The expression of heat shock proteins(HSPs) can protect against acute lung injury(ALI).However,HSPs are restrained from clinical application due to the toxicity of most of the former inductors.Glutamine,which also has the ability to induce the expression of HSPs,can protect against ALI and sepsis,and may serve as a candidate for clinical application.

Objective To investigate the in vivo effects of glutamine(Gln) on inflammatory cytokine release in murine peritoneal macrophages during sepsis. Methods Sixty Kunming mice were randomly divided into sham-operation group(Sham group,n=20),operation control group(Con group,n=20) and Gln-treatment group(Gln group,n=20).Sepsis was induced by cecal ligation and puncture in Gln group and Con group,and Gln(0.75 g/kg) or saline was immediately administered via single tail vein injection.Serum was collected and macrophages were harvested from peritoneal lavage at 6 h in these three groups.Intracellular and serum cytokines of tumor necrosis factor-?(TNF-?),interleukin(IL)-6 and IL-10 were detected by ELISA.The expression of TNF-? mRNA in macrophages was analyzed by RT-PCR,and the expression of heat shock protein(HSP) 72 in macrophages was evaluated by Western blotting. Results Gln group demonstrated significantly lower intracellular TNF-? and IL-6 levels than Con group(P0.05).The serum TNF-? level was significantly lower in Gln group than in Con group(P

Animals ; Arrestins ; metabolism ; Hypoxia ; metabolism ; Male ; Prefrontal Cortex ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; metabolism ; beta-Arrestin 1 ; beta-Arrestins

Endophytic fungi were isolated from Macleaya cordata growing in Dabie Mountain by agar-block method, and then the endophytic fungi were grouped into different types based on their morphological characteristics, and thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) were employed to determine whether the metabolic substances contained sanguinarine or not, and then preliminarily identified by morphological method. The results showed that the leaves hosted the largest number of endophytes (96 isolates) followed by the stems (57 isolates) and finally the roots (28 isolates), respectively. Based on morphological characteristics the endophytic fungi were grouped into 26 types in our study. TLC and HPLC results showed that there was sanguinarine in the metabolic substances of BLH 51 strain. According to the morphological characteristic, the BLH 51 strain was identified as Fusarium proliferatum. All these indicated that the medicinal plant M. cordata harbors abundant endophytes, which could be a new source for the search of active secondary metabolites.
Benzophenanthridines ; metabolism ; Endophytes ; isolation & purification ; Fungi ; isolation & purification ; Isoquinolines ; metabolism ; Papaveraceae ; metabolism ; microbiology ; Plant Leaves ; microbiology ; Plant Roots ; microbiology ; Plant Stems ; microbiology

Objective To optimize and compare four in vitro cytotoxicity assays in order to find a relatively rapid assay that can replace the traditional 51Cr release assay to evaluate the cytotoxicity of immune cells prepared for immunotherapy.Methods Four assays including BATDA, CAM (calcein acetoxymethyl ester), CytoTox-Glo and PKH were optimized and used to measure the in vitro cytotoxicity of NK-92 cells to K562 cell line.Intra-and inter-assay reproducibility of these assays and their correlation with 51Cr release assay were analyzed.Results After optimization, all of the four cytotoxicity assays showed good correlation with effector to target (E/T) ratio in a certain range.Compared with the other three assays, CytoTox-Glo assay showed obvious hook effect at a high E/T ratio of 40∶1.BATDA assay could detect the significant cytotoxicity of NK-92 cells to K562 cells after incubating them for one hour and that was the shortest time taken by the four assays to detect in vitro cytotoxicity.Both CAM and PKH assays took about four hours and CytoTox-Glo assay took six to eight hours to detect the significant cytotoxic potency.All of the four assays, especially BATDA and CAM assays, showed good intra-and inter-assay reproducibility.Among these assays, BATDA assay showed the best correlation with the traditional 51Cr release assay.BATDA assay, as compared with the other three assays, could be used to detect the cytotoxicity of Caov3 cells, an adherent cell line, and showed good results in evaluating the cytotoxic potency of autologous primary NK cells and CD19-CAR T cells.Conclusion Compared with the other three assays, BATDA assay shows the best linear correlation with 51Cr release assay and has the advantages of time-saving and good reproducibility.Besides, it is a better assay for detecting the cytotoxicity of adherent cells.BATDA assay is a promising substitute for 51Cr release assay in evaluating the in vitro cytotoxic potency of NK cells and other immune cells.

Objective To evaluate the short term effect of bipolar arthroplasties in aged cases with intertrochanteric fractures.Methods There were 23 males and 15 females(at age of 70-93 years, average 76 years)with unstable intertrochanteric fractures treated with the third generation cementing techniques and bipolar arthroplasties that were followed up for average 2.4 years.The clinical effect was evaluate with Harris scale,X-ray films and complications.Results Of all,34 cases(89.5%)could walk freely.The average Harris scale was 84.2 points.The average period until walk with full-weight load was 5.6 weeks.The greater trochanters was united in 35 cases(92%)during average 4.2 months.No peri-prothetic ossteolysis and loosening or subsidence occurred.Conclusions Cemented bipolar arthro- plasty has good advantages of reduced laying up period and few complications.The short-term outcome is satisfactory hut the long-term outcome needs deeper observation.

In order to study genetic variation diversity of porcine circovirus type 2 (PCV2) strains in Shanxi,the genomic sequences of nine PCV2 strains including SXQX,SXCZ,SXTY2,SXJC,SXJX,SXLL,SXPY,SXPG and SXXY recently isolated from some areas of Shanxi from 2013 to 2016,was cloned,sequenced and received by GenBank.The amplified PCV2 genomic sequences,ORF2 sequences and Cap protein amino acid of these nine strains were analysed and compared with those of published 28 PCV2 strains by DNAStar,drawing phylogenetic tree.The results showed that the genomic sequences of SXJX,SXJC and SXXY PCV2 strains were 1 768 bp,and the others were 1 767 bp,which accounted for 33％ and 67％,respectively.The homologies of nucleotide sequences of the nine strains were 94.7％-99.8％,the homologies of nucleotide sequences of the nine strains with the 28 isolates from different regions of the world PCV strain were 93.9％-99.9％,and the homologies of nucleotide sequences of the nine strains with the domestic vaccine strains were 95.1％-99.8％.The phylogenetic analysed that SXJX,SXJC and SXXY belonged to genotype PCV-2D,SXLL,SXPY and SXCZ belonged to genotype PCV-1C,and SXTY14,SXPG and SXQX belonged to genotype PCV-1A/1B.Thus it proved that the epidemic strain of PCV2 was mainly PCV-2b in Shanxi.The homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains were 90.0％-100.0％ and 87.1 ％-100.0％ respectively,the homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains with the 28 isolates from different regions of the world PCV strain were 87.6％-100.0％ and 84.1％-100.0％ respectively,and the homologies of ORF2 nucleotide sequences and Cap amino acid of the nine strains with the domestic vaccine strains were 91.0％-100.0％ and 89.3％-100.0％ respectively.The Cap amino acids of SXQX,SXJX,SXTY14,SXPG,SXJC and SXXY PCV2 were 233,ORF2 of SXQX,SXTY14 and SXPG located at 1 033-1 734 bp,ORF2 of SXXY,SXJX and SXJC located at 1 033-1 734 bp,and the Cap amino acids of SXCZ,SXLL and SXPY PCV2 were 234,ORF2 of them located at 1 030-1 734 bp,in addition,the positions of 1 030-1 734 bp were more three bases TCA than other ORF2 genome sequence of 1 767 bp,resulting in increasing a K (Lys) of amino acid sequencein at the 234 position.Also Cap protein of 9 PCV2 strains showed more amino acid variation in addition to the only high-ly conserved glycosylation sites (NYS) (pp.143-145 amino acid).It provided theoretical basis for the PCV2 immune prevention of research in Shanxi,and the data of basic theory of molecular pathogenesis of PCV2.

OBJECTIVETo observe the clinical efficacy of acupuncture at Hegu (LI 4) on central facial nerve paralysis after ischemic stroke, and explore dose-effect relationship among different stimulation intensities of acupuncture at Hegu (LI 4) as well as its optimal treatment plan.

METHODSAccording to different acupuncture stimulation intensities which were based on treatment time and needle insertion direction, fifty patients were randomly divided into a Hegu 1 group, a Hegu 2 group, a Hegu 3 group, a Hegu 4 group and a control group, ten cases in each one. Different stimulation intensities of acupuncture at Hegu (LI 4) combined with facial paralysis acupoints, including Yingxiang (LI 20), Dicang (ST 4), Jiache (ST 6) and Quanliao (SI 18), were applied in Hegu 1 to 4 groups; meanwhile acupuncture at stroke acupoints, including Neiguan (PC 6), Shuigou (GV 26) and Sanyinjiao (SP 6), and medication treatment were adopted. Except acupuncture at Hegu (LI 4), the treatment of the control group was identical as Hegu groups. The treatment duration lasted for 14 days. The House-Brackmann facial never grading systems (H-B), Toronto facial grading system (TFGS), degrees of facial never paralysis (DFNP), facial disability index (FDI) and clinical efficacy were compared among groups.

RESULTS(1) Compared before the treatment, H-B, TFGS, DFNP and physical function score in FDI were all improved significantly in the Hegu 1 to 4 groups (all P < 0.05), but social function score in FDI was not obviously improved (all P > 0.05); all the scores in the control group were not evidently changed (all P > 0.05). (2) Compared with the control group, differences of H-B before and after treatment in the Hegu 1 to 4 groups, differences of TFGS in the Hegu 2 group and differences of DFNP in the Hegu 1 and Hegu 2 group were significantly improved (all P < 0.05). The differences of any scale among Hegu 1 to 4 groups were not significant (all P > 0.05), in which the most evident change was found in Hegu 2 group. (3) The total effective rate was 90.0% (9/10), 100.0% (10/10), 90.0% (9/10) and 80.0% (8/10) in Hegu 1 to 4 groups, which were significantly higher than 60.0% (6/10) in the control group (all P < 0.05).

CONCLUSIONAcupuncture at Hegu (LI 4) has affirmative clinical efficacy on central facial nerve paralysis after ischemic stroke, in which oblique insertion along the opposite direction of meridian for 5 s of twirling manipulation has the best clinical effect.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Facial Paralysis ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications

Yeast strains with improved ethanol yield are important for efficient bioconversion of lignocellulosic biomass for fuel ethanol.Candida shehatae CICC1766 was adapted to 4%(v/v)ethanol,and then subjected to UV mutagenesis.One respiration deficient mutant Rd-5 with improved xylose fermentation capability was selected.Protoplasts of Rd-5 were inactivated by UV treatment,followed by the PEG-mediated protoplast fusion with a Saccharomyces cerevisiae strain with good ethanol-fermenting capability.The xylose fermenting capability of the fusants was investigated,and the fusant F6 demonstrated good ethanol fermentation performance,producing 18.75g/L ethanol from 50g/L xylose with an ethanol yield of 0.375 or 73.4% of its theoretical value of 0.511.Comparing with its parent Candida shehatae strain,the ethanol yield of F6 was increased by 28%.

BACKGROUNDHeat shock protein 70 (HSP70) is expressed highly in epithelial tumours associated closely with human papillomavirus 16 (HPV16) infections. However, evidence about the direct relationship between HSP70 expression and HPVs infections are still lacking. In the present study, we examined the expression of HSP70 in keratinocytes introduced with HPV16 E6/E7 oncogenes.

METHODSStable transfected cells were established by transfection of the plasmids pLXSN16E6/E7 into cultured primary keratinocytes and subsequently selected by plasmid specific selection antibiotic (G418) at the required concentration. The expression of HSP70 in pLXSN16E6/E7 transfected keratinocytes was determined by Western blot. The correlation of HSP70 expression and E6/E7 transfection was further confirmed by doubly labelled immunofluorescent staining.

RESULTSCompared to non-transfected keratinocytes, there was a significant trend for higher levels of HSP70 in pLXSN16E6/E7 transfected keratinocytes. Doubly labelled immunofluorescent staining experiment showed that the co-localization of HPV16 E6/E7 and HSP70 in transfected keratinocytes was observed and increased expression of HSP70 was strongly associated with the transfection of HPV16 E6/E7.

CONCLUSIONSOur studies demonstrated increased levels of HSP70 proteins in keratinocytes stably transfected by HPV16 E6/E7 oncogenes. It suggests that the expression of HSP70 is modulated by HPV16 E6/E7 proteins, which may be involved in HPV16 E6/E7 induced immortalization.

Cells, Cultured ; HSP72 Heat-Shock Proteins ; biosynthesis ; Humans ; Keratinocytes ; metabolism ; Oncogene Proteins, Viral ; genetics ; Papillomavirus E7 Proteins ; Repressor Proteins ; genetics ; Transfection