A rapid detection method of active pharmaceutical ingredients( API) in weak API signal drugs by surface enhanced Raman scattering ( SERS) technology combined with paper substrate was established in this work. By soaking the filter paper in silver nanoparticles solution ( Ag NPs) to synthesize Ag NPs-paper as the substrate, and then the sample solution was dropping on the substrate with SERS detection. On the basis of strengthen ability of Ag NPs-paper, result of SERS detection and optimal preparation conditions, the fast identification method of weak API signal drugs was established. In this case, the SERS spectra of weak API signal drugs and their standards SERS spectra were obtained, where the correlation coefficient of weak API signal drug SERS spectra and its standard was more than 0. 9. The result showed that by this method, the low content API in weak API signal drugs could be well investigated, and the deficiencies of the normal Raman spectroscopy efficiently was also overcome. In conclusion, the synthesize method of Ag NPs-paper was simple, and the strengthen effect of this Ag NPs-paper on the intensity was obviously observed. Paper substrate-SERS method was simple, rapid and sensitive, and could be used to detect weak API signal drugs, presenting broad application prospects in the rapid detection of weak API signal drugs.

OBJECTIVE: To investigate the characteristics of syndromes of phlegm and blood stasis in patients with coronary heart disease by multiple statistical methods of matching matrix, factor analysis and clustering analysis, and to provide some references for classification and normalization of diagnosis of syndromes of phlegm and blood stasis of coronary heart disease. METHODS: The correlations among 46 kinds of symptoms in syndrome of non-phlegm and non-blood stasis, syndrome of blood stasis, syndrome of phlegm and syndrome of phlegm-blood stasis blocking in 200 patients with coronary heart disease were analyzed by matching matrix, factor analysis and clustering analysis. RESULTS: The manifestations of tongue and pulse in syndromes of phlegm and blood stasis were significantly different from those in syndrome of non-phlegm and non-blood stasis. The pathogenesis of viscera in syndromes of phlegm and blood stasis lied in the heart and kidney, and the syndrome of deficiency of heart qi was the most common one while the syndrome of deficiency of kidney qi took the secondary place. The syndrome of phlegm was often accompanied by syndrome of deficiency of spleen qi. Only 15 ones of 46 clinical symptoms showed high frequency in concomitant appearance in syndrome of blood stasis, syndrome of phlegm and syndrome of phlegm-blood stasis blocking. Apart from having the common symptoms in syndrome of deficiency in origin, the syndrome of phlegm especially showed white and greasy fur and slippery pulse as well as distention and fullness of chest and abdominal distension; the syndrome of blood stasis showed purplish tongue and ecchymosis on tongue as well as fixed pain; and the syndrome of phlegm-blood stasis blocking showed the main symptoms of both syndrome of phlegm and syndrome of blood stasis. CONCLUSION: The statistical methods of matching matrix, factor analysis and clustering analysis are convenient, and can definitely indicate the clinical characteristics and syndrome differentiation of viscera of different syndromes of phlegm and blood stasis, which are beneficial to further research of diagnosis and differentiation of such syndromes of coronary heart disease.

OBJECTIVETo investigate the protective effect of hypothermia combined with dexamethasone on spermatogenesis and the expression of intercellular adhesion molecule 1 (ICAM1) after testicular torsion-detorsion.

METHODSWe made unilateral testicular torsion models in 100 pubertal male Sprague-Dawley rats by 720 degree torsion of the left testis and then randomly divided them into four groups of equal number to be treated with normal temperature + physiological saline (group A), hypothermia + physiological saline (group B), normal temperature + dexamethasone (group C), and hypothermia + dexamethasone (group D). After 48 hours, we collected the testes, observed pathological changes of the testicular tissue by HE staining under the light microscope, detected the apoptosis of spermatogenic cells by TUNEL, and determined the expression of ICAM1 by Western blot.

RESULTSHE staining showed different degrees of testicular tissue injury in the four groups of rats, most obvious in group A, but mild in the other three. The ICAM1 protein expression was significantly higher in group A (0.68 +/-0. 03) than in B (0. 49 +/- 0. 06, P <0. 05) , C (0. 46 +/- 0. 09, P < 0.05) , and D (0.17 +/- 0.08, P <0.01). The nuclei were deep brown or brown. Lots of apoptotic spermatogenic cells were seen in the torsion testis of group A, with a significantly higher apoptosis index ( [33. 13 +/- 3.21 ]%) than in B ( [ 17. 12 +/-5.23 ]%, P < 0.05), C ([14.13 +/- 2.03]%, P <0.05), and D ([9.05 +/- 1.03]%, P <0.01).

CONCLUSIONHypothermia combined with dexamethasone can protect the testis from injury as well as the reproductive function of the testis after testicular torsion-detorsion and reduce the expression of ICAM1.

Animals ; Dexamethasone ; pharmacology ; Disease Models, Animal ; Hypothermia, Induced ; Intercellular Adhesion Molecule-1 ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Spermatic Cord Torsion ; metabolism ; physiopathology ; Spermatogenesis ; drug effects

BACKGROUND:Adiponectin possess functions of lowering blood glucose and blood lipids, and improve insulin sensitivity. But, controversy results about the effect of adiponectin on skeletal muscle have been reported.OBJECTIVE:To study the effects of eukaryon expressed adiponectin on the glycogen synthesis and glucose oxidation in skeletal muscle cell strain C2C12 myotubes by transfecting plasmids carrying mouse adiponectin.DESIGN: A controlled experiment.SETTING: The Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: PcDNA3.0 plasmid with mouse adiponectin cDNA, pcDNA3.0-mad (generously presented from Dr. Gong,University of Maryland), C2C12 cell strain (purchased from ATCC, GRL-1722), DMEM high glucose (Gibco), MEM (Hyclone), fetal bovine serum (Hanagzhou Sijiqing), equine serum (Hyclone), lipofectamine 2000 (Invitrogene), G418 (Gibco), rabbit anti-mouse adiponectin IgG (ACRP303-A, Alpha Diagnostic International), chemiluminescence kit (ECL+PLUS,Amersham), SABC instant immunohistochemistry kit (Boster), D-[U-14C] glucose (specific activity 9.25-13.32 GBq/mmol,NEC), scintillation fluid POP, POPOP (SIGMA), liquid scintillation counter (LS3801, Beckman, USA).METHODS:This study was carried out in the Central Laboratory of the Second Affiliated Hospital of Sun Yat-sen University from March to August, 2003. ① After extraction of plasmid, double digest with Xho Ⅰ and Xba Ⅰ and identification with HindⅢ digest were carried out. ② Plasmid pcDNA3.0-mad and pcDNA3.0 blank vector were transfected using liposome to C2C12 cells, and the stably transfected cells were screened by 500 mg/L G418 for 3 weeks, G418 resistant C2C12 cells were thereafter harvested, therefore stable transfected pcDNA3.0-mad and pcDNA 3.0 C2C12 cell strains were established.③ Adiponectin protein expression was determined by Western blot analysis and immunohistochemistry. ④ Glucose oxidation and glycogen synthesis detections were divided into control, vector and pcDNA3.0-mad (mad)group. Each group was further divided into 4 subgroups with 0, 0.5, 5 and 100 nmol/L insulin (n =6), respectively. Detection of glucose oxidation and glycogen synthesis was carried out with 14C-labeled glucose by counting radioactivity of 14CO2 or 14C labeled glycogen with scintillation, respectively.MAIN OUTCOME MEASURES:Changes of glycogen synthesis and glucose oxidation in skeletal muscle cell strain C2C12myotubes.RESULTS: ① Results of plasmid transfection and restrict digest: After plasmid extraction, double digest with Xba Ⅰ and Xho Ⅰ was carried out along with HindⅢ digest identification.Digest fragments were in accordance with expectation.Length of adiponectin cDNA fragment was 781 bp, plasmid fragment was 5 446 bp, adiponectin cDNA was inserted between digest sites (Xho Ⅰ and Xba Ⅰ ) of eukaryotic expression vector pcDNA3.0. ② Plasmid transfection of C2C12 cell and positive clone screening: On the 10th day of G418 media culture screening after transfection, most C2C12 cells died.Positive clone appeared at the 2nd week. G418 resistant C2C12 colonies were harvested at the 3rd week. ③ Western blot and immunohistochemical identifications: Both confirmed that adipoenctin gene was stably transfected into cells in the Mad group, with successful adipoenctin expression. ④ Effect of stably transfected adiponectin gene to myocyte glucose metabolism:The myocyte glycogen synthesis and glucose oxidation increased along with the increasing of insulin concentration. The linear regression analyses of measured myocyte glucose oxidation amount showed that the regression coefficients of the control group, blank vector group and mad group were 23.34, 2;3.23 and 26.06 respectively. This result indicated that in C2C12 cell stably transfected with adiponectin gene, when insulin concentration increased, the acceleration rate of glucose oxidation increasing was higher than other 2 groups. However, no significant difference could be observed in glycogen synthesis and glucose oxidation of C2C12 cells under basic status without insulin stimulus and treatment status with different insulin concentrations between control group, blank vector group and mad group (P＞ 0.05).CONCLUSION: ① We have successfully established stably adiponectin gene transfected C2C12 cell strain with adiponectin protein expression ability. ② Transfection with adiponectin cDNA had no significant effect on the glucose oxidation and glycogen synthesis of C2C12 myotubes.③ The glucose oxidation and glycogen synthesis of C2C12 myotubes increased with the increasing of insulin concentration. ④ Adipoenctin may coordinate with insulin in improving myocyte glucose oxidation and increasing myocyte glucose uptake.

One hundred and fifty-one type 2 diabetic patients with coronary heart disease ( T2 DMC) and 142 cases of type 2 diabetes mellitus were included for analyzing the influence of different glucose-lowering rates on MB isoenzyme of creatine kinase (CKMB) and muscle hemoglobin level changes to search for the rational glucose-lowering rate.The level of CKMB in type 2 deabetes mellitus group was significantly lower( P＜0.05 ) at follow-up than that before and after intensive therapy.In type 2 diabetes mellitus group,when the fasting or postprandial glucose-lowering rate was not greater than 6 mmol· L-1 · d-1,the level of CKMB and muscle hemoglobin were significantly lower at follow-up than that before intensive therapy ( P＜0.05 ).When the fasting glucose-lowering rate is greater than 6 mmol· L-1 · d-1,the level of CKMB is significantly higher after intensive therapy than that before glucose-lowering ( P＜0.05 ).In T2DMC group,when the fasting or postprandial glucose-lowering rate was not greater than 4 mmol· L-1 · d-1,the level of CKMB and muscle hemoglobin was significantly lower at follow-up than that before intensive therapy(P＜0.05 or P＜0.01 ),buthigher at follow-up when the fasting glucose-lowering rate was greater than 4 mmol· L-1 · d-1(P＜0.05).

Objective To compare the changes of high sensitive-C reactive protein (hs-CRP) and cardiac troponin Ⅰ ( cTn Ⅰ ) levels before and after intensive therapy in patients with type 2 diabetes,and to find out the reasonable glucose-lowering rate.Methods One hundred and thirty-two cases of type 2 diabetes( T2DM group) and 135 cases of type 2 diabetes with coronary heart disease( T2DM+CHD group) received intensive therapy.After testing hs-CRP and cTn Ⅰ levels,the variations were analyzed.Results The ranges of the change in cTn Ⅰ and hs-CRP levels were different under four glucose-lowering rates in the T2DM+CHD group( P＜0.05 ).cTn Ⅰ and hs-CRP levels were higher than those before intensive therapy in the T2DM+CHD group with glucose-lowering rate greater than 4.0mmol· L-1 · d-1.The other two subgroups with glucose-lowering rate less than 4.0 mmol· L-1 · d-1 showed decreased cTn Ⅰ and hs-CRP levels.While at the end of 3 months follow-up,cTn Ⅰ and hs-CRP levels were all significantly lower than those before intensive therapy in four subgroups ( P＜0.05 ).Conclusions The increase of cardiovascular events after intensive therapy may be due to excessively fast glucose-lowering rate.The reasonable glucose-lowering rate for patients with type 2 diabetes should depend on whether there is accompanying coronary heart disease.For type 2 diabetes with coronary heart disease,excessively fast glucose-lowering rate could lead to acute rise ofcTn Ⅰ and hs-CRP levels,which causes myocardial injury.The mechanism of myocardial injury resulted from excessively fast glucose-lowering rate may be due to activation of the inflammatory pathway.In type 2 diabetes with coronary heart disease,long-term good control of blood glucose could alleviate inflammatory response and cardiac damage resulted from excessively fast glucose-lowering rate.

Objective To investigate the clinical features, diagnosis and treatment of malignant pancreatic endocrine tumor. Methods The clinical data of 38 patients with malignant pancreatic endocrine tumor who had been admitted to First Affiliated Hospital of Nanjing Medical University from January 1969 to December 2008 were analyzed retrospectively. Of all patients, 6 were with insulinoma, 23 with pancreatic polypeptide tumor, 4 with glucagonoma and 5 with pancreatic carcinoid. Results All patients except 1 with insulinoma were found with pancreatic lesion by imaging examination. The resection rate was 87% (33/38). Pathological examination found 7 patients with liver metastasis, 5 with lymph node metastasis, 1 with tumor thrombus in vessels and lymphatic vessels, and 28 with local invasion. Twenty-four patients were followed up, and neither recurrence nor metastasis was found except 1 patient with insulinoma who received reoperation for local recurrence and 1 patient with pancreatic carcinoid who received radiofrequency ablation for liver metastasis. Conclusions The diagnosis of pancreatic endocrine tumor mainly depends on imaging examination. The malignancy of pancreatic endocrine tumor is determined after the comprehensive analysis of preoperative imaging findings, intraoperative examination, post-operative pathological examination and the data obtained during follow-up. The malignant pancreatic endocrine tumor should be managed actively by resection because of its relatively low malignancy, high operative resectability and relatively good prognosis.

Objective To evaluate the clinical value of ultrasonography detecting the poster urethrovesical angle(PUVA) in diagnosis of stress urinary incontinence(SUI).Methods From Jan.2006 to Dec.2008,the PUVA in resting phase (PUVA-r) and stress phase( PUVA-s) between with SUI and 100 healthy women were measured by color Doppler ultrasound,including 57 cases in SUI degree Ⅰ,22 cases in SUI degree Ⅱ,5 cases in SUI degree Ⅲ.Results (1) The PUVA-r and PUVA-s were (130±29) and (158±36)in SUI patients,which were significantly higher (113±19)and (115±23)in control group (P<0.01).(2) Correlation analysis showed that PUVA-s was positively correlated with the severity of SUI ( P < 0.01).(3) The area under the receiver operating characteristic curves (ROC) of PUVA-s was 0.82 (95% CI:0.76-0.89) between SUI patients and healthy women.When PUVA-s ≥ 140 ° was chosed as the cut-off value,the specificity,accuracy rate and positive predictive value in diagnosis of SUI were 84%,79%,81%,which were significantly higher than those when cut-off value of PUVA-s ≥ 120(54%,68%,62%; P < 0.05 ).Conclusion The PUVA-s in SUI patients are significantly increased and is positively correlated with the severity of SUI,which are indicated that PUVA-s ≥ 140should be used as cut-off value in diagnosis of SUI by ultrasonography.

Objective To evaluate the expression and relationship of miR-100 and FZD-8, one of the major compo?nents of Wnt signaling pathway, and the correlation of their expressions with lymph node metastasis in patients with breast cancer. Methods The expression of miR-100 was determined in 50 samples of human breast cancer tissues and adjacent normal tissues by in situ hybridization. The correlation of miR-100 expression with lymph node metastasis was analyzed by Mann-Whitney U test. The expression of FZD-8 was measured in 50 samples of human breast cancer tissues and adjacent normal tissues by immunohistochemistry. The correlation of the miR-100 expression with the protein expression of FZD-8 was evaluated by Pearson rank analysis. Results The expression of miR-100 was significantly lower in human breast can?cer tissues than that in adjacent normal breast tissues [2.00 (1.00, 3.00) vs. 6.00 (3.50, 8.00)]. The miR-100 expression was lower in patients with lymph node metastasis than that in patients without lymph node metastasis [1.50 (1.00, 2.75) vs. 3.00 (2.00, 4.00)]. The expression of FZD-8 was significantly higher in human breast cancer tissues than that in adjacent normal breast tissues [8.00 (6.00, 9.00) vs. 6.00 (3.75, 9.00)]. The miR-100 expression was negatively correlated with the FZD-8 pro?tein expression in human breast cancer tissues (rs=-0.592, P<0.001). Conclusion The miR-100, as an anti-metastasis-miRNA, may involve in the metastasis of breast cancer, which may be related with the regulation of the expression of FZD-8.

OBJECTIVE:To evaluate the effects of different doses of flurbiprofen axetil on analgesia effects of patients after laparoscopic cholecystectomy. METHODS:120 patients undergoing laparoscopic cholecystectomy were selected and randomly divid-ed into group A,B and C,with 40 cases in each group. Group A,B and C were given the mixture 100 ml of flurbiprofen axetil 100,150 and 200 mg combined with tramadol 600 mg and ondansetron 4 mg respectively and 0.9% Sodium chloride injection for patient controlled intravenous analgesia(PCIA)at the end of operation. Mean arterial pressure(MAP),heart rate(HR)and static and dynamic visual analogue scale(VAS)scores were observed in 3 groups at the end of operation,4,8,24 and 36 h after sur-gery. The incidence of incision pain,neck-shoulder pain and hypochondrium,the occurrence of ADR were recorded 36 h after oper-ation. RESULTS:After operation,There was no statistical significance in comparison of 3 groups with MAP,HR,static and dynam-ic VAS(P>0.05),4,8,24,and 36 h after operation,MAP,HR,static and dynamic VAS score of group B and C decreased sig-nificantly,there was statistical significance,compared with group A(P<0.05);there was no statistical significance in above indi-cators between group B and group C(P>0.05). After operation,the incidence of incision pain,neck-shoulder pain and hypochon-drium in group A were significantly higher than group B and C,with statistical significance(P<0.05),but there was no statistical significance between group B and group C(P>0.05). After operation,the incidence of ADR in group A and B were significantly lower than in group C,with statistical significance(P<0.05),but there was no statistical significance between group A and group B(P>0.05). CONCLUSIONS:Flurbiprofen axetil 150 mg combined with tramadol 600 mg and ondansetron 4 mg can improve he-modynamics and patient controlled intravenous analgesia in patients underwent laparoscopic cholecystectomy with lower incidence of ADR.