Objective To establish a new determination method for the measuring of alkaline phosphatase activity (ALP) with p-acetyl phenyl phosphace (PAP-PNa_2) as substrate.Methods With the help of Vital semiautomatic analyzer,researched a continuous-monitoring procedure and set up experimental parameters.Results When using this assay,the wavelength of PAP's absorption was 325 nm and the Km of ALP was 0.376 mmol/L.The molecular extinction coefficient of PAP at 340 nm was 23 390 L?mol~(-1)? cm~(-1) and the concentration of citrate buffer was 0.438 mol/L.During the process,we found that the optimum pH of enzyme was 10.4,and the concentration of substrate was 5.0 mmol/L.The time of linear reaction was 900 seconds,and the linear range was 0-1 110 U/L.Serum total ALP were 63.1-118.3 U/ L(male) and 52.5-89.0 U/L(female),based on results from 60 heath adults.Conclusions The method is practical in its repetition and convenience,saves time and is not liable to be affected by bilirubin in serum.It is especially suited to the use of automatic analyzers.

Objective To explore the association between fibrinogen gene polymorphism(rs1049636) and serum γ′ fibrinogen level and ischemic stroke (IS) .Methods 421 IS patients and 421 age‐and gender‐ matched healthy controls ,including 283 males and 138 females ,were recruited in this assay .The plasma γ′fibrinogen concentration was measured by enzyme‐linked immunosor‐bent assay (ELISA) .Fibrinogen gene polymorphism(rs1049636) were genotyped by using PCR‐LDR assay .Results γ′fibrinogen concentrations in IS patients[(159 .4 ± 97 .4)U/dL] were significantly higher than that in control group[(114 .2 ± 73 .0)U/dL] with statistically significant difference(P<0 .001) .Single nucleotide polymorphism(SNP) analysis showed that rs1049636 C allele was significantly associated withγ′fibrinogen level ,but not associated with increased risk of IS(P=0 .077) .Conclusion An associ‐ation between increasedγ′fibrinogen level and IS existed in Chinese Han population .However ,no association between rs1049636 C allele and IS risk was observed in our study .

OBJECTIVETo predict the exported proteins of the novel bacterium Phenylobacterium zucineum HLK1(T) using genome-wide computational identification by searching the export signals including N-terminal signal peptides and alpha-transmembrane helices.

METHODSThe computational identification of signal sequences was based on a consensus between multiple predictive tools, including SignalP V3.0, LipoP V1.0, Phobius and TMHMM 2.0. Type IV signal peptides and proteins exported via TAT machinery were searched manually based on the conservative motifs. All the predicted proteins were classified according to the Cluster of Orthologous Group (COG) standard.

RESULTIn the total 3861 proteins encoded by P. zucineum HLK1(T) 1 378 (35.7%) were predicted to be exported proteins, most of which (totally 735, 19.0% of the proteome and 53.3% of all the exported proteins) were uncleavable transmembrane helices. In addition, 499 type I signal peptides (12.9%, 36.2%), 101 lipoproteins (2.6%, 7.3%) were also identified. Four Type IV signal peptides and 12 TAT proteins were detected as well. According to the COG classification standard, most of these exported proteins were P proteins related to inorganic ion transport and metabolism and S proteins whose functions were unknown.

CONCLUSIONThe genome of HLK1(T) coded various types of exported proteins which may play an important role in the interaction between P. zucineum and the host cell, and facilitate the strain to invade into the cell.

Amino Acid Sequence ; Bacterial Proteins ; genetics ; metabolism ; Caulobacteraceae ; genetics ; metabolism ; Computational Biology ; methods ; Consensus Sequence ; genetics ; Genome, Bacterial ; Molecular Sequence Data ; Protein Sorting Signals ; genetics ; Protein Transport ; genetics ; Proteome ; metabolism

Chronic enteritis can produce an excess of reactive oxygen species resulting in cellular damage. Stanniocalcin-1(STC-1) reportedly possesses anti-oxidative activity, the aim of this study was to define more clearly the direct contribution of STC-1 to anti-oxidative stress in cattle. In this study, primary intestinal epithelial cells (IECs) were exposed to hydrogen peroxide (H2O2) for different time intervals to mimic chronic enteritis-induced cellular damage. Prior to treatment with 200 microM H2O2, the cells were transfected with a recombinant plasmid for 48 h to over-express STC-1. Acridine orange/ethidium bromide (AO/EB) double staining and trypan blue exclusion assays were then performed to measure cell viability and apoptosis of the cells, respectively. The expression of STC-1 and apoptosis-related proteins in the cells was monitored by real-time PCR and Western blotting. The results indicated that both STC-1 mRNA and protein expression levels positively correlated with the duration of H2O2 treatment. H2O2 damaged the bovine IECs in a time-dependent manner, and this effect was attenuated by STC-1 over-expression. Furthermore, over-expression of STC-1 up-regulated Bcl-2 protein expression and slightly down-regulated caspase-3 production in the damaged cells. Findings from this study suggested that STC-1 plays a protective role in intestinal cells through an antioxidant mechanism.
Animals ; Animals, Newborn ; Blotting, Western/veterinary ; Caspase 3/*genetics/metabolism ; Cattle ; Cattle Diseases/etiology/*genetics/metabolism ; Duodenum/metabolism ; Enteritis/etiology/genetics/metabolism/*veterinary ; Epithelial Cells/metabolism ; *Gene Expression Regulation ; Glycoproteins/*genetics/metabolism ; Hydrogen Peroxide/pharmacology ; Male ; Proto-Oncogene Proteins c-bcl-2/*genetics/metabolism ; RNA, Messenger/genetics/metabolism ; Real-Time Polymerase Chain Reaction/veterinary

OBJECTIVETo explore the risk factors for irritable bowel syndrome (IBS) among school adolescents in China.

METHODA stratified, randomized study by cluster sampling was conducted, which recruited 51,956 students from high and primary schools in Chinese cities. All students were requested to fill in a questionnaire.

RESULT(1) Factors including class (odds ratio 1.12), excessive intake of pepper (odds ratio 1.17), fried (odds ratio 1.08) and starch-based foods (odds ratio 1.06), gastrointestinal tract infection (odds ratio 2.66), abuse of analgesic (odds ratio 1.49), inheritance (odds ratio 1.83), fatigue (odds ratio 1.32) and repression (odds ratio 1.45) were significantly associated with the presence of IBS (P < 0.05). High protein food (odds ratio 0.90) was a protective factor.

CONCLUSIONDifferent food intake, gastrointestinal tract infection, abuse of analgesic, inheritance and psychological factors might be related to development of IBS in the students of the cities involved in this study.

Adolescent ; Child ; China ; epidemiology ; Humans ; Irritable Bowel Syndrome ; epidemiology ; etiology ; Risk Factors ; Sampling Studies ; Students ; Surveys and Questionnaires

Objective To determine the prevalence and risk factors of functional constipation (FC) by using Rome Ⅲ criteria in the local adult communities.Methods A stratified randomized and community-based study by multi-stage cluster sampling was employed.A household survey was conducted from April to May 2010.All of the participants were interviewed face-to-face by filling out the self-administered questionnaires which based on Rome Ⅲ criteria for the diagnosis of FC.Self-rating anxiety scale (SAS),self-rating depression scale (SDS) and Athens insomnia scale (AIS) were carried out to evaluate the psychological characteristics and qualities of sleep.Results A total of 7648 subjects fulfilled the questionnaires,with the response rate as 90.0％.211 patients met the Rome Ⅲ criteria,including 90 males and 121 females.The adjusted prevalence rates of FC were 2.5％ in males,3.3％ in females and with an overall rate as 2.9％.The ratio of men to women was 1∶1.32,with significant difference between males and females (P=0.043).The most common group was in the 18-29 year-olds (x2=37.359,P=0.000).FC patients were more likely to be detected in the group with normal BMI (x2=16.087,P=0.002),having received high education (x2=27.604,P=0.000),being intelectuals ( x2=6.922,P=0.031 ) and divorced ( x2=22.000,P=0.000) than in other groups. Multivariate analysis showed that excessive intake of high-fat food was significantly associated with the presence of FC (odds ratio as 1.253,P=0.000),whereas foods with high-fiber (odds ratio as 0.854,P=0.029) might serve as protective factors.Significant differences between FC groups and control groups were found in the incidence of anxiety (with odds ratio as 2.583,P=0.000) and insomnia (odds ratio as 2.443,P=0.000).Conclusion The prevalence of FC in adult communities in Shanghai Songjiang district was not higher than that in other parts of the communities.Excessive intake of high-fat food,anxiety and insomnia might be risk factors for FC and foods with high-fiber contents might serve as protective factors.

Objective To systematically evaluate the compliance and correct rate of hand hygiene(HH) among doctors and nurses in secondary and above hospitals in mainland China,and provide data support for HH promotion.Methods Literatures about study on HH in mainland China after 2015 were retrieved from Chinese and English databases,researchers evaluated the literatures according to the relevant standards,screened the qualified literatures and extracted the data strictly according to the inclusion and exclusion criteria,Meta analysis was adopted to analyze the compliance and the correct rate of HH of doctors and nurses in different levels of hospitals and at different departments.Results The overall compliance rate of HH of nurses was higher than doctors,compliance rates of HH of doctors and nurses were 55.9％ (95 ％CI:45.9％,65.8％) and 66.0％ (95％CI:58.8％,73.1％) respectively;HH correct rates of doctors and nurses were 65.2％ (95％CI:53.6％,76.9％) and 67.9％(95％CI:56.9％,78.9％) respectively.Compliance rates of HH of doctors and nurses in tertiary hospitals were both higher than secondary hospitals,compliance and correct rates of HH of doctors and nurses in intensive care unit were both higher than general departments.Conclusion The compliance and correct rate of HH in mainland China need to be further improved,especially the promotion in primary hospitals and general departments.

Objective To investigate the occurrence of healthcare-associated infection(HAI)in different weight groups of neonates in neonatal intensive care unit (NICU)and general neonatal wards,and find out the high risk factors for HAI.Methods Targeted monitoring on hospitalized neonates in the neonatal department of a hospital in 2016 and January-November 2017 was prospectively surveyed,HAI rate,device usage rate and device-associated in-fection rate in different weight groups of neonates in NICU and general neonatal wards were compared.Results From 2016 to November 2017,a total of 3 872 neonates were monitored,the total hospitalization days were 42 427 days,56 patients had 62 cases of infection,HAI rate and HAI case rate were 1.45% and 1.60% respectively.HAI rate and HAI case rate in NICU were 5.28% and 6.03% respectively;HAI rate and HAI case rate in general neo-natal wards were both 0.46%,constituent ratio of septicemia ranked first of infection in both NICU and general neonatal wards,accounting for 43.75% and 50.00% respectively,followed by lower respiratory tract(LRT)infec-tion,accounting for 27.08% and 21.43% respectively.HAI rate of different weight groups of neonates in NICU and general wards were both significantly different(χ2＝107.78,46.65,respectively,both P＜0.0001),the lower weight of the neonates,the higher HAI rate;ventilator usage rate in neonates was positively correlated with HAI (r＝0.973,P＝0.027),the higher device usage rate,the higher HAI rate.Fungal septicemia rate per 1 000 days in rainy season was higher than that of non-rainy season (0.53‰ vs 0.37‰).A total of 70 specimens were detected, 45 strains of bacteria and fungi were isolated,pathogens were all from LRT and blood system,Pseudomonas aeru-ginosa was the main pathogen from LRT,Klebsiella pneumoniae and Candida krusei were the main pathogens from blood system.Conclusion Targeted monitoring on neonatal HAI can timely find out high risk factors for neonatal HAI,analyze the causes of infection,formulate corresponding prevention and control measures,reduce HAI rate, and ensure the medical safety of neonates.

OBJECTIVETo investigate the mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test (IRP).

METHODSTwenty-one iron overloading, 26 non-iron overloading IRP patients and 10 normal controls were enrolled in this study. The expressions of ROS, Bcl-2, Caspase-3 and apoptosis of BMMNC were analyzed by flow cytometry (FCM). Antioxidants were added to iron overloading IRP BMMNC, and then the changes of indices above were detected by FCM. The number and apoptosis of T lymphocytes of IRP patients were also detected.

RESULTSROS and apoptosis of BMMNC, myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones and normal controls (P < 0.05). The expressions of Bcl-2 on BMMNC, erythrocytes and stem cells of iron overloading IRP patients were significantly lower than those of non-iron overloading IRP ones (P < 0.05). The levels of Caspase-3 on myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones and normal controls (P < 0.05). After treatment with antioxidants, the expressions of ROS, Caspase-3 and apoptosis of iron overloading IRP BMMNC significantly decreased, but opposite for Bcl-2. The percentages of CD4(+) lymphocytes [ ( 40.86 ± 8.74）%] and CD4(+)/CD8(+) (1.44 ± 0.36) in PB of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones [(35.96 ± 7.03)% and 1.14 ± 0.37] and normal controls [(28.00 ± 6.73)% and 0.79 ± 0.21], respectively (P < 0.05), as opposite for CD8(+) lymphocytes (P < 0.05). The apoptosis of CD8(+) lymphocytes [(27.35 ± 10.76)%] and the ratio of CD8(+) apoptosis/CD4(+) apoptosis (2.51 ± 0.81) in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones [(15.47 ± 8.99)%] and normal controls (1.39 ± 0.47), respectively (P < 0.05). The apoptosis of erythrocytes and stem cells coated with auto-antibodies in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP and normal controls.

CONCLUSIONMechanisms underlying bone marrow damage by iron overload might be through the follows: ①The increased ROS induced by excessive iron deposition affected the expressions of Caspase-3 and Bcl-2, which caused more BMMNC apoptosis; ②The abnormal number and ratio of T lymphocytes caused by iron overload aggravated the abnormality of immunity of IRP; ③Iron overload may increase the damage to erythrocytes and stem cells coated with auto-antibodies.

Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Case-Control Studies ; Caspase 3 ; metabolism ; Coombs Test ; Female ; Humans ; Iron Overload ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; physiopathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; Young Adult

Objective: To evaluate the impact of KIT D816 mutation on the salvage therapy in relapsed acute myeloid leukemia (AML) with t(8;21) translocation. Method: The characteristics of the first relapsed AML with t(8;21) translocation from 10 hospitals were retrospectively collected, complete remission (CR(2)) rate after one course salvage chemotherapy and the relationship between KIT mutation and CR(2) rate was analyzed. Results: 68 cases were enrolled in this study, and 30 cases (44.1%) achieved CR(2). All patients received KIT mutation detection, and KIT D816 mutation was identified in 26 cases. The KIT D816 positive group had significantly lower CR(2) compared with non-KIT D816 group (23.1% vs 57.1%, χ(2)=7.559, P=0.006), and patients with longer CR(1) duration achieved significantly higher CR(2) than those with CR(1) duration less than 12 months (74.1% vs 31.9%, χ(2)=9.192, P=0.002). KIT D816 mutation was tightly related to shorter CR(1) duration. No significant difference of 2 years post relapse survival was observed between KIT D816 mutation and non-KIT D816 mutation group. Conclusion: KIT D816 mutation at diagnosis was an adverse factor on the salvage therapy in relapsed AML with t(8;21) translocation, significantly related to shorter CR1 duration, and can be used for prediction of salvage therapy response. KIT D816 mutation could guide the decision-making of salvage therapy in relapsed AML with t(8;21) translocation.
Antineoplastic Combined Chemotherapy Protocols ; Cytarabine ; Humans ; Leukemia, Myeloid, Acute/therapy* ; Prognosis ; Retrospective Studies ; Salvage Therapy