Objective To investigate changes in the expression of apurinic/apyrimidinic endonuclease (APE) and the oxidative DNA damage marker 8 OHdG in distant hippocampus regions of the rat brain after focal cerebral ischemia of the middle cerebral artery.Methods SD rats were divided into the sham surgery group and the pMCAO group (induced by middle cerebral artery occlusion).Pathological changes in brain tissues were examined at 2 h,6 h,12 h,24 h,48 h and 72 h.The expression of APE and 8-OHdG was measured by immunohistochemical staining methods.TUNEL staining was performed to detect apoptosis.Results Reduction of APE expression in the CA1 region of the hippocampus on the ischemia side appeared at 2 h in the pMCAO group and continued as ischemia persisted (F=11.91,P＜0.05).The expression of 8OHdG and TUNEL immunoreactivity in the CA1 region of the hippocampus on the ischemia side were first observed at 6h in the pMCAO group and intensified during the remainder of induced ischemia (F=9.23 and 10.46 respectively,P＜0.05 for both).Compared with the sham group,8-OHdG expression and TUNEL immunoreactivity in the pMCAO group were at nearly the same levels from 24 h to 72h.Conclusions Oxidative DNA damage occurs in hippocampus regions of the rat brain after experimentally induced focal cerebral ischemia of the middle cerebral artery.APE expression declines in regions distant from focal cerebral ischemia.Development and accumulation of oxidative DNA damage can induce apoptosis in certain brain regions.

This article proposes the "Efficacy Theory" hypothesis of the traditional Chinese medicines (TCMs): TCMs take effects and weaken toxicities through the additive effects of numerous effective forms (including their constituents or/and metabolites) on a same target, the synergistic effects based on the overall action of the additive effects on individual targets and their toxicities scattering effects. A TCM may include approximately 1000 constituents and each constituent may produce about 100 metabolites in vivo after oral administration. Numerous effective forms of incalculable constituents and their metabolites could work like a "army group" together. When the quantity of a specific target molecule is larger than the pharmaceutical molecules, the molecules of different kinds of effective forms could combine with the target molecules successively, to exert the additive effects. When the target molecules are mostly occupied ("target most spaces occupied"), this TCM begins to work. The additive effects maybe exert not only in concentration but also in a time order way, which gives a sustained efficacy of TCM. The additive effects and the toxicities scattering effects are resulted from the same effective groups and not identical toxic groups among different effective form molecules. The "toxicities scattering effect" can be used to explain the non-toxic TCMs, but not fit for toxic TCMs. The efficacy theory showed that the variety of constituents and metabolites may participate in the process of pharmacodynamic actions, including the additive effects, synergy effects and toxicities scattering effects, which may be useful for explaining and developing the characteristic advantage of the TCMs. The questions we need to study or confirm are as follows: What are the TCMs' pharmacodynamic substance basis and mechanism made up of Why are toxicities of most TCMs' smaller How is the TCMs' "Efficacy Theory" which reflects characteristic advantage of TCMs applied in the research and development of new drugs.
Animals ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Humans ; Medicine, Chinese Traditional ; Pharmaceutical Preparations ; chemistry

Objective To establish a big animal model of secondary infection of acute necrotizing pancreatitis (ANP). Methods Thirty young pigs were allocated to experiment group ( n = 20 ) or control group (n = 10). The ANP model was induced by retrograde injection of a mixture solution of 5% sodium taurocholate and 5% trypsin (0. 5 ml/kg body weight) into the main pancreatic duct and ligation of the proximal end of the main pancreatic duct, and then the second step was injecting 3 ～ 4 ml living Escherichia coli (E coli) suspension (108/ml) to the necrotic area of the pancreas by fine needle aspiration technique under CT guidance in the experiment group, and by injecting 3 ～ 4 ml inactivated E coli in the control group using the same method. Multi-slice spiral CT dynamic enhanced scan was performed in both groups 1 day and 2 or 3 days after ANP modeling and 5 days after bacterial injection to calculate the CTSI score. Serum amylase, blood WBC count and blood bacterial culture was performed in both groups. 5 days later, the animals were scarified to observe the infected or necrosis foci, and perform smear, bacterial culture and pathologic examinations of the tissue around the infected or necrosis foci. Results The ANP secondary infection model was successfully established in 16 of the 20 animals in the study group, with a success rate of the 80.0% (16/20). There were 17 foci where the positive rate of bacterial culture was 100% (17/17 foci), and the success rate of blood bacterial culture was 68.8%(11/16). In the control group, the ANP model was established successfully in 7 of 10 animals (70%), except for one case of contamination, only one foci was identified;the positive rate of bacterial culture and the success rate of b|ood bacterial culture was 14.3% (1/7). Serum amylase and white blood WBC count increased with similar trends, WBC count in the study group was significantly higher than that in the control group (P<0.01). The mean CT severity index(CTSI) was all ≥4 in beth groups, indicating the severity was moderate to severe. Conclusions A stable and reliable model of secondary infection of ANP in big could be established satisfactorily by injecting active E. coli into the pancreatic necrosis tissue under CT guidance, which helps further pathogenic mechanism studies and clinical studies, especially imaging studies.

Acute Disease ; Adult ; Chromatography, Gas ; Female ; Humans ; Lung ; chemistry ; Phosphines ; analysis ; blood ; poisoning

To enhance the immunogenicity of DNA and adenoviral vector vaccines expressing HIV-1 subtype B gp160, human interleukin 15 (hIL15) DNA adjuvant (pVR-hIL15) was constructed. BALB/c mice received DNA prime/protein boost immunization with pVR-HIVgp160/Ad5-HIVgp160 alone or combined with pVR-hIL15. Cellular and humoral immune responses were evaluated by IFN-gamma enzyme-linked immunosorbent spot assay and enzyme-linked immunosorbent assay, respectively. Compared with those immunized with vaccines alone, the mice immunized with vaccines combined with pVR-hIL15 had significantly increased specific cellular response and antibody titer (P < 0.05). It suggests that the IL15 DNA adjuvant can enhance the immune responses induced by prime-boost regimen using DNA and adenoviral vector encoding HIV-1 subtype B gp160.
Adenoviridae ; genetics ; Adjuvants, Immunologic ; Animals ; Antibodies, Viral ; immunology ; Antibody Specificity ; Female ; Genetic Vectors ; genetics ; HIV Envelope Protein gp120 ; immunology ; HIV Envelope Protein gp160 ; genetics ; immunology ; HIV Envelope Protein gp41 ; immunology ; Humans ; Immunity, Cellular ; Immunity, Humoral ; Interleukin-15 ; genetics ; Mice ; Mice, Inbred BALB C ; Vaccines, DNA ; genetics ; immunology

To investigate me material basis of Mahuang Fuzi Xixin decoction (MFXD) for anti-inflammation and immune-suppression based on the combined method of serum chemical and serum pharmacological. The LC-MS/MS fingerprints of MFXD, drug-containing serum and blank serum were compared to define the components in plasma. Histamine, β-hexosaminidase released from RBL-2H3 cell infulenced by drug-containing serum at different time points were measured by ELISA. The effect of drug-containing serum on lipopolysaccharide-induced splenocyte proliferation at different time points were determined by MTT. A correlation analysis was made on components of MFXD and pharmacological indexes based the stepwise regression method. After the intragastrical administration with MFXD, 32 components were discovered in rat serum, including 27 prototype components (10 from Mahuang, 13 from Fuzi and four from Xixin) and five unknown components. Compared with blank serum, drug-containing serum could reduce the release of histamine from RBL-2H3 induced by antigen at different time points (P < 0.05); except the 4-hour drug-containing serum, all of the remaining drug-containing serums could inhibit the RBL-2H3 mastocyte degranulation induced by antigen at different time points (P < 0.05). Drug-containing serum could significantly lipopolysaccharide-induced mouse splenocyte proliferation at 15 and 30 min (P < 0.05). A regression analysis was made on the chemical data of components absorbed into blood and pharmacological indexes, i. e. release rate of histamine, release rate of β-hexosaminidase and inhibition rate of splenocyte. This suggested the close correlations among methyl pseudo-ephedrine, pseudoephedrine and histamine released from RBL-2H3 induced by antigen; pseudoephedrine, hypaconine, methyl pseudoephedrine and β-hexosaminidase released from RBL-2H3 induced by antigen; as well as benzoyl hypaconine, benzoylaconine, 14-benzoyl-10-OH-mesaconine, mesaconine and lipopolysaccharide-induced mouse splenocyte proliferation. Methylpseudoephedrine, pseudoephedrine, benzoyl hypaconine, benzoylaconine and mesaconine may be part of material basis of MFXD on anti-inflammation and immune suppression.
Animals ; Anti-Inflammatory Agents ; chemistry ; pharmacology ; Cell Degranulation ; drug effects ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Female ; Histamine ; immunology ; Immunosuppressive Agents ; chemistry ; pharmacology ; Male ; Mass Spectrometry ; Mast Cells ; drug effects ; immunology ; Mice ; Rats ; Rats, Wistar ; Serum ; chemistry

OBJECTIVETo develop a new injectable biomaterial carrageenan/nano-hydroxyapatite/collagen (nHAC/Carr) for bone surgery and characterize it.

METHODSnHAC/Carr was developed by filling carrageenan with nano-hydroxyapatite/collagen (nHAC) granules.

RESULTSIt was found that nHAC/Carr had similar X-ray diffraction patterns with that of nHAC. Fourier transform infrared spectroscopy showed that carrageenan did not change its structure in nHAC/Carr. The rheological behavior of nHAC/Carr was the same as carrageenan. The scanning electron microscopy and porosity analysis showed that nHAC/Carr had porous structure and its porosity was about 90%.

CONCLUSIONThe biomaterial nHAC/Carr may be used as an injectable bone substitute.

Bone Substitutes ; chemistry ; Carrageenan ; Collagen ; Durapatite ; Injections ; Microscopy, Electron, Scanning ; Nanoparticles ; Porosity ; Surface Properties ; X-Ray Diffraction

Objective To estimate the uncertainty of measurement in detecting of hepatitis B virus DNA(HBV DNA)by method of fluorescence quantitative polymerase chain reaction(FQ-PCR),and discuss the application value.Methods The process of the detection of HBV DNA by FQ-PCR was analysed to confirm and simplify the sources of uncertainties of measurement,which were obtained by disposing the data of methodology validation,internal quality control(type A evaluation of uncertainty)and external quality assessment(type B evaluation of uncertainty);combined uncertainty and expanded uncertainty were obtained by statistical methods.Results The main sources of uncertainties of measurement were:precision within laboratory,precision between laboratory,method bias.The expanded uncertainty of HBV DNA by FQ-PCR was U=0.62(k=1.96,n=2).The uncertainty caused by method bias was found mostly.Conclusion Expanded uncertainty can be compared in different results of HBV DNA by FQ-PCR,and it provides guide significance for observing the cure effect of anti-HBV and choosing the concentration of quality control.

OBJECTIVETo establish a method for GC fingerprint determination of the chemical constituents in Herba Asari.

METHODGC and GC-MS were used to optimize the fingerprint determination method, and identify the main peaks in the GC fingerprint.

RESULTA preferable method for GC fingerprint determination of the chemical constituents in Herba Asari was established.

CONCLUSIONA general acquaintance of the chemical constituents in Herba Asari can be obtained by using the preferable GC fingerprint determination method, which is useful for quality evaluation of the crude drug of Herba Asari.

Anisoles ; analysis ; Asarum ; chemistry ; classification ; Gas Chromatography-Mass Spectrometry ; methods ; Monoterpenes ; analysis ; Plants, Medicinal ; chemistry ; Quality Control ; Safrole ; analysis

This study is to develop an HPLC method for the simultaneous determination of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoisoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in Smilacis Glabrae Rhizoma. Samples of Smilacis Glabrae Rhizoma, Heterosmilacis Chinensis Rhizoma and Heterosmilacis Yunnanensis Rhizoma were separated on an Agilent Zorbax SB-C18 column with gradient elution of acetonitrile-0.05% phosphoric acid at a flow rate of 1.0 mL · min(-1). The detected wavelength was set at 230 nm and the column temperature was maintained at 35 °C. The contents of (-)-epicatechin, 5-O-caffeoylshikimic acid, neoastilbin, astilbin, neoisoastilbin, isoastilbin and engeletin in 84 Smilacis Glabrae Rhizoma samples were in the range of trace-1.381, trace-9.913, trace-3.673, 0.6706-27.08, trace-1.181, trace-4.833 and trace-2.754 mg · g(-1), respectively. The established method was used for analysis of common adulterants. The results demonstrated that the contents of (-)-epicatechin in Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma were 0.01163-0.06007 mg · g(-1) and 0.01583-0.08585 mg · g(-1), respectively, while the other six constituents were not detected. The method is simple and accurate, and can be used for the quality control of Smilacis Glabrae Rhizoma. The constituents of Heterosmilacis Yunnanensis Rhizoma and Heterosmilacis Chinensis Rhizoma are significantly different from Smilacis Glabrae Rhizoma, and they are not suitable for using as Smilacis Glabrae Rhizoma.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Liliaceae ; chemistry ; Rhizome ; chemistry