OBJECTIVE:To prepare ornidazole-?-cyclodextrin inclusion compound and establish its quality control method.METHODS:The ornidazole-?-cyclodextrin inclusion compound was prepared by saturated water solution method and the content was determined by ultraviolet spectrophotometry.RESULTS:The finished preparation was confirmed to be ornidazole-?-cyclodextrin inclusion compound under the microscope.The detection concentration range of ornidazole was5.1～20.5?g/ml(r=0.9999);the average recovery rate was101.52%,RSD=1.4%.CONCLUSION:This preparation method is practicable;the finished preparation is stable in quality and the established quality control method is simple,reliable.

Objective To establish a spontaneous abortion rat model for a syndrome in traditional chinese medicine, kidney deficiency, and observe the changes of physiological indicators and related cytokine expression in the model. Methods 40 female and 20 male rats were used in this study. The female and male rats were mated (mating ratio 2:1). The day of vaginal smear with a large number of sperm was considered as the first day of pregnancy. The rats were randomly divided into control group and model group. The model group received 450 mg/kg hydroxyurea every day. Mifepristone was given on the eighth day in a dose of 3.75 mg/kg. The diet amount, the diameter index of kidney, ovary and embryos were analyzed. The mRNA expression of Th1/Th2 cytokine was detected by RT-PCR, and the expression of co-stimulating factors CD80, CD86, CD28, CTLA-4 were determined by flow cytometry.Results Comparing the model group with control group on the eighth day, there were significant differences between the model and control groups in quantity of food and water intake, and weight increase (P<0.05), and also in the embryonic diameter index, average of abortion rate, Th1/Th2 type cytokines, co-stimulating factor CD80, CD86, CD28, and CTLA-4 (P<0.05). Conclusion A rat model of spontaneous abortion due to kidney deficiency can be successfully established with hydroxyurea and mifepristone. The high expression of Th1 (TNF-α, IFN-γ) may cause abortion and be harmful to pregnancy. Th2 type (IL-4, IL-10) may facilitate pregnancy. The expression co-stimulating factor CD80, CD86, CD28, CTLA-4 may be relevant to the spontaneous abortion.

Engineering and redesign of enzymes are important to industrial biocatalysis. Fusion enzyme technology, based on fusion protein design, is frequently used in multifunctional enzyme construction and enzyme proximity control. Here, we reviewed the recent progress in molecular design strategy and application studies of fusion enzymes. The concept and features of fusion enzymes were introduced, followed by a systematical summary of the design strategy of fusion enzymes. In particular, the effects of different linker properties on fusion enzymes and their possible mechanisms were discussed. In addition, recent studies on fusion enzyme applications were also discussed. Finally, based on our own studies on fusion enzymes and the current research progress, the key problems in fusion enzyme technology and perspectives of this field were discussed.
Biocatalysis ; Biotechnology ; Enzymes ; chemistry ; Protein Engineering ; Recombinant Fusion Proteins ; chemistry

This study investigates the metabolic regulatory effects of exogenous lactate on obesity mice induced by high-fat diet.We established obesity and metabolic disorder C57 mice model using a synthetic high-fat forage containing 60% fat.Some mice were fed with high-fat diet for 4 weeks to establish the model, being given 500 mg/(kg?d) lactate with ip for 4 weeks at the same time; the others were fed with high-fat diet for 8 weeks to establish the model, being given 500 mg/(kg?d) lactate 4 weeks after 4 weeks of modeling.During the trial period, the change of body weight and food intake, as well as serum glucose, lactate, triglycerides, insulin, and liver glycogen levels of both groups of mice were measured.Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT) were used to assess glucose metabolism and insulin resistance in the body.At the end of the experiment, adipose tissue was dissected for weighing and histopathological examination.The expression of lipid synthesis and lipolysis genes in adipose tissue was detected by real-time PCR.The results showed that: (1) in the 4-week preventive medication trial, lactate had no significant effect on the body weight of normal and high-fat diet (HFD) mice, yet it increased the subcutaneous fat/visceral fat weight ratio; lactate could significantly reduce fasting blood glucose and liver glycogen levels in HFD mice while increasing blood lactate levels, significantly improving impaired glucose tolerance in HFD mice; lactate could improve the size and arrangement of adipocytes in the HFD group while significantly down-regulating the expression of fatty acid synthesis and lipolysis genes in adipose tissue; (2) in the 8-week treatment, both routes of lactate administration could partially reduce body weight in HFD group mice and reduce food intake, with the improvement trend for fat weight; both routes of lactate administration could significantly reduce fasting blood glucose levels in HFD mice, while significantly improving glucose and insulin tolerance, with some improvement of fasting insulin levels and insulin resistance index; both routes of lactate administration showed different degrees of improvement effect on adipocyte morphology in obese mice while significantly down-regulating lipolysis gene expression in adipose tissue.Therefore, for high-fat diet-induced obese mice with metabolic imbalance, exogenous lactate can stimulate glucose metabolism, inhibit adipose tissue lipolysis, and prevent adipocyte hypertrophy, thereby improving glucose tolerance and insulin sensitivity and reducing sugar-lipid metabolic disorder.

As the COVID-19 pandemic worsens, early case detection is vital to limiting community spread. We describe our experiences with four COVID-19 cases at the polyclinics in January and February 2020. This retrospective case series highlights the challenges primary care clinicians face in the early identification of suspect cases based on clinical criteria only. To improve case detection, clinicians can sharpen their clinical acumen by keeping abreast with the latest COVID-19 developments and by maintaining a high state of vigilance.
Adult ; Aged ; COVID-19/epidemiology* ; Female ; Humans ; Male ; Pandemics ; Primary Health Care ; Retrospective Studies ; SARS-CoV-2 ; Singapore/epidemiology*

BACKGROUNDCombined emphysema and pulmonary fibrosis, including idiopathic pulmonary fibrosis (IPF), is a distinct disorder described with upper-lobe emphysema and lower-lobe fibrosis on chest computed tomography. Smoking appears to be the predominant risk factor for this disorder. We aimed to compare clinical features, smoking history, physiological and radiological findings between IPF with and without emphysema.

METHODSA sample of 125 IPF patients over a period of 48 months were evaluated. High resolution CT scans were reviewed blinded to clinical data. The IPF patients with or without emphysema were classified accordingly.

RESULTSThe prevalence of emphysema in this IPF sample was 70/125. IPF with emphysema was significantly associated with smoking status (OR 63; 95% CI 4.4 to 915; P = 0.002) and smoking pack year (OR 1.1; 95% CI 1.05 to 1.13; P = 0.000). The patients with IPF and emphysema had a higher decrease in carbon monoxide diffusing capacity adjusted for alveolar volume ((58±19)% pred vs. (66±21)% pred; P = 0.021) and a higher prevalence of pulmonary hypertension (24/70 vs. 7/55; P = 0.006). The two groups of patients had similar forced and residual volumes. No significant differences were found in cell differentials of bronchoalveolar lavage or the scores of fibrosis on chest CT. Survival of the patients with emphysema was significantly less than that of patients with IPF alone.

CONCLUSIONSCigarette smoking induces IPF combined with emphysema. Emphysema further impairs physiological function and increases the prevalence of pulmonary hypertension that leads to poor prognosis. The inclusion of the patients with combined pulmonary fibrosis and emphysema in IPF clinical trials may lead to under evaluation of the effect of treatment in patients.

Aged ; Female ; Humans ; Idiopathic Pulmonary Fibrosis ; etiology ; physiopathology ; Male ; Middle Aged ; Pulmonary Emphysema ; complications ; physiopathology ; Smoking ; adverse effects

The outbreak of coronavirus disease (COVID-19) caused by SARS-CoV-2 virus continually lead to worldwide human infections and deaths. Currently, there is no specific viral protein-targeted therapeutics. Viral nucleocapsid protein is a potential antiviral drug target, serving multiple critical functions during the viral life cycle. However, the structural information of SARS-CoV-2 nucleocapsid protein remains unclear. Herein, we have determined the 2.7 Å crystal structure of the N-terminal RNA binding domain of SARS-CoV-2 nucleocapsid protein. Although the overall structure is similar as other reported coronavirus nucleocapsid protein N-terminal domain, the surface electrostatic potential characteristics between them are distinct. Further comparison with mild virus type HCoV-OC43 equivalent domain demonstrates a unique potential RNA binding pocket alongside the -sheet core. Complemented by binding studies, our data provide several atomic resolution features of SARS-CoV-2 nucleocapsid protein N-terminal domain, guiding the design of novel antiviral agents specific targeting to SARS-CoV-2.