Objective:To isolate monocytes from human peripheral blood mononuclear cells( PBMC) ,induce macrophages,and identify the function of macrophages.Methods:Monocytes were isolated from PBMC using magnetic activated cell sorting( MACS) anti-CD14 microbead.Sorted CD14+and CD14-cells were checked by flow cytometer to evaluate the efficiency of sorting.The sorted CD14+cells were cultured in IMDM media with 10%human AB serum and 10 ng/ml M-CSF for 7 days to generate macrophages,which were identified by morphological features and phagocytosis function.Results:A high purity of monocytes was obtained by MACS anti-CD14 microbead.The percentage of CD14+cells was 10% and 85.8% before and after sorting, respectively.The macrophages were approximately 40-45 μm in maximum diameter and had the fried egg colony morphological features after 7 days culture.The lymphoma ( Raji) cells were efficiently engulfed by macrophages.Conclusion: The high purity of CD14+monocytes is isolated from PBMC and monocyte-derived macrophages efficiently engulfed lymphoma cells.

Objective To investigate the expression of serum amyloid A (SAA) in patients adipose tissue with gestational diabetes mellitus (GDM) and the correlations between SAA and insulin resistance (IR) and body mass index (BMI).Methods A total of 60 single full-term pregnant women underwent cesarean section from June 2013 to December 2013 was enrolled in this study (GDM group,n =30;control group,n =30);serum SAA level was detected with Enzyme-Linked Immunosorbent Assay (ELISA);and mRNA expression of SAA1 in adipose tissue was determined by reverse transcription PCR (RT-PCR);SPSS software was used to compare these markers,and the correlations between SAA and HOMA-IR,BMI were analyzed with Pearson correlation method.Results SAA,mRNA expressions in omental and subcutaneous fat in GDM group (0.447 ± 0.069,0.291 ± 0.067) were significantly higher than those in control group (0.194 ± 0.070,0.231 ± 0.068,P ＜ 0.01).Serum SAA levels [(21.038 ± 6.648) mg/L] and homeostasis model assessment of insulin resistance(HOMA-IR) (4.168± 2.416) in GDM group were significantly higher than those in control group [(14.384 ± 12.770) mg/L,2.045 ± 1.008,P ＜ 0.05];SAA1 mRNA expression levels in omental and subcutaneous fat were positively correlated with serum SAA (r =0.353,0.342,P ＜ 0.01).SAA1 mRNA expression levels in omental were positively correlated to pregestational BMI,late gestational BMI,weight gain in pregnancy and HOMA-IR (r =0.543,0.644,0.340,0.473,P ＜ 0.01),and SAA1 mRNA expression levels in subcutaneous fat were positively correlated to pregestational BMI,late gestational BMI,and HOMA-IR (r =0.788,0.693,0.504,P ＜ 0.01),but was no correlation with weight gain in pregnancy(r =0.013,P ＞ 0.05).Conclusions SAA mRNA expressions in omental and subcutaneous fat in GDM group and serum SAA levels increase,which is positively correlated with BMI and the degree of insulin resistance,SAA may participate in the formation of GDM by increasing insulin resistance.SAA may be used as a new monitor of GDM.

Objective To explore the optimum flow shear stress and mass transport for the construction of tissue-engineered bone.Methods The β-tricalcium phosphate (β-TCP) scaffolds seeded with human bone marrow-derived mesenchymal stem cells (HBMMSCs) were cultured in perfusion bioreactor.When the same flow rate was applied,the flow shear stress was separately 1×,2× and 3×.When the same flow shear stress was applied,the flow rates were separately 3 ml/min,6 ml/min and 9 ml/min.Cell proliferation was measured by MTT method.The construction of tissue-engineered bone was evaluated by measuring alkaline phosphatase (AKP) activity,secretion of osteopontin (OP) and osteocalcin (OC),and the mineralization of extracellular matrix (ECM).The flow shear stress and the mass transport were obtained using computational fluid dynamics.Results When the flow rate was same,the most cell proliferation was found in 2× group.The AKP activity and secretion of OC was higher in 2× and 3× groups than in those in 1× group.After 28days,the highest amount of mineralization of ECM was found in 3× group.When the flow shear stress was same,the AKP activity was highest in 6 ml/min group.After 28 days,secretion of OC and formation of mineralized ECM was highest in 3 ml/min group.When the flow rate was same,the flow shear stress was separately 0.004-0.007 Pa,0.009-0.013 Pa and 0.013-0.018 Pa.When the flow shear stress was same,the flow rate was separately 0.267-0.384 mm/s,0.521-0.765 mm/s and 0.765-1.177 mm/s.Conclusion When the tissue-engineered bone was constructed,0.013-0.018 Pa flow shear stress and 0.267-0.384 mm/s mass transport velocity could improve the construction of the tissue-engineered bone in vitro.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has spread worldwide and threatened human's health. With the passing of time, the epidemiology of coronavirus disease 2019 evolves and the knowledge of SARS-CoV-2 infection accumu-lates. To further improve the scientific and standardized diagnosis and treatment of maternal SARS-CoV-2 infection in China, the Chinese Society of Perinatal Medicine of Chinese Medical Association commissioned leading experts to develop the Recommendations for the Diagnosis and Treatment of Maternal SARS-CoV-2 Infection under the guidance of the Maternal and Child Health Department of the National Health Commission. This recommendations includes the epidemiology, diagnosis, management, maternal care, medication treatment, care of birth and newborns, and psychological support associated with maternal SARS-CoV-2 infection. It is hoped that the recommendations will effectively help the clinical management of maternal SARS-CoV-2 infection.

Female ; Humans ; Middle Aged ; Propofol ; therapeutic use ; Pulmonary Embolism ; complications ; Ventricular Fibrillation ; drug therapy ; etiology

Recombinant PRRSV △2ORF5 gene was constructed using DNA shuffling from four genetically different strains of PRRSV to study its heterologous cross-neutralizing ability. The △2ORF5 mutant gene was cloned into the vector pET-32a and transferred into E. coli BL21. SDS-PAGE confirmed that the molecular weight of the recombinant △2ORF5 was about 42 kDa, consistent with the predicted result. Then the purified recombinant protein was injected into BALB/c mouse to obtain polyclonal antibody. Western blotting analysis with mouse-anti-△2ORF5 polyclonal serum indicated that the parental virus recombinant GP5 protein reacted with the specific antibodies. Four parental viruses could be inhibited by the anti-△2ORF5 polyclonal antibody and the inhibition rates were higher than 53%. This work has laid a foundation for further development vaccine for PRRSV.