Objective To investigate the reliability of the integrated method for kinetic assay of substrate in the presence of product inhibition using glutathione-S-transferase(GST) as model.Methods Purified GST from pig liver was used to catalyze the conjugation of reduced glutathione(GSH) to 1-chloro-2,4-dinitrobenzene(CDNB)(final concentration at 1.0mmol/L),and reaction curve was monitored by product absorbance at 340nm.Maximal product absorbance after the completion of reaction was predicted by the integrated method.Results The optimization of product inhibition constant conferred to resistance to the variation of residual substrate concentration on the estimation of maximal product absorbance.This integrated method for kinetic substrate assay was also resistant to common source of errors.The recovery for extra GSH in rat liver homogenate was above 98% with linear response ranged from 2.0?mol/L to 90 ?mol/L.The concentration of GSH in rat liver was consistent to previous reports.Conclusion The integrated method is valid for kinetic assay of substrate when there is product inhibition,and it exhibits some universality as a kinetic method for enzymatic analysis with obvious advantages.

Objective To relate the results of surface electromyography (sEMG) of the femoris among patients with hemiplegia after a brain lesion with scores on the modified Ashworth scale (MAS),and to decide whether sEMG can be used as a quantitative index for evaluating spasm.Methods The affected quadriceps femoris of sixty hemiplegic stroke survivors were assessed by the same therapist using the MAS.sEMGs were also recorded and the root mean square and average sEMG readings from the vastus medialis (VM),rectus femoris (RF) and vastus lateralis (VL) were extracted.The correlation between these sEMG indexes and the MAS grading was analyzed.Results The average sEMG signals from the VM,RF and VL were strongly correlated with the MAS grading,and they increased with increasing MAS grade.Spearman correlation analysis demonstrated highly significant correlation (P ≤ 0.001) when the quadriceps femoris was moved passively.Conclusions Surface EMG signals from the VM,RF and VL correlate well with MAS grades,rising when the affected quadriceps femoris of a stroke survivor is moved passively.sEMG can present the electrical activity during passive movements of the quadriceps femoris and can be useful in spasticity assessment and clinical treatment.

Objective To design a kind of aseptic drainage device with safe practice and accurate measurement for clinical nurses. Methods The experiment group included 63 cases using drainage device with accurate measurement,and control group included 63 cases using disposal drainage bag with scale of visual measurement.The accuracy of drainage liquid measurement was compared between two groups,and bacterial culture was made for the drainage liquid of two groups.Results The measurement error of experiment group and control group was(3.31±1.8)mL and(56.0±5.8)mL,respectively.The difference was significant(t=-4.593,P<0.01).The bacterial culture results showed no obvious difference between two groups(positive rate were 15.9%vs 17.5%,χ2=0.057,P>0.05). Conclusion The accurately measured aseptic drainage device is convenient for nurses to practice in clinics,and its application can reduce workload of nurses and decrease the occurrence of pollution.

To improve the water solubility of daidzein, solid inclusion complexes of daidzein with two amino-modified β-cyclodextrins (ACDs), i.e., mono-6-amino-6-deoxy-β-cyclodextrin (NCD) and mono-6-ethylenediamino-6-deoxy-β-cyclodextrin (ENCD), were prepared by the saturated solution method in water under the preparation conditions as follows: the ratio of daidzein/ACD was 3∶1 and the stirring time was 72 h (83% and 67% yields, respectively).The formation of two inclusion complexes was confirmed by x-ray diffractometry (XRD) and themogravimetric (TG) analysis.The inclusion stoichiometry of the inclusion complexes was 1∶1 from the Job plot and their complexation stability constants (KS) were 899.2 and 203.8 L/mol from fluorescence titration, respectively.After formation of inclusion complexes with NCD and ENCD, the water solubility of daidzein was dramatically raised from 8.31 μg/mL to 15.2 and 13.2 mg/mL at 25℃, increasing by 1800-fold and 1500-fold.

Objective To further understand the diagnosis,clinical features and prognosis of myeloid neoplasms with erythroblast more than 50％ of bone marrow(BM) nucleated cells in the WHO Classification(2016) by analyzing the clinical data,diagnosis and prognosis of 3 patients with myeloid leukemia.Methods The ages,medical histories,symptoms and signs,and laboratory examinations from 3 patients with myeloid neoplasms whose erythroblast cells were more than 50％ of BM nucleated cells when newly diagnosed were collected.Then,they were diagnosed with the WHO Classification criteria(2008) and the WHO Classification criteria(2016),respectively,and their prognosis was evaluated with the revised International Prognostic Scoring System(IPSS-R).Results According to the WHO Classification criteria(2008),all of 3 patients were diagnosed as acute erythroid leukemia(AEL).However,according to the WHO Classification criteria(2016),2 patients were diagnosed as myelodysplastic syndrome with excess blasts-2(MDS-EB-2),and 1 was diagnosed as acute myeloid leukemia(AML) with maturation.Moreover,their prognostic scores were also different.The former two patients were older men with significant dysplasia and complex genetic abnormalities,and had poor prognosis,while the latter was a middle-aged woman with no obvious dysplasia and genetic abnormalities,and had medium prognosis.Conclusion The WHO Classification(2016) is more reasonable than the WHO Classification(2008),which tends to focus more on the different biological characteristics of diseases,and may better distinguish two types of diseases with different clinical features and prognosis.

Objective To explore the effects of interstitial cells of liver cancer and normal liver cells co‐cultured on the biological function of liver cancer malignancy so as to understand the signal pathway involved by the interaction between these cells and confirm the role of interstitial cells in cancer progression in tumor microenvironment .Methods We co‐cultured interstitial cells or hepatocyte growth factor (HGF ) and human normal liver cell L‐02 ,and then detected the expressions of the tumor‐suppressing gene PTEN and the oncogene K‐RAS and changes of cell proliferation .The downstream signaling pathways were detected by Real‐time PCR and Western blot .Results The expression of PTEN was downregulated at the transcription level by 1 .15 times and translation level by 10 times (P<0 .05) ,while the transcription level and translation level of K‐RAS increased by 1 .4 times and more than 9 times , respectively ( P< 0 .05 ) in normal liver cells co‐cultured with liver cancer mesenchymal cells .The proliferation ability was increased by more than 2 times .ELISA experiment results showed that the medium from co‐culture contained HGF over 3 times more than the control group ( P<0 .05 ,1 085+108 vs .387+23) .At the same time ,cells in the experimental group expressed more than four times of cMET than the control group cells (P< 0 .05) .Exogenous HGF consistently promoted liver cell proliferation and viability (P<0 .05) .Conclusion Our study shows that liver cancer interstitial cells activate the HGF/cMET signaling pathway by secreting HGF and promote the proliferation of normal liver cells ,suggesting a new way to explore the molecular mechanism of tumor microenvironment in tumor development and treatment of hepatocellular carcinoma .

OBJECTIVE:To improve the level of rational drug use,and to reduce the workload of clinical pharmacists. METH-ODS:Established“Intelligent Management and Solution for Clinical Rational Drug Use”of our hospital was introduced,including post-prescription review(all prescriptions review)system, self-maintenance system for rational drug use knowledge base,pre-pre-scription review (rational drug use real-time review) early warning system. The effects of intelligent system were evaluated. RE-SULTS & CONCLUSIONS:After the application of the system,the following aspects could be achieved,including“all prescrip-tions review”,“multi-prescriptions/longitudinal review”,system knowledge base real-time update and autonomous definition of re-view rules,effective communication and intervention between physicians and pharmacists before printing prescription by Yaozhidao instant messaging system. Through related trial evaluation,the proportion of false positive prescriptions decreased by 17.65% after 4 months of the application of the system;drug consultation prescriptions review average score of outpatient pharmacists increased from 81 to 94.85;the time of prescription review decreased by 4.72-5.46 times. Through mandatory interception of unqualified pre-scriptions,severe warning,timely prompt and pharmacists’intervention,the rate of unqualified prescriptions pre-reviewed by the system in respiration department decreased from 32% in Jan. 2015 and 21% in May to 4.62% in Dec. So,the application of the system promotes rational drug use in outpatient department,reduces the workload of pharmacists and ensures the safety of drug use comprehensively.

Objective To investigate the correlated pathogenetic factors and vaginal local immunity in vulvovaginal candidiasis (VVC). Methods A case control study was conducted to compare VVC group (60 cases) with normal group (60 cases). All of the women filled up the specific questionnaires. Routine examination, pH test and bacterial culture were done on the vaginal discharge. Cytokines of the vaginal lavage were measured by enzyme linked immanosorbent assay. Results (1) Outcomes of the questionnaires: there was no significant difference between the two groups in educational background,knowledge of gynecologic infection, history of gynecologic infection, hygienic habit, sex life, or use of medicine (P >0. 05). The incidence of chronic cervicitis in normal group (43%, 26/60) was higher than in VVC group (22%, 13/60; P <0.05). (2) There was no difference in vaginal pH between the two groups ( P > 0. 05 ). (3) Detection rate of candida albicans by vaginal discharge routine examination was 72% (43/60). (4) The concentrations of interleukin (IL) 2, and IL-4 in vaginal lavage did not show significant difference between the two groups ( P > 0. 05 ), but the concentrations of human defensin 5,human beta-defensin (HBD) 1, and HBD2 in WC group [(0.94±0.44) mg/L, (3.1±0.4) μg/L,(10±6) μg/L] were higher than normal group (P < 0.05). Conclusions VVC is a common vulvovaginitis. There is no significant correlation between the incidence of VVC and educational background,knowledge of gynecologic infection, history of gynecologic infection, hygienic habit, sex life, or use of medicine in the child-bearing period. Human defensin may be closely correlated with the pathogenesis of VVC.

Objective To investigate the profile of local immunity of vagina and the immune defense mechanisms against lower genital tract infections. Methods Vaginal lavage was collected from healthy women and patients of vulvovaginal candidiasis, bacterial vaginosis, Trichomonol vaginitis, human papilloma virus infection(VVC), and chlamydia trachomatis infection. Each group included 60 cases. The level of interleukin (IL) 2,4,5, 13,8 and human defensin 5 (HD5) were detected by enzyme linked immunosorbent assay(ELISA). Results (1) Cytokine of helper T cell 1(Th1): the level of IL-2 between healthy women and VVC/ bacterial vaginosis (BV)/ trichomonol vaginitis (TV)/ chlamydia trachomatis (CT) patients had no significant difference. The IL-2 level(96±33 )×10-3 pg/L of human papilloma virus (HPV) infection patients was significantly higher than that of healthy women( P<0.05). (2)Cytokine of helper T cell 2 (Th2) : the level of IL-4 between healthy women and VVC/CT patients had no significant difference. The level of IL-5 between healthy women and BV patients had no significant difference. The IL-13 level(42±15)×10-3 pg/L of TV patients was significantly higher than that of healthy women (30±29)×10-3 pg/L (P<0.05). The IL-4 level (103±28)×10-3 pg/L of HPV infection patients was significantly higher than that of healthy women (36±22)×10-3 pg/L (P<0. 05 ). (3) IL-8 : the IL-8 level (5.8± 2.7) pg/L of TV infection patients was significantly higher than that of healthy women (2. 6±2.4) pg/L (P<0.05). The level of IL-8 between healthy women and BV patients had no significant difference. (4)HD5:the HD5 level of TV , BV, VVC, HPV and CT infection patients were significantly higher than that of healthy women (P<0.05). Conclusions (1) HD5 plays an important role in the defence of vaginal epithelial cell. (2) Th2 may be more important than Thl in lower genital tract infections.(3) IL-8 plays an important role in extrinsic source infections.