Objective To evaluate the performance of the Beckman Coulter ACT-5DIFF AL automated hematology analyzer and to verify whether it meets the clinical requirement.Methods The residual contamination rate,accuracy,precision,uncertainty,measurement range,reference interval,and sample injection pattern of detecting system were evaluated.Results The residual contamination of each parameter was less than or equal to 0.18％.According to room between qualitative evaluation results,compared to the target value,bias ranged from 0.32％ to 2.29％.Different concentrations of laboratory variation coefficient (namely precision) of each parameter ranged from 0.35％ to 4.46％,and both of which were less than a third of the CLIA'88 ability verification analysis quality requirements.The expanded uncertainty of each parameter was Uwhite blood cell (WBC) low =7.4％,UWBC high =3.8％,Ured blood cell (RBC) low =3.4 ％,U RBC high =2.8 ％,Uhemoglobin (HGB) low =3.9 ％,UHGB high =2.2 ％,Uplatelet (PLT) low =9.8 ％,UPLT high =7.6％,UMCV low =2.6％,and UMCV high =2.5％.Analysis had a wide measuring range:WBC (0.2 ～ 137.3) × 109/L,RBC(0.72 ～ 7.66) × 1012/L,HGB (20 ～ 231)g/L,PLT(25 ～983) × 109/L,and hematocrit (HCT) (6.1 ～68.0)％.All of them had a linear relationship,and the correlation coefficient of linear regression was close to 1.0.The reference interval quoted was suitable.Both of the automatic and the hand sample injection pattern had no significantly difference in result detection.Five categories of WBC were verified up to standard.Conclusions Under the circumstance of indoor quality control approved,each performance indicator approximately reached the laboratory quality requirements,and it also met the clinical requirements.

CYP2D6 is one of the cytochrome P450 isozymes which are involved in the metabolism of various drugs with wide use. Polymorphism at the CYP2D6 locus is one of the most widely known causes for pharmacogenetic variability in humans beings. This review focuses on the importance of CYP2D6 polymorphism in the metabolism of tamoxifen, relationships between the genetic polymorphism and prognosis of patients who have underwent endocrine therapy, and evidences indicating that CYP2D6 may be used as a predictive marker for choosing optimal endocrine therapy for patients with breast cancer.

Chromosomal integration enables stable phenotype and therefore has become an important strategy for breeding of industrial Saccharomyces cerevisiae strains. pAUR135 is a plasmid that enables recycling use of antibiotic selection marker, and once attached with designated homologous sequences, integration vector for stable expression can be constructed. Development of S. cerevisiae strains by metabolic engineering normally demands overexpression of multiple genes, and employing pAUR135 plasmid, it is possible to construct S. cerevisiae strains by combinational integration of multiple genes in multiple sites, which results in different ratios of expressions of these genes. Xylose utilization pathway was taken as an example, with three pAUR135-based plasmids carrying three xylose assimilation genes constructed in this study. The three genes were sequentially integrated on the chromosome of S. cerevisiae by combinational integration. Xylose utilization rate was improved 24.4%-35.5% in the combinational integration strain comparing with that of the control strain with all the three genes integrated in one location. Strain improvement achieved by combinational integration is a novel method to manipulate multiple genes for genetic engineering of S. cerevisiae, and the recombinant strains are free of foreign sequences and selection markers. In addition, stable phenotype can be maintained, which is important for breeding of industrial strains. Therefore, combinational integration employing pAUR135 is a novel method for metabolic engineering of industrial S. cerevisiae strains.
Genetic Engineering ; methods ; Genetic Vectors ; Metabolic Engineering ; Plasmids ; genetics ; Saccharomyces cerevisiae ; genetics ; Xylose ; metabolism

Objective To investigate therapeutic effects of phlorizin on the early stage of diabetic retinopathy(DR)in diabetic db/db mice.Methods The 24 db/db male mice were divided into 3 groups of phlorizin-treated diabetic group(20mg/kg of phlorizin by intragastric gavage)(DMT,n =8),diabetic group(normal saline solution by intragastric gavage)(DM,n=8),with C57BLKS/J db/ m mice as the control group(CC,n=8)for ten weeks.After ten weeks,all mice were sacrificed and the retina was isolated.The specimen was embedded in paraffin wax and sliced.The sections were dyed with Sudan red and observed for retinal morphological change.Meanwhile,the expressions of GSK-3β and phospho-GSK-3β in retinas were determined by western blot.Results During the study,the CC group showed good condition.The DM group featured with polydipsia,polyphagia and a rapid increase of body weight.Phlorizin-treated diabetic mice displayed much better than the DM group.Compared with CC group,DM group showed cells proliferation in capillary endothelia and obvious thickening in basement membrane.After administration of phlorizin,the vascular lesion of central regions was remarkably improved.The expression level of phospho-GSK-3β protein was lower in DM group than in control group(76.2±15.8 vs.255.4 ± 10.7,t =16.30,P＜0.01)and was higher in Phlorizin-treated DM group(188.5±18.4)than in DM group(76.2± 15.8,t=8.05,P＜0.01).Conclusions Phlorizin decreases phospho-GSK-3β3expression in retina in db/db diabetic mice and suppresses the GSK-3βactivity,which leads to the protection of DR.

Objective To explore the influence of the clinical pathway to control on hospital stay and medical expenses of pediatric rotavirus enteritis.Methods 245 infants with IRE were randomly assigned into two groups,109 in research group,136 in control group.All were treated with conventional therapy,for research group with clinical pathway.136 patients with previous medical routine therapy patients in the control group were compared.Compared two sets of average day in hospital,cure rate and cost,expenses for medicine and checking expenses proportion.Results The results shwed that the observation group of children's average hospital stay was lower than the control group,the cure rate higher than that in the control group,each time of total cost,expenses for medicine,each day medicine reducing( P ＜ 0.05 ).Checking expenses proportion was increased ( P ＜ 0.05),as to therapeutic effect,there was significant difference between two groups( P ＜ 0.05 ).Both expenses of drug ratio and ratio of the average daily expenses of drug are reduced.Conclusion Clinical pathway is applied in pediatric rotavirus enteritis can improve recovery rate,reduce hospitalization time and lower hospitalization fees and expenses,charges,medicine and inspection charge proportion.And can improve health education effect and patient satisfaction.

Objective To investigate the bacterial resistance in nationwide and understand the distribution of bacterial and resistance trend.MethodsThe 6507 clinical isolates were collected from 19 hospitals in 17 cities.The susceptibility tests were performed using agar dilution method recommended by Clinical and Laboratory Standards Institute (CLSI) in central laboratory.The values of MIC50,MIC90 and MICrange were calculated by SPSS 17.0 and the susceptibilities of isolates to antimicrobial agents were determined by using CLSI (2010) guideline.Of all 6507 isolates,4691 strains were collected from target wards and 1816 were isolated from others wards.ResultsAmong 4691 strains,1156 were Gram-positive (24.6％ ) and 3535 were Gram-negative (75.4％).Based on the minimum inhibitory concentration results,the prevalence of methicillin resistant Stapylococcus aureus and methicillin resistant Stapylococcus epidermidis are 51.6％ ( 325/630 ) and 87.0％ ( 228/262 ) respectively.Staphylococci showing intermediate or full resistance to vancomycin were not observed. Coagulase negative Staphylococci showed 2.5％ (16/642)intermediate rate and 1.6％ ( 10/642 ) full resistance rate to teicoplanin,and showed 0.5％ ( 3/642 )resistance rate to linezolid.Antibiotic resistance rate of Enterococcus faecalis to ampicillin was 17.1％(19/111),while the resistance rate of Enterococcus faecium to ampicillin reached up to 85.0％(164/193).Three Enterococcus faecium were resistant to glycopeptides.The prevalence of penicillin resistance Streptococcus pneumoniae and penicillin intermediate Streptococcus pneumoniae were 41.2％ ( 145/352) and 37.2％ (131/352) respectively based on oral penicillin criterion,while the prevalence were 0.0％ (0/352) and 6.0％(21/352) based on vein to non-meningitis criterion.A vast majority of Enterobacteriaceae maintained high susceptibility to carbapenems,with resistance rate less than 2.0％.In addition,tigecycline,moxalactam,fosfomycin and amikacin displayed desirable antibacterial activity against Enterbacteriaceae,and resistance rates to these drugs were all less than 10.0％.For non-fermenting Gramnegative isolates,resistance rate of Pseudomonas aeruginosa and Acinetobacter baumannii to imipenem were 23.1％ ( 139/601 ) and 53.5％ (419/784) respectively.Resistance rate of Acinetobacter baumannii was much higher than that during the period 2007 - 2008.Colistin,tigecycline,minocycline and fosfomycin demonstrated good antibacterial activity against Acinetobacter baumannii in vitro.Conclusions Compared with MOHNARIN 2007 -2008year surveillance results, significant increase in resistance rate of Acinetobacter baumannii was demonstrated.Resistant strains to linezolid and tigecycline were found.Bacterial resistance has been a widespread problem in our country,which requires much more attention.

Objective: To investigate the pathogen spectrum distribution and drug resistance of febrile neutropenic patients with hematological diseases in Shanghai. Methods: A retrospective study was conducted on the clinical isolates from the febrile neutropenic patients hospitalized in the departments of hematology in 12 general hospitals in Shanghai from January 2012 to December 2014. The drug susceptibility test was carried out by Kirby-Bauer method. WHONET 5.6 software was used to analyze pathogenic bacteria and drug susceptibility data. Results: A total of 1 260 clinical isolates were collected from the febrile neutropenic patients. Gram-positive bacteria accounted for 33.3% and Gram-negative bacteria accounted for 66.7%. Klebsiella pneumoniae (12.5%) , Stenotrophomonas maltophilia (9.5%) , Escherichia coli (9.1%) , Pseudomonas aeruginosa (8.7%) , Acinetobacter baumannii (6.6%) , Staphylococcus aureus (5.6%) and Enterococcus faecium (5.0%) were ranked in the first 7 of all pathogens. In the respiratory tract secretions specimens, non-fermented strains accounted for 56.2%. Stenotrophomonas maltophilia accounted for 15.2%. Enterobacteriaceae and coagulase-negative Staphylococci accounted for 42.3% (104/246) and 32.6% (85/246) respectively in blood samples. Enterobacteriaceae and Enterococcus bacteria accounted for 39.4% (76/193) and 28.5% (55/193) respectively in pus specimens. The detection rates of methicillin resistant Staphylococcus aureus (MRSA) and methicillin resistant coagulase negative Staphylococci (MRCNS) were 54.3% and 82.5%, respectively. Staphylococcus bacterial strain was not found to be resistant to linezolid, vancomycin and teicoplanin. The detection rate of Enterococcus vancomycin-resistant strains was 8.9%. Enterococcus was not detected resistance to oxazolidinone strains. Enterobacteriaceae bacteria were highly sensitive to carbapenems. The resistance rate of Pseudomonas aeruginosa to imipenem and meropenem was 34.1% and 15.8%, respectively. Stenotrophomonas maltophilia was more sensitive to minocycline hydrochloride, levofloxacin and sulfamethoxazole. The resistance rate of Acinetobacter baumannii only to cefoperazone-sulbactam was less than 10.0%. The antibiotic resistance rate of Klebsiella pneumoniae, Stenotrophomonas maltophilia, Pseudomonas aeruginosa and Acinetobacter baumanii to most of common antibiotics was lower than that of the CHINET surveillance. Conclusions The pathogenic strain distribution in common infection sites of febrile neutropenic patients was characterized. Bacterial resistance surveillance was better than the CHINET nationwide large sample surveillance in China.

Various c-mesenchymal-to-epithelial transition (c-MET) inhibitors are effective in the treatment of non-small cell lung cancer; however, the inevitable drug resistance remains a challenge, limiting their clinical efficacy. Therefore, novel strategies targeting c-MET are urgently required. Herein, through rational structure optimization, we obtained novel exceptionally potent and orally active c-MET proteolysis targeting chimeras (PROTACs) namely D10 and D15 based on thalidomide and tepotinib. D10 and D15 inhibited cell growth with low nanomolar IC₅₀ values and achieved picomolar DC₅₀ values and >99% of maximum degradation (D_max) in EBC-1 and Hs746T cells. Mechanistically, D10 and D15 dramatically induced cell apoptosis, G1 cell cycle arrest and inhibited cell migration and invasion. Notably, intraperitoneal administration of D10 and D15 significantly inhibited tumor growth in the EBC-1 xenograft model and oral administration of D15 induced approximately complete tumor suppression in the Hs746T xenograft model with well-tolerated dose-schedules. Furthermore, D10 and D15 exerted significant anti-tumor effect in cells with c-MET^Y1230H and c-MET^D1228N mutations, which are resistant to tepotinib in clinic. These findings demonstrated that D10 and D15 could serve as candidates for the treatment of tumors with MET alterations.

Objective: To carry out the 4^th round of third-party evaluation on the implementation and effect of the 1^st year of the 2^nd Phase National Healthcare Improvement Initiative(abbreviated as Initiative)since 2015. Methods: The 4^th round of the evaluation survey adopted the same methods, organization and execution, and technical roadmap as the former three rounds of evaluations. Results: The 4^th round of evaluation was carried out from 18 March to 9 April, 2019 at 185 public hospitals in 31 provinces(autonomous regions, municipalities directly under the Central Government)and Xinjiang Production and Construction Corps.Facility survey, health professional survey and patient survey were conducted at each of the sample health facilities. A total of 120 782 valid questionnaires were collected from 144 non-psychiatric health facilities, 16 246 valid questionnaires were obtained from 41 psychiatric health facilities, and 252 cases of outstanding departments/hospitals in healthcare improvement were also collected. The average overall scoring of the 12 dimensions to assess Initiative implementation at 144 non-psychiatric health facilities was 84.4%. The overall outpatient satisfaction scoring was 91.1%, 96.7%for the inpatients. The overall inpatient satisfaction(family members inclusive) at 41 psychiatric health facilities was 93%. Areas remaining to be improved include day-surgery, telemedicine and medical social work. Compared with technical services, non-technical care should be further strengthened. The compensation, workload and work environment of the healthcare providers are still to be improved. Conclusions The implementation of the Initiative by health facilities has been greatly improved. The percentage of health facilities and patients who had positive perceptions of improved doctor-patient relationship has been increasing. Patient care experiences at public hospitals have been generally improved, and the implementation of promoting traditional Chinese Medicine practices also made progress. However, work satisfaction of healthcare providers was found to be rather low, compared to the high level of patient satisfaction.

Thoracic aortic dissection (TAD) without familial clustering or syndromic features is known as sporadic TAD (STAD). So far, the genetic basis of STAD remains unknown. Whole exome sequencing was performed in 223 STAD patients and 414 healthy controls from the Chinese Han population (N = 637). After population structure and genetic relationship and ancestry analyses, we used the optimal sequence kernel association test to identify the candidate genes or variants of STAD. We found that COL3A1 was significantly relevant to STAD (P = 7.35 × 10
Aneurysm, Dissecting/genetics* ; Case-Control Studies ; Cluster Analysis ; Cohort Studies ; Collagen Type III/genetics* ; Computational Biology ; Genetic Predisposition to Disease ; Humans