Objective To investigate the optimal extraction technique and antioxidant activity of curcuminoids from Curcuma longa L. Methods The optimized extraction process of curcuminoids from Curcuma longa L. with ethanol as extracting solvent were studied based on orthogonal test.The effect of each factor such as the ethanol concentration, the amount of solvent, extraction temperature and extraction time were investigated based on a single-factor experiment.The effects of extraction yield of curcuminoids under different processing conditions were compared by HPLC method. Content of curcuminoids and antioxidant activity of the extract were used as comprehensive survey index. Free radical scavenging DPPH method was used to assess the antioxidant effect. And the extraction technique parameters were optimized. Results The condition of optimal extraction technique was: ethanol concentration 60%, amount of solvent 20 folds, extraction time 2 h, extraction temperature 90 ℃. Under these conditions, the extraction yield of curcuminoids was 3. 65%. DPPH free radical scavenging yield of the curcumin extract was 79.79%. Conclusion This optimized technology is simple and have a good reappearing rate, and the extracts of Curcuma longa L.show good antioxidant activity.

The method was established for determination of geniposidic acid, chlorogenic acid, pinoresinoldiglucoside, which are three kinds of constituents of Eucommia ulmoides absorbed into the blood components. LC-MS/MS technique was applied to determine the blood components of the bloodstream after administration of E. ulmoides extract. At the same time, HPLC was used for detection of the ingredients content of the blood sample from 23 batches of E. ulmoides. The results showed that geniposidic acid, chlorogenic acid and pinoresinoldiglucoside are prototype into the blood in rats after oral administration of E. ulmoides extract, The linear range of geniposidic acid, chlorogenic acid and pinoresinoldiglucoside was good, and the average recoveries geniposidic acid, chlorogenic acid and pinoresinoldiglucoside were 98.69%, 100.8% and 98.39%, respectively. The method is simple and feasible with good reproducibility.
Animals ; Chlorogenic Acid ; blood ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Eucommiaceae ; chemistry ; Glycosides ; blood ; Iridoid Glucosides ; blood ; Lignans ; blood ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

AIMTo observe the differences of hemodynamics and nitric oxide synthase(NOS) activity of ventricular cardiac muscle in two septic shock models and explore the possible mechanism.

METHODSTwo rat models of septic shock[lipopolysaccharide(LPS)-induced and cecal ligation and puncture (CLP)-induced septic shock] were used. The hemodynamic parameters and nitric oxide synthase activity of ventricular cardiac muscle were measured.

RESULTSThe hemodynamic parameters in CLP-induced model were increased in the early stage and decreased in the late stage while in LPS-induced model the parameters showed the same change of the CLP late stage. Both LPS model and CLP model (late stage) showed significant increase in NOS activity, but there was no difference between the two models. After treatment of the NOS inhibitor N-nitro-L-arginine methyl ester (L-NAME), the parameters of CLP-late stage and LPS model increased significantly. The NOS activity reached the highest level in the CLP-middle stage. The production of nitrite/nitrate decreased significantly in LPS model and CLP model(late stage) after treatment of L-NAME, but the nitrite/nitrate produced by constitutive NOS in LPS model was higher than CLP model(late stage).

CONCLUSIONThe increase of the NOS activity may be the main reason to lead to the depression of the hemodynamic parameters. Inducible NOS may play the leading role in the LPS model while cNOS and iNOS have the same effect in the CLP model.

Animals ; Hemodynamics ; Lipopolysaccharides ; Male ; Myocytes, Cardiac ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rats ; Rats, Sprague-Dawley ; Shock, Septic ; classification ; metabolism

0.05).There was significant difference in the 3-year survival rate between A and C group. Conclusions The 3-year survival rate was dramatically increased with combined therapy mainly by cisplatin, the dose of 60～80mg is tolerant for the elderly aged above seventy years, and perioperation complications can be cured.

Objective To investigate SLC25A13gene mutation in neonatal intrahepatic cholestasis caused by citrin defi-ciency (NICCD). Method A total of 17 children with NICCD were collected. PCR-RFLP method was used to analyze the most common eight mutations of SLC25A13 gene in Chinese populations and results were analyzed together with routine laboratory examinations. Results In the 17 NICCD patients, there were six cases of homozygous mutation, three cases of compound heterozy-gous mutation and eight cases of single heterozygous mutation in SLC25A13 gene. Three kinds of mutations detected were 851del4 (73.1%), 1638ins23 (11.5%) and IVS6+5G>A (15.4%). The seventeen cases showed classical NICCD symptoms of low birth weight, pathological jaundice. And laboratory data suggested liver dysfunction, hyperbilirubinemia, hyperbileacidemia, hy-poproteinemia, hypoglycemia, coagulation disorders, hyperlactacidemia and hyperammonemia. Conclusions 851del4, 1638ins23 and IVS6+5G>A are hot spots of SLC25A13 gene mutation in Chinese populations. PCR-RFLP is a rapid, convenient and reliable technology for NICCD molecular diagnosis.

Objective To observe prospectively and systematically the effect and safety of rhendostati injection (endostar) combined with FOLFOX as a second line chemotherapy for advanced/metastatic colorectal cancer. Methods 23 patients with histological confirmed advanced/metastatic colorectal cancer after first line chemotherapy failure were observed. The dosage of 15 mg/time of endostar solved in 500ml normal saline was slowly intravenously dropped 4 h from day 1 to day 14. Oxaliplatin 85 mg/m~2 iv 2-3 h dl, d15. CF 200 mg/m~2 iv 2 h followed by 5-Fu 400 mg iv bolus and 5-Fu 600 mg/m~2 iv 22 h dl-2, d15-16 were given, every 4 weeks as one cycle. Efficacy was evaluated after 2 cycles according to RECIST criteria. Results 23 cases had been completed totally 56 cycles. Among 23 cases, 8 cases were PR, 12 cases SD, and 3 cases PD. The objective response rate (RR) was 34.8 % (8/23), and the disease control rate (DCR) was 87.0 % (20/23). The median time to progression was 7 months. The 1-year survival rate were 50.0 %. The 2-year survival rate was 40.0 %. The occurrence rate of G3/4 toxicities was low, including neutropenia(21.7 %), anemia(4.3 %), thrombocytopenia (13.0 %). Those toxicities were mainly related with the chemotherapy agents. Meanwhile transient electrocardiogram changes mild ST-T of changes occurred in 3 cases. 2 cases were mild hypertension and were symptomatically controlled. Conclusion There are better efficacies of endostar combined with FOLFOX chemotherapy for advanced/metastatic colorectal cancer, and it is low toxic and tolerable. It is worth of further clinical observation. More experiences need to be accumulated.

Objective To investigate the differences in clinical efficacy and safety between thrombolysis followed PCI (percutaneous coronary intervention) and primary PCI in patients with acute STEMI (ST elevation myocardial infarction). Methods A total of 215 STEMI patients who visit our clinic within 12 h since onset of their symptoms from May 2013 to January 2015 were enrolled. All eligible patients were divided into Early PCI group(n=68) and pPCI group (n=147) based on whether or not they received injection of recombinant human prourokinase thrombolytic therapy before their visit. Immediate TIMI (Thrombolysis In Myocardial Infarction) flow grade of infarct-related artery (IRA) before and after PCI treatment, post?operative CTFC (Corrected TIMI Frame Count) and TMPG (TIMI myocardial perfusion grade) were compared between these two groups. The incidence of bleeding during hospital stay , left ventricular function at 6 month after intervention and major adverse cardiac events (MACE) were all observed. Rusults There is no obvious difference between the baseline of two groups. Before PCI, the proportion of TIMI grade 2-3 was higher in Early PCI group (77.9%vs 20.4%,P<0.05)than that in pPCI group;but there was no significant difference in the proportion of TIMI grade 2-3 between these two groups after PCI (P>0.05). CTFC and peak value of serum CK-MB were lower [(27.7 ± 5.0) vs (32.6 ± 7.1), P<0.05;(225.8 ± 108.3) U/L vs (283.4 ± 110.6) U/L, P<0.05] and rate of TMPG 3 is higher (82.4%vs 68.7%, P<0.05)in Early PCI group than those in pPCI group. No significant difference was found in the incidence of bleeding and MACE during hospital stay and Left ventric?ular function at 6 months after operation between these two groups. By contrast, LVEFs were higher while LVEDds (LVED diameter) were lower after 3 and 6 months of the intervention compared to those before intervention in both groups (P <0.05). Conclusion It is a safe and effective reperfusion strategy for STEMI patients to receive rhPro-UK thrombolytic thera?py followed early PCI as an alternative way to those who failed to receive pPCI on time. It didn′t increase the occurrence of bleeding complications and MACE, and at the same time it presented the same benefit in improving recent cardiac function as pPCI did.

Objective To investigate mechanism involved in the differentiation of FBL-3 cells(eryth-roleukemia cells)induced by dendritic cells(DC). Methods To cultural the FBL-3 cells together with the DC supematant of different concentration for 72 hours,then we used the Wright's staining methed to record the ma-ture monocyte cell population, the transmission electron microscope to observe the ultramicrostructure, the flow eytoraetry to detect the expression rate of the surface molecular CD14. Results The Wright's staining methed, the transmission electron microscope and the flow cytometry all presented that after being induced by the super-natant of the DC ,the FBL-3 cells can partly differentiate into monocytes,inversion monocyte consistent with the personal characteristics. And the intensity of the DC supernatant was connected with interleukin-12. Conclusion The DC supernatant can induce the FBL-3 cells into monocytoid cell, the differentiated cells correspond to monocyte in macro-appearance,uhramicrostructure and phenotype. The competence of the DC supematant are partly concerned with interleukin-12.

Objective To develop a nylon membrane chip for rapid and systematic detection of the diabetes-associated 45 mutant loci in mitochondrial DNA(mtDNA).Methods The mutant-and wild-type probes were designed for detection of 45 mutant loci in mtDNA with Primer Premier 5.0 and NCBI BLAST softwares and the 90 probes with 8 poly T were immobilized on the Hybond N~+ nylon membranes which were treated with 5?SSC Buffer by UV-crosslinking;Then asymmetric PCR was employed to obtain the target single strand DNA(ssDNA).The PCR products were labeled with biotin after purification.NBT/BCIP was used as substrate that yields a very intense purple signal followed by AP-avidin,and the signals were observed in 24 samples with known sequences to evaluate the chips,each sample was repeatedly measured three times.Results The specific target fragments of 45 loci can be amplified under the same condition with nine sets of primers.The annealing temperatures of the wild-type [(59.01?1.42)℃] and mutant-type [(59.34?1.29)℃ ] probes are so close(t=1.046,P =0.301)that hybridization can be performed at the same temperature.The spots on the membrane chip are distinct,regular and well-distributed.The results of positive-and negative-control are perfect.The signals of negative probes and the background are similar.The results of chip were nearly concordant with that of DNA sequences(?~2=113.132,Kappa value =0.888,P = 0.000)except for T16189C mutant.Conclusions We have successfully developed a nylon membrane chip for rapid and systematic detection of the diabetes-associated 44 mutant loci in mtDNA.It could be used for screening for diabetic patients and high-risk people.

Objective To investigate the relationship between the genetic predisposition to type 2 diabetes mellitus(T2 DM)and mitochondrial DNA base variants in elderly patients.Methods PCR restriction fragment length polymorphism(PCR-RFLP)analysis was used to screen base variants at position 3243 and 3426 of mitochondrial DNA in 186 elderly cases with T2 DM and 170 healthy controls,and DNA sequence was confirmed.Results No carrier of 3243 A→G variant or 3426 A→G variant was found in both groups,however there was 1 case with 3290 T→C variant in diabetic group.Conclusions No significantly association was found between mitochondrial DNA 3243 or 3426 base variants and the predisposition of type 2 diabetes mellitus in elderly patients.