The research and quality control of traditional Chinese medicine has meaningful importance, because it has influence not only in the health and treatment of patients, but also in the solid growth and development of phar-maceuticals companies. In some cases, for the complex of TCM, the common QC method on single or multi-target compounds can't really and truly disclose the quality of the Chinese materia medica. Therefore, a lot of researchers do plenty of works to make clear the effectiveness basis, to improve the quality and realize the modernization of TCM. All of these works close together with modern analysis and separation technology. In this article, a novel analy-sis technology-UltraPerformance Convergence Chromatography (UPC2) based its characters and applications should be introduced. It should be a helpful technology for the TCM researchers to facilitate the study and QC works.

ObjectiveIn order to investigate anti-ageing mechanisms of the notoginsenoside Rg1,we used Aβ_(1-42) and D-galactose to establish aging rat model. Methods Ninety rats were divided into three groups at random: sham group, model group, treatment group. Aging rat models were established by injecting peritoneally D-galactose (100 mg/kg) to the rats for 56 days and after 35 days aggregated Aβ_(1-42)(μg) was injected to the right lateral ventricle of rats. Meantime, rats were treated by intragastric administration the notoginsenoside Rg1. Then spatial memory of experimental rats was examined with the Morris water maze(MWM). The thiol antioxidants including glutathione reductase (GR) and glutathione peroxidase (GSH-Px) activities were examined by colorimetric method. The concentration of the pro-caspase-3 and Bcl-2 were examined by the immunohistochemistry and Western blotting method. Results In aging model rats escape latercies were significantly prolonged (P<0.05), while decreases were seen in the time of staying the third quadrants of platform, the number of crossing over a platform, the concentration of the GR, GSH-Px, and pro-caspase-3 as compared with the sham group(P<0.05). After treatment of the notoginsenoside Rg1, the aging model rats exhibited significant increases in the time of staying the third quadrants of platform, the number of crossing over a platform, the concentration of the GR, GSH-Px, and pro-caspase-3(P<0.05), while a decrease was observed in escape latercies as compared to control group(P<0.05). Moreover there was no significant difference in the expression of the Bcl-2(P>0.05). Conclusion The results from our study indicate that the notoginsenoside Rg1 could improve the oriented learning and memory capacity and prevent the neurodegeneration of central nervous systems in aging model rats by up-regulating the expression of the thiol antioxidants(including GR and GSH-Px) and resisting the cleavage of the pro-caspase-3.

This study was aimed to develop the method for determination of mannitol and meglumine in diterpene lactones of Ginkgo Injection. The HPLC-ELSD was used to determine mannitol and meglumine in diterpene lactones of Ginkgo Injection. The analysis was carried out on Waters XBridge Amide (4.6 mm í 150 mm, 3.5 μm) column. The mobile phase was the mixture of acetonitrile, and 50 mmol·mL-1 ammonium acetate with a linear gradient elu-tion at a flow rate of 1.0 mL·min-1. The column temperature wad maintained at 30℃. The operating conditions of the ELSD were a nebulizer nitrogen with the flow rate of 2.0 L·min-1 and an evaporator tube at the temperature of 90℃. The results showed that mannitol had a good linear relationship in a range of 1.9665 μg - 19.665 μg. The average recovery rate was 100.57% (RSD = 0.92%, n = 9). The meglumine showed a good linear relationship in a range of 0.4838 μg - 4.638 μg. The average recovery rate was 100.67% (RSD = 0.72%, n = 9). It was concluded that this method was simple and accurate with good reproducibility. It met with the requirement of the determination of the contents of mannitol and meglumine in diterpene lactones of Ginkgo Injection.

Objective To evaluate the incidence and to investigate risk factors of supraventricular arrhythmia (SVAs) in postoperative cancer patients in intensive care unit ( ICU ). Methods Data of 570 patients consecutively admitted to oncologic surgical ICU of Cancer Hospital of Chinese Academy of Medical Sciences from Nov. 2008 to Oct. 2009 were retrospectively collected. Univariate and multivariate logistic analysis were conducted for potential factors that influenced SAVs. Results Thirteen patients with a history of atrial fibrillation (AF) were excluded and 557 patients were eligible for the study. SVAs occurred in 72 patients ( 12. 93% ). Multivariate analysis showed four independent predictors of SVAs including age ( OR = 1. 066,95%CI: 1. 034 - 1. 099,P ＜0. 001 ) ,a history of coronary heart diseases ( OR = 2. 644,95% CI: 1. 459 - 4. 790,P ＜ 0. 05), sepsis ( OR = 2. 374,95% CI: 1. 098 - 5. 135, P ＜ 0. 05 ) and intra-thoracic procedure ( OR =2. 322,95 % CI: 1.061 - 5.084, P ＜ 0. 05 ) . ICU length of stay, severity ( APACHE Ⅱ scores in SVAs patients) were significantly greater in patients who were not affected by SVAs ( ICU stay: [2 ( 1 ～ 77 )]vs [3 ( 1 ～ 40 )]days,P ＜ 0. 001; APACHE Ⅱ score: [9 (0 ～ 37 )] vs [11 (3 ～ 38 )], P = 0. 001 ). Nine cases died in SVAs patients ( 12. 5% ) and 19 died in the non-SVAs patients (3.9%), with significant difference between the two groups( x2 = 9. 673, P = 0. 002). Conclusion In oncologic surgical ICU, the incidence of SVAs is high. Age,history of coronary heart diseases, sepsis and intra-thoracic procedure were independent rsik factors of SVAs. SVAs prolong ICU length of stay. SVAs is a marker of critical illness severity.

Objective To explore the activition and polarization of microglia in the epileptic rats induced by lithium chloride-pilocarpine. Methods One hundred male SD rats were randomly divided into five groups: control group and different time points model groups including 1d,3d,7d and 14d. Epilepsy models were established by lithium chloride-pilocarpine intraperitoneal injection. The control group was given the same dosage of normal saline. The morphology change was detected by immunofluorescence,and the expressions of iNOS and Arg-1 were determined by IHC at respective time points. Results Compared the model groups with control group,microglia was activated,synapsis was shorten,volume got bigger,most of them seemed as amoebocyte,the expression of iNOS increased and Arg-1 decreased,especially at 3d.ConclusionThe results from this study indicated that microglia was activated and polarized in epileptic rats induced by pilocarpine.

This study was aimed to identify pET21b-HPV16E2/BL21(DE3) strain and to optimize the expression of human papillomavirus type 16 (HPV16) E2 protein by orthogonal analysis. Four influence factors on two levels were selected to increase the target protein quantity. The four factors were induction time, induction temperature, inductor concentration and cell density. The quantity of HPV16 E2 protein was used as the evaluation parameter. Induced by IPTG, HPV16 E2 protein was analyzed by SDS-PAGE and Western Blot. Target protein was analyzed by GIS imaging system to quantify the protein level. SPSS13. 0 software was applied to analyze the result. Data showed that the expression strain pET211rHPV16 E2/BL21(DE3) was identified correctly. HPV16 E2 protein expressed mainly at insoluble form. The 42KD protein band was identified by SDS-PAGE and Western blot. Orthogonal test was applied on influence factor analysis and expression optimization successfully. Main influence factors were inductor concentration and induction temperature. The optimimum condition of maximum expression quantity was 37 degrees C, 7h, 1.0 mmol/L IPTG and OD600 1.0. In this experiment, orthogonal test could not only be used to analyze the influential factors and promote the target protein expression, but also be used to provide a better experiment method for molecular biological study.
DNA-Binding Proteins ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Human papillomavirus 16 ; metabolism ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Papillomavirus Infections ; virology ; Recombinant Proteins ; biosynthesis ; genetics

Objective : To investigate the effect and mechanism of exosome ( Exo) transported miR⁃223 on brain tissue injury and microglial activation in rats with traumatic brain injury ( TBI) . Methods : The miR⁃NC plasmid and miR⁃223 mimic plasmid were transfected into HEK293 cells by liposome method , and the expression level of miR⁃223 in the cells was determined by quantitative real⁃time PCR . Exo was extracted from transfected HEK293 cells and identified by transmission electron microscopy , nanoparticle tracking analysis and Western blot , the expression level of miR⁃223 in Exo was determined by quantitative real⁃time PCR . Forty SD rats were randomly divided into sham group , model group , NC⁃Exo group and miR⁃223 ⁃Exo group , with 10 rats in each group , TBI model was prepared by modified Feeney free fall method in all groups except sham group , rats in NC⁃Exo group and miR⁃223 ⁃Exo group were injected with cell⁃derived Exo transfected with miR⁃NC plasmid and cell⁃derived Exo transfected with miR⁃223 mimic plasmid via tail vein , respectively . Two weeks later , hematoxylin⁃eosin (HE) staining was used to observe the pathological changes of brain tissue in each group , Nissl staining was used to detect the changes and distribution of Nissl bodies in each group , enzyme⁃linked immunosorbent assay (ELISA) was used to measure the serum levels of tumor necrosis factor⁃α (TNF⁃α ) , interleukin⁃1β (IL⁃1β) and interleukin⁃6(IL⁃6) , immunofluorescence double staining was used to observe the expression of nod⁃like receptor family pyrin domain containing 3(NLRP3) and ionized calcium binding adaptor molecule 1 (Iba⁃1) , Western blot was used to detect the protein expression of NLRP3 , apoptosis⁃associated speck⁃like protein containing( ASC) and Caspase⁃1 . Results: After transfection , compared with control group and miR⁃NC group , the relative expression of miR⁃223 in miR⁃223 group significantly increased (P < 0. 05) . The isolated particles had typical Exo morphology , the peak particle size was about 120 nm , the Exo marker proteins CD9 , CD63 and CD81 were significantly overexpressed , and the relative expression of miR⁃223 significantly non of brain tissue in the miR⁃223 ⁃Exo group was improved , the morphology and number of Nissl bodies were re⁃increased (P < 0. 05) . Compared with the model group , the damage phenome stored , the levels of TNF⁃α , IL⁃1β and IL⁃6 in serum decreased ( P < 0. 05) , the intensity of NLRP3 and Iba⁃1 fluorescence staining in brain tissue decreased (P < 0. 05) , the relative protein expressions of NLRP3 , ASC and Caspase⁃1 in brain tissue were down⁃regulated (P < 0. 05) . Conclusion Exo operation of miR⁃223 can significant ly improve brain tissue injury and inhibit microglial activation in TBI rats , which may be related to the inhibition of NLRP3 .

Chronic constipation is a common gastrointestinal disease severely affecting the patient׳s quality of life. The traditional treatment of constipation is the use of laxatives. Recently, several new drugs including lubiprostone, linaclotide and prucalopride have been approved for treatment of chronic constipation. However, a significant unmet medical need still remains, particularly among those patients achieving poor results by current therapies. The 5-HT4 receptor modulators velusetrag and naronapride, the guanylate cyclase C agonist plecanatide and the ileal bile acid transporter inhibitor elobixibat are recognized as the most promising drugs under investigation. Herein, we give a comprehensive review on the pharmacological therapeutics for the treatment of chronic constipation, with the purpose of reflecting the drug development trends in this field.

OBJECTIVETo identify risk factors associated with overall survival (OS) for patients undergoing primary hepatic resection for metastatic colorectal cancer.

METHODSThe clinical and pathological data were prospectively collected from 191 consecutive patients undergoing primary hepatic resection for colorectal liver metastases from January 2000 to August 2012. The survival curve was drawn by Kaplan-Meier method, and the survival rates were analyzed by Log-rank test. Parametric survival analysis was used to identify predictors of cancer-specific survival.

RESULTSThe 5-year overall survival were 38.4% and median survival time was 33 months; 5-year disease-free survival were 23.6%, and the median disease-free survival time was 10.0 months. 5-years survival rate was significantly lower in patients with synchronal metastasis than in patients with heterochronia metastasis (27.4% vs. 51.8%, χ(2) = 6.527, P < 0.05). In overall survival, univariate analysis found 7 risk factors: gender (χ(2) = 5.219), N stage of the primary tumor (χ(2) = 5.591), bilobar metastases (χ(2) = 4.269), number of metastases ≥ 2 (χ(2) = 5.051), disease-free interval ≥ 6 months (χ(2) = 6.527), carcinoembyonic antigen level ≥ 30 µg/L (χ(2) = 4.454), and extrahepatic disease (χ(2) = 5.158). On multivariate analysis, 3 risk factors were found to be independent predictors of poor survival: N stage of the primary tumor (RR = 2.198, 95%CI: 1.146-4.216), disease-free interval ≥ 6 months (RR = 1.840, 95%CI: 1.139-2.973), carcinoembyonic antigen level ≥ 30 µg/L(RR = 1.854, 95%CI: 1.056-3.255).

CONCLUSIONSResection of liver metastases provides good long-term cancer-specific survival benefit. N stage of the primary tumor, disease-free interval, carcinoembyonic antigen level are important prognostic factors for colorectal liver metastasis.

Adult ; Aged ; Aged, 80 and over ; Carcinoembryonic Antigen ; blood ; Colorectal Neoplasms ; pathology ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Liver ; surgery ; Liver Neoplasms ; secondary ; surgery ; Male ; Middle Aged ; Prognosis ; Risk Factors ; Survival Rate ; Young Adult

In clinical application, a microneedle system that continuously delivers drugs is of great value for the delivery of some vaccines and hormone drugs. In this study, a controlled-release chitosan-based microneedle array (PVA/CS-MN) was designed, combining microneedle patches with drugs for controlled-release of drugs. Here we report the optimization of the preparation process of PVA/CS-MN. The appearance, morphology, mechanical properties, dissolution and swelling properties, and in vitro penetration properties of the MN arrays were characterized. The PVA/CS-MN prepared by the optimal process showed good morphology and mechanical properties. PVA/CS-MN can smoothly open microchannels on the skin and achieve controllable dissolution and swelling functions. Ascorbic acid (l-ascorbic acid) was used as a model drug to prepare a Vc-PVA/CS-MN. In vitro transdermal diffusion experiments showed that the Vc-PVA/CS-MN released about 57% of the drug within 1 h. About 66.7% of the drug was slowly released within 12 h, and a total of 92% of the drug was released after 7 days. The controllable sustained-release properties and excellent drug delivery efficiency of PVA/CS-MN provide a new option for sustained transdermal drug delivery.
Ascorbic Acid ; Chitosan ; Delayed-Action Preparations ; Drug Delivery Systems ; Hormones ; Vaccines