OBJECTIVE:To improve the detection method for the dissolution of dihydroartemisinin in Dihydroartemisinin and piperaquine phosphate tablets. METHODS:The dissolution experiment adopted paddle method using 0.1 mol/L hydrochloric acid so-lution 500 mL as solvent with rotating speed of 75 r/min and sampling time of 45 min. In sample pre-treatment,the volume of 3.6% sodium hydroxide solution was increased from 5 mL to 20 mL,and that of phosphoric acid was increased from 0.2 mL to 0.7 mL. HPLC was adopted to determine the dissolution of dihydroartemisinin. The determination was performed on YMC-Pack ODS-A column with mobile phase consisted of 0.02 mol/L disodium hydrogen phosphate solution(pH adjusted to 2.4 using phosphoric ac-id)-acetonitrile(65 : 35,V/V)at the flow rate of 1.0 mL/min. The detection wavelength was set at 237 nm,and column temperature was 30 ℃. The sample size was 20 μL. RESULTS:The linear range of dihydroartemisinin were 7.802-117.03 μg/mL(r=0.9999). The limit of quantitation was 2.0 ng,and the limit of detection was 0.6 ng. RSDs of precision and reproducibility tests were all low-er than 1.0%. The recoveries were 99.18%-100.46%(RSD=0.45%,n=9). Average dissolutions of dihydroartemisinin in 3 batch-es of samples were 94.9%,77.9%,89.6%,respectively. CONCLUSIONS:Improved method enhance the accuracy for the limit of sensitivity,dissolution and detection results.

Objective To find a proper way for accurate results on completing blood counts.Methods Based on the results from automatic hematology analyzer XE-2100,set up the criteria for blood cell microscopic examination.1368 blood specimen were detected and the results were analyzed according to the criteria.Statistics on the data were made to evaluate the accordance between warnings of analyzer and manual examination,likewise the reliability of the criteria.Results Comparing with microscopic examination,analyzer warning on low PLT has good accordance,Kappa value was 0.95,U value was 35.19,P

Objective To study the benzofuran compounds from roots and rhizomes of Ligularia caloxantha, which is a folk medicine used in the Naxi Nationality in Yunnan Province. Methods Compounds were separated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were elucidated by physicochemical properties and spectral analysis. Results Eighit compounds are isolated from ethanolic extracts of the roots and rhizomes. They were identified as euparin (Ⅰ), 6-methoxy-euparin (Ⅱ), 4, 5-dimethoxy-2-hydroxybenzaldehyde (Ⅲ), 2-acetyl-5, 6-dimethyoxybenzofuran (Ⅳ), 4-hydroxyacetophenone (Ⅴ), 2-isopropenyl-5, 6-dimethoxy-2, 3-hydrocumaran (Ⅵ), 8?-hydroxy-7(11)-eremophilen-12, 8?-olide (Ⅶ), lupeol (Ⅷ). Conclusion All the eight compounds were obtained from L. caloxantha for the first time as benzofurans. Compounds Ⅰ and Ⅱ are two major ones with insecticide and insect food refusal-induced activities. That is the relative reason of L. caloxantha used for folk anti-insect.

Objective To observe the effect of interaction in children speech therapy.Methods 90 cases with children aphasis were assessed with evaluation for delayed speech and language development by Sign-significance (S-S) method made by Chinese Rehabilitation Research Center and dysarthria evaluation, and treated.Results After treatment, all cases had different improvment on their own level and were in the process of rehabilitation care.Conclusion The speech therapy interaction is easy, economical and can stimulate children' interest to learn speech and get the training goal.

To enhance the specificity of anti-TNF-α single chain Fv antibody (TNF-scFv) to inflamed site, we constructed a bispecific antibody BsDb that targets TNF-α and ED-B-containing fibronectin (B-FN) by covalently linking TNF-scFv and the anti-ED-B scFv L19 at the gene level via a flexible peptide linker deriving from human serum albumin. BsDb was successfully secreted from Pichia pastoris as functional protein, identified by immunoblotting, and purified to homogeneity with affinity chromatography. BsDb retained the immunoreactivity of its original antibodies TNF-scFv and L19, and showed a marked gain in antigen-binding affinity and in TNF-α-neutralizing ability, when compared to TNF-scFv and L19 that were produced in Escherichia coli. In the adjuvant-induced arthritis (AIA) mice model, BsDb showed selective accumulation and retention in the inflamed paws but rapid clearance from blood, resulting in high arthritic paw to blood ratios. These data indicate that BsDb is endowed with high specificity to inflamed site and low toxicity to normal tissues and holds great potential for in vivo application for the targeted therapy of RA and other chronic inflammatory diseases.
Animals ; Antibodies, Bispecific ; biosynthesis ; immunology ; Antibodies, Neutralizing ; biosynthesis ; immunology ; Escherichia coli ; Fibronectins ; chemistry ; immunology ; Humans ; Mice ; Single-Chain Antibodies ; biosynthesis ; immunology ; Tumor Necrosis Factor-alpha ; immunology

Objective To investigate the clinical and pathological characteristics between types 1 and 2 endometrial uterine cancers.Methods The clinical materials of 9 437 patients with uterine cancer were collected with retrospective analysis from 62 hospitals during 2000 to 2010.Results The mean age of type 1 endometrial cancers was less than type 2.There were more young patients in type 1 endometrial cancers.The mean menopause age of type 2 endometrial cancers was greater than type 1.The mean age of menarche,obesity,diabetes,hypertension,infertility,and nulliparous were not significant differences between types 1 and 2 endometrial cancers.There were more patients with advanced tumor,deep myometrium invasion,estrogen receptor (ERs) negative,progesterone receptor (PR) negative,P53 positive,lymph vascular space involvement,cervical stromal invasion,adnexal metastasis,and lymph node metastasis in type 2 endometrial cancers.Conclusions Type 2 endometrial uterine cancers occurred in elder people with more pathological risk factors and more malignant biological activities.

The successful separation of breast cancer stem cells (BCSCs) is the foundation of BCSC research.At present,people could gather BCSCs to some extent in a number of ways.However,as more and more biomarkers are found in BCSCs separation,the heterogeneity becomes a hot spot.How to gather BCSCs as much as possible and how to explain the heterogeneity could provide new insights in the treatment of breast cancer.

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.

Objective To determine the diagnostic and assessment value of soluble urokinase plasminogen activator receptor (suPAR) level in septic patients.Methods Totally 82 septic patients in the Department of Intensive Care Unit of The First Affiliated Hospital,Sun Yat-Sen University were prospectively analyzed from June 2013 to March 2014.Another 29 patients with systemic inflammatory response syndrome (SIRS) and 15 healthy subjects served as controls.Septic patients were divided into sepsis group (n =27),severe sepsis group (n =27) and septic shock group (n =28) according to the severity,and there was no significant difference in age and sex among these groups.Measurement of plasma suPAR,serum procalcitonin (PCT) and C-reactive protein (CRP) levels,and calculation of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) and sequential organ failure assessment (SOFA) score were performed.Comparison of group differences for continuous variables was done by oneway ANOVA or nonparametric Kruskal-Wallis test.Spearman rank correlation analysis was applied to establish the relation between variables.Receiver operating characteristics (ROC) curve was created and area under curve (AUC) was calculated to determine the diagnostic value of these variables in sepsis.Results The levels of plasma suPAR in SIRS group,sepsis group,severe sepsis group,septic shock group,and healthy control group were (8.22±0.61),(11.45±1.12),(12.99±1.28),(15.75± 1.23) and (4.65 ±0.30) ng/mL,respectively.Plasma suPAR levels in SIRS group and sepsis group were higher than that in healthy control group (P ＜ 0.01),and elevated plasma suPAR was accompanied by increased severity of sepsis (P ＜ 0.05).PCT levels of sepsis group (17.66 ± 8.42) ng/mL,severe sepsis group (9.67 ±3.56) ng/mL and septic shock group (29.19 ± 10.78) ng/mL were greater than that in SIRS group (1.10 ± 0.78) ng/mL,P ＜ 0.01.CRP levels elevated in all groups,but there were no significant differences among them.When suPAR and CRP were applied to distinguishing sepsis from SIRS,the AUC values from suPAR and combination of suPAR and PCT were 0.817 (P ＜ 0.01,95 ％ CI:0.714-0.921) and 0.927 (P＜0.01,95％ CI:0.870-0.985),respectively.Using9.52 ng/mL suPAR as the best cut-off to distinguish sepsis from SIRS,there were 71.93％ sensitivity and 95.46％ specificity.The levels of plasma suPAR positively correlated with PCT levels (r =0.326),APACHE Ⅱ score (r =0.492) and SOFA score (r =0.386),P ＜ 0.01.Conclusions Plasma suPAR levels significantly elevated in septic patients and correlated with the severity of sepsis.Sepsis and SIRS may be discerned by plasma suPAR levels.Joint use of suPAR and PCT could greatly increase the specificity of diagnosis of sepsis.

[ ABSTRACT] AIM:To investigate the role of protease activated receptor-2 ( PAR-2 ) in the process of tryptase mediated IEC-6 cell injury.METHODS:The rat intestinal epithelial cell line IEC-6 was treated with tryptase at different concentrations (1 μg/L, 10 μg/L, 100μg/L and 1 000μg/L) in the presence or absence of PAR-2 antagonist FSLLRY-NH2 for 12 h respectively.The cell survival rate was detected by MTT assay.The protein levels of PAR-2 and cleaved-caspase 3 were determined by Western blotting.The LDH activity was also measured.RESULTS:Compared with control group, the cell survival rates were significantly decreased in 100 μg/L and 1 000 μg/L tryptase treated groups, the LDH activities were significantly increased in 10 μg/L to 1 000 μg/L tryptase treated groups, and the protein levels of PAR-2 and cleaved caspase 3 were significantly increased in 100μg/L and 1 000μg/L tryptase treated groups (P<0.05).Com-pared with 1 000 μg/L tryptase treated group, the LDH activity and cleaved caspase 3 protein level were dramatically de-creased while the survival rate was significantly increased in the presence of PAR-2 antagonist FSLLRY-NH2 (P<0.05). CONCLUSION:Tryptase induces IEC-6 cell injury in a dose-dependent manner by activating PAR-2.