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【Objective】 To establish gas chromatography for the determination of tributyl phosphate(TBP) residues in human prothrombin complex and then verify it. 【Methods】 Acid modified polyethylene glycol(PEG)(20M) capillary column was used with n-hexane as solvent. The chromatographic parameters were as follows: gasification chamber temperature at 220 ℃, column temperature at 155 ℃, detector temperature at 220 ℃, column flow rate at 2.0 mL/min, carrier gas as N2, detector as FID, and collection time for 10min. The accuracy, repeatability, linearity, specificity, intermediate precision, detection limit, quantitative limit, range and durability were verified. 【Results】 The verification results showed that the method had good specificity. The linear regression correlation coefficient of standard curve was 0.999 90. The recovery rate were 98.4%, 97.5% and 95.7% when the concentration at 50%, 100%(30μg/mL) and 150%, respectively, with an average recovery of 97.2% and a relative deviation of 2.15%. When the concentration was 100%, the repeatability was 2.08%, and the relative deviation of intermediate precision was 1.63%. The detection limit was 0.255 μg/mL, and the quantitative limit was 0.511 μg/mL. After changing capillary chromatographic columns with different batch numbers but the same types and manufacturer, the applicability test of the system met the requirements, and the method had good durability. 【Conclusion】 This method can be used for the determination of TBP residues of human prothrombin complex in laboratory.
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【Objective】 To establish and verify the vacuum decay method for the tightness inspection of blood products. 【Methods】 The method for inspecting the tightness of blood product was established, and the detection limit, linearity, range, accuracy, precision and durability were verified according to the requirements of methodological verification.The validated method was used to check the tightness of blood product packaging. 【Results】 The detection limit of this method was 2.5 μm, linear correlation coefficient was r=1, the differential pressure of positive sample was within the allowable range of accuracy, and the durability met the requirements.The RSD of results of 6 repeatability tests and 12 intermediate precision tests were both less than 10%, and all validation items met the verification standards. 【Conclusion】 Vacuum decay method can be used to check the tightness of blood products.
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Objective:To investigate the role of peroxisome proliferator-activated receptor γ (PPARγ)/CD36 signaling pathway in macrophage lipid metabolism after Mycobacterium tuberculosis ( Mtb) infection. Methods:THP-1-derived macrophages were infected with Mtb. Four groups were included in this study, which were control group, Mtb infection group, Mtb+ rosiglitazone (ROZ, PPARγ agonist) group and Mtb+ GW9662 (PPARγ antagonist) group. Western blot and RT-PCR were used to detect the expression of PPARγ in macrophages at protein and mRNA levels, respectively. The lipids in cells were detected by oil red O staining. The concentrations of total cholesterol (TC), triglycerides (TG), low density lipoprotein (LDL-C) and high density lipoprotein (HDL-C) in the supernatant of cell culture were detected by automatic biochemical analyzer. The expression of CD36 was detected by immunohistochemistry. CCK-8 was used to detect the proliferation rate of macrophages. Results:Mtb infection significantly increased the expression of PPARγ in macrophages ( P<0.001), promoted intracellular lipid aggregation and CD36 expression and decreased the levels of TC, TG, LDL-C and HDL-C in the supernatant of cell culture ( P<0.001) and cell proliferation rate ( P<0.001). PPARγ agonist significantly enhanced the intracellular lipid accumulation and CD36 expression that were induced by Mtb infection and down-regulated the lipid level in the supernatant of cell culture and cell proliferation rate, while PPARγ antagonist reversed the above effects. Conclusions:PPARγ played a role in lipid metabolism in Mtb-infected macrophages through affecting CD36 expression.
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【Objective】 To study the quality control of human albumin intermediates content in production process using High Performance Liquid chromatography (HPLC) method. 【Methods】 In accordance with the general principles of The Chinese Pharmacopoeia (2015 edition), 3121 human albumin polymer was used to determine the component V precipitation solution, human albumin stock solution, human albumin semi-finished products and human albumin products in the production process of human albumin. The chromatographic retention time and the chromatographic peak area percentage corresponding to the retention time were analyzed. 【Results】 The polymer content of component V precipitation solution, human albumin stock solution, human albumin semi-finished products, and human albumin products were was 0.2% vs 0.0% vs 1.9% vs 1.7%, 0.2% vs 0.0% vs1.8% vs1.5% and 0.1% vs 0.0% vs 1.6% vs 1.5%, respectively. 【Conclusion】 HPLC can be used as a monitoring method for human albumin production.
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Objective To evaluate the applicationof the double-disk synergy test(DDST) and combined disk test(CDT) in clini cal metal enzyme phenotype deteetion.To evaluate the value of multiplex PCR in detecting the metallo-β-1actamase in clinical.Methods 56 strains of metallo-β-1actamase-positive strains were identified [NDM-1(n=9),VIM(n=32),IMP(n=15)]for appraise the two methods.By optimizing the design of PCR primers,3 pairs of primers were designed and detected (IMP,VIM,NDM-1) in one tube for evaluate the method.Results The DDST was 80.36％(45/56),and 100.00％(56/56) in the CDT.The accuracy of multiplex PCR was 100.00 ％ (56/56),and the size of the amplified fragment was used to distinguish three types of metallo-β-lactamase.Conclusion The CDT is more suitable for clinical application than DDST.Multiplex PCR has the characteristics of simple,rapid and accurate in detection of metallo-β-1actamase.It is suitable for daily use of clinical microbiological laboratory,which will help the clinical timely and effective administration.
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Objective To identify the drug resistance-related genes in a clinically isolated strain of Enterobacter cloacae.Methods A strain of Enterobacter cloacae was isolated from sputum of a patient with chronic obstructive pulmonary disease from the Affiliated Hospital of Xuzhou Medical University in March 2013.Modified Hodge test and metal enzyme inhibition test were performed for drug-resistant phenotype screening.Carbapenemase genes blaMUS-1, blaVIM-1, blaVIM-2, blaIMP, blaKPC-2, blaNDM-1, blaOXA-48 and blaGESwere amplified by polymerase chain reaction (PCR), and the positive products were sequenced and analyzed.Plasmid conjugation and transformation experiments were used to confirm that the resistance gene mediated by plasmids.Agar dilution method was used for antibiotic susceptibility test.Results Both modified Hodge test and metal enzyme inhibition test were positive in this strain of Enterobacter cloacae.blaNDM-1 gene and blaKPC-2 gene were detected by PCR, and further confirmed by sequencing.blaNDM-1 gene was carried by IncX plasmid with 54×103 bp, KPC-2 gene was carried by untyping plasmid with 42×103 bp.The strain was only sensitive to tetracycline (MIC=2 μg/mL) and tigecycline (MIC=1 μg/mL).The symptoms were improved after the patient was treated by tigecycline combined with Piperacillin/Tazobactam.Conclusion blaNDM-1 and blaKPC-2 genes in Enterobacter cloacae can be mediated by plasmids, and appropriate therapy for its infection should be based on the result of antibiotic susceptibility test.
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Objective To evaluate the applicationof the double-disk synergy test(DDST) and combined disk test(CDT) in clini cal metal enzyme phenotype deteetion.To evaluate the value of multiplex PCR in detecting the metallo-β-1actamase in clinical.Methods 56 strains of metallo-β-1actamase-positive strains were identified [NDM-1(n=9),VIM(n=32),IMP(n=15)]for appraise the two methods.By optimizing the design of PCR primers,3 pairs of primers were designed and detected (IMP,VIM,NDM-1) in one tube for evaluate the method.Results The DDST was 80.36％(45/56),and 100.00％(56/56) in the CDT.The accuracy of multiplex PCR was 100.00 ％ (56/56),and the size of the amplified fragment was used to distinguish three types of metallo-β-lactamase.Conclusion The CDT is more suitable for clinical application than DDST.Multiplex PCR has the characteristics of simple,rapid and accurate in detection of metallo-β-1actamase.It is suitable for daily use of clinical microbiological laboratory,which will help the clinical timely and effective administration.
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Objective To investigate the mechanism related to reduced antibiotic sensitivity of Acinetobacter baumannii inducted by meropenem in vitro.Methods Three strains of clinically isolated carbapenems-sensitive Acinetobacter baumannii were induced by meropenem in vitro, and the mutant strains (MS1, MS2 and MS3) were obtained.Minimal inhibitory concentrations (MICs) of antimicrobial agents to strains before and after induction were determined by automatic drug sensitivity analyzer .The homology of strains was analyzed by Enterobacterial repetitive intergenic consensus -polymerase chain reaction ( ERIC-PCR).Modified Hodge test and EDTA-Na2-double disk synergy test were used to detect carbapenemase and metallo-β-lactamase (MBL), respectively.Main carbapenemase genes were detected by PCR and followed by DNA sequencing.Expressions of adeB and outer membrane proteins in strains before and after induction were detected with fluorescence quantitative PCR and SDS -polyacrylamide gel electrophoresis , respectively.t test was used for data analysis .Results The sensitivity of mutant Acinetobacter baumannii strains to meropenem and most antibiotics was reduced , except to imipenem, amikacin and polymyxin; and the reduced sensitivity to meropenem in MS2 and MS3 was of genetic stability.ERIC-PCR showed 100%homology between the mutant strains and parental strains .Both carbapenemase and metallo -β-lactamase were negative in mutant strains and parental strains , and only OXA-51 gene was found.The expressions of adeB gene in mutant strains were 24.26 ±0.91, while those in parental strains were 22.81 ±0.38, and the difference was not significant (t =2.534, P >0.05).Outer membrane protein with molecular weight 54 000 was missing in MS1, while that with molecular weight 47 000 was missing in MS2 and MS3.Conclusion Reduced antibiotics sensitivity in meropenem -induced Acinetobacter baumannii may be correlated with the deficiency of outer membrane protein with molecular weight 47 000.
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Objective The aim of this study was to analyze the effect of gender and age on shoulder and hip range of motion (ROM) and to determine the differences of ROM of normal side shoulder and hip joints between the data we collected and the published standards (Guideline of Examination for Body Impairment in Clinical Forensic Medicine SF/ZJD0103003-2011) in China. Methods We collected cases with unilateral injury of shoulder or hip from clinical forensic medicine. The differences of ROM of normal side joint between the data we collected and the published standards were studied. Descriptive statistics was calculated between male and female subjects in ifve age groups including 0~25, 26~40, 41~50, 51~60 and >60 years, and the ROM changes with age were also studied. Results The data collected in this study was signiifcantly different from the published standard. With age growth, the ROM of shoulder and hip were decreased and a significant difference between genders in partial activity direction of shoulder and hip was found in the individuals who were over 50 years. Conclusion Gender and age appear to be inlfuential factors to determine the normal ROM of the shoulder and hip joint, and there is signiifcantly difference of ROM of normal side shoulder and hip joints between the data we collected and the published standards (Guideline of Examination for Body Impairment in Clinical Forensic Medicine SF/ZJD0103003-2011).
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Advances in embryo culture technology and cryopreservation have led to a shift in in vitro fertilization [IVF] from early fresh or frozen-thawed cleavage embryo transfer to fresh or frozen-thawed blastocyst stage transfer. To compare the clinical outcomes of fresh embryo transfers and frozen-thawed embryo transfers. In this retrospective case control study, patients undergoing IVF cycles from January 2012 to December 2012 were enrolled in Assisted Reproduction of Wuhan Union Hospital were enrolled. A total of 1891 cycle contains 1150 fresh embryo transfers and 741 frozen-thawed embryo transfers were studied. All data were transferred directly to SPSS 18 and analyzed. Clinical pregnancy rates of fresh cleavage-stage embryo transfers compared with fresh blastocyst transfers, frozen-thawed cleavage-stage embryo transfers, post thaw cleavage-stage extended blastocyst culture transfers and frozen-thawed blastocyst transfers were 52.7%, 35.88%, 35.29%, 47.75%, 59.8% in patients under 35 years of ages and 41.24%, 26.92%, 11.32%, 46.15%, 55.8% in patients older than 35 years old, respectively. The multiple pregnancy rates, abortion rates and ectopic pregnancy rates did not differ significantly among the five groups. The clinical pregnancy rates were not different significantly between fresh cleavage-stage embryo transfers and fresh blastocyst transfers. But the clinical pregnancy rate of frozen-thawed blastocyst transfer was the highest among fresh/frozen-thawed embryo transfers
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Objective To compare the efficacy of laparoscopic gallbladder bed dissection(LGBD)between retrograde plus anterograde and anterograde only manners.Methods All of 575 cases of laparoscopic cholecystectomy(LC)were randomly divided into two groups(group A and group B).Patients underwent LGBD with retrograde plus anterograde mannerin group A(297 cases)oronly with anterograde manner in group B(278 cases)respectively.The efficacy and safety were compared between two groups.Results The operative time were(10.0±3.7)min and(13.0±3.9)min in group A and group B respectively (P<0.05).There were 33 cases of bleeding volume more than 5 ml in group A with 85 cages in group B (P<0.05).Three cases of gallbladder rupture were found in group A compared with 10 cases in group B (P<0.05).Postoperative fluid collection in gallbladder bed occurred in 5 cases in group A compared with 19 cases in group B(P<0.05).The conversion rate was 3.70%(11/297)and 3.96%(11/278)in group A and group B respectively(P>0.05).One cage of abdominal fluid collection was observed in group A whichwas relieved after conservative therapy.No other complications occurred.Conclusions LGBD applying retrograde plus anterograde manner may shorten the operative time,decrease the bleeding during operation and minimize the postoperative complications.Surgeons might benefit from its advantage of easier manipulation.
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Objective: To evaluate feasibility of using European Nutritional Risk Screening 2002(NRS2002) for nutritional status assessment in primary hospital.Methods: 799 patients were enrolled in this study.697 patients who were according NRS request were assessed using NRS method in hospitalization day.The patient whose NRS scores were higher than 3 were justified under nutritional risk.102 patients who were not according NRS request were assessed by exeamining the serum albumin(ALB) in the next day.The patients whose serum ALB was lower than 35 g/L were justified malnutrition.Results: There were 697 patients could be evaluated by NRS method.Body Mass Index(BMI) was lower than 18.5 in 159(22.8%) cases and NRS scores were ≥3 in 240 cases(34.4 %) among 697 cases.Serum albumin was lower than 35 g/L in 31 (30.4%) cases among 102 cases who could not be evaluated by NRS method because of incapable of standing up(66/102,64.7%) and ascites or hydrothorax(17/102,16.7%) and concomitance of above-mentioned causes(11/102,10.8%) and coma(8/102,7.8%).Conclusion: Assessment of nutritional status of hospitalized patients in primary hospital using NRS2002 method is feasible.The patients who could not be evaluated by NRS 2002 method can be assessed by assaying serum ALB.