OBJECTIVE: To establish an HPLC method for simultaneous determination of the contents of Chlorogenic Acid and Coffeic Acid in the Fruit of Chaenomeles. METHODS: HPLC was applied to determine and compare the contents of 11 batches of Chaenomeles medicinal materials collected at different time, prepared with different process and stored for different length of time, using Coffeic Acid and Chlorogenic Acid standard substances. The chromatogram conditions were as follows: Kromasil C18( 250mm? 4. 6mm, 5? m) column; mobile phase of ( A) acetonitrile, ( B) 0. 05% phosphoric acid; gradient el-ution ( 0～ 10min, 0% A～ 15% A, 100% B～ 85% B; 10～ 40min ( 15% A～ 30% A , 85% B～ 70% B) ; flow rate at 0. 8 mL? min- 1; detection wavelength at 325nm. RESULTS: The content of Chlorogenic Acid in batches 1 to 9 were respectively 0. 020% , 0. 035% , 0. 043% , 0. 051% , 0. 260% , 0. 104% , 0. 081% , 0. 056% , 0. 034% ; that of Coffeic Acid were respectively 0. 005% , 0. 006% , 0. 008% , 0. 010% , 0. 051% , 0. 024% , 0. 020% , 0. 015% , 0. 007% . The contents of Chlorogenic Acid and Coffeic Acid in batch 8’ ( burn- prepared) were respectively 0. 045% and 0. 010% ; those of Chlorogenic Acid and Coffeic Acid in batch 9’ ( stored for 1a) were respectively 0. 023% and 0. 005% . CONCLUSIONS: The method is simple, reproducible and suitable for the quality control of Fruit of Chaenomeles. The best time for harvesting batch 5 is on July 12, and the best process for this batch is solarization. The storage time has certain influence on the content of Chlorogenic Acid.

To improve the precision of image registration based on corner detection, a relative position function between multiple points to determine matching points accurately. First the corners in images are detected using Harris detector, and clustering method is used to eliminate most wrong matches after coarse screening. Then the proposed relative position function is used as a criterion of precise matching. Finally the image registration process is accomplished by affine transformation. Results show that the proposed algorithm is more effective and accurate than conventional registration algorithm.

Objective To establish a HPLC method for the determination of geniposide in Mengning Tablets and to investigate the thermal stability of geniposide. Methods The content of geniposide in Mengning Tablets was determined on a E- clipse XDB-C_(18) column with a mobile phase consisting of 0.2% triethylamine (containing 0.1% phosphate) and acetonitrile (85: 15) at a flow rate of 1.0 mL?min~(-1) and detected at a wavelength of 240 nm. The thermal stability of geniposide was investigated by hyperthermal accelerated test. Results A linearity was obtained from 0. 10 ?g to 0.52 ?g of geniposide (r=0.9997, n=5) and the recovery was 98.92%(n=5). The variation of geniposide in Mengning Tablets was in first-order reaction and the expiry date of the medicine at normal temperature was 2.83 years when geniposide content was used as criteria. Conclusion This is a fast method for the determination of the expiry date of a Chinese patent medicine and this method is useful infor the development of a new medicine.

The anti-hepatitis B virus (HBV) effects and its mechanisms of the ethanol extracts of Hypericum perforatum L. (EHP) in vitro were explored. HepG2 2.2.15 cells, a stable HBV-producing cell line, were cultured as the model system to observe the anti-HBV effect. The viral antigens of cellular secretion, HBsAg and HBeAg, were determined by enzyme linked immunosorbent assay (ELISA). The quantity of HBV-DNA released in the supernatant was assayed by real-time PCR. In order to understand the mechanisms of the suppression of HBV replication, all HBV promoters (Cp, Xp, S1p, S2p and Fp) with luciferase reporter gene were transfected into HepG2 cells respectively. Then the activities of viral promoters were examined by luciferase reporter assay. It was found EHP effectively suppressed the secretion of HBsAg and HBeAg from HepG2 2.2.15 cells in a dose-dependent manner, as well as the extracellular HBV DNA. And EHP could selectively inhibit the activity of HBV promoter Fp. Our data suggest that EHP exerts anti-HBV effects via inhibition of HBV transcription, which helps to elucidate the mechanism underlying the potential therapeutic value of EHP.

Objective To explore the possible mechanism of Jianyi Recipe for improving the function of islet cells from the aspects of synthesis,secretion and inactivation of glucagon-like peptide-1 (GLP-1).Methods The diabetic rat model was established by feeding with high-lipid food combined with injection of streptozotocin (STZ).The rats were randomly divided into model group,Jianyi Recipe group,and normal group.The treatment for the rats lasted for 4 weeks.The blood glucose level was detected by the rapid blood glucose meter.The plasma levels of GLP-1 and insulin were detected by Luminex liquid phase protein chip technology.Pancreatic duodenal homeobox-1 (PDX-1) mRNA expression level was detected by quantitative real-time fluorescence polymerase chain reaction (PCR).The level of GLP-1 in ileum L cells was detected by immunohistochemistry,and dipeptidyl peptidase Ⅳ (DPP-Ⅳ)level was detected by enzyme-linked immunosorbent assay.Results Jianyi Recipe could decrease the levels of fasting blood glucose and postprandial glucose (P ＜ 0.05),promote the secretion of insulin (P ＜ 0.05),and increase PDX-1 mRNA expression level in the pancreas of the diabetic rats.Compared with the model group,plasma GLP-1 level,and ileal GLP-1 positive expression area and integrated optical density were increased (P ＜ 0.05) in Jianyi Recipe group,while the differences of serum DPP-Ⅳ levels were insignificant between the two groups (P＞ 0.05).Conclusion Jianyi Recipe maybe regulate the synthesis and secretion of GLP-1 to promote PDX-1 gene expression and insulin secretion,so as to reduce blood glucose in diabetic rats.

Objective To study the association between transforming growth factor-β1 (TGF-β1) polymorphism and type 2 diabetes mellitus in Han population of Shanghai. Methods In this case-control study , 1 234 cases of T2DM patients were recruited and 1 272 healthy individuals were selected as control. Five ml of blood sample was collected from each subject ,from which the whole genomic DNA was extracted.The polymorphism was detected by the Taqman technology. Result Significant association was observed in TGF-β1 T896C genotypes and alleles with T2DM (P = 0.0001 and P = 0.004, OR = 1.18 [1.05 ~ 1.33], respectively). Conclusion The polymorphism of T896C in TGF-β1 gene may be associated with T2DM in Han population from Shanghai.

Objective:To investigate the expressions and clinical significances of SMO , STAT3 and MMP-9 in triple negative breast cancer ( TNBC) .Methods:The expressions of SMO , STAT3 and MMP-9 were immunohistochemically detected in 33 cases of TNBC, 30 cases of mammary hyperplasia and 18 cases of normal breast tissue , and the relationships among SMO , STAT3 and MMP-9 and clinicopathological parameters and prognosis of TNBC patients were analyzed .Results:In TNBC and mammary hyperplasia tissue , the positive expression rates of SMO were 90.9%and 60.0%,the positive expression rates of STAT3 were 96.9% and 73.3%, the positive expression rates of MMP-9 were 90.9% and 36.7%, respectively, however, expressions of them were completely absent in normal breast tissue .The significant correlations were observed between expressions of SMO , STAT3 and MMP-9 in TNBC, mammary hyperplasia and normal breast tissue (P<0.05).The expression of SMO and STAT3 was correlated with superior histologic grade and tumor stage .The expression of MMP-9 was correlated with metastasis of lymph node .The positive correlation was manifested between SMO and STAT3(r=0.361,P<0.05),between SMO and MMP-9 (r=0.633,P<0.01),between MMP-9 and STAT3 (r=0.803,P<0.01) in TNBC.The survival time of TNBC patients was correlated with SMO expression and pTNM (P<0.05).Conclusion:SMO, STAT3 and MMP-9 may play important roles in development of TNBC and may be an important therapeutic target in TNBC .

Objective To survey cervical myoelectric signals during craniocervical flexion, neutral and extension postures, and to explore the evidence that proper head position can alleviate cervical muscle fatigue in a lateral recumbent position. Methods Surface electromyography (sEMG) signals were detected from the sternocleidomastoid, upper trapezius and erector spinae muscles of 30 young subjects bilaterally during craniocervical flexion,neutral and extension postures in the left lateral recumbent position. The integrated trace area (IEMG) and median frequency (MF) were estimated. Results The average IEMG of the sternocleidomastoid muscles was significantly lower in flexion than in extension bilaterally. The average IEMG of the erector spinae muscles was lower in extension than in flexion bilaterally, and the difference was again significant. The IEMGs of the upper trapezius muscle showed no significant difference on average in the three postures bilaterally. There was no significant MF difference in any of the muscles. Conclusions The muscles in the cervical back were less activated during craniocervical extension in a lateral recumbent position. A little cranicocervical extension is optimal while resting in a lateral recumbent position.

Objective To investigate the ultrasonograms of biliary sludge and its relationship with the prognosis of patients after liver transplantation.Methods Ultrasonograms of 36 patients with biliary sludge after liver transplantation were retrospectively analyzed.These patients were divided into two groups according to the different treatment outcomes:poor prognosis group and favorable prognosis group.The characteristics of initial ultrasonograms of biliary sludge when they were found at first time compared between two groups .Results Biliary sludge combined with ischemic-type biliary lesions accounted for 95% ( 19/20 ) in poor prognosis group while 13%(2/16)in favorable prognosis group.There was significant difference between two groups (P<0.05).In favorable prognosis group , biliary sludge was found locating in hepatic hilar region in all 16 cases (16/16) and the sludge had clear boundaries dissociated with bile duct walls were observed in 14 cases (14/16).There was no incrassation of bile duct walls in hepatic hilar region , nor dilation of intrahepatic bile ducts observed in 14 cases ( 14/16 ).In poor prognosis group , biliary sludge was observed widespread not only in hepatic hilar region but also in intrahepatic biliary ducts in 10 cases ( 10/20 ) , and the boundaries between sludge and bile duct walls were vague in 15 cases (15/20).Obvious incrassation of bile duct walls in hepatic hilar region was observed in 16 cases (16/20), and dilation of intrahepatic bile ducts was observed in 19 cases (19/20).There were significant differences in the above 4 ultrasound features between two groups (all in P<0.05).Conclusions There are differences in the ultrasonograms of biliary sludge between different prognosis groups.Biliary sludge combined with ischemic-type biliary lesions suggests a poor prognosis , which may need more active interventional treatments.

Objective To investigate the changes in neutrophil immunophenotypes in neonates with late-onset sepsis and their clinical significance.Methods A total of 42 neonates with late-onset sepsis were enrolled prospectively as sepsis group from Children's Hospital of Fudan University from January 2015 to October 2016,which included 26 preterm infants and 16 term infants.Another 33 neonates without infectious diseases consisting of 20 preterm infants and 13 term infants were selected as control group.According to the severity of sepsis,neonates in the sepsis group were further divided into severe (n=11)and mild sepsis (n=31) subgroups.Expression of CD16 and CD62L on neutrophils was measured by flow cytometry to determine the distribution of neutrophil subsets in neonatal peripheral blood.Differences in the distribution of neutrophil subsets between the two groups and two subgroups were compared using Wilcoxon rank-sum test.Multivariate logistic regression analysis was used to investigate the relationship between neutrophil subsets and the severity of neonatal sepsis.Receiver operating characteristic (ROC) curves were used to assess the diagnostic value of neutrophil subsets for the severity of sepsis.Results Four neutrophil subsets were identified in neonatal peripheral blood,including early-stage neutrophils (CD16-/CD62L-),immature neutrophils (CD16-/CD62L+),mature neutrophils (CD16+/CD62L+) and activated neutrophils (CD16+/CD62L-).Activated [preterm:61.1％ (20.2％-79.4％),term:47.6％ (15.2％-70.1％)] and mature neutrophils [preterm:35.7％ (19.9％-75.8％),term:52.0％ (25.6％-82.8％)] were the dominant subsets in the control group.In preterm infants,the proportion of early-stage [3.5％ (1.7％-9.4％) vs 1.9％ (0.6％-4.0％),Z=-2.501,P=0.012] and immature neutrophils [6.3％ (0.7％-45.5％) vs 0.4％ (0.3％-0.7％),Z=-3.878,P＜0.001] were higher,but that of activated neutrophils [8.3％ (2.3％-49.2％) vs 61.1％ (20.2％-79.4％),Z=2.991,P=0.002] were lower in the sepsis group than those in the control group;same differences were found in the absolute counts of each neutrophil subsets.Among term infants,more immature neutrophils were found in the sepsis group than those in the control group [49 (18-200) vs 13 (5-36)/μl,Z=-2.193,P=0.028].The proportion and the absolute counts of early-stage and immature neutrophils in the severe sepsis subgroup were all higher than those in the mild cases [early-stage neutrophils:5.8％ (3.4％-17.8％) vs 3.0％ (1.4％-7.3％),304 (137-1478) vs 158 (53-321)/μl;immature neutrophils:23.0％ (6.3％-47.0％) vs 0.9％ (0.5％-6.8％),1003 (487-2818) vs 85 (18-275)/μl;all P＜0.05].Multivariate logistic regression analysis showed that the proportion of early-stage neutrophils was associated with the severity of sepsis (OR=1.2,95％CI:1.0-1.4,P=0.012).In addition,the diagnostic value of the proportion of early-stage neutrophils for severe sepsis was the highest when the cut-off value was 3.3％,with the area under the ROC curve was 0.7 (95％CI:0.6-0.9),sensitivity of 81.8％ (95％CI:48.2％-97.7％) and specificity of 62.3％ (95％CI:42.2％-78.2％).Conclusions There are four neutrophil subsets in the peripheral blood of neonates and autoactivation of neutrophils may exist.With the onset of sepsis,neutrophil subsets react differently between preterm and term infants.The proportion of early-stage neutrophils may be correlated with the severity of neonatal sepsis,which may have a predictive value for severe sepsis.