ABSTRACT
@#The corona virus disease 2019 (COVID-19) caused by the new coronavirus (SARS-CoV-2) has spread rapidly around the world,posing a serious threat to the public"s health. As of September 30,2020,the number of infected people in the world has reached 33 million,causing more than 1 million deaths. Normalized nucleic acid detection methods based on lab have long turnaround time and high cost. Therefore,there is an urgent need to develop a convenient method to detect SARS-CoV-2,so as to achieve rapid testing and timely control of the epidemic when resources are limited.This review summarizes the point-of-care testing (POCT) methods developed for SARS-CoV-2 in terms of extraction,amplification and detection,and briefly introduces commercial POCT instruments that integrate these three steps,in order to provide references for emergency response and rapid deployment of COVID-19 and other emerging infectious diseases.
ABSTRACT
Objective To establish an ELISA for quantitative determination of decoy receptor 3 (DcR3) in human plasma.Methods A solid phase double antibody sandwich method was established for quantitative determination of DcR3.The anti-DcR3 antibody was immobilized onto ELISA plate.DcR3 in samples was captured by anti-DcR3 on ELISA plate and then detected by biotin-anti-DcR3 and subsequent peroxidase-labeled streptavidin,and the color was developed by adding substrate.The standard DcR3 samples on the same plate were detected simultaneously to calculate the DcR3 concentrations in unknown samples.The sensitivity,specificity,precision,recovery,linearity and DcR3 range in normal human adults were assessed.Results The sensitivity of the developed assay was 0.051 ng/mL.The intra-coefficient of variation (CV) was less than 10％ and inter-CV was less than 15％.The average recovery rate was 90.50％.When 2-fold amount of anti-TNF-α was added into the coated antibodies,10-fold amount of biotin-labeled anti-LIGHT,antiFAS or anti-TNF-α was added into the detection antibodies,or 10 fold amount of purified LIGHT protein was added into the standard DcR3 samples as competitor,no disturbing effects on standard curve were found.The linear range of the assay was from 0.25 to 16 ng/mL (r≥0.98).The concentration of DcR3 tested in 128 plasma samples from healthy adults was (0.21 ± 0.05) ng/mL with 95％ CI ranged from 0.14 to 0.28 ng/mL and no difference of age and sex was found.Conclusion The established ELiSA for determining plasma DcR3 exhibited high specificity,sensitivity,precision,fine linearity and wide detecting range.This method could be used for quantification of DcR3 in plasma.
ABSTRACT
Objective To explore the clinical effect of amiodarone combine with irbesartan on arrhythmia patients with heart failure ,to observe its effect on cardiac function and its mechanism .Methods 124 patients with chronic heart failure combine with arrhythmia were selected in the hospital from September 2013 to November 2014 ,they were randomly divided into the control group and the observation group ,62 patients in each group .The control group was treated with amiodarone on the basis of routine thera-py ,the observation group was treated with amiodarone combine with irbesartan .The clinical efficacy of the two groups were as-sessed after treated 6 months ,the maintenance of sinus rhythm were observed after 1 ,3 and 6 months .By ultrasound heartbeat to assess heart function :cardiac stroke volume (SV) ,left ventricular ejection fraction (LVEF) ,left ventricular end systolic diameter (LVESD) and left ventricular end diastolic diameter (LVEDD) ,and to observe the adverse reactions .Results The efficiency rate of the observation group was 88 .71% (55/62) ,higher than that of the control group 62 .90% (39/62) ,the difference was statistically significant (P 0 .05) .Conclusion Amiodarone combine with irbesartan has good clinical efficacy in the treatment of heart failure combine with arrhythmia ,the maintenance rate of sinus rhythm can be significantly enhanced ,and improve heart function ,it has a higher se-curity .
ABSTRACT
Objective:To establish the method validation of microbial limit tests for hospital paste preparations, including com-pound salicylic acid paste, zinc oxide paste and compound pine tar paste. Methods:According to the microbial limit test described in China pharmacopoeia 2010 edition, the method validation of count of bacteria, fungi and yeasts and tests for specified microorganisms was established. Results:Medium dilution method could be used in bacteria, fungi and yeasts count and specified microorganisms ex-amination for compound salicylic acid paste and zinc oxide paste. For compound pine tar paste, bacteria, fungi and yeasts count and the pseudomonas aeruginosa examination could use medium dilution method, while the staphylococcus aureus examination should employ membrane filtration method. Conclusion:The methods of microbial limit tests for the three hospital paste preparations are established, which can be used to control the quality of hospital preparations effectively.
ABSTRACT
Objective To study the influence of valsartan combined with indapamide on the UmAlb level and blood pressure in patients with hypertension and diabetes mellitus.Methods 90 patients with hypertension and diabetes mellitus were selected and randomly divided into three groups by number table.30 cases of the control group A were treated with indapamide,30 cases of the control group B were treated with valsartan,30 cases of the observa-tion group were treated with indapamide combined with valsartan.After treatment,the clinical parameters and effect of the three groups were compared.Results After treatment,the systolic blood pressure and diastolic blood pressure of the three groups were decreased(all P <0.05),and the observation group decreased obviously(all P <0.05).The total effective rate of the observation group was 96.7%,which was higher than 86.6% of the control group A and 83.3% of the control group B(χ2 =4.1 2,4.22,all P <0.05).Conclusion In the treatment of hypertension and diabetes,the antihypertensive effect of indapamide combined with valsartan is obvious and can protect the renal func-tion,which should be promoted in clinical use.
ABSTRACT
Limulus Factor C, a serine protease zymogen from the amoebocytes of the limulus, has high affinity for endotoxin. When Factor C is activated by endotoxin, it hydrolyses artificial tripeptide substrate and measurable products are released, so it can be used as an alternative reagent for endotoxin analysis. Factor C gene of Tachypleus tridentatus was obtained through RT-PCR and the recombinant protein was expressed by Bac-to-Bac/BmNPV baculovirus expression system in silkworm larvae. The activity of Factor C was detected with diluted serum of silkworm larvae, and the sensitivity of endotoxin detected was 0.2 EU/mL when the serum was diluted at 1:500. The silkworm larvae expressed limulus Factor C could be used to develop a new low-cost endotoxin test reagent.
Subject(s)
Animals , Arthropod Proteins , Baculoviridae , Bombyx , Metabolism , Enzyme Precursors , Genetic Vectors , Larva , Metabolism , Recombinant Proteins , Serine EndopeptidasesABSTRACT
Pyrosequencing is a tool based on bioluminescence reaction for real-time analyzing DNA sequences. The sensitivity of pyrosequencing mainly depends on luciferase in reaction mixture. However, the instability of pyrosequencing reagents caused by fragile wild Photinus pyralis luciferase (PpL) in conventional pyrosequencing usually leads to unsatisfied results, which limits the application of pyrosequencing. In order to improve the stability of pyrosequencing reagents, the coding sequences of mutant thermostable Luciola lateralis luciferase (rt-LlL) was synthesized, and inserted into the plasmid of pET28a(+) to express the thermostable rt-LlL with a 6 x His-tag in the N terminal. The purified rt-LlL with the molecular mass of 60 kDa was obtained by Ni-affinity chromatography. The specific activity of rt-LlL was determined as 4.29 x 10(10) RLU/mg. Moreover, the thermostability of rt-LlL was investigated, and the results showed that rt-LlL had activity at 50 degrees C, and remained 90% of activity after incubated at 40 degrees C for 25 min. Finally, rt-LlL was used to substitute commercial Photinus pyralis luciferase in conventional pyrosequencing reagent to get thermostable pyrosequencing reagent. Comparing with conventional pyrosequencing reagent, the thermostable pyrosequencing reagent is more stable, and it's activity would not lose when incubated at 37 degrees C for 1 h. This study laid foundation of establishing reliable and stable pyrosequencing system which would be applied in Point-of-Care Testing.
Subject(s)
Animals , Enzyme Stability , Escherichia coli , Genetics , Metabolism , Fireflies , Luciferases , Genetics , Recombinant Proteins , Genetics , Sequence Analysis, DNA , MethodsABSTRACT
ObjectiveTo analyze the effect of lowering the upper limit of normal fasting glucose on the distribution of impaired gheose regulation(IGR)in the population in Wuhan urban area.MethodsThe health profiles of 1896 people after physical examinations were analyzed by X2 test.ResultsThe low limit for impaired fasting glucose(IFG)was decreased from 6.1 to 5.6 mmol/L,IFG increased 271 patients.the prevalence of IFG increased from 13.1%to 27.4%(X2=62.68,P<0.05),and the value of normal glucose test decreased from 60.3%to 47.5%(X2=120.03,P<0.05).Overweisht or obesity,hypetention,fatty liver,hyperlipidemia,ECG abnormality were significantly different(P<0.05)in groups of fasting glucose 5.6-6.0 mmol/L to<5.6 mmol/L and to 6.1-6.9 mmol/L ConclusionThe new cut point of IFG has significantly influenced the distribution of IGR in the population of Wuhan urban areas.In the newly IFG.increased subjects,metabolic abnormalities of glucose and lipid occur,and an intervention of life-style is needed for them.
ABSTRACT
The modern large-scale pyrosequencing technology is a revolution of DNA sequencing.One of the key points in this technology is to get an ATP sulfurylase immobilized on the surface of magnetic beads and with a high activity.Biotinylated ATP sulfurylase can be immobilized on magnetic beads coated with streptavidin through the specific conjunction between biotin and streptavidin, but using chemical modification method to biotinylate ATPS will affect the activity of the enzyme.ATP sulfurylase fused with the carboxyl terminal 87 residues of Escherichia coli biotin carboxyl carrier protein(BCCP87) was expressed in E.coli using fusion expression strategy.Results from Western blot analysis and SDS-PAGE analysis showed that the fusion protein could be biotinylated in vivo, and the molecular mass of the fusion protein was about 64 ku.The biotinylated ATP sulfurylase could be immobilized on the surface of magnetic beads coated with strepavidin, and the immobilized ATPS could be used for quantification of PPi and pyrosequencing.An effective enzyme for the large-scale chip-based pyrosequencing system was supplied.
ABSTRACT
Objective To study the mating behaviour and mode of Oncomelania hupensis hupensis. Methods The mature snail, Oncomelania hupensis hupensis was observed under light microscopy, and all the course of the copulation behavior was recorded. Results Oncomelania hupensis hupensis's copulation behavior included calling and mating. There were two basic types of mating behaviour inclouding four subtypes. Conclusion The mating behavior and mode of Oncomelania hupensis hupensis are described.