Objective To evaluate the viability of adipocytes after they are subjected to different compressive stress in vitro for improvement of autologous fat graft and its clinical application.Methods Fat grafts were harvested from healthy patients who underwent liposuction for body contouring.Then the fat grafts were randomly divided into 5 experimental groups,including control group without any compressive stress,and continuous compressive stress groups (0,25,50,75,and 100 kPa),and the glucose transport test was performed at days 1,2,3 and 4.An MTT assay was also performed after 4 days with continuous compressive stress in each group with the same experimental protocol for control.Routine histological examination was done in all groups to examine possible structural destruction after compressive stress.Results As compared with the control group,the glucose transport test showed transported glucose concentration decreased with an increase in compressive stress in all of the other four groups (P＜0.01),and this effect would increase with action time.MTT assay showed absorbance (A492nm) also decreased with an increase in compressive stress in four days (P＜0.05),the similar fndings of MTT assay on the viability of fat grafts in all five groups and correlated with the glucose transport test (r=0.838,P＜0.01).Histology showed significantly distorted and fractured adipocytes and cell injury rate was to raise with an increase in compressive stress.Conclusions Our study demonstrates the harmful effect on the viability of fat grafts with an increase in compressive stress and therefore we should reduce this effect in clinical application of fat grafts.

Objective: To investigate the prevalence and resistance changes of carbapenem-resistant Enterobacteriaceae (CRE) strains isolated from children patients of Chinese Bacterial Resistance Surveillance Network (CHINET) from 2005 to 2017. Methods: Antimicrobial susceptibility testing was carried out by disk diffusion method (KB method) and automated systems. Results were analyzed according to the Clinical and Laboratory Standards Institute (CLSI) 2017 edition standards. Results: Among the 4 481 CRE clinical strains, the overall prevalence of CRE in children was 6.4%, including 8.8% in neonatal period, 7.3% in infancy, 3.8% in early childhood, 4.0% in preschool, 4.7% at school age and 7.4% of puberty. The CRE prevalence of citrobacter spp. remained stable in 2005-2017, whereas other bacteria showed an upward trend, which was higher than that of the adult group (P<0.01). Among the 4 481 CRE strains, there were 2 905 strains of Klebsiella spp. (64.8%), 813 strains of Escherichia coli (18.1%), 549 strains of Enterobacter spp.(12.3%), and 65 strains of Citrobacter spp.(1.5%). Among the 4 481 CRE strains, 20.7%, 13.3%, and 11.8% were from the intensive care unit (ICU), neonatal department and internal medicine wards, respectively. Specimens were distributed as respiratory (42.8%), urine (26.3%), and blood (14.9%). The results of antimicrobial susceptibility testing exhibited that the CRE strains were highly resistant to most commonly used antimicrobial agents in clinical practice, such as imipenem, meropenem and ertapenem, as well as penicillins and cephalosporins, etc. Conclusion The prevalence of CRE strains in children is increasing year by year, and their antimicrobial resistance to common antibacterial agents in clinical practice is extremely serious, to which serious attention needs to be paid. According to the results of antimicrobial susceptibility testings, the antibacterial agents should be rationally selected to effectively control the spread of CRE.

IFT46 is one of the important components of intraflagellar transport complex B in Chlamydomonas reinhardtii, and plays important roles in the assembly, movement and perception of ciliary. To study its functional mechanism, a GST-tagged and an MBP-tagged prokaryotic expression plasmid, pGEX-2T-ift46 and pMAL-C2X-ift46 were constructed, respectively, by inserting ift46 into the pGEX-2T and pMAL-C2X vector, and then transformed into Escherichia coli BL21 (DE3) for protein expression. SDS-PAGE (15%) analysis results showed that the molecular weights of the fusion protein GST-IFT46 and MBP-IFT46 were 70 kDa and 86 kDa, respectively. We used the fusion protein GST-IFT46 purified by affinity adsorption purification (more than 95% purity) for immunity to New Zealand white rabbits. The 5th immune serum was collected and the antibody titer was determined to be 256 000 by ELISA. The antiserum was purified by Protein A affinity adsorption purification and immobilized MBP-IFT46 purification, and the specificity of polyclonal antibodies was evaluated by Western blotting and immunofluorescence. Results showed that the polyclonal antibody prepared could specifically and precisely bind IFT46 in C. reinhardtii, and IFT46 was mainly concentrated at basal body regions and few localized along the entire length of the flagellum as punctuated dots, which will make a foundation to further study the mechanism of IFT46 in cilia related diseases such as obesity, diabetes and polycystic kidney disease.
Algal Proteins ; biosynthesis ; immunology ; Animals ; Antibodies ; chemistry ; Blotting, Western ; Chlamydomonas reinhardtii ; chemistry ; genetics ; Electrophoresis, Polyacrylamide Gel ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; Fluorescent Antibody Technique ; Intracellular Signaling Peptides and Proteins ; biosynthesis ; immunology ; Plasmids ; Rabbits ; Recombinant Fusion Proteins ; biosynthesis

Objective To investigate the antimicrobial resistance profile of the clinical isolates collected from selected hospitals across China. Methods Twenty-nine general hospitals and five children's hospitals were involved in this program. Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems. Results were interpreted according to CLSI 2017 breakpoints. Results A total of 190 610 clinical isolates were collected from January to December 2017, of which gram negative organisms accounted for 70.8％ (134 951/190 610) and gram positive cocci 29.2％ (55 649/190 610). The prevalence of methicillin-resistant strains was 35.3％ in S. aureus (MRSA) and 80.3％ in coagulase negative Staphylococcus (MRCNS) on average. MR strains showed much higher resistance rates to most of the other antimicrobial agents than MS strains. However, 91.6％ of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole, while 86.2％ of MRCNS strains were susceptible to rifampin. No staphylococcal strains were found resistant to vancomycin. E. faecalis strains showed much lower resistance rates to most of the drugs tested (except chloramphenicol) than E. faecium. Vancomycin-resistant Enterococcus (VRE) was identified in both E. faecalis and E. faecium. The identified VRE strains were mainly vanA, vanB or vanM type based on phenotype or genotype. The proportion of PSSP or PRSP strains in the non-meningitis S.pneumoniae strains isolated from children decreased but the proportion of PISP strains increased when compared to the data of 2016. Enterobacteriaceae strains were still highly susceptible to carbapenems. Overall, less than 10％ of these strains (excluding Klebsiella spp.) were resistant to carbapenems. The prevalence of imipenem-resistant K. pneumoniae increased from 3.0％ in 2005 to 20.9％ in 2017, and meropenem-resistant K. pneumoniae increased from 2.9％ in 2005 to 24.0％ in 2017, more than 8-fold increase. About 66.7％ and 69.3％ of Acinetobacter (A. baumannii accounts for 91.5％) strains were resistant to imipenem and meropenem, respectively. Compared with the data of year 2016, P. aeruginosa strains showed decreasing resistance rate to carbapenems. Conclusions Bacterial resistance is still on the rise. It is necessary to strengthen hospital infection control and stewardship of antimicrobial agents. The communication between laboratorians and clinicians should be further improved in addition to surveillance of bacterial resistance.