Objective A HPLC method was established for determination of the content of ATST in the aqueous humor of rabbits. MethodsThe mobile phase was consisting of methanol-1％ triethylamine(57:43) and omeprazole (OMZ) as internal standard. The detection was carried out with an ultraviolet detector operated at 235nm. ResultsThere was linearity over the range of 2. 056～41.12 ug/ml in the humor aquosus, r=0.9997. The average recovery of ATST was 94.58 ％. Intra-day and in- ter-day RSD were less than 5 ％ and 10 ％ ( n = 5), respectively. Conclusion The method is reliable. It can be used for the study on the pharmacokinetics of ATST.

AIM: To observe the role of confocal microscopy in infectious keratitis management.
METHODS:Totally 466 patients (467 eyes) diagnosed as infectious keratitis from January 2010 to December 2013 were retrospectively studied. the corneas were examined early by in vivo confocal microscopy. The characteristics of their images and clinical features were studied and summarized.
RESULTS:All patients were recorded, the average age was 54. 4±13. 0 years, in which 264 cases (56. 7%) were male, and 202 cases ( 43. 3%) were female. In the 466 patients, 190 (40. 8%) were fungal keratitis, 148 (31. 8%) were viral keratitis, 125 (26. 8%) were bacterial keratitis and 3 ( 0.6%) were acanthamoeba keratitis. There were fungal hyphae in the images of fungal keratitis. Amebic cysts were found in acanthamoeba keratitis.
CONCLUSION:Confocal microscope can help the early diagnose and treatment of infectious keratitis. It is a noninvasive imaging technique that provides high resolution images of ocular structures at a cellular level and infectious keratitis represents one of its most important clinical uses.

Objective To study the anatomical location and pathology of membranous obstruction of the inferior vena cava(IVC)in Budd-Chiari syndrome(BCS)with research on the etiologic mechanism and pathology.Methods Analysis of 100 normal adults was performed including the gross anatomy of IVC segment from the level of diaphragm up to right atrium.The conventional,microscopic pathologic examination of the biopsy sampling IVC obstruetice mambrane tissue in 70 cases toghther with the complete resected membrane from the radical therapy for 20 cases of BCS,were collected and under investigation.Results The macroscopic examination revealed the obstructive membrane in one case(1%)localizing at the diaphragmatic level,approximately 28 mm,away from the IVC entrance into the right atrium and a newly found valvula was seen on the left lateral wall of the upper part of the hepatic vein orifice.Simultaneously,47% adults showed Eustachiun valve existing in IVC near the entrance to right atrium.Microscopy confirmed all the forementioned membranes consisting of vascular valvular structures.Among them(21/70),30% showed additional organized thrombus formations,and 9%(6/70)with a few amount of inflammatory cellular infiltrations.The total intact resection membrane was continuous with the vascular wall under microscopic examination.Conclusions The first newly report of the existence of a special valvula at the diaphragmatic level of IVC reveals the possibility of high correlation with the occurance of IVC membranous obstruction type in BCS.

This paper briefly analyzed and discussed the current status and major scientific challenges of traditional Chinese medicine (TCM) formulaology research. To promote formulaology research, a new strategy and corresponding technology, network formulaology, were proposed to reveal the complex interaction between functional chemome and biological responses network. The research framework and directions of network formulaology were also summarized and prospected.
Chemistry, Pharmaceutical ; methods ; standards ; Drugs, Chinese Herbal ; chemistry ; Internet ; Medicine, Chinese Traditional ; standards

Human erythropoietin receptor (hEPOR) plays an important role in regulating the red blood cell production by promoting the proliferation and differentiation of RBC from erythroid precursors, hEpoR is a transmembrane protein, and its extrocellular domain (sEpoR) is of great importance in Epo signal transduction pathway. We cloned the gene of sEpoR by RT-PCR from the total RNA of human fetal liver and expressed it in E. coli after insertion of the gene in the expression vector pBV220. The cloned gene was confirmed by sequencing analysis and gene product was confirmed by both Western blot and its first 11 amino acid residues sequence of the N-terminal. In vitro bioassay showed that the purified gene product can repress the growth of TF-cells in the presence of Epo.

A set of central composite design experiments were designed by using four factors which were ethanol amount, ethanol concentration, refrigeration temperature and refrigeration time. The relation between these factors with the target variables of the retention rate of schizandrol A, the soluble solids content, the removal rate of fructose and the removal rate of glucose were analyzed with Bayesian networks, and ethanol amount and ethanol concentration were found as the critical process parameters. Then a network model was built with 2 inputs and 4 outputs using back propagation artificial neural networks which was optimized by genetic algorithms. The R2 and MSE from the training set were 0.983 8 and 0.001 1. The R2 and MSE from the test set were 0.975 9 and 0.001 8. The results showed that network analysis method could be used for modeling of Schisandrae Chinensis Fructus ethanol precipitation process and identify critical operating parameters.
Bayes Theorem ; Chemical Precipitation ; Cold Temperature ; Cyclooctanes ; chemistry ; Ethanol ; chemistry ; Fructose ; analysis ; Fruit ; chemistry ; Glucose ; analysis ; Lignans ; chemistry ; Neural Networks (Computer) ; Polycyclic Compounds ; chemistry ; Reproducibility of Results ; Schisandra ; chemistry ; Time Factors

L9 (3(4)) orthogonal experiment was used to design the extraction technology of compound Clematidis Radix spray. Weight coefficients of active ingredients and dry extract rate were solved by information entropy. Support vector machine (SVM) was established and the model parameters were optimized through the genetic algorithm. Grid search algorithm was used for optimization of extraction technology of Clematidis Radix spray. The optimal extraction technology was to extract Clematidis Radix spray in water with 6 times the weight of herbal medicine for 3 times, with 2 h once. Bias of value between real and predicted by SVM was 1.23%. SVM was compared with traditional intuitive analysis of orthogonal design. It indicates that the new method used to optimize the extraction parameters of compound Clematidis Radix spray is more accurate and reliable.
Clematis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Support Vector Machine ; Technology, Pharmaceutical

Objective To develop a multiplex PCR-based reverse line blot(mPCR/RLB) hybridization assay to detect,simultaneously,seven genes encoding AR-erm(A/TR),erm(B),mef(A/ E),tet(M),tet(O),aphA-3,aad-6 and two AR-related genes,int-Tn and mreA in group B streptococcus.Methods Nine pairs of specific primers and Oligonucleotide probes targeting erm(A/TR), erm(B),mef(A/E),tet(M),tet(O),aphA-3,aad-6 int-Tn and mreA respectively were modified according to former studies or designed in this study.The primers and probes were labeled with biotin and amino,respectively.The nine genes were amplified simultaneously in the same tube.PCR product hybridized with the probes labeled in the BiodyneC nylon membrane to detect the nine genes.To detect the sensitivity and specificity of the method developed,PCR with single pair of primer targeting each gene were tested in 318 isolates tested and the results were compared with the one abtained by RLB.Results The nine resistance-related genes could be successfully detected by mPCR/RLB assay developed in this study.Based on sequencing,21 of 22 isolates with mef had mef(E)and eight of 353 with int-Tn had an atypical sequence.Except for the above 29 genes,all the others corresponded well with the results obtained by single pair primer PCR.Conclusion The mPCR/RLB assay developed in this study is simple,rapid and suitable for surveillance of antibiotic resistance in GBS.

Objective To investigate the immunoregulatory effect and compare the different regula- tions of bone marrow mescnchymal stem cells(MSCs)derived from both lupus(NZBWF1/J)and normal(BALB/ c)mice on T lymphocytes in vitro.Methods MSCs from NZBWF1/J and BALB/c mice bone marrow were iso- lated and expanded,and identified by the surface phenotypes.CD3~+ T lymphocytes isolated by nylon wool columns were stimulated by phorbol myristate acetate(PMA)and co-cultured with or without the two strains of MSCs for 24 h.Intracellular eytokines of T cell,such as interferon(IFN)-?,interleukin(IL)-4,IL-12,IL-6, were analyzed by flow cytometry and quantification of transcription factors T-box expressed in T cells(T-bet) and GATA-binding protein 3(GATA-3)were detected by reverse transcriptase PCR(RT-PCR).T cell apop- tosis was assessed by flow cytometry using rhodamine123.Results The results showed that a decrease of CD3~+ T cell apoptosis was seen when NZBWF1/J MSCs or BALB/c MSCs were added to T cells stimulated by PMA(P＜0.05),and an increase of TH2 cytokines by NZBWF1/J MSCs and TH1 eytokines by BALB/c MSCs were observed in the CD3~+ T cells eo-cuhured with MSCs(P＜0.05).Conclusion It is suggested that the al- teration of T subsets caused by MSCs may interfere with the systemic lupus erythematosus(SLE)development and normal MSCs may be effective in the improvement of SLE.NZBWF1/J MSCs have defective immunoregula- tory function when compared with MSCs from healthy mouse strains.

Objective To obtain the recombinant corticotropin releasing hormone (CRH) protein with soluble, high purity protein through optimizing prokaryotic expression condition and purifying glutathione thiol transferase (GST)-CRH protein. Methods To detect the expression of soluble CRH protein through grope of the host strain GST-CRH temperature of induction expression, the host strain concentration (OD600), IPTG concentration and induction time, the purification of GST-CRH was performed by GST-CRH agarose gel. Western Blot assay was used for the expression identification of the target protein. Results The optimal conditions for the induction of CRH protein were determined: temperature of 30 ℃, IPTG induced concentration 0.1 mmol/L, bacteria density (OD600) 0.8, the induction time of 8 hours, purified GST-CRH>95% fusion protein was obtained. Conclusion The optimal expression conditions of GST-CRH are obtained, and the soluble protein of high purity GST-CRH is also obtained.