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Objective Apolipoprotein E (APOE) is mainly involved in lipid metabolism and cholesterol transport, which is important for health. To establish a pyrosequencing based method for detection of 112T>C and 158C>T single nucleotide polymorphisms (SNPs) in Apolipoprotein E gene. Methods Three amplification systems including Taq enzyme buffer from TaKaRa company (T buffer), La Taq enzyme buffer specified for G-C rich regions (L buffer) and Trans Taq enzyme buffer from TransGen company (TT buffer) were chosen to optimize PCR system and temperature by annealing at 60 °C and gradient annealing from 62 to 68 °C respectively. The specificity of this method was evaluated by comparing its results with those of Sanger sequencing. The sensitivity of this method was evaluated by gradient diluting human genomic DNA as detection template. Results According to the concentration and specificity of the products, the optimum condition was L buffer with 60℃ annealing programs. Pyrosequencing results of 20 samples were completely consistent with those of Sanger sequencing. The sensitivity of this method could be as low as 0.16 ng genomic DNA. Conclusion A method based on pyrosequencing detecting 112 and 158 polymorphisms in APOE gene was established, which can be applied in clinical personalized medicine.
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@# Objective To analyze the epidemiological characteristics of mumps in Jiamusi city, and predict the incidence trend, so as to provide evidence for taking targeted prevention and control measures in the cold areas in Heilongjiang Province. Methods The mumps cases were collected from China Information System for Disease Control and Prevention in Jiamusi city from 2004 to 2017.Descriptive epidemiologic method was used for analyzing the epidemic of mumps in Jiamusi city during 2004-2017, and the autoregressive integrated moving average (ARIMA) was used to forecast the incidence of mumps in 2018. Results A total of 1 586 cases of mumps were reported in Jiamusi city during 2004-2017, the average annual incidence rate was 4.52/100 000. The incidence of mumps increased year by year from 2015 to 2017.The ratio of male to female was 1.67 :1,those aged from 3 to 19 years accounted for 86.32% of the total cases, the higher incidence rates were found at the age of 5-9 and 10-14 years.The incidence of mumps presented obviously seasonal characteristics.Most cases concentrated from April to July and from November to January. The incidence of urban disease was higher than that of other counties; The established finally mode was ARIMA(4,1,2)(2,0,0)12 and the predicted incidence from January 2018 to June 2018 was consistent with the actual one.From July 2018 to December 2018,predictive mumps incidence were:0.30/100 000, 0.35/100 000, 0.38/100 000, 0.39/100 000, 0.35/100 000, 0.33/100 000. Conclusions ARIMA model could predict the trend of mumps in Jiamusi city.To reduce the incidence of children, it is recommended to develop a second dose of mumps vaccine at the preschool age(3-6 years)or primary school;At the same time, surveillance and control should be continually strengthened in kindergartens and schools.
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Preimplantation genetic diagnosis (PGD) was developed to play a supporting role in assisted reproductive technology. With this kind of detection method, embryos with copy number variations, chromosome translocations or single mutations were excluded and the normal embryos were chosen and implanted. Theoretically, the application of these procedures could improve the implantation and pregnancy rate and help to delivery healthy offspring. PGD was considered to be more precise, higher specific and non-invasive with the appearance of microarray hybridization technology, the next generation sequencing and time-lapse monitoring technology. This paper presented a review of new Methods used in PGD, including fluorescence in situ hybridization, array comparative genomic hybridization, SNP array, next generation sequencing, MicroSeq-PGD, MaReCs, time-lapse monitoring and cfDNA-based method, and their advantages and disadvantages as well as efficacy in the procedures in which they are used.
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The aim of this study is to apply 3D printing technology to hospital drug dosing operations, and explore its feasibility and scalability. Drugs often dosed in hospitals are selected as models. The commercially available drug was ground into powder, diluted with medicinal excipients and then mixed with 75% ethanol and binder to prepare a paste for 3D printing. The dose and physicochemical properties of divided tablets were controlled by setting print parameters and printing models in computer software. Different 3D printers were employed to evaluate the impact of the device on the dosing tablet. Two drugs were dosed in this study to explore the scalability of 3D printing technology between different drugs. The drug content of the three divided dose tablets (warfarin sodium 1 mg, 2 mg, hydrochlorothiazide 5 mg) was 1.02±0.03, 1.96±0.01, 5.19±0.06 mg. The content uniformity was 1.0, 5.3, 2.6, respectively. The drug dissolution rate was (99.3±1.2)%, (101.5±0.3)%, (98.1±0.8)% in 45, 45 and 30 min. The mechanical properties of the three sub-doses and the stability within 30 days were in line with the Chinese Pharmacopoeia (2015) requirements. At the same time, it was found that the printing parameters and prescriptions can affect the properties of the divided dose tablets. By controlling the dilution ratio of commercial drug and printing parameters, the drug release rate can be customized to achieve individualized treatment. Both different modes of 3D printers can produce qualified sub-doses, and 3D print dispensing technology was also versatile between the two drugs. 3D printing can prepare small-volume, high-precision, high-repetition dosing tablets, with all properties in compliance with pharmacopoeia regulations. Thus, this method can be used as a new and scalable sub-dosing method.