Purpose Semi-macro-chromogenic substrate methods have been adopted by British Pharmacopoeia to determine the anti-F Ⅹ a activity and anti-F Ⅱ a activity of low molecular weight heparin. Based on the above methods, the micro-chromogenic substrate methods were established to determine the anti-F Ⅹ a and anti-F Ⅱ a activity of heparins. Methods After the adjustment of the concentration of reagents and the whole volume of the test, 96-well microplates and the microplate spectrophotometer have been used to determine the absorbance. Results Six samples of heparin's potencies of anti-F Ⅹ a and anti-F Ⅱ a have been determined. The test results show that the errors of the tests are small and the repeatability is good. Conclusion Micro-chromogenic method can be used to determine the activity of anti-F Ⅹ a and anti-F Ⅱ a of heparins. This method improved the efficiency of the test and decreased the cost of the test. Mow the chromogenic substrate methods are being considered as the pharmacopoeia methods to substitute the anticoagulant methods.

Objective To establish a method to quantify the residual cross-linker vinyl sulfone and its hydration product 1 ,4-Thioxane-1 ,1-Dioxide.Methods The polysaccharide was precipitated by ethanol,however vinyl sulfone and its hydration product were soluble,and can be analyzed by direct injection.The analysis was carried on Agilent DB-wax capillary column (30m ×0.53 mm,1.0 μm).The flame ionization detector (FID)and flame photometric detector (FPD)were used to detect samples and the efficiency were compared. Results The linear range of vinyl sulfone and its hydration product were separately 0.5 ~20μg/mL and 2~100μg/mL detected by FID.The Limit of Quantity (LOQ)were 0.8μg/mL and 2.3μg/mL, respctively.The Limit of Detection (LOD)were 2.6μg/mLand 7.6μg/mL,respectively,and the average recoveries (n=9)of them were 104.3%and 92.4%,respectively.Conclusion FID could meet the needs of the test,and this method is simple and accurate with high sensitivity and good repeatability,which can be used for quality control of trace vinyl sulfone and its hydration products 1 ,4-Thioxane-1 ,1-Dioxide in cross-linked sodium hyaluronate injection.

Objective To compare the clinical value of fluid attenuated inversion recovery imaging (FLAIR),magnetic resonance diffusion-weighted imaging(DWI) and magnetic resonance angiography(MRA) in diagnosis of early cerebral infarction.Methods 70 cerebral infarction patients were chosen.They underwent FLAIR,DWI and MRA examination.Results 70 patients with early cerebral infarction mostly were multiple lessions accouting for 60％.Supper actue period had 22 cases and acute period had 48 cases.DWI had the highest detection rate of cerebral infarction,particularly in the hyperacute period the detection rate reached 100％,which was significantly higher than the FLAIR and MRA(x2 =12.4,P ＜ 0.05).Conclusion Especially in the early diagnosis of hyperacute cerebral infarction,DWI is superior to FLAIR and MRA.FLAIR showed better on the cortex and periventricular lesions,MRA can check out the blood vessel lesions in the infarction area.

Objective To establish the chromogenic substrate method to determine the activity of cytochrome C.Methods Used TMB as the chromogenic substrate, reacted at 37 ℃ for 15 min, generated the yellow products, and detected the absorbance at 450 nm.The experimental design method is the 4 ×4 parallel line quantitative analysis.ResuIts The activities of cytochrome C injection samples have been determined.The linear regression equation was Y=0.9875 X -1.0221,R2 =0.9996.The accuracy and repeatability were 1.1 % and 3.6 %.ConcIusion The chromogenic substrate method was simple operation, sensitive and can be used to determine the activity of cytochrome C.

Objective To investigate the correlation between liver-type fatty acid binding protein (L-FABP) and nonalcoholic fatty liver disease (NAFLD) extent and other clinical parameters.Methods Ninety-six patients of NAFLD (NAFLD group) and 100 cases of healthy controls (control group) were selected.The levels of serum L-FABP and blood biochemical parameters were measured by enzyme-linked immunosorbent assay.The lesion degree was assessed by ultrasound.The body mass index (BMI),waist to hip ratio (WHR) and homeostasis model assessment insulin resistance index (HOMA-IR) were calculated.Results The WHR,BMI,fasting plasma glucose (FBG),triglycerides (TG),alanine aminotransferase (ALT),HOMA-IR and L-FABP in NAFLD group were higher than those in control group,there were statistical differences (P ＜ 0.05).The result of correlation analysis showed L-FABP level was positively related with ALT,TG,FBG,WHR,BMI,HOMA-IR (r =0.735,0.728,0.681,0.713,0.699,0.673 ;P ＜0.05),and negative correlation with HDL-C (r =-0.607,P ＜ 0.05).The L-FABP level in control group was (15.42 ± 2.51) g/L,mild NAFLD was (15.96 ± 2.92) g/L,moderate NAFLD was (17.48 ± 3.91) g/L,serious NAFLD was (25.14 ± 5.37) g/L.There was statistical difference in L-FABP level between serious NAFLD and control group,mild NAFLD,moderate NAFLD (P ＜ 0.05).Conclusion Serum L-FABP level of serious NAFLD patient significantly increases,and L-FABP level is related with biochemical parameters of liver function.

OBJECTIVE To explore the clinical and etiological characteristics in patients with ventilator-associated pneumonia(VAP) in emergency intensive care unit(EICU),and improve the prevention and treatment level of VAP.METHODS To analyze the clinical information of 54 patients with VAP,and to compare the clinical information with the incidence rate of VAP.Bacterial culture of airway secretion and drug sensitive test were used to analyze etiology.RESULTS Totally 148 patients were enrolled in this study,of which 54 were diagnosed as VAP.The incidence rate of VAP group was 36.5%.Respirator used time of VAP group was obviously longer than that of non VAP one.Antibiotic prophylaxis could not lower the incidence rate.A total of 59 pathogen strains were isolated by bacteria culture.The major pathogenic bacteria in VAP were Pseudomonas aeruginosa(22.0%),Acinetobacter baumannii(16.9%),Staphylococcus aureus(15.3%),Escherichia coli(10.1%),and Klebsiella pneumonia(10.1%).CONCLUSIONS VAP is one of the significant reasons for failed mechanical ventilation treatment.To prevent and treat VAP should pay attention to disinfection and aseptic manipulation,and make the rational use of antimicrobial agents.

Objective To investigate the influencing factors and improve potency methods of chymosin, to verify the stability and applicability of the national standard of chymosin.Methods The effects of different formula milk powder substrate and enzyme concentration on the determination of the activity of chymosin were studied.3 ×3 dose-response parallel line method was established.The results were compared with the different methods of absolute and relative methods.Results The different formula milk powder had a significant effect on the determination of the absolute potency of the activity of chymosin.The concentration of the enzyme was a power function relationship with the milk clotting time.Compared with the absolute potency, reproducibility of the relative potency of the results was better in different laboratories.The suitable doses in 3 ×3 dose-response parallel line method were 0.35,0.44,0.55U/mL.The confidence limit rate was less than 5%.The potency of the national standard of chymosin (140712-201302) was not significantly different between 2013 and 2015.In a certain dose range, the dose-response of the national standard of chimosin and gastropylor complex or lamb'tripe extract was linear, and the two lines were parallel.Conclusion A lot of factors can affect on the potency of chymosin.Relative potency is determinate by reference standard which can eliminate the influence of different substrates, different operators and endpoint judgment on the determination in order to make results have comparability between laboratories.The test design of 3 ×3 dose-response parallel line can control the test deviation better than the single point determination.The stability of the national standard of chymosin(140712-201302) is good, and is suitable for the potency of chymosin of the products of gastropylor complex and the extract of the lamb.

Objective To evaluate the clinical effect of ultrasound-guided percutaneous transhepatic cholangial drainage (PTCD) combined with endoscopic retrograde cholangio-pancreatography (ERCP) in treating malignant obstructive jaundice, and to discuss its technical points. Methods A total of ten cases with malignant biliary obstruction were enrolled in this study. After the failure of ERCP treatment, the patients had to be treated with ultrasound-guided PTCD immediately. The guide-wire was inserted into the duodenum through intrahepatic bile duct and common bile duct to connect with ERCP, which was followed by the biliary stent implantation or the removal of physical factors causing obstruction. The clinical results were analyzed. Results Technical success was obtained in all 10 cases. In the patients who underwent a successful guide-wire docking with ERCP the postoperative serum bilirubin was significantly decreased. The main complications were fever, elevation of amylase and transient bloody bile. Conclusion With the help of docking technique the combination use of ultrasound-guided PTCD and ERCP is a new tentative treatment for malignant obstructive jaundice after the failure of initial ERCP treatment. This technique carries promising value in clinical practice as it can significantly increase the success rate of ERCP.

The oxidation-reduction properties,relative to the Fe(Ⅲ)/Fe(Ⅱ) conversion,of horseradish peroxidase(HRP)in surfactant films at edge-plane pyrolytic graphite(EPG)electrode have been investigated by using direct electrochemical methods.Cyclic voltammograms for DDAB/HRP/CME(chemically modified eelctrode),DMPC/HRP/CME,DHP/HRP/CME at 2V/s in phosnhate buffer solution (pH 7.O) show one pair of reversible,reproducible redox peaks. The redox potentials (E1/2) in DDAB/HRP/CME,DMPC/HRP/CME and DHP/HRP/CME are-0.212V,-0.330V,-0.381V respectively. Experimental result indicated that the membranes,especially the positively charged one,providing special film environment,might interfere in the protein-electrode electron transfer process. The electron transfer processes of HRP were described in the absence and in the presence of hydrogen peroxide.There are good linear relationships between the currents and the H2 O2 concentration using all the three CMEs.The results obtained are of potential value for basic and applied bioelectrochemistry and represent an opportunity to build the third generation of biosensors.

Objective To understand the incidence of hemolytic disease in newborn (HDN)among the newborns with jaundice and the coincidence degree of the blood group serological results and the clinical diagnosis in HDN.Methods The microcolumn gel method was adopted to detect the 3 serological indexes in 276 jaundice newborns of maternal fetal blood group incompatibility,in-cluding the direct antiglobulin test,free antibody test and antibody release test.Results 108 cases of HDN were clinically diagnosed with the positive rate of 39.13%.The positive detection rate in newborns with 0-2 d old was highest(50.00%).Conclusion The serological test can provide the basis for the early diagnosis and treatment of HDN.Collecting the specimen as early as possible can improve the positive diagnosis rate of HDN.