TT genotype accounted for one case (1.85%),AT +TT genotype accounted for 16 cases (29.63%).Low expression and high expression of T gene mutation occurs no significant ( Z=0.323 , P=0.747 ) .Conclusion: MKK4 protein expression correlated with -1044 A/T gene promoter polymorphisms was no significant correlation .

Objective To evaluate the value of DNA quantitative analysis in diagnosis of breast tumor.Methods The data of 128 cases diagnosed with breast disease from Feb.2005 to Jan.2006 in Department of Pathology in Wuhan Center Hospital were collected,among whom 72 cases had benign breast lesion,10 cases had boundary breast lesion and 46 cases had malignant breast disease.Automated DNA imaging cytometry was used to identify DNA indexes of the 128 cases.Results The positive rate was 1.39％ (1/72),30.00％ (3/10) and 78.26％ (36/46) respectively in benign breast lesion,boundary breast lesion and malignant breast disease.The difference had statistical significance (P ＜ 0.05).In invasive ductal breast cancer,the positive rate was 57.14％ (4/7),76.47％ (13/17) and 100％ (18/18) respectively for breast cancer of grade Ⅰ,Ⅱ and Ⅲ.Conclusion Automated DNA imaging cytometry is of some value in analyzing the malignant grade of breast cancer and predicting the prognosis of breast diseases.

Objective:To evaluate the effects of intranasal administration of glial cell line-derived neurotrophic factor (GDNF) on postoperative cognitive dysfunction in aged rats.Methods:Forty healthy Sprague-Dawley rats of both sexes, aged 21-23 months, weighing 480-600 g, were divided into 4 groups ( n=10 each) using a random number table method: sham operation group (group S), operation group (group O), intranasal administration of low-dose GDNF group (group G1) and intranasal administration of high-dose GDNF group (group G2). Rats underwent exploratory laparotomy under anesthesia with chloral hydrate in O, G1 and G2 groups, while the rats in group S only received sham operation.The rats in group G1 and group G2 were intranasally treated with GDNF 25 and 50 μg (in 25 μl of PBS), respectively, and PBS 25 μl was nasally administered in group S and group O every day for 3 consecutive days after operation or sham operation.Morris water maze test was performed on days 3-7 after surgery, and then the rats were sacrificed, and hippocampal tissues were removed for determination of the expression of GDNF, high mobility group box 1 (HMGB1), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), activated caspase-3 and Bax (by Western blot). Results:Compared with group S, the escape latency was significantly prolonged on days 5-7 after operation, the number of crossing the platform was reduced, time spent in the target quadrant was shortened, expression of GDNF was down-regulated, and expression of IL-1β, TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was up-regulated in group O, and the number of crossing the platform was reduced, time spent in the target quadrant was shortened, and expression of IL-1β and TNF-α was up-regulated in G1 and G2 groups ( P<0.05). Compared with group O, the escape latency was significantly shortened on days 5-7 after operation, the number of crossing the platform was increased, time spent in the target quadrant was prolonged, expression of GDNF was up-regulated, expression of TNF-α, HMGB1, activated caspase-3 and Bax in hippocampi was down-regulated in G1 and G2 groups, and IL-1β in hippocampi was down-regulated in group G1 ( P<0.05). Compared with group G1, the expression of TNF-α in hippocampi was down-regulated ( P<0.05), and no significant change was found in the other parameters mentioned above in group G2 ( P>0.05). Conclusions:Intranasal administration of GDNF can improve postoperative cognitive dysfunction, and the mechanism may be related to inhibiting neuroinflammatory responses and neuroapoptosis in aged rats.

Objective: To compare the pharmacodynamic action of Radix Notoginseng after fermented by different strains bioconversion.Methods:The models of cerebral ischemia and hypoxia were established,thrombosis in vivo were induced by collagen-Adr,ventricular fibrillation of mice by chloroform,and perfusion experiment of the ex vivo heart of cavia cobaya was used.Results:100,200mg/kg doses of all samples had protective effects on ischemia and hypoxia mice,1# sample can inhibit thrombosis induced by collagen-adrenaline obviously after intraperitoneal injection in dosage of 200mg?kg-1?d-1 for 5days.All groups significantly reduced the incidents of ventricullar fibrillation induced by chloroform in mice except 100mg/kg dose of 1#,furthermore,compared with control high dose group(5#),the incidents of high dose groups of 3、4# deceased significantly.All samples had the effect of dilating coronary artery,but there effects on myocardial contractile force and cardiac preload were different.Conclusion:The primary cardiovascular related pharmacological researches demonstrated the vary action changes of 1-4# samples compared with the sample 5#.This result has revealed that the chemical structure of Radix Notoginseng transformed and that is coincide with initial analysis of phytochemical analysis.

Objective To investigate the ability of magnetic resonance imaging (MRI) in tracking magnetically labeled mesenchymal stem cells (MR-MSCs) in a swine myocardial infarction (MI) model.Methods Adult Chinese mini-pigs (n = 6) were subjected to open-chest experimental MI operation.Their antegeneic bone marrow-derived mesenchymal stem cells ( MSCs ) was cultured and doubly labeled with ferumoxides and DAPL On the 14 th day after MSCs transplantation, the size and location of the myocardial infarction were assessed by using delayed-enhancement MRI (DE-MRI). Then the labeled MSCs were injected intramyocardially into peri-infarct zone and normal myocardium. At 24 hrs and 3 weeks after injection, the contrast and the volume of the MR-MSCs hypointense lesion from the MR images were acquired, and the contrast was determined using the difference in signal intensity between the hypointense and normal myocardium divided by signal intensity of the normal region.After humane euthanasia, the heart was excised and histology corresponding to MRI slices that demonstrated MR-MSCs lesions was performed.Repeated-measures ANOVA and a paired t test were used for comparison of the contrast and the volume of the MR-MSCs hypointense lesion at different time points. Comparisons between independent groups were performed with the standard Student t test.Results The labeling efficiency of ferumoxides and DAPI was 100% . On the 14 th day after the MI operation, the average percentage of infracted myocardial area was (33.6±8.9)% .Twenty- four hours after MSCs transplantation, MSCs injection sites appeared as ovoid hypointensive lesions with sharp border on T2 * images. At 24 h after injection, the signal contrast [(67.00±5.48)% vs (61.92 ±7.76)%,t = 1.65,P =0.1158] and the size [(0.56 ±0.24) cm2 vs (0.52 ± 0.25 ) cm2, t = 0.39, P = 0.7044 ] of the lesions showed no statistical difference between the peri infarct zone and the normal myocardium.At 3 weeks after injection, the signal contrast decreased and the size diminished both in the peri-infarct zone and in the normal myocardium. Moreover, the contrast of the lesions in peri-infarct zone decreased more significantly than that in normal myocardium [(26.88 +7.27)%vs (15.00 :t:4.51)%, F =20.08, P =0.0003].Post mortem analysis found the fluorescontly labeled MSCs demonstrated on histological sections.There were much more dense fluorescently labled MSCs per high power fields at injection sites of normal myocardium than at injection sites of peri-infarct zone [ (106 ±25 )/HPF vs ( 143 ± 31 )/HPF, t = - 2.47, P = 0.0293 ].In MSCs injection sites of the peri-infarct zone,the capillary density was significantly higher than that in control sites [ (13.4 ± 4.0 )/HPF vs (9.4 ±3.1 )/HPF, t = 2.49, P = 0.0229].At 3 weeks after injection, ferumoxide was contained within partial original MSCs.Conclusion Magnetic resonance imaging of MSCs is a feasible method for the in vivo tracking of transplanted stem cells and could reflect the tendency of the local stem cell quantity, but there still has limitation for the semi-quantitation of the transplanted stem cells.

Objective To investigate the present status and influencing factors of psychological distress among psychiatric nurses, and provide theoretical basis for intervention measures. Methods Totally, 812 psychiatric nurses in six psychiatric hospitals of Shandong province were selected to complete the Kessler Psychological Distress Scale (K10), Perceived Organizational Support(POS), Simplified Coping Style Questionnaire(SCSQ) and Emotion regulation questionnaire(ERQ). Results The mean scores of psychological distress among psychiatric nurse was (21.29±8.11) points,70.3%of psychiatric nurses had psychological distress. There were significant differences in psychological distress among psychiatric nurses with different age (F=6.886,P=0.001), working years (F=4.403,P=0.004), professional title (F=6.597,P=0.000) and experience of nurse-patient conflict (F=7.228,P=0.000).Psychological distress was negatively related to cognitive reappraisal(r =- 0.093), positive coping style (r =- 0.163) and perceived organizational support(r =- 0.379), while psychological distress was positively related to expressive suppression(r=0.160) and negative coping style(r=0.258)(all P<0.01). Regression analysis revealed that the factors impacting psychological distress were positive coping style, perceived organizational support, expressive suppression, negative coping style and nurse-patient conflict (R2=0.224,F=46.611,P=0.000). Conclusions Psychiatric nursing managers should pay attention to the cultivation of emotional management ability of nurses and the ability to deal with conflict,encouraging them apply positive thinking cognition and positive coping style to deal with negative events in the work,creating a supportive work environment in order to improve the level of mental health.