The bioactivity of a working reference standard was determined by replicate bioassays with calibration against a primary reference standard. In this study the number of bioassay replicates needed for calibration first was calculated theoretically, and if the mean value of the experimental bioassay replicates fell within the predefined bioactivity level the bioactivity of the working reference was defined as 100%. Our results showed that when the total intermediate precision of the bioassay method was at 11.66% and the predefined bioactivity level was set at 95%-105% with a confidence level of 95%, 21 bioassay replicates should be carried out for calibration. The average value of the 22 experimental bioassay replicates was 101.96%, so the bioactivity of the working reference standard was consistent with that of the primary reference standard at 100%. The results suggest that a strategy of first calculating the number of bioassay replicates needed for calibration and then determining whether the resulting experimental mean value is within the predefined bioactivity level will be of value to the biopharmaceutical industry.

Asthma ; diagnosis ; metabolism ; Biomarkers ; metabolism ; Breath Tests ; Child ; Exhalation ; Humans ; Nitric Oxide ; metabolism ; Predictive Value of Tests ; Severity of Illness Index

Adolescent ; Child ; Female ; Humans ; Hypertension ; blood ; Insulin ; blood ; Insulin Resistance ; Male

Objective To study the construction of the lentiviral-mediated RNA interference(RNAi) vector targering rat suppressors of cytokine signaling 3(SOCS3) gene.Methods Three target sequences were selected by on-line designer software on Ambion according to rat SOCS3 mRNA sequence(NM053565),the complementary DNA contained both sense and antisense oligonucleotides were designed and synthesized.After annealing,these double strands DNA were cloned to pRNA-Lenti-green fluorescent protein(GFP),which contained U6 promoter and GFP.The resulting Lentiviral vector containing SOCS3 shRNA was named pRNA-Lenti-SOCS3-GFP.After the rat glioma cells(C6)were transduced with the constructed 1entiviral vectors,real-time polymerase chain reaction was used to evaluate the level of SOCS3 expression(including siRNA1 group,siRNA2 group,siRNA3 group,vacuity group and siRNA-Negative group).The pRNA-Lenti-SOCS3-GFP and Lentivector Pakaging plasmid mix were cotransfected into 293T to package Lentivirus particles.Culture supematant was harvested,then the virus titer was determined by serial dilution assay.Results The SOCS3 mRNA sequence was successfully cloned to pRNA-Lenti-GFP,which was proved by PCR and DNA sequence.Compared with control group,the SOCS3 mRNA expressions were obviously suppressed in all 3 experimental groups,especially the expression rate in siRNA1 group was reduced by 80%.The Lentiviral particle titer was determined by serial dilution assay with 1.0?1010 TU?L-1.Conclusion The lentiviral-mediated RNAi vector of rat SOCS3 gene has been constructed successfully,this may provide a potential tool for studying and treating SOCS3-related diseases.

OBJECTIVETo discuss the hemostasis of the different polarities of Platycladi Cacumen Carbonisatum (PCC) on the blood heat and hemorrhage syndrome rat model induced by dry yeast.

METHODThe SD rats were divided into seven groups. Yunnan Baiyao was taken as the positive control drug. The rats in the control group and model group were fed with CMC-Na for 7 days, and the rats in other groups were fed with corresponding drugs simultaneously. On day 7, the blood heat and hemorrhage syndrome rat model was established. Indexes including the whole blood viscosity, plasma viscosity, thrombin time (TT), activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen content (FIB), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), blood platelet count (PLT), thrombocytocrit (PCT), mean platelet volume (MPV), platelet distribution width (PDW) and the rate of platelet aggregation induced by ADP were detected. Additionally, the pathological examinations of lungs among each group were compared.

RESULTCompared with the control group, the RBC, HGB and HCT of rats in the model group increased significantly, with distinct increase in high, middle and low whole blood viscosity and plasma viscosity of rats in the model group; TT and APTT were notably prolonged, while PT was notably shortened, with significant increase in FIB content; PLT, PCT, MPV and PDW remarkably increased; Additionally, the rate of platelet aggregation induced by ADP significantly decreased. After ig administration of the ethyl acetate extract of PCC, the low whole blood viscosity and plasma viscosity remarkably decreased; TT and APTT were significantly shortened, with notable reduction in PDW and in FIB content Additionally, the rate of platelet aggregation induced by ADP significantly increased. The injury of lungs was also improved in ethyl acetate extract group. The rate of platelet aggregation induced by ADP of n-butanol extract group notablly increased. Plasma viscosity of water extract group remarkably decreased, with TT being significantly shortened. But the effects of n-butanol extract or water extract were weaker than that of ethyl acetate extract. And the effect of petroleum ether extract was the weakest.

CONCLUSIONEthyl acetate extract is the active part of PCC, showing the effect of hemostasis by reducing the low whole blood and plasma viscosity, improving coagulation function mainly by acting on the endogenous coagulation, and ameliorating the function of platelet aggregation.

Animals ; Blood Coagulation ; drug effects ; Blood Viscosity ; drug effects ; Cupressaceae ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Filtration ; Hemostatics ; isolation & purification ; pharmacology ; Male ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Thrombin Time

Objective To improve the methods of making myocardial ischemia-reperfusion model and observe the changes of electrocardiogram and ultrastructure of myocardium in rats.Methods The chest of young male SD rats through the fourth intercostal space was opened,and the left coronary artery was tied with a silicagel tube,after 30 minutes,untied to perfuse.Changes of electrocardiogram were observed and recorded.After reperfusion,the levels of AST,LDH,and CK-MB were measured and the tissue samples of the infarct areas were examined by transmission electronic microscope.Results The 90 percent of total rats were made myocardial ischemia reperfusion model successfully.In myocardial ischemia reperfusion rats:the QRS wave of myocardial ischemia-reperfusion rats was much higher than that of control group;the level of cardiac enzymes increased;myocardial and vascular endothelial cells ultrastructure was damaged seriously.Conclusions The improvement of modus operandi is right.Ischemia reperfusion can cause evident damage of myocardial and vascular endothelial cells ultrastructure in rats,and damage of myocardial cells is more severe than vascular endothelial cells.

The pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a serious impact on global public health and the economy. SARS-CoV-2 infiltrates host cells via its surface spike protein, which binds to angiotensin-converting enzyme 2 on the host cell membrane. As a result, small molecules targeting spike protein have emerged as a hotspot in anti-SARS-CoV-2 drug research. Activity screening is an important step in seeking small molecule drugs. Therefore, this article aims to review the biological activity evaluation methods of small molecule inhibitors targeting SARS-CoV-2 spike protein, with the goal of laying the foundation for the discovery of new anti-SARS-CoV-2 drugs.

Chronic Disease ; Humans ; Nasal Polyps ; classification ; Sinusitis ; classification

Adolescent ; Child ; Diagnosis, Differential ; Dystonic Disorders ; diagnosis ; drug therapy ; physiopathology ; Female ; Humans ; Male

Otolaryngology ; Periodicals as Topic ; Quality of Life ; Sickness Impact Profile