Objective To establish homogeneous immunoassay for detecting serum cardiac troponin I (cTnI) by using light induced chemiluminescent immunoassay (LiCA). Methods Polyclonal antibodies of cTnI were coated on the receptor particles, monoclonal antibodies of cTnI were biotinylated, and the donor particles were coated with streptavidin, all of which were composed of LiCA reagents. The optimal test conditions and analytical performance of the detection method were studied. Results The method was rapid, sensitive, and detection time was 17.5 min.The analytical sensitivity was 0.045 ng/mL and the functional sensitivity was 0.053 ng/mL.The recovery rate was 104.96%-108.21%;The within-run and the between-run coefficients of variation were 3.88%-5.53%and 7.60%-8.75%, respectively. The interference rates for the endogenous substances were less than 10%. The reference value of cTnI was less than 1.05 ng/mL;Results of cTnI LiCA correlated well with direct chemiluminescence detection (r2 =0.979). Conclusions This approach can be used for the quantitative detection of serum cTnI, and it is homogeneous and is free of clean separation. It provides a convenient, highly sensitive detection platform for clinical practice.

OBJECTIVETo explore activity laws of mitochondrial complex II in patients of deficiency-cold syndrome (DCS) and deficiency-heat syndrome (DHS) under various ambient temperatures.

METHODSSubjects were recruited by questionnaire and expert diagnosis from grade 1 - 3 undergraduates at Henan College of Traditional Chinese Medicine in November 2012, and assigned to a normal control group, the DCS group, and the DHS group, 20 in each group. Their venous blood samples were collected at two different temperature conditions. Activities of mitochondrial complex II were measured by spectrophotometry.

RESULTS(1) Comparison of mitochondrial complex It under various ambient temperatures: Compared with room temperature in the same group, activity values were all increased in the normal control group at cold temperature with significant difference (P <0.05), but there was no significant difference in the DCS group and the DHS group (P >0. 05). Compared with the normal control group, activity values of complex H were reduced in the DCS group at cold and room temperatures with significant difference (P <0.05). Compared with the DCS group, activity values of complex It were increased in the DHS group with significant difference (P <0. 05). (2) Changes of adjustment rates: Compared with room temperature, the adjustment rate all rose at cold temperature in the normal control group and the DHS group with significant difference (P <0.05), but with no significant difference found in the DCS group (P >0. 05). Compared with the normal control group at the same temperature, the adjustment rate in the DHS group and the DCS group was all reduced at cold and room temperatures with significant difference (P <0. 05). There were no significant difference in the adjustment rate between the DHS group and the DCS group (P > 0. 05).

CONCLUSIONSEnvironment temperature can affect the activity of mitochondrial complex II with different influence degrees on different syndrome types of people, but its change trend are basically identical.

Cold Temperature ; Electron Transport Complex II ; metabolism ; Hot Temperature ; Humans ; Medicine, Chinese Traditional ; Syndrome ; Temperature

OBJECTIVETo purify the extract of fructus gardeniae for injection by macroreticular resins in purification process of traditional Chinese medicine (TCM) Injections.

METHODUsing fructus gardeniae as sample, on base of obtaining the extract by employing macroreticular resin, quality evaluation and rationality of purification methods had been studied by the quantitative analysis of active ingredients and the characteristics of micromeritics, safety and stability of the extract.

RESULTThe experiment showed the extract of fructus gardeniae for injection had been produced successfully by macroreticular resin.

CONCLUSIONUsing macroreticular resins is a promising purification way of TCM injections, whereas a more consummate method of quality evaluation must be established to ensure safety, efficiency and stability of the preparation in the process.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; toxicity ; Fruit ; chemistry ; Gardenia ; chemistry ; Injections ; Iridoids ; isolation & purification ; Mice ; Plants, Medicinal ; chemistry ; Pyrans ; isolation & purification ; Quality Control ; Resins, Synthetic ; Technology, Pharmaceutical ; methods

OBJECTIVETo detect the expression of the peripheral blood P2X5 receptor at various ambient temperatures, and to explore its relationship with deficiency-cold syndrome and deficiency-heat syndrome.

METHODSSubjects were selected by questionnaire and expert diagnosis, and assigned to the normal control group, the deficiency-cold syndrome group, and the deficiency-heat syndrome group, 20 in each group. 5 mL venous blood was collected at room temperature (25 °C) and cold temperature (-4-5 °C) respectively. Then the expression of P2X5 receptor was relatively quantified by real-time fluorescence quantitative PCR, and compared at room temperature and cold temperature respectively.

RESULTSThe expression of P2X5 receptor in deficiency-cold syndrome and deficiency-heat syndrome groups was lower than that in the normal control group at room temperature (P < 0.05). It decreased more at cold temperature in the deficiency-cold syndrome group than in the normal control group (P < 0.01) as well as in the deficiency-heat syndrome group (P < 0.05). The expression of P2X5 receptor showed no difference in all groups at two different temperatures (P > 0.05).

CONCLUSIONSThe expression of P2X5 receptor was different in different syndrome groups at various ambient temperatures. Ambient temperatures had insignificant effect on the expression of P2X5 receptor of the population with the same syndrome.

Cold Temperature ; Hot Temperature ; Humans ; Medicine, Chinese Traditional ; Receptors, Purinergic P2X5 ; metabolism ; Syndrome

OBJECTIVETo explore the immune mechanism of moxibustion on protecting gastric mucosa injury.

METHODSForty healthy SD rats were randomly divided into four groups: a blank group, a model group, a moxibustion acupoint group and a moxibustion non-acupoint group, 10 rats in each one. Eight days before model establishment, moxibustion at "Zusanli" (ST 36), "Zhongwan" (CV 12), "Guanyuan" (CV 4), "Pishu" (BL 20) and "Weishu" (BL 21) was applied in the moxibustion acupoint group while these acupoints' controlled points were selected in the moxibustion non-acupoint group, and no treatment was given in the model group, once a day in three groups for continuous 16 days. The helicobacter pylori (Hp) model was established by intragastric administration of Hp. HE staining microscopic examination was used to observe inflammation severity in gastric mucosa, and enzyme-linked immunosorbent assay (ELISA) was adapted to measure content of heat shock protein (HSP) 72, TNF-alpha and IL-1beta, and real-time quantitative PCR was used to measure the expression of TLR2 mRNA, TLR4 mRNA, CD14 mRNA and MyD88 mRNA in peripheral blood mononuclear cells, and western blot method was used to measure content of NFkappaB and IkappaBalpha in peripheral blood mononuclear cells.

RESULTSCompared with the blank group, the expression of HP could be seen in the smear of gastric mucosa by Gram's staining in the model group; the inflammation severity score was obviously increased as well as content of serum HSP 72 and TNF-alpha and IL-1beta in gastric tissue; and expression of TLR2, 4 mRNA, CD14 mRNA, MyD88 mRNA, NFkappaB was increased (P < 0.01), but the expression of IkappaBalpha was reduced (P < 0.05). After the moxibustion, the inflammation severity score was reduced in the moxibustion acupoint group, and the content of serum HSP 72 was increased, and the expression of TNF-alpha and IL-1beta in gastric tissue and expression of TLR2 mRNA, TLR4 mRNA, CD14 mRNA, MyD88 mRNA and NFkappaB were reduced (P < 0.01), but the expression of IkappaBalpha was increased (P < 0.05). The differences between the moxibustion non-acupoint group and the model group were not significant (P > 0.05).

CONCLUSIONThe pretreatment of moxibustion at acupoints could induce the over expression of serum HSP 72. By combining TLR 2 and 4 receptors to trigger receptor signal transduction pathways, the releases of downstream signal substances are regulated; as a result, the releases of related immune substances are regulated to relieve the gastric mucosa injury of rats with HP gastritis.

Acupuncture Points ; Animals ; Female ; Gastritis ; immunology ; therapy ; Helicobacter Infections ; genetics ; immunology ; therapy ; Helicobacter pylori ; physiology ; Humans ; Interleukin-1beta ; genetics ; immunology ; Male ; Moxibustion ; NF-kappa B ; genetics ; immunology ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; immunology

OBJECTIVETo investigate the feasibility of local drug delivery into the inner ear using solid lipid nanoparticles (SLN) and evaluate its potential for inner ear disease treatment in terms of the pharmacokinetics of the delivered drug in the inner ear.

METHODSDexamethasone acetate (DA)-loaded SLN was prepared with Compritol 888 ATO as the matrix by means of hot dispersion-ultrasonic technique. A high-performance liquid chromatography (HPLC) was established for determining DA and dexamethasone (Dex). The pharmaceutical properties of DA-loaded SLN including the particle size, entrapment ratio and in vitro release were estimated. DA-loaded SLN was administered via intratympanic injection or intravenous injection in guinea pigs and Dex concentration in the perilymph was measured with HPLC for estimation of the pharmacokinetic parameters.

RESULTSThe mean diameter of the DA-loaded SLN was 106.8 nm with entrapment ratio of 83.8%, and the in vitro DA release from the nanoparticles well conformed to Weibull distribution, with sustained-release of DA from the SLN exceeding 6 days. After intravenous injection of DA-loaded SLN in guinea pigs, Dex failed to be detected in the perilymph. Compared with Dex-loaded in situ gel following intratympanic injection, the relative bioavailability of Dex in the perilymph was 504% following intratympanic injection of DA-loaded SLN, which also resulted in increased t(1/2) and mean residence time (MRT) by 0.5 and 1.9 folds respectively.

CONCLUSIONNanoparticles can be a promising tympanic drug delivery system for topical drug administration in the treatment of inner ear diseases.

Administration, Topical ; Animals ; Anti-Inflammatory Agents ; administration & dosage ; pharmacokinetics ; Dexamethasone ; administration & dosage ; pharmacokinetics ; Drug Delivery Systems ; Ear, Inner ; metabolism ; Female ; Guinea Pigs ; Male ; Nanoparticles ; administration & dosage ; Round Window, Ear ; metabolism

OBJECTIVETo investigate the evaluation method for self-emulsifying drug delivery system of volatile oil from rhizome of Ligusticum chuanxiong (VOC SEDDS).

METHODThe self-emulsifying ability, the efficiency of self-emulsification, the properties of emulsion, the dissolution of volatile oil from Rhizome of Ligusticum Chuanxiong and the stability of the emulsion were determined.

RESULTThe optimized formulation can fully emulsify in 5 min and the particle sizes were around 102 nm. Zeta potential was about -30 mV. The O/W emulsions were stable through centrifugation with high reproducibility. In vitro dissolution test indicated that over 80% of drug dissolved in 30 min and VOC SEDDS was stable under light and high temperature in 10 d.

CONCLUSIONVOC SEDDS has strong self-emulsifying ability, fine stability and high dissolution rate in vitro.

Drug Compounding ; methods ; Drug Delivery Systems ; Drug Stability ; Emulsions ; Light ; Ligusticum ; chemistry ; Oils, Volatile ; administration & dosage ; chemistry ; isolation & purification ; Particle Size ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry ; Solubility ; Surface-Active Agents ; chemistry ; Temperature ; Time Factors

Novel solid lipid nanoparticle (SLN) system is prepared with Compritol ATO 888 and tricaprylic glyceride. DSC, XRD, SAXS and NMR are employed to study the novel carrier property and microstructure. When the peak melting point decreased from 70.8 degrees C to 61.4 degrees C, the enthalpy sharply decreased. It could be concluded that the regular crystal lattices in the novel carriers are broken out for the oil joined in them. Melting behavior is occurred at -17.7 degrees C while novel SLN is composed of oil and solid lipid mixture from the DSC measurement. Most alpha phase and least beta' phase are in the nano carrier system whether drug loading or not from the XRD investigation. There is only 0.1 nm change of long space among the novel SLN made of mixture and the lipid matrix and traditional SLN; therefore, it is impossible of the oil molecular insert into the solid glyceride structure. Since the different melting behavior (DSC measurements) and molecular move state (NMR investigations), two lipid matrix are still in two state of liquid and solid lipid in the novel SLN carrier. Presume the microstructure of the novel SLN prepared by our experiment would be that liquid oil has formed superfine nano accommodation encapsulated with solid lipid, but the whole particle is still in nano size range.
Calorimetry, Differential Scanning ; Caprylates ; chemistry ; Diterpenes ; administration & dosage ; chemistry ; Drug Carriers ; chemistry ; Drug Delivery Systems ; Epoxy Compounds ; administration & dosage ; chemistry ; Fatty Acids ; chemistry ; Magnetic Resonance Spectroscopy ; Nanoparticles ; Particle Size ; Phenanthrenes ; administration & dosage ; chemistry ; Triglycerides ; chemistry ; X-Ray Diffraction

OBJECTIVETo obtain the optimized extraction technology of Paeonia suffruticosa by comparing several extraction method.

METHODExtract P. suffruticosa by ethanol circumfluence, distillation-decoction, CO2-SFE and traditional decoction, and analyse the results according to the total extraction rate, extraction rate of paeono, extraction of other ingredients and production feasibility.

RESULTTotal extraction rates of which are 12.66%, 13.51%, 7.28%, 7.56% respectively; extraction rates of paeonol are 2.45%, 2.26%, 0.31%, 1.15% in turn; Phenolic glycosides can be extracted by ethanol circumfluence, distillation-decoction, traditional decoction, but not by CO2-SFE.

CONCLUSIONDistillation-decoction is the most proper extraction technology of P. suffuticosa at present.

Acetophenones ; isolation & purification ; Drugs, Chinese Herbal ; isolation & purification ; Paeonia ; chemistry ; classification ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods

This paper introduced an experimental study of the preparation of polylacticacid (PLA) nanoparticles of cucurbitacin (CuC) using a precipitation method. The residual acetone, ratio of CuC PLA precipitates, and the relationships between the ratios of two precipitates and drug incorporation rates were measured. It appeared that the nanoparticles with 60% of PLA incorporated with 5.5% of CuC were formed when acetone was injected into the aqueous phase. As the acetone gradually evaporated, drug incorporation/encapsulation continued, with most of CuC (about 70%) formed new crystalline cores and suspended in the form of microcrystals in the medium, resulting a suspension containing both nanoparticles and microcrystals. We also concluded that this system may not necessarily be suitable for all lipophilic drugs to be prepared to PLA nanoparticles with good incorporation rate. The drug incorporation depended on the interactions among drug, PLA, and organic solvents, in addition to the solubility of the drug.
Antineoplastic Agents, Phytogenic ; administration & dosage ; Chemical Precipitation ; Cucurbitaceae ; chemistry ; Cucurbitacins ; Delayed-Action Preparations ; Drug Compounding ; methods ; Drug Delivery Systems ; Lactic Acid ; Microspheres ; Nanotechnology ; Particle Size ; Plants, Medicinal ; chemistry ; Polyesters ; Polymers ; Triterpenes ; administration & dosage ; isolation & purification