To establish a UPLC-MS/MS method for the simultaneous determination of geniposide, genipin 1-O-beta-D-gentiobioside and geniposidic acid in rat brains and study the brain pharmacokinetics of the three iridoid glycosides in stroke rat after the oral administration of Xingnaojing. In this experiment, brain samples were precipitated with protein for twice. Acquity BEH C18 column was adopted, with acetonitrile-0.1% formic acid-water as the mobile phase for gradient elution. ESI source was adopted for mass spectra; multiple reaction monitoring (MRM) was conducted to detect negative ions. The time for sample analysis was 3.5 min. the results showed good linear relations among the three iridoid glycosides, with the extraction recovery between 99.6% and 114.3%, good intra- and inter-day precisions and accuracies and stability in line with the requirements. The t1/2 and MRT in the three components were similar in brains of stroke rats. Geniposide and genipin 1-O-beta-D-gentiobioside showed double peaks; where as geniposidic acid showed a single peak. In conclusion, the method is so specific, sensitive, accurate and reliable that it can be used to study the brain pharmacokinetics of Xingnaojing oral preparation.
Animals ; Brain ; metabolism ; Brain Chemistry ; Chromatography, High Pressure Liquid ; methods ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Iridoids ; chemistry ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Suitable pretreatment of biological samples can truly reflect the role of law of the measured components played in the body and will provide experimental evidence for the studies on metabolic process, material basis of efficacy, mechanism of action, pharmacology, toxicology and the others. Biological samples include blood, urine, hair, tears, etc. There are also many samples processing methods, such as the direct protein precipitation, liquid-liquid extraction and solid phase extraction and so on. These methods could be used alone or combined.
Animals ; Body Fluids ; chemistry ; Chemical Precipitation ; Chemistry Techniques, Analytical ; methods ; Humans ; Liquid-Liquid Extraction ; Proteins ; isolation & purification ; Solid Phase Extraction

Objective To evaluate the effects of diagnosis and treatment of acute pancreatitis com- plicating pseudoaneurysm by CT scan and transcatheter embolization.Methods Eighteen cases of acute panereatitis complicating pseudoaneurysm were performed abdominal angiography and transcatheter em- bolization.The CT and digital substraction angiography(DSA)findings and clinical data were retrospec- tively analyzed.The location of pseudoaneurysm and hemostasis were observed.Results CT scan re- vealed regional or diffuse slight high density fluid collections in abdominal cavity of all patients.En- hanced CT showed contained extravasation of contrast which indictated ruptured the pseudoaneurysm. DSA revealed 22 pseudoaneurysm,and 20 had evidence of bleeding.All pseudoaneurysms were success- fully embolized with coils,and hemostasis was achieyed in 18 pseudoaneurysm immediately.Conclusions The acute pancreatitis complicating pseudoaneurysm can be examined with CT scan and transcatheter embolization is safe and effective therapeutic method.

AIM:To observe the effect of fresh amniotic membrane transplantation ( FAMT) in acute ocular chemical burns.
●METHODS:A prospective study of 25 consecutive cases (36 eyes) with acute ocular chemical burns were treated with FAMT. The clinical efficacy was observed such as the time of amniotic membrane absorbed, corneal epithelialization & transparency, visual acuities and complications.
●RESULTS: With follow-up ranged from 3 to 6mo, 31 eyes′ amniotic membrane were dissolved in 2wk (86%). A total of 33 eyes showed corneal epithelialization in 4wk ( 92%) , 3 eyes showed persistent corneal epithelial defects and need secondary limbal stem cell transplantation or corneal transplantation ( 8%) . A total of 10 eyes showed superficial corneal vascularization (28%), 6 eyes′ cornea were opacity in part (17%), and one eye was symblepharon (3%).
●CONCLUSION:Early FAMT is an effective treatment in the management of acute ocular chemical burns to support epithelial healing, restore ocular surface integrity with potential to improve vision and reduce the incidence of complications. Furthermore, FAMT has advantages of easily obtain and convenient usage, which is suitable in local hospital of our country.

Cognitive impairment is a common complication of diabetes. Hippocampus plays an important role in cognitive function. In hyperglycemia, synaptophysin, a crucial synaptic vesicle membrane protein in hippocampus neuron is found to be down-regulated. Recent evidences have shown that angiotensin IV can facilitate memory acquisition and recovery. However, whether it can also improve cognitive functions of diabetic rats with cognitive disorder, and the possible mechanisms are uncertain. Hence, the objectives of this study. Forty fi ve Sprague Dawley male rats were randomly divided into three groups: Control, diabetic group and diabetes with angiotensin IV treatment group. The cognitive functions, mainly learning and memory of the rats were evaluated using Morris water maze task. The synapses ultrastructure, relative mRNA concentrations and protein expression levels of synaptophysin in hippocampus CA1 area were estimated using transmission electron microscope, RT-PCR, immunohistochemistry and western blotting, respectively. Our study showed that in the diabetic rats with angiotensin IV treatment, the cognitive impairment as measured by Morris water maze task improved, the ultrastructure of synapses in hippocampus reversed, the relative mRNA concentrations and protein levels of synaptophysin in hippocampus signifi cantly increased, when compared with diabetic rats. We conclude that angiotensin IV plays an important role in improving cognitive function of diabetic rats. The possible mechanisms are up-regulating the expression of synaptophysin and normalizing the ultrastructure of synapses in hippocampus.

Objective To investigate the molecular epidemiology of methicillin-resistant Staphylococcus aureus in China in 2005.Methods From January to December 2005,395 consecutive and non-repetitive isolates of methicillin-resistant Staphylococcus aureus were collected from 17 teaching hospitals in 14 cities.The genotypes of SCCmec were determined by multiplex PCR pulsed-field gel electrophoresis (PFGE)was used to type the chromosome DNA of MRSA.Muhilocus sequence typing(MLST)was used to type the housekeeping genes.Fifty-three strains were selected for MLST typing according to the antimicrobials susceptibility patterns,PFGE types,SCCmec types and the distribution of the regions.The toxin gene was detected by PCR.Results Among 395 isolates of MRSA,SCCmec Ⅲ,untypeable type and type Ⅱ accounted for 61.5%(243/395),24.3%(96/395)and 14.2%(56/395)respectively.In Shenyang,60.7%(17/28)of the isolates were SCCmec Ⅱ,which was significantly higher than other areas. Twenty-four different types and 42 subtypes were found by PFGE typing.Clone A accounted for 50.1%, existing in 13 teaching hospitals in 12 cities and clone R accounted for 23.5%,existing in 9 teaching hospitals in 8 cities.Six sequence types(ST)were found in these isolates with ST239 and ST5 accounting for 75.5% and 17.0% among these isolates,respectively.The prevalence of pvl gene was 2.5% among 395 isolates of MRSA.Conclusions The most types of SCCmec in China were Ⅲ and Ⅱ,and distribution of SCCmec types differed among regions.MRSA outbreaks caused by epidemic multiple-drug resistant clones occurred in big teaching hospitals in China.Meanwhile,the same PFGE pattern may spread among areas. Several international epidemic MRSA clones may exist in China.

Objective To demonstrate the carbonic anhydrase Ⅲ (CAⅢ) for 25 000 protein decreased in skeletal muscle of myasthenia gravis (MG). Methods The protein molecular properties responsible to antibodies against 25 000 protein and CAⅢ were analyzed by a combination method of two-dimensional electrophoresis and immuno-Western blot. Competitive binding reactions of the antibodies to the purified 25 000 protein and muscular homogenate were observed by using immuno-Dot blot and immuno-Western blot, respectively. The expression of CAⅢ from normal and MG muscles was detected by immuno-Western blot. Results Combination analysis of two-dimensional electrophoresis and immuno-Western blot showed that the protein of immunological responsible to antibodies against 25 000 protein and CAⅢ had an identical molecular mass and isoelectric point. Competitive binding reactions proved that 25 000 protein and CAⅢ were the same substance, either by immuno-Dot blot or by immuno-Western blot. In addition, a much similar result was obtained when the levels of 25 000 protein from normal and MG muscles were detected by antibodies against 25 000 protein and (CAⅢ) by immuno-Western blot. Conclusion 25 000 protein decreased in the MG skeletal muscle was proved to be just a known protein CAⅢ, which made a basis for further exploring the relationship of CAⅢ deficiency and MG pathogenesis.

To investigate the osteogenesis characteristics of cultured rat mesenchymal stem cells (MSCs) under bone induction condition.

Objective To report a three-generation Chinese family with freckle and to make a genetic linkage analysis in this family.MethodsGenetic linkage analysis was carried out in this family using microsatellite markers distributed over chromosome 4q and 1.Two-point logarithm of odds(LOD)scores were calculated using the Linkage program package(version 5.1),and haplotype was analyzed with Cyrillic version 2.01 software.Results Freckle was inherited in an autosomal dominant pattern with a penetrance of99.9% in this family;linkage to chromosome 4q was ruled out however,supportive evidence was obtained for linkage to microsatellite markers D1S2635 and D1S2844 in chromosome 1q with a maximum LOD score of 1.50.Haplotype analysis in this family localized the locus of freckle to a 12 Mb region flanked by D1S2624 and D1S2799.Conclusions Freckle is a genetically heterogeneous disorder.The causative gene may be located in a 21.2 cM region on chromosome 1q22-24.

OBJECTIVEGas chromatography (GC) was used to investigate the cellular fatty acid (CFA) composition of 141 Acinetobacter baumannii and 32 A. calcoaceticus isolates from different locations in China and to find chemical markers to differentiate these two closely related bacteria.

METHODSWhole cell fatty acid methyl esters (FAMEs) were obtained by saponification, methylation, and extraction for GC analysis, followed by a standardized Microbial Identification System (MIS) analysis.

RESULTSAll A. baumannii and A. calcoaceticus strains contained some major fatty acids, namely, 18:1 ω9c, 16:0, Sum In Feature 3, 12:0, 17:1ω8c, 3-OH-12:0, 17:0, Sum In Feature 2, 2-OH-12:0, and 18:0 compounds. Although most of the total CFAs are similar between A. baumannii and A. calcoaceticus strains, the ratios of two pairs of CFAs, i.e., Sum In Feature 3/18:1 ω9c versus 16:0/18:1 ω9c and Sum In Feature 3/18:1 ω9c versus unknown 12.484/18:1 ω9c fatty acids, could differentiate these two closely related bacteria. A. baumannii could be easily classified into two subgroups by plotting some ratios such as Sum In Feature 3/16:0 versus 17:0 and Sum In Feature 3/2-OH-12:0 versus 17:0 fatty acids.

CONCLUSIONThe ratios of some CFAs could be used as chemical markers to distinguish A. baumannii from A. calcoaceticus.

Acinetobacter baumannii ; classification ; cytology ; metabolism ; Acinetobacter calcoaceticus ; classification ; cytology ; metabolism ; Biomarkers ; metabolism ; Fatty Acids ; metabolism ; Species Specificity