OBJECTIVE: To develop an HPCE method of tetramethylpyrazine hydrochloride in rat plasma using pyramide as the internal standard. METHODS: The separation was performed on a fused-silica uncoated capillary with an inner diameter (ID) of 75 μm, an effective length of 50 cm and a total length of 60 cm. The optimum HPCE conditions were as follows: 100 mmol·L-1 SDS-30 mmol·L-1 sodium tetraborate solution (40:60) was used as the background electrolyt, the samples were injected at the anodic end at 0.5 psi for 6 s, separation voltage was 20 kV at positive power, capillary temperature was 20°C, and detection wavelength was 295 nm. RESULTS: The linear range was 1.12-286.72 mg·L-1(r=0.9996), and the inter-day and intra-day precisions expressed as RSDs were less than 4.21% and 4.74%, respectively. The average recoveries were 91.22%-97.48%, and the limit of detection was 0.16 mg·L-1. CONCLUSION: This method is rapid and accurate to quantify tetramethylpyrazine hydrochloride in rat plasma.

Objective To investigate whether leukotriene D4 (LTD4) regulates eotaxin-3 (Eot-3) expression in bronchial epithelial cells, and study effect of pranlukst on the regulation.Methods BEAS-2B cells and normal human bronchial epithelia cells were pre- treated with LTD4 for 1 hour,stimulated with interleukin-4, the cells were incubated for 24 hours. Eot-3 protein in supernatant were measured by enzyme linked immunosorbent assay(ELISA). The cells were pretreated with pranlukast in different concentration, then the above procedure was repeated. Results The untreated bronchial epithelial cell expressed Eot-3 protein on a very low level. After stimulating with IL-4 and incubating for 24 hours, Eot-3 production increased significantly. Pretreating the cells with LTD4 enhanced the inducing effect of IL-4. Pranlukast inverted the upregulation of LTD4. Conclusions Upregulating the expression of Eot-3 induced by IL-4 on bronchial epithelial cells may explain partially the mechanism of leukotrienes involving airway allergic inflammation of asthma. The invertion impact on upregulation of LTD4 by pranlukast may be one of mechanisms that leukotrienes receptor antagonist cure asthma.

Background Many eye diseases such as central retinal artery occlusion,glaucoma and ischemic optic neuropathy,etc.lead to retinal ischemia-reperfusion injury (RIRI) and furthmore visual functional damage.It is neeessary to study the treatment of RIRI.Objective This study was to observe and discuss the influence of aminoguanidine on the retina morphological changes and its mechanism after RIRI.Methods Eighty clean healthy male Japanese white rabbits were randomly divided into normal injury group,RIRI group and aminoguanidine (AG)treated group.The model of RIRI was established by infusing saline solution into the anterior chamber to elevate intraocular pressure (IOP) in both RIRI group and AG group.AG was intraperitoneally injected in the models of the AG group,and normal saline solution was used at the same method in tbe normal group and the RIRI group.The fundus photography and fundus fluorescein angiography(FFA) were pertormed on the rabbits at the moment of retina ischemia and 6,24 and 72 hours after reperfusion.The parts of rabbits were sacrificed 1,6,24 and 72 hours after reperfusion,followed by the enucleation of the eyeballs.Retinal section was prepared for TUNEL examination to evaluate the apoptosis of retinal cells.Nitric oxide (NO) concentration in retina was detected with nitrate reductase,and the activity of inducible nitric oxide synthase (iNOS) was measured by colorimetric detection.The use of the animals followed the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The fundus photography and FFA showed that the retinal edema was more mild,and the percentage of vascular occlusion was lower in the AG treatment group than that in RIRI group and the amount and area of fluorescein leakage were also smaller than the treatment group.The numbers of TUNEL positive cells in the AG treatment group were less than those in the RIRI model group at 1,6,24 and 72 hours after experiment (F分组 =2762.37,P =0.00 ; F时间 =894.24,P =0.00).Numbers of TUNEL positive cells between adjacent time points were significantly different in both RIRI model group and AG treatment group (RIRI group:q =24.475,36.591,-20.37,P＜0.05;AG group:q =20.94,16.79,-6.92,P＜0.05),with the peak value at 24 hours after experiment.NO contents were significantly higher in the RIRI model group compared to AG group at various time points(q =3.84,4.01,8.91,3.75,P＜0.05),and those between adjacent time points showed significant differences (RIRI group:q=4.77,13.403,-10.29,P＜0.05;AG group:q=4.55,9.05,-5.08,P＜0.05).iNOS activity was significantly elevated in the RIRI model group compared with AG group(q=-3.74,-4.94,-6.53,-3.98,P＜0.05),and obvious differences also were seen between the adjacent time points in both two groups(RIRI group:q =8.43,6.71,-6.39,P＜0.05 ;AG group:q =4.16,5.08,-3.93,P＜0.05).Conclusions Aminoguanidine can protect the retinal function and morpbology from oxidative stress damage after RIRI by reducing the NO level and inhibiting the iNOS activity in retina.

Objective To investigate whether the protein kinase C inhibitor can promote the apopto- sis of multidrug resistance tumor cell lines which are induced by chemotherapy drugs.Methods Choose the KB/S(oral squamous cancer cell line)and KB/VCR(its multidrug resistant cell line)to compare the Adri- amycin-induced apoptosis with or without staurospolin(protein kinase C inhibitor).The apoptosis is stained with acridine orange,tested by flow cytometry,and approved by electron microscope.Results 36 hours after the treatment with 0.04 ?g/ml adriamycin,apoptotic cells of KB/S are 96.68%,and after 48 hours,the apop- totic cells of KB/VCR are 64.99%.When the concentration of adriamycin are augmented to 0.4?g/ml and 2.0?g/ml,the apoptotic cells of KB/VCR are 69.74% and 37.18% respectively.When treated with stau- rospolin together,the apoptotic cells of KB/VCR increased to 72.58%(?~2=4.5,P0.05)respectively.These results were testified by electron microscope and acridine orange-stain.Conclu- sion The resistance to apoptosis may be one of the mechanisms of multidrug resistance and the protein ki- nase C inhibitor may reverse this resistance by promoting the apoptosis of multidrug resistance tumor cells.

Near-infrared spectroscopy ( NIR ) is widely used in the area of food quantitative and qualitative analysis.Variable selection technique is a critical step of the spectrum modeling with the development of chemometrics.In this study, a novel variable selection strategy, automatic weighting variable combination population analysis (AWVCPA), was proposed.Firstly, binary matrix sampling (BMS) strategy that gives each variable the same chance to be selected and generates different variable combinations, was used to produce a population of subsets to construct a population of sub-models.Then, the variable frequency ( Fre) and partial least squares regression ( Reg) , which were two kinds of information vector ( IVs) were weighted to obtain the value of the contribution of each spectral variables, the influence of two IVs of Rre and Reg was considered to each spectral variable.Finally, it used the exponentially decreasing function ( EDF) to remove the low contribution wavelengths so as to select the characteristic variable.In the case of near infrared spectrum of beer and corn, the prediction model based on partial least squares ( PLS ) was established.Compared with other variable selection methods, the research showed that AWVCPA was the best variable selection strategy in the same situation.It had 72.7% improvement compared AWVCPA-PLS with PLS and the predicted root mean square error (RMSEP) decreased from 0.5348 to 0.1457 on beer dataset.It had 64.7% improvement compared AWVCPA-PLS with PLS and the RMSEP decreased from 0.0702 to 0.0248 on corn dataset.

The statement of"qi transformation leading to the removal of pathogenic dampness"was recorded in Wen Bing Tiao Bian(Analysis on Epidemic Febrile Diseases)written by the Qing Dynasty physician WU Ju-Tong.Dampness in the triple energizer is caused by the dysfunction of qi transformation,and the treatment of dampness must be based on the activation of qi movement and focused on the promotion of qi movement and the restoration of the qi transformation in the triple energizer.For the treatment of dampness attack in the upper energizer,therapies of dispersing lung to smooth qi and resolving dampness to relieve the obstruction are recommended.For the treatment of dampness obstruction in the middle energizer,therapy of activating spleen qi by strengthening spleen and moving qi is stressed for helping the removal of dampness and for the eradication of the source of dampness.For the treatment of dampness stagnation in the lower energizer,therapy of draining dampness with sweet-light medicines and activating yang can be used according to the illness status.The three methods of treating dampness,namely dispersing the upper energizer,activating the middle energizer and draining the lower energizer,all embody the mechanism of"qi transformation leading to the removal of pathogenic dampness",and the therapies of dispersing lung with light medicines,inducing perspiration by opening striated layer,eliminating dampness with aromatics and draining dampness with sweet-light medicines should be used in accordance with the syndromes.The elucidation of the academic thoughts of"qi transformation leading to the removal of pathogenic dampness"can provide theoretical reference for the fundamental research of dampness syndrome and clinical application of therapies for resolving dampness in Chinese medicine.

OBJECTIVETo develop a method for analyzing the chemical compositions of Gegen and Fenge extracts using high-performance capillary electrophoresis (HPCE).

METHODSUsing HPCE/DAD, the chemical composition of the extracts was analyzed with the buffer solution of 40 mmol/L borax containing 16.7% methanol, with injection pressure at 137.9 kPa for 5 s, separation voltage at 25 kV in 0-5 min time range and at 22 kV in 5-25 min time range, and the temperature of the capillary of 20 degrees celsius.

RESULTS AND CONCLUSIONThe method for analysis of Gegen and Fenge extracts was established, which identified puerarin and daidzein as the two major components. This simple and rapid analysis method can be used for Gegen and Fenge extract fingerprinting.

Dendritic Cell Sarcoma, Follicular ; pathology ; Dendritic Cell Sarcoma, Interdigitating ; pathology ; Female ; Humans ; Middle Aged ; Stomach Neoplasms ; pathology

Objective To observe the characteristics of acute graft-versus-host disease (aGVHD) confined solely to gastrointestinal tract after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Clinical data of 307 patients who received allo-HSCT in our hospital between November 2007 and December 2008 were retrospectively analyzed. Clinical features, risk factors, therap(e)utic effects and prognosis of 38 patients whose aGVHD confined solely to gastrointestinal tract after allo-HSCT were analyzed.Results The incidence of aGVHD was 56. 7% (174/307). aGVHD confined solely to gastrointestinal tract occurred in 38 out of 174 patients (21.8%). The incidence of aGVHD confined solely to gastrointestinal tract was affected by human leukocyte antigen (HLA)-compatible level. The patients transplanted from HLA-matched donor had a higher probability of aGVHD confined solely to the gastrointestinal tract. After treatment with glucocorticoids or anti-CD25 monoclonal antibody, 2 patients who had aGVHD confined solely to the gastrointestinal tract died of uncontrolled aGVHD and concurrent infection. The total effective rate was 94.7%. But for patients who had aGVHD between Ⅱ-Ⅳ degree and not confined solely to gastrointestinal,therapeutic effective rate was 85.2%. There was a difference in therapeutic effect between the two groups (P =0. 015). Anti-CD25 monoclonal antibody was used in 12 patients who had aGVHD confined solely to the gastrointestinal tract and all patients' conditions of aGVHD were controlled. Conclusions aGVHD confined solely to gastrointestinal tract after HSCT was not rare. The incidence of aGVHD confined solely to gastrointestinal tract was higher in HLA-matched sibling donor than in mismatched transplantation.Therapeutic efficacy for the aGVHD confined solely to gastrointestinal tract was good. The favorable outcome might be related with the application of anti-CD25 monoclonal antibody.

To investigate the antitumor activities of the immunoconjugates composed of anti-type IV collagenase monoclonal antibody Fab' fragment and lidamycin (LDM) prepared with different linkers. The immunoconjugates were prepared by linking Fab' to lysine-69 of LDM apoprotein by SPDP, LCSPDP, SMBS or SSMPB as the intermediate drug linkers. Immunoreactivities of the conjugates were determined by ELISA. The cytotoxicities of the conjugates were examined by clonogenic assay. In vivo antitumor effects of the conjugates were evaluated in nude mice bearing subcutaneously implanted HT-1080 tumor. ELISA assay showed that the conjugates retained part of the immunoreactivity of 3G11 against the antigen. The cytotoxicities of the Fab'-SMBS-LDM and Fab'-SSMPB-LDM to HT-1080 cells were significantly potent, compared with Fab'-SPDP-LDM, Fab'-LCSPDP-LDM and free LDM. In animal models at the same condition, free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM inhibited the growth of HT-1080 tumor by 70.9%, 74.8% and 72.3%, while Fab'-SMBS-LDM and Fab'-SSMPB-LDM reached 78.0% and 87.7%, respectively. The median survival time of the mice treated with free LDM, Fab'-SPDP-LDM and Fab'-LCSPDP-LDM were prolonged by 71.9%, 82.2% and 107.5%, respectively, compared with that of untreated group. Whereas, the median survival time of Fab'-SMBS-LDM and Fab'-SSMPB-LDM were prolonged by 145.2% and 165.8%, respectively, indicating that Fab'-SSMPB-LDM was more effective than Fab'-SMBS-LDM in tumor suppression and life span prolongation. Fab'-SSMPB-LDM has more marked selective antitumor efficacy and lower toxicity, and might be a novel candidate for cancer therapy.
Aminoglycosides ; pharmacology ; Animals ; Antibiotics, Antineoplastic ; pharmacology ; Antibodies, Monoclonal ; immunology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Collagenases ; immunology ; Enediynes ; pharmacology ; Fibrosarcoma ; pathology ; Humans ; Immunoconjugates ; pharmacology ; Immunoglobulin Fab Fragments ; immunology ; Matrix Metalloproteinase Inhibitors ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Tumor Burden ; drug effects