AIM: HPLC-ELSD method was established to investigate the content of oleanolic acid and ursolic acid in Compound Isodon Rernifolia(D.Don) Kudo Tablets. METHODS: The determination of oleanolic acid and ursolic acid was conducted by HPLC-ELSD using a Agilent Eclipse XDB-C_(18)(4.6 mm?250 mm,5 ?m) column;The mobile phase consisted of methanol-0.5% ammonium acetate solution(82∶18).The flow rate was 1.0 mL/min.The column temperature was set at 30 ?C.The drift tube temperature was set at 85 ?C.The gas flow rate was 2.0 L/min. RESULTS: The linear ranges of oleanolic acid and ursolic acid were 0.45-2.69 ?g(r=0.999 9) and 0.63-3.75 ?g(r=0.999 3),respectively.The average recoveries of oleanolic acid and ursolic acid were(100.3%)(RSD=1.32%) and 100.6%(RSD=1.66%),respectively. CONCLUSION: The method is simple,accurate,and specific which is better than the original one.

Objective:To improve previous method of primary rat cortical neuron culture to get purer and more long-lasting cells for study.Methods:Timed-pregnant Wistar rats at a gestational age of 16 or 17 days(16-17 d) were used.Fetal brains were removed and the cerebral cortices were dissected out.Papain digestion and mechanical dissociation were combined to conduct mono-cell suspending media.Four to six hours(4-6 h) post-plating,all plating media were removed from cultures and replaced with Neurobasal medium supplemented with B27.On the third day,10 ?mol/L cytosine arabinoside(Ara-C) was added to the culture for 24 h to inhibit the outgrowth of glial cells.Half of the culture medium was changed every week.The morphological changes of neuron cells were observed by light microscope.Double immuno-staining of microtubule-associated protein 2(MAP2) and karyon were applied to assess the culture purity.Evaluation of synapse formation was processed by immunocytochemical analysis using antibodies against both pre-and postsynaptic protein markers.Results:The improved method could remarkably increase the cell number and reduce neuronal damnification.The primary culture was characte-rized by high uniformity,purity,normal synapse formation and longtime livability.Conclusion:This is a simple and reliable technique for the in vitro primary culture of rat cortical neurons.

AIM:To study the role of peroxisome proliferator-activated receptor-? (PPAR-?) signal transduction pathway in cardiac hypertrophy induced by high glucose and insulin (HGI). METHODS:The cultured neonatal rat cardiomyocytes were used to observe the effect of fenofibrate (FF),a selective PPAR-? agonist,on cardiomyocyte hypertrophy induced by HGI (glucose at concentration of 25.5 mmol/L and insulin at 0.1 ?mol/L). The cardiomyocyte hypertrophic responses were assayed by measuring the cell surface area,protein content,and mRNA expression of atrial natriuretic factor (ANF). The expressions of mRNA and protein were assayed by real -time PCR and Western blotting. RESULTS:In cultured cardiomyocytes,HGI induced profound change of hypertrophic morphology,the significant increase in cell surface area,protein content and ANF mRNA expression compared to those in vehicle control (P

AIM: To study the role of peroxisome proliferator-activated receptor-α (PPAR-α) signal transduction pathway in cardiac hypertrophy induced by high glucose and insulin (HGI). METHODS: The cultured neonatal rat cardiomyocytes were used to observe the effect of fenofibrate (FF), a selective PPAR-α agonist, on cardiomyocyte hypertrophy induced by HGI (glucose at concentration of 25.5 mmol/L and insulin at 0.1 μmol/L). The cardiomyocyte hypertrophic responses were assayed by measuring the cell surface area, protein content, and mRNA expression of atrial natriuretic factor (ANF). The expressions of mRNA and protein were assayed by real -time PCR and Western blotting. RESULTS: In cultured cardiomyocytes, HGI induced profound change of hypertrophic morphology, the significant increase in cell surface area, protein content and ANF mRNA expression compared to those in vehicle control (P<0.01), but the expressions of PPAR-α mRNA and protein decreased significantly (P<0.05). At the same time, the expression of cyclooxygenase 2 (COX-2), one of the PPAR-α downstream effectors was obviously elevated (P<0.05). However, FF (0.1, 0.3 and 1 μmol/L) inhibited the cardiomyocyte hypertrophy induced by HGI in a concentration-dependent manner (P<0.01). FF at concentration of 0.3 μmol/L increased the expressions of PPAR-α in both mRNA and protein levels (P<0.05) and inhibited the expressions of COX-2 (P<0.05), which were abolished by MK 886 (0.3 μmol/L), a selective PPAR-α antagonist (P<0.05). CONCLUSION: PPAR-α signal transduction pathway and its downstream effector COX-2 might involve in the cardiomyocyte hypertrophy induced by HGI.

Objectives To analyze the clinical characteristics and signiflcance of Mycoplasma pneumoniae pneumonia (MPP) combined with secondary thrombocytosis in infants and young children. Methods Clinical features, laboratory and imaging data of the infants and young children with MPP were collected, and compared between the two groups of children with and without secondary thrombocytosis. Results Sixty-seven (67) infants and young children with secondary thrombocytosis (28 males and 39 females, with onset at 2 years and 2 months after birth in average) and 269 infants and young children with normal platelet counts (138 males and 131 females, at 2 years and 3 months after birth in average) were included. Signiflcant longer durations of hospitalization(P =0.018) and fever(P =0.000), higher temperature peak(P =0.000), as well as higher morbidity of refractory MPP(P =0.001) and more complications(P =0.000)were observed in the group of MPP with secondary thrombocytosis. Moreover, the laboratory data of white blood cell count(P =0.000), C-reactive protein(P =0.000),procalcitonin (P =0.000), erythrocyte sedimentation rate(P =0.000)and higher morbidity of multiple pathogen infection(P =0.033) were observed between the two groups.Conclusions More severe clinical manifestations and higher complication morbidity could be observed in the group of MPP combined with secondary thrombocytosis, implicating the value of platelet count in clinical assessment of MPP in infants and young children at early stage.

Objective To examine the effects of carbon dioxide (CO2) pneumoperitoneum on local pancreas pathological changes,serum levels of amylase,IL-1,IL-6,and the positive rate of dissolubility adhesion molecule (CD11a/CD18 and CD11b/CD18) expression in rats with severe acute pancreatitis (SAP). Methods Fifty healthy male Sprague-Dawley (SD) rats were randomly divided into 3 groups:CO2 pneumoperitoneum group (n=20):SAP was induced by injecting 5% sodium taurocholate through retrogradely common biliopancreatic ducts via duodenal papilla,and then CO2 pneumoperitoneum was established at a pressure of 12 mm Hg (1 mm Hg=0.133 kPa) for 30 min; SAP group (n=20):The rats were treated as same as CO2 pneumoperitoneum group,except CO2 pneumoperitoneum; Simple operation group (n=10):Laparotomy was performed and nothing was done to duodenum and pancreas except for moving them softly. The blood samples were collected for examining serum levels of amylase,IL-1,IL-6,and the positive rates of CD11a/CD18 and CD11b/CD18 expression,and histopathologic examination of pancreas was performed. Results Compared with simple operation group,the pancreatic pathologic histology score,serum levels of amylase,IL-1,IL-6,and the positive rates of CD11a/CD18 and CD11b/CD18 expression were significantly higher in CO2 pneumoperitoneum group and SAP group (P=0.000). The levels of IL-1 and IL-6 were significantly lower in CO2 pneumoperitoneum group as compared to SAP group (P=0.000). There was no significant difference between CO2 pneumoperitoneum group and SAP group in pancreatic pathologic histology score (P=0.294),the level of serum amylase (P=0.073),the positive rates of CD11a/CD18 (P=0.155) and CD11b/CD18 expression (P=0.201). Conclusion CO2 pneumoperitoneum has inhibitory effect on the levels of IL-1 and IL-6,rather than the positive rates of CD11a/CD18 and CD11b/CD18 expression in SD rats with SAP.

Aim To investigate the effect of CD44 anti-body-A3 D8 on the expression of IL-3 Rα and down-stream PI3K/Akt in NB4 cells. Methods The ex-pression of IL-3 Rα mRNA was detected by real-time quantitative RT-PCR, the IL-3Rα protein expression and changes of PI3 K/Akt signal pathway in NB4 cells treated with A3D8 were analyzed by Western blot. An-nexin-V-FITC/PI double staining flow cytometry was u-tilized to detect the apoptotic cells. The inhibitor of PI3 K/Akt signaling LY294002 combined with A3 D8 was used to inhibit the PI3K/Akt in NB4 cells. Re-sults After treated with A3 D8 , both the transcription-al level and translational level of IL-3 Rα were remark-ably reduced, and the PI3K/Akt pathway was inhibi-ted. LY294002 improved the inhibitory and apoptotic effects of A3D8 on NB4 cells. Conclusion CD44 antibody A3 D8 can downregulate the expression of IL-3Rα and inhibit the downstream PI3K/Akt pathway.

Objective To probe into the reliability and validity of the questionnaire on clinical pathways and patients' satisfaction.Methods Investigating the hospital patients' satisfaction with the questionnaire designed by the former Ministry of Health and analyzing the internal consistency and constructive validity through statistical methods.Results Cronbach' s α coefficient of the adjusted questionnaire was 0.947 and those of the each subscale were 0.563,0.968 and 0.820.The split-half reliability of the questionnaire was 0.869.The number of factors is 3 as same as the designed assuming structure,and the cumulative contribution rate of 3 factors was 74.178％.The factor loading of each item was above 0.4,each item of a common factor with a higher load factor.Conclusion Although there are differences from hospital to hospital,this questionnaire is applicable to investigating patients' satisfaction of the clinical pathway management mode in view of the statistical and clinical practice.

Objective To investigate the clinical value of the type and the steepest slope of tumor's time-intensity curve (TIC) in assessing the pathologic response of locally advanced breast cancer treated with neoadjuvant chemotherapy (NAC). Methods Thirty-six patients with pathologically confirmed locally advanced breast cancer who finished four courses of neoadjuvant chemotherapy underwent preoperative breast MRI three times during the NAC. Pathologic response was assessed according Miller-Payne grading system, grade 4 and 5 were defined as major histological response ( MHR, n = 16) group, and grade 1 to 3 as nonmajor histological response( NMHR,n = 20)group. The type and the steepest slope of tumor's TIC were compared between two groups before NAC, after the second cycle and after the fourth cycle of NAC. ROC analysis was carried out to assess the clinical value of the TIC parameters. Results After the second cycle of NAC, the steepest slope of TIC and its first change rate were different between the MHR group [ ( 1.93 ±0.88) %/s, median 35.6%] and NMHR group [(2.73 ± 1.22) %/s, median - 11.4%] (P =0.045 and 0. 01,t=1. 09,Z= -3.64). After the fourth cycle, the proportion of type Ⅰ in MHR group (62.5% ,10/16) was significantly higher than that in NMHR group (10.0%, 2/20, P = 0.01, Z=-2. 02), and the proportion of type Ⅲ in MHR group (6. 2% ,1/16)was significantly lower than that in NMHR group (60. 0% ,12/20,P =0. 01 ,Z = -1.48). The steepest slope and its second change rate were different between the MHR group [ ( 1.33 ± 0. 52) %/s, median 56. 8% ] and NMHR group [ (2. 33 ±0. 94) %/s, median 15. 8% ] ( P ＜ 0. 01, t = 1.82, Z = - 3. 58 ). After the second cycle, the area under curve of the steepest slope of TIC and its first change rate were 0. 70 ( P = 0. 04 ), 0. 80 ( P = 0. 01 ),respectively. After the fourth cycle, the area under curve of the type Ⅰ, the type Ⅰ + Ⅱ, the steepest slope and its second change were 0. 78 ( P = 0. 03 ), 0. 69 ( P = 0. 06), 0. 82 ( P = 0. 01 ), 0. 92 ( P = 0. 01 ),respectively. Conclusion The steepest slope of TIC and its first change rate could assess the NAC response after the second cycle, and the type Ⅰ, the steepest slope and its second change could assess the NAC response after the fourth cycle.

Objective To investigate the clinical value of tumor's longest diameter and volume in assessing pathologic response in locally advanced breast cancer treated with neoadjuvant chemotherapy(NAC).Methods A prospective study was undertaken in women undergoing NAC for locally advanced breast cancer in order to determine the ability of quantitative MRI to assess the final pathologic response. 36 cases with pathologically confirmed locally advanced breast cancer who had been undergone four courses of NAC underwent preoperative breast MRI three times during the NAC. Pathologic response was assessed according Miller & Payne grading system, of which grade 4 and 5 defined as major histological response (MHR), and grade 1 to 3 as non-major histological response (NMHR). The tumor's longest diameters and volumes in MHR were compared with those in NMHR by Mann-Whitney U test before, after the second and fourth cycle of NAC. Concordance correlation coefficient (CCC) were assessed to evaluate the agreement between the two method. Receiver operating characteristic curve (ROC) analysis was carried out to assess the clinical value of tumor size and the change rate.Results Before,after the second and the fourth cycle of NAC, the difference of tumor's longest diameters and their first change rate between MHR [(2.75±1.16) cm,(2.19±1.07) cm ,(1.58±0.75) cm and (21.70±15.09)%]respectively,and NMHR [(2.71±1.10) cm,(2.33±0.90) cm,(2.01±0.94) cm and (11.68±10.27)%] respectively were not significant(P>0.05). The second change rate of tumor's longest diameter in MHR [(39.00±15.38)% ] was significantly higher than that in NMHR[ (25.83±21.77)% ](P=0.04). Before, after the second and the fourth cycly of NAC, the differences of tumor volumes and their first change rate between MHR [ median 14.00 cm~3 ( range 2.96～83.41 cm~3 ) , median 7.31 cm~3 (range 0.05～55.35 cm~3), median 2.69 cm~3 (range 0～33.40 cm~3 ) , median 48.65% (range 33.64%～98.48%) ] and NMHR [median 4.25 cm~3 (range 4.78～106.55 cm~3), median 10.53 cm~3 (range 1.72～42.85 cm~3), median 7.56 cm~3 (range 0.68～156.58 cm~3), median 52.04% (range-35.79%～78.31%) ] were not significant. The second changing rate of tumor's volume in MHR [median 85.37% (ranged 27.52%～100%)] was significantly higher than that in NMHR [median 68.80% (ranged -289.60%～94.24%)](P=0.01). CCC was computed before and (0.82), after the second cycle (0.67) and after the fourth cycle (0.55), in all examinations pooled together (0.78).The second change rates of tumor's longest diameter and volume were equal to predict the final pathologic response, and the area under curve were 0.75 and 0.80, respectively (P=0.61). Conclusion The agreement between the tumor's longest diameters and tumor's volumes is good in all in breast carcinoma. The assessment efficacy of the change rate of tumor's longest diameter and that of volume were low.