Objective: To establish the determination method of adenosine in Compound Houtou Capsule. Methods: HPLC was performed to determine adenosine on DISCOVERY column, using methanol-0.05 mol/L potassium dihydrogen phosphate solution (13∶87,v∶v) as a mobile phase and detection wavelength at 259nm.Results: The linearity of this method was good, and the average recovery was 100.72%, RSD =0.22%. Conclusion: This method was simple and accurate, and available for the quality control of Compound Houtou Capsule.

Objective To investigate the association between genetic polymorphisms of exon3 of nicotinic acetylcholine receptor ?4 subunit (CHRNA4) and sporadic Alzheimer's disease(SAD).Methods In 23 SAD patients and 30 people of normal control, exon3 of CHRNA4 was screened by PCR-DGGE and DNA sequencing.Results Three new genetic polymorphisms were found in the exon3 of CHRNA4: C104T, A136G, G169A.The frequnces of these three genes in SAD group were 35%,46%,61% respectively and in control group were 13%,13%,33% respectively. There were significant difference between two groups (all P

BACKGROUND:Compared with traditional hydrogels, intel igent hydrogels can appear to exhibit different responses to different stimuli such as temperature, pH value, light, and magnetic field, produce two-stage structure and alter chemical structure to generate the ordered supramolecular structure by self-assembling, and ultimately cause the formation of the gel with three-dimensional structure.
OBJECTIVE:To review the research status of intel igent hydrogels and its application in tissue engineering.
METHODS:A computer-based search of CNKI and PubMed databases was performed to retrieve articles addressing intel igent hydrogels in tissue engineering published before 2014. The keywords were“hydrogel, tissue engineering”in Chinese and English.
RESULTS AND CONCLUSION:Intel igent hydrogels are classified into temperature sensitivity, pH sensitivity, photosensitivity, magnetic susceptibility and temperature/pH dual responsive hydrogels. The change of the external environment can be automatical y detected and responded. Intel igent hydrogels exhibit a series of outstanding performances in drug delivery systems, drug delivery, repair and improvement of defected tissues, which are not possessed by traditional materials. In particular, the intel igent hydrogels show considerable superiorities in tissue engineering, including low immunogenicity that reducing inflammation and rejection, biodegradability, realizing the three-dimensional scale simulation of cel microenvironment that is conducive to cel adhesion, growth, migration and differentiation.

Object To study the different fingerprints of hydrophilic and hydrophobic components in roots of Salvia miltiorrhiza Bge. collected from different habitats, and discovery the relationship between the two types of components. Methods The fingerprints were detected by RP-HPLC and the results were analyzed by SPSS software. Results Roots of S. miltiorrhiza collected from different habitats showed different fingerprints of hydrophilic and hydrophobic components, there is no obvious relationship between the two types of components. Conclusion In order to control the quality of 5. miltiorrhiza roots that is used to manufacture injections, we must assay the hydrophilic components, such as salvianolic acid B should be determinded.

Objective To prepare a thrombus-targeted ultrasonic contrast agent and to investigate its targeted ability to fresh blood clots. Methods We first synthesized FITC-KGDS-Palm compound, and then prepared thrombus-targeted microbubbles using "ultrasound & high speed shearing method".Fluorescence labeling thrombus-specific peptides and KGDS,directed at the activated glycoprotein(GP)Ⅱb/Ⅲa receptor of platelets were attached to the surface of lipid microbubbles. The concentration and size of TUCA were measured by Malvern Zeta Sizer Nano-ZS590 and Coulter counter.Immunofluorescence was applied to confirm the conjugation.The conjunct ratio was assessed by flow cytometer (FCM).Results The KGDS-TUCA was straw yellow turbid liquor,and the concentration was 1.5×10~9/mL,and the average size was 1.5 μm. The targeted microbubbles conjugated with the thrombus-specific peptides showed bright green rings by fluorescence microscope.FCM demonstrated that the wavelength of shell of KGDS-TUCA changed greatly,and the conjunct ratio was 90.04%.In vitro study showed KGDS-TUCA remained stable for 48 h at 4 ℃ and target-attached to blood clots and showed good stability.Conclusion The ultrasound & high speed shearing method to prepare TUCA is easy and in favor of purification.KGDS-TUCA has high specific biological activity.The conjunct ratio and stability of KGDS-TUCA are excellent.

Objective To investigate specific immune responses in mice induced by recombinant major outer membrane protein(rMOMP)of C.trachomatis serovaf E.Methods Thirty-six female BALB/cmice aged 3 to 4 weeks Were divided into three groups.i.e.,adjuvant group vaccinated、with purified rMOMP and Freund's adjutant,solitary group vaccinated with rMOMP only and control group vaccinated with phosphate buffered saline(PBS).All the mice were intramuscularly vaccinated on week 0,2 and 4.Blood samples and vaginal washes were obtained from these mice on week 6,then,mice were challenged with elementary body(EB)of C.trachomatis serovar E at the footpad followed by the observation of delayed hypersensitivity.On week 7.mice were genitally infected with C.trachomatis EB;one week later,blood samples and vaginal washes were obtained again;six weeks later,spleen lymphocytes were isolated from the mice and stimulated bv C.trachomatis or ConA followed by the detection of cell proliferation with MTT assay.In vitro neutralization assay was also performed.ELISA was used to determine the titers of Chlamydia-specific IgO antibody in sera and IgA antibody in vaginal washes,as well as the level of IFN-γ in culture supernatant of lymphocytes and sefa of mice.Vaginal swabs were collected after genital challenge and subjected to C.trachomatis culture.Results The absorbance at 405 ms of Chlamydia-specific IgG antibody and proliferation index of lymphocytes were 0.641±0.059 and 5.085±1.291.respectively,in mice immunized with rMOMP and Frennd's adjuvant.significantly higher than those in mice immunized with rMOMP only(0.424±0.015 and 3.123 ±0.840.both P<0.05).The thickness of right hind footpad increased by 0.324±0.054 mm and 0.272±0.064 mm,respectively,in solitary group and adjuvant group,respectively,with significant difference between the two groups(P<0.05).A significant increase was also observed in the adjuvant group compared with the control group in the above three parameters(all P<0.01).Conclusion The rMOMP of C.trachomatis could efficiently induce Chlamydia-specific humoml and cellular immune responses in mice.

Objective To disscuss and analyze the clinical effects of whole body mild hypothermia on MODS caused by cerebral hemorrhage.Methods One hundred and eighty patients with MODS caused by cerebral hemorrhage during the period of hospitalized from January 2013 to December 2015 in ICU of our hospital were divided into two groups (observation group and control group) randomly,90 cases patients in each group.Patients in control group were treated with conventional treatment intervention,patients in control group were treated with whole body mild hypothermia on the base of conventional treatment intervention,the score of MODS,the ratio of,VO2 and DO2,the time during the period of hospitalized in ICU,the time of mechanical ventilation,epilepsy incidence and mortality,NIHSS score and PADL score of the patients in these two group after treatment were compared.Results After the comparion,the score of MODS and the state of oxygen consumption and oxygen supply of the patients in observation group were better than the patients in control group,there was significant differences,and had statistical significance (P＜0.05);the time during the period of hospitalized in ICU and the time of mechanical ventilation of the patients in observation group were shorter than the patients in control group,there was significant difference,and had statistical significance (P＜0.05);the mortality were reduce significantly of the patients in observation group than the patients in control group,there was significant difference,and had statistical significance (P＜0.05);the epilepsy incidence of the patients in observation group and control has no significant difference,has no statistical significance (P＞0.05);the NIHSS score and PADL score of the patients in observation group were higher than the patients in control group,there was significant difference,and has statistical significance (P＜0.05).Conclusion The clinical effects of whole body mild hypothermia on MODS caused by cerebral hemorrhage have important significance,can effectively improve the balance of multiple organ dysfunction syndrome patients,can delay the progression of multiple organ dysfunction syndrome patients,improve the prognosis,it is worthy of clinical application and promotion.

Objective To analyze the epidemiological characteristics of Klebsiella pneumoniae car-bapenemase(KPC)-producing Escherichia coli(E. coli)strains isolated in Hangzhou,China. Methods A total of 25 KPC-producing Escherichia coli strains were collected from four hospitals in Hangzhou from July 2012 to January 2014. Antibiotic susceptibility of the isolates to 22 common antimicrobial agents was deter-mined by using Kirby-Bauer(K-B)disk diffusion method. PCR analysis and gene sequencing were used for bla KPC gene screening. The modified Hodge test was performed to detect the production of carbapenemase. Pulsed-field gel electrophoresis(PFGE)and multi-locus sequence typing(MLST)were used for homology analysis. Results All of the 25 clinical isolates were confirmed to be KPC-producing E. coli strains,harbo-ring the blaKPC-2 gene. These KPC-producing isolates showed high drug resistance rates and were resistant to almost all β-lactam antibiotics. PFGE typing classified the 25 isolates into three main homologous clone groups,including clone group A(4 isolates),clone group B(5 isolates)and clone group C(2 isolates), and some single clones(14 isolates). MLST typing classified the isolates into eight ST types,including ST131(14 isolates),ST167(3 isolates),ST2003(3 isolates),ST410(1 isolate),ST457(1 isolate), ST1463(1 isolate),STnew1(1 isolate)and STnew2(1 isolate). The typing results of PFGE and MLST were consistent with each other. Conclusion The prevalent KPC-producing E. coli strains in Hangzhou, China were ST131 type,which were resistant to multiple antibiotics and had been detected in several hospi-tals. The epidemic of KPC-producing E. coli strain often occurred at some special wards,such as Intensive Care Unit(ICU)and emergency ICU.

Objective To extract,isolate and identify the polysaccharide from ginseng,and to investigate its anti-tumor activity in vitro. Methods The ginseng polysaccharide was obtained through water extraction and ethyl alcohol deposition method. Use the Sevage method to remove the protein in the crude polysaccharide. The structural characteristics of the polysaccharide were determined by FT-IR spectra.The RM-1 and HeLa cells were divided into control group and different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )of ginseng polysaccharide groups. The survival rates of the cells in various groups were detected by MTT method. The indirect killing effects of ginseng polysaccharide with different concentrations (0,50,100,200,300,400 and 500 mg·L-1 )on the cancer cells were determined by CTL test. Results The extraction rate of ginseng polysaccharide was 8.7 6% and the structural characteristics demonstrated that the main component of the extract was polysaccharide.The result of MTT showed that there were no significant differences of the survival rates of RM-1 and HeLa cells between different concentrations of ginseng polysaccharide groups after treated for 24 h compared with control group (P>0.05).The result of CTL test showed that the cytototic LHD release rates in different concentrations of ginseng polysaccharide groups were increased significantly (P<0.05)compared with control group;with the increase of ginseng polysaccharide concentration, the cytotoxic LDH release rates were increased firstly and then were decreased.When the concentration of ginseng polysaccharide was 100 mg·L-1 ,the cytotoxic LDH release rate was the biggest.Conclusion Ginseng polysaccharide can indirectly inhibit the growth of the tumor cells by activating T cells and play an anti-tumor effect.

Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.