Effects of three different first-aid training methods on knowledge retention of caregivers and teachers: a randomized and longitudinal cohort study in China.

OBJECTIVES: The purpose of this study was to assess the effects of pediatric first-aid training methods on caregivers' and teachers' knowledge retention. STUDY DESIGN: This was a randomized longitudinal cohort study. METHODS: A stratified random sampling method was used to select 1282 caregivers and teachers with the help of local education authorities in 18 districts and 1 county of Shanghai, China. The selected caregivers and teachers were randomly allocated into groups that were exposed to 3 models of training, including an interactive training model (group A), lecture-based training model (group B), and video instruction training model (group C), for pediatric first-aid training for caregivers and teachers (PedFACTs). Before and after the training, a descriptive questionnaire composed of demographic information and 37 simple-choice questions about first aid was administered. During the follow-up, 120 caregivers and teachers from each of the three methods were randomized and retested 9 months after their training and 120 caregivers and teachers were randomly reselected in each of the three methods and retested 4 years after their training. RESULTS: Immediately after training, there was a significant difference in the postassessment results between groups A and B (P = 0.002) as well as between groups A and C (P < 0.001). The average interactive training model score was the highest, followed by the instruction training model and video instruction training model. There was no significant difference among the three groups in the reassessment scores at 9 months and 4 years after training (P = 0.744, P = 0.595). The difference in passing the assessment among the three groups at 9 months or 4 years after training was not maintained at a significant level. CONCLUSION: The three training methods did not affect knowledge retention of the caregivers and teachers at nine months or four years after training completion. Video instruction may be an effective, convenient, and feasible method to train caregivers and teachers.

[Effects of two standards on the overweight trend of infants and toddlers in urban Shanghai].

Objective: To observe and compare the effects of two standards on the overweight trend in urban Shanghai infants and young children. Methods: A cluster randomized controlled trial was conducted in 19 communities in two districts of Shanghai, and the subjects (n=15 019) were divided into S-group and W-group by sealed envelope randomization. The subjects were newborns born between November 2013 and December 2014. The 2005 Shanghai growth standard was applied in the S-group and the 2006 WHO growth standard was used in the W-group. At each follow-up time point age of 1, 2, 4, 6, 9, 12 and 18 months, the outpatient physician assessed the length and weight of the infants according to the standard adopted by each group and provided feeding guidance. The weight-for-age Z scores (WAZ), length-for-age Z scores (LAZ) and weight-for-length Z scores (WLZ) were calculated according to the WHO standard. Weight, length, WAZ, LAZ, WLZ and overweight ratio (WLZ≥2) were compared between the two groups using t test, Wilcoxon test and χ(2) test. Results: A total of 6 509 infants (3 391 were boys, 3 118 were girls) were in the W-group, and 8 510 infants (4 374 were boys, 4 136 were girls) were in the S-group. Among the boys, the weight values at the age of 4, 6, 9, 12, 18 months in the W-group were all lower than those in the S-group ((7.5±0.8) vs. (7.7±0.8) kg, (8.6±0.8) vs. (8.7±0.8) kg, (9.6±0.9) vs. (9.7±0.9) kg, (10.4±1.0) vs. (10.5±1.0) kg, (11.5±1.1) vs.(11.7±1.1) kg; t=4.329, 2.422, 3.739, 2.451, 2.736; P<0.01, 0.015,<0.01, 0.014, 0.009). The length had no significant difference between two groups at all months of age(all P>0.05). The overweight ratio in the W-group was lower than that in the S-group at the age of 9, 12, 18 months(3.3% (71/2 170) vs. 4.9% (143/2 927), 2.5% (51/2 037) vs. 4.5% (126/2 818), 0.8% (7/832) vs. 3.1% (39/1 266); χ(2)=6.520, 14.209, 12.350; P=0.011,<0.01,<0.01).Among the girls, except at the age of 2 months (W-group (5.6±0.6) vs. S-group (5.7±0.6), t=2.935, P=0.003), weight values had no significant difference between the two groups at other age months (all P>0.05).The length in the W-group was higher than that in the S-group at 12 and 18 months of age ((75.6±2.4) vs.(75.5±2.3)cm, (82.4±2.9) vs.(82.2±2.7) cm; t=2.351, 2.197; P=0.019, 0.028). The ratio of overweight in the W-group was lower than that of S-group at the age of 12 and 18 months (1.8% (33/1 871) vs.3.0% (80/2 658), 0.6% (5/790) vs.1.7% (20/1 178); χ(2)=6.764,4.276; P=0.009, 0.039). Conclusions: The application of WHO growth standard can help to reduce the weight gain rate of boys, promote the linear growth of girls, and thus alleviate the overweight trend of infants within 18 months. It suggested that 2006 WHO growth standard should be applied to infants within 1 year of age in Shanghai.

Comparing the effectiveness of pharmacist-managed warfarin anticoagulation with other models: a systematic review and meta-analysis.

WHAT IS KNOWN AND OBJECTIVE: Anticoagulation management services are well known to improve the quality of patient care and to reduce the rates of hospitalization and emergency department visits following adverse events related to anticoagulation therapy. The complexity of managing warfarin has led to the development of a variety of specialized models managed by pharmacists, physicians, nurses, and self-managed care. The aim of the study is to compare the effectiveness of pharmacist-managed anticoagulation control of warfarin with other models. METHODS: We performed a systematic literature search of the PubMed, Medline@Web of Knowledge, EMBASE, Cochrane Library and Cumulative Index to Nursing and Allied Health Literature to identify randomized controlled trials (RCTs) from database inception up to July 2015. The search terms used for the study were 'warfarin', 'pharmacists', 'Vitamin K antagonist', 'anticoagulation' and 'management model.' We used the Cochrane Collaboration's tool from the Cochrane Handbook to assess the risk of bias of RCTs. We performed statistical analyses using RevMan 5.3 and used the Grading of Recommendations, Assessment, Development, and Evaluations profiler to rate the quality of evidence of the outcomes. The anticoagulation control outcomes were the percentage of time within the standard and expanded therapeutic range and thrombosis events; the safety outcomes were bleeding events and mortality, and patients' satisfaction of anticoagulation service. RESULTS AND DISCUSSION: Eight RCTs from 981 potentially relevant publications with a total of 1493 patients were included. Meta-analysis of the RCTs showed that a significant difference existed between pharmacist-managed care and other models for satisfaction (mean difference (MD) = 0·41, 95% CI, 0·01-0·81, P = 0·04, low-quality evidence) with heterogeneity, and the percentage of time within the standard therapeutic range (MD = 3·66, 95% CI 2·20-5·11, P < 0·00001, high-quality evidence) without heterogeneity. However, the pharmacist-managed group demonstrated no significant improvement on the percentage of time within the expanded therapeutic range (MD = 2·85, 95% CI -0·56 to 6·26, P = 0·10, moderate-quality evidence) with heterogeneity, mortality [odds ratio (OR) = 0·97, 95% CI, 0·44-2·11, P = 0·09, high-quality evidence] without heterogeneity, the prevention of bleeding events (OR = 0·89, 95% CI, 0·56-1·44, P = 0·64, high-quality evidence) without heterogeneity, and thrombosis events (OR = 0·81, 95% CI, 0·34-1·92, P = 0·64, high-quality evidence) without heterogeneity. WHAT IS NEW AND CONCLUSION: The advantage of pharmacist-managed warfarin anticoagulation therapy in terms of anticoagulation control, safety and mortality are unclear, but resulted in significantly better patient satisfaction. Compared with other models, the superiority of pharmacist-managed warfarin anticoagulation needs to be further evaluated and validated in future research.

[Effects of Pro229-->Ser and Glu243-->Gly on the characters of thermostable catechol 2, 3-dioxygenase].

In order to investigate the effects of amino acid replacement on the characters of thermostable catechol 2, 3-dioxygenase, two mutants (Pro229Ser and Glu243Gly) of this enzyme were obtained by using the method of PCR random mutagenesis. The wild type thermostable catechol 2, 3-dioxygenase and these two mutants (Pro229Ser, Glu243Gly) were over expressed in E. coli TG1 and purified. The enzymatic characters and thermostability of the wild type enzyme and the two mutants (Pro229Ser, Glu243Gly) were analyzed. The results revealed that the optimum enzymatic temperature of the two mutants were the same as that of the wild type enzyme (60 degrees C) and the Kcat/Km value of Pro229Ser and Glu243Gly (4.89 +/- 0.01 x 10(6) mol-1 s-1 and 5.88 +/- 0.01 x 10(6) mol-1 s-1, respectively) were reduced compared with the wild type enzyme (6.97 +/- 0.01 x 10(6) mol-1 s-1). However, the thermostability of Pro229Ser extremely decreased 10.2 degrees C and the thermostability of Glu243Gly slightly increased 1.5 degrees C. It was proposed that Pro229 played an important role on the thermostability of thermostable catechol 2, 3-dioxygenase.

Purification, crystallization and preliminary X-ray studies of thermostable alkaline phosphatase from Thermus sp. 3041.

Thermostable alkaline phosphatase from Thermus sp. 3041 has been expressed in Escherichia coli, purified and crystallized. The crystals belong to space group P2(1)22(1), with unit-cell parameters a = 57.7, b = 69.9, c = 111.5 A. Diffraction data were collected to 2.54 A with a completeness of 91.1% (87.8% for the last shell), an R(merge) value of 0.105 (0.312) and an I/sigma(I) value of 9.5 (3.6).

[A study of a mutant of thermostable alkaline phosphatase].

To reveal the mechanism of protein thermostability, we used in vivo random mutagenesis to generate variants of pTAP503F which contained thermostable alkaline phosphatase (FD-TAP). After screening about 5,000 clones, we obtained 4 temperature-sensitive mutants. The study of enzymatic properties of one mutant (TAPM3) showed that the thermostability of the mutant enzyme descended a lot, compared to the wild type, while the thermoactivity remained stable. DNA sequencing showed that the G-A transition in position 1,239 resulted in the substitution from glysine to serine in position 427. This mutation conspicuously affected thermostability, Michaelis constant and energy of activation. This suggests that only one substitution of amino acid will make great changes in thermostability and other properties, meanwhile, side-chain size, charge of residues and so on, which loosen the structure of protein, will result in the descent of thermostability.

[Studies of the active site, thermostability and thermophilicity of the thermostable alkaline phosphatase by site-directed mutagenesis].

Through PCR-mediated mutagenesis, three mutants E68S, S70A and E68SS70A around active site S(69) were obtained. Their enzymatic characteristics was determined. It was found that the specific activity of E68S ascended 8 times while its optional reactive temperature climbed 20 degrees C and its Tm descended 3 degrees C; the specific activity of S70A ascended 1 time while its optional reactive temperature climbed 5 degrees C and its Tm descended 2 degrees C; the specific activity of E68SS70A descended 50% while its optional reactive temperature climbed 5 degrees C and its Tm descended 19 degrees C. These result implied that the amino acids, beside the active site, were contributed not only to enzymatic activity but also to its thermostability and thermophilicity. The work provided the direction for mutation to improve enzymatic specific activity and studying the mechanism of thermostability and thermophilicity.

[Thermostable alkaline phosphatase from Thermus sp. FD3041: cloning of the gene and expression in Escherichia coli].

A genomic library of Thermus sp. FD3041 which produces thermostable alkaline phosphatase (FD-TAP) was constructed with the vector pUC118 and the host E. coli TG1. 3-10kb inserted fragments of foreign DNA were identified in 90 percent of the 12,000 clones thus obtained. Five positive clones were detected after screening the plated library by the method of colony coloration for TAP in situ. Preliminary analysis of the enzyme expressed from one recombinant plasmid pTAP362 showed that the properties of the recombinant enzyme, such as the thermal stability and optimal temperature of reaction, were identical to those of the native enzyme. The gene of FD-TAP was located on the 2.0kb BamHI-HindIII fragment of the pTAP362, determined by its physical map and the change of enzyme activity in different partially deleted plasmids. Results of thermostability experiment in PCR thermal cycle showed that the FD-TAP would be suitable for labelling of primers and detection of PCR amplified products.

[Synthesis and antimalarial activities of fluorenemethanols].

For the purpose of improving the oral antimalarial activities of the fluorenemethanols (reported by us in previous articles) which were less effective by oral than by subcutaneous administration, 24 alpha-(alkylaminomethyl)-2, 7-dichloro-9-substituted benzylidene-4-fluorenemethanols (III) were synthesized. The results of preliminary screenings demonstrated that five compounds (No. 1-4, 8) exhibited significant antimalarial activities against Plasmodium berghei NK65 strain in mice by oral administration, at dose of 6.25 mg.kg-1.d-1 x 3 with suppressive rate of 100%. Further evaluation of these 5 compounds showed that 4 of them (No. 1-4) were superior to chloroquine in parallel tests, their ED50 and ED90 were 1.0, 1.6; 0.6, 0.9; 0.7, 1.5 and 0.8, 1.6 mg.kg-1.d-1 x 3 respectively, while the ED50 and ED90 of chloroquine were 1.9 and 2.9 mg.kg-1. d-1 x 3 respectively; one compound (No 8) was equal to chloroquine, its ED50 and ED90 were 1.5 and 3.2 mg.kg-1.d-1 x 3 respectively. Further assessment of these 4 compounds are in progress.

Cloning of DNA fragments with promoter function from temperate phage of Bacillus licheniformis.

Phage Blp7 DNA was digested with restriction enzyme and ligated to the restriction enzyme digested vector pTG402. The ligated mixture was used to transform competent cells of E. coli MC1061. Plasmid DNA was extracted from pooled transformants and competent cells of B. subtilis were transformed. By selecting yellow colonies upon spraying with catechol solution, 22 clones containing DNA fragments with promoter function were obtained. The promoter activity of 15 clones was determined by the color reaction of catechol-2,3-dioxygenase. The inserted fragment of the most potent promoter was mapped with restriction enzymes. CatO2 ase activity of two clones was measured in cells of B. subtilis of all growth phases and was found to increase rapidly at the end of the log-phase. It is inferred that these two promoters might be recognized by sigma 37.

Transient expression of the cloned mouse c-Ha-ras 5' upstream region in transfected primary SENCAR mouse keratinocytes demonstrates its power as a promoter element.

The mouse Ha-ras oncogene is activated by point mutation and overexpressed in developing papillomas during two-stage skin carcinogenesis in SENCAR mice. One of our research aims is to characterize the factors regulating Ha-ras gene expression at the transcriptional level in SENCAR mouse epidermis. Towards this goal, we sequenced 1400 bp of the 5' upstream region of the mouse Ha-ras gene so as to characterize various cis-regulatory elements present in the gene. We identified seven sites with the proper consensus sequence for binding the SP1 transcription factor and three potential binding sites for the CTF-1 factor. In addition, we located a 13-base sequence with 92% homology to the consensus sequence for an estrogen response element and two hexamers with consensus sequences identical to the core sequence of the glucocorticoid response element. A series of transient gene expression vectors was constructed in which various regions of the mouse Ha-ras 5' upstream region were fused to the chloramphenicol acetyltransferase (CAT) gene. These expression plasmids were transfected into newborn and adult primary SENCAR epidermal cells, the epidermal cell population that presumably contains the stem cells involved in two-stage skin tumorigenesis. Transient gene expression assays carried out after 48-72 h indicated that a 2.3-kb Ha-ras 5' fragment produced CAT activity comparable to that produced by pSV2CAT and pdolCMVCAT, both of which are plasmids with strong viral promoters and enhancers driving CAT gene expression. Maintenance of transfected keratinocytes under both nondifferentiating (0.05 mM calcium) and differentiating (1.2 mM calcium) culture conditions demonstrated that the mouse Ha-ras upstream region was relatively unresponsive to changes in calcium concentration in transient expression assays carried out in either newborn or adult keratinocytes. Our results demonstrated the power of the cloned mouse Ha-ras promoter and upstream region in driving transient gene expression after transfection into primary keratinocytes.

Effect of central RAS on one-kidney Grollman hypertension in rats and its mechanism.

The effect of central renin-angiotensin system (RAS) on one-kidney Grollman hypertension during the maintaining phase and its mechanism were investigated in rats. The arterial blood pressure (ABP) and the content of angiotensin II (A II) and norepinephrine (NE) in brain regions was measured respectively. 4 weeks after operation the ABP was elevated significantly, and it sustained at high level 8 weeks post-operatively. However, ABP in the control group underwent no significant changes at the same period. The A II and the NE content in the brain regions of the operated group were significantly higher than in those of the age-matched control group. During the maintaining phase of hypertension captopril (150 micrograms/10 microliters) was injected into the lateral cerebroventricle at 0.5 h, 1.0 h and 1.5 h respectively, and ABP and content of A II and NE were determined at the corresponding time. The results showed that the above three parameters decreased consistently at 0.5 h and 1.0 h, and increased gradually at 1.5 h, suggesting that the central RAS might play an important role in the maintaining phase of one-kidney Grollman hypertension in rats.

Triiodothyronine amplifies norepinephrine stimulation of uncoupling protein gene transcription by a mechanism not requiring protein synthesis.

Transcription of the uncoupling protein (UCP) gene in rat brown adipose tissue has been evaluated by a nuclear run-on transcription assay. Nuclei from euthyroid rats treated with norepinephrine (NE) exhibited a 3-4-fold increase in transcription 2 h after the injection, whereas the UCP mRNA abundance increased 2-3-fold during the same interval. In contrast, neither UCP mRNA nor the transcription rate increased in response to the same treatment in hypothyroid rats, but the transcriptional response was recovered 2 h after the injection of a receptor saturating dose of 3,5,3'-L-triiodothyronine (T3). When injected to rats given T3 4 h before, NE increased UCP gene transcription and mRNA abundance by 4-5 and 3-4-fold, respectively, by 2 h. Neither cycloheximide nor emetine prevented this effect of T3. In euthyroid rats, the UCP gene transcription rate increased within 15 min of exposure to 4 degrees C. In contrast, in hypothyroid rats exposed to 4 degrees C overnight the transcription rate remained unchanged, but it started to increase steeply 2 h after the intravenous injection of a receptor saturating dose of T3 with UCP mRNA lagging approximately 1 h behind. Unilateral denervation of interscapular brown adipose tissue in hypothyroid rats caused a 60-70% ipsilateral reduction in transcription rate and UCP mRNA. When T3 was given to these rats, it only stimulated UCP gene transcription and UCP mRNA accumulation in the intact side, but there was a symmetrical stimulation when NE was injected following the T3. We conclude that the UCP gene is under dual control by T3 and NE. The primary signal seems to be generated by NE. This signal increases UCP mRNA by stimulating the transcription rate of the UCP gene. From the denervation experiments and the lack of response of UCP to NE in hypothyroid rats, we estimate that T3 amplifies 4-5-fold the transcriptional response to NE. No stabilization of UCP mRNA needs to be invoked to explain the acute effect of T3. The T3-dependent signal amplifying the NE effect is consistent with being the T3-receptor complex or another T3-derived signal that is neither a newly synthesized protein nor a rapidly turned-over pre-existing protein, activated by T3.