Metagenomic analysis of symbiotic microorganism in sailors with long sea voyage / 国际检验医学杂志

Objective To investigate the influence of long sea voyage working environment on the symbiotic microorganisms and their relationship with their hosts .Methods The periumbilical microbial sam-ples from the operating workers of long sea voyage before and after operation were collected .Then 16S rRNA V4 section amplification ,sequencing and whole metagenome shotgun high-throughput sequencing were per-formed .Moreover the bacterial community structure ,kinds and microorganism metabolic function change were analyzed .The peripheral blood was collected from the workers of long sea voyage operation and shore-based operation for conducting the blood routine analysis .Results After 105 d ocean sailing ,the diversity of perium-bilical microbial community in the workers with long sea voyage operation decreased and the relative abun-dance of Firmicutes increased ,w hile w hich of Proteobacteria decreased ;w hich of Staphylococcus increased , while which of Corynebacteria decreased ,the differences were statistically significant (P<0 .05) ,the relative a-bundance of pathogenic bacteria or conditional pathogenic bacteria ,especially Staphylococcus epidermidis and Staphylococcusaureus aureus ,increased significantly .T he functional gene analysis indicated that the expres-sion of periumbilical microbial infection related genes increased after the long sea voyage operation .Compared with shore-based operation workers ,the proportion of workers with peripheral blood lymphocytes abnormal elevation in the long sea voyage group increased significantly ,the difference was statistically significant (P<0 .05) .Conclusion The periumbilical skin symbiotic microorganisms may reflect the health conditions in the workers with long sea voyage operation .

Observation of the curative effect of two kinds of mini perforator free flap for digital injuries reconstruction / 中华显微外科杂志

Objective Discuss the clinical efficacy by using two kinds of mini perforator free flap for digital injuries reconstruction. Methods From August, 2014 to February, 2017, 45 patients were managed randomly with either radial artery superficial palmar branch(RASPB)perforator free flap or digital artery(DA)perforator free flap for digital skin defects reconstruction, and they were therefore divided into two groups according to the flap type. There were 24 patients in RASPB group, with an average wound dimensions ranged from 1.8 cm×1.5 cm to 4.0 cm×2.5 cm, and an average harvested flap size ranged from 2.0 cm×1.7 cm to 4.2 cm×2.6 cm. Another 21 patients were in DA group, with an average wound dimensions ranged from 2.0 cm×1.5 cm to 3.8 cm×3.0 cm, and an average harvested flap size ranged from 2.2 cm×1.6 cm to 3.9 cm×3.2 cm. The survival rate, sensory function, donor site complications, hand function recovery and aesthetic outcomes of two groups were compared by the SPSS22.0 statistical software after surgery. Results The mean follow up period was 15 months. All flaps were primary survived without vascular crisis. The flaps were soft in texture,trimness in appearance and none of them overtop the normal skin for more than 0.5 cm. Both groups had a favorable sensory recovery.All cases recovery to S3+or better.In Group RASPB,the mean two point discrimination(2 PD)was 7.85±1.15 mm(ranged from 7.0 mm to 9.0 mm). And it was 6.02±0.94 mm(ranged from 6.0 mm to 8.0 mm)in DA group. The difference between two groups was statistically significant(P <0.05). Then we synthetically analyzed flap texture and sensory function,and calculated the qualified ratio of each group.There was no significant difference between two groups(P > 0.05). The degree of scar contracture demonstrated donor site compli cations in RASPB group was lesser than that in DA group(P<0.05).The range of motion of interphalangeal joint was used to reflect the hand function. And we calculated the ratio of repaired and contralateral sites. The difference of the mean ratio between two groups was not statistically significant(P>0.05). Conclusion On account of the characteristics of invariant anatomy position, sufficient blood supply, favorable aesthetic outcome and minimal donor site mobility, both RASPB perforator flap and DA perforator flap were optimal for digital skin defects reconstruction.Besides,incorporated with nerve and tendon,the RASPB perforator flap can also be used for complex tissue transplantation,and the surgery field was only on the arm.While the DA perforator free flap had an advantage of better sensory recovery and appearance.

Expression of stathmin and its influence on taxol sensitivity in esophageal squamous cell carcinoma / 医学研究生学报

Objective Stathmin, a microtubule-destabilizing protein , has high expression in esophageal squamous cell carci-noma(ESCC), while taxol is a common chemotherapy microtubule-targeted drug for esophageal cancer .This study aimed to investigate the impact of stathmin expression and its influence on taxol sensitivity in ESCC . Methods We established 2 cell models with ST-MN1 gene overexpression in KYSE 510 and KYSE 170 cell lines, including KYSE 510-Stathmin, KYSE 170-Stathmin, KYSE 510-Control and KYSE 170-Control.MTT assay and colony formation were applied to compare the taxol sensitivity between experimental group and control group .Flow cytometry was used to measure the apoptosis of KYSE 510-Stathmin and KYSE 510-Control after taxol treatment.Western blot was used to test the changes of related factors to apoptosis and autophagy . Results ①Stathmin protein ex-pressions in KYSE 510-Stathmin and KYSE 170-Stathmin cells were higher than those of control cells (P<0.01).② The percentages of inhibition were significantly decreased in KYSE 510-Stathmin and KYSE 170-Stathmin cells 24 h after 50, 100,250 nmol/L taxol treat-ment compared with KYSE 510-Stathmin cells(P <0.01).③The percentages of inhibition were significantly reduced in KYSE 510-Stathmin and KYSE 170-Stathmin cells after 250 nM taxol treatment for 24, 48, 60 h (P<0.01).④After taxol treatment,the number of colony formation in KYSE 510-Stathmin cells was higher com-pared with KYSE 510-Control cells (P<0.01).⑤The percentage of cell apoptosis in KYSE 510-Stathmin was significantly lower than that of KYSE 510-Control cells by flow cytometry (11.90%±0.78%vs 29.63%±3.26%, P<0.05).Western blot showed the ap-optosis of associated proteins such as the activation of Caspase 8 and Caspas9. Conclusion The result indicates that overexpression of stathmin inhibits taxol sensitivity in ESCC cell lines .

Short gel method for pretreatment of protein samples with high concentration of detergent / 生物工程学报

In proteomic research, to improve protein solubility of membrane proteins and nuclear proteins, buffers containing high concentration of detergent, such as 4% SDS, were widely used. However, high concentration of detergent might severely interfere with the downstream proteomic analysis, including protein quantitation and trypsin digestion. To improve the proteomic compatibility of buffers with high concentration of detergent, we used short gel method to pretreat buffers containing detergent. Protein samples were first separated by a short (2-2.5 mm) SDS-PAGE electrophoresis, and proteins were quantitated by comparing with bovine serum albumin standards via optical density analysis. The gel was then cut and peptides were recovered using in-gel digestion. The quantitative linearity range of this method was 1 to 8 μg. The quantitation was accurate and reproducible. After short gel analysis, recovered peptides generated high mass spectrometry signals. In conclusion, short gel method eliminated the interference of high concentration detergent in the proteomics analysis, and it was suitable for protein samples' pretreatment, and was worth to apply in proteomic research.

Detection of circulating tumor cells in patients with hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the detection efficiency of circulating tumor cells (CTCs) in patients with hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>Immunomagnetic negative enrichment by nanometer magnetic beads and label-free capture with Captor(TM) system were used to isolate and enrich CTCs from peripheral blood of HCC patients, and epithelial and HCC markers were applied to identify CTCs by immunofluorescence staining. CTCs were detected in 50 HCC patients before and after hepatectomy to test the method for isolation, enrichment and identification. The dynamic changes of pre- and post-operative CTCs' numbers were compared. The clinical data were analyzed using SPSS 19.0 software.</p><p><b>RESULTS</b>Negative enrichment methods by nanometer magnetic beads and label-free capture using Captor(TM) system were both suitable for CTCs isolation and enrichment in HCC patients. The positive detection rate of CTCs in HCC patients via negative enrichment was 96.0% (48/50), the preoperative median number of CTCs was 16 per 7.5 ml blood, and the postoperative median number was 17 per 7.5 ml blood.</p><p><b>CONCLUSIONS</b>Both negative enrichment and Captor(TM) system are suitable for isolation and enrichment of CTCs in HCC patients. There is a significant difference in the numbers of CTCs before and after operation, and dynamic detection of CTCs will provide helpful prognostic information for HCC patients in clinics.</p>

Recent advance in high accuracy iTRAQ for quantitative proteomics / 生物工程学报

Nowadays, proteomics focuses on quantitative analysis rather than qualitative. In the field of quantitative proteomics, Isobaric tags for relative and absolute quantitation (iTRAQ) is one of the most widely used techniques. The advantage of iTRAQ is high throughput, high stability and free of the restriction of sample property. iTRAQ is suitable for almost all kinds of samples, and up to 8 samples can be analyzed simultaneously by commercially available kit. Along with the development of techniques, more and more mass spectrometry (MS) platforms are used in iTRAQ experiments and the accuracy of iTRAQ has been improved. iTRAQ has been applied to studies of microorganism, animal, plant, medical and protein post-translational modification. Here we review the recent progress in the development of iTRAQ and its applications in quantitative proteomics.

Outcome of free flap vascularized by the dorsal artery of big toe in treating finger skin defect / 中华显微外科杂志

Objective To evaluate the outcome of the procedure of using the free flap vascularized by the dorsal artery of big toe to treat finger skin defect.Methods From August 2010 to April 2013,21 cases of finger shin defect were treated with the free flap vascularized by the dorsal artery of big toe in which emergency surgeries were conducted in 9 cases and sub emergency surgeries were conducted in 12 cases.The age of the 21 patients was 21 to 48 years old and 15 of them were males and 6 were females.Thumb was involved in 6 patients and index finger was involved in 15 patients.The skin defect occurred at dorsal in 7 patients and palm in 14 patients.The area of the flaps ranged from 2.2 cm × 1.6 cm to 4.0 cm × 3.2 cm.Observed the restoration of the affected fingers' appearance and function,investigated the clinical results and concluded the indication and advantages and disadvantages of this procedure by following up.Results All of the 21 flaps survived at the last office visit.The follow-up period ranged from 3 to 18 months.The shapes and the fingerprints of the flaps were satisfied.The color and texture of the flaps were similar to those of the finger skin.The 2 points discriminations of the flaps was 6-8 mm.No function deficits were found in the donor feet.Conclusion This free flap is satisfied in the shape,easy to harvested and the blood supply was constant in its location,and recommend it in treating the small of middle area skin defect in the finger.

The study progress of HSP-105 in tumor on applied and basic research fields / 肿瘤研究与临床

Heat shock protein (HSP) 105 is a 105x103 stress protein that belongs to the HSP-110 family. It is released by tissues in response to a wide variety of stresses including infection and ischaemia. Studies have shown that the molecule is involved as a biochemical mediator of heat induced apoptosis by binding to p53 at scrotal temperatures and dissociating from it at suprascrotal temperatures in testicular germ cells. Recent studies have shown that HSP-105 is over-expressed in various malignancies besides in normal testicular tissue. HSP-105 may be a candidate of testis antigen.

Genome-wide Association Analysis Based on Copy Number Variations / 生物化学与生物物理进展

Copy number variations (CNVs) refer as a DNA segment that is 1 kb or larger and is presented at a variable copy number in comparison with a reference genome. Classes of CNVs include insertions, deletions, duplications and their complex combinations. Because they widely distributed in the genome with some important characteristics, such as heritable, relative stable and heterogeneity, CNVs are considered as novel genomic polymorphism markers. And the alteration of gene dosage which resulted from CNVs could change phenotype, so a novel CNV genome-wide association analysis (CNV-GWAS) strategy appeared recently and began to used for identifying susceptible genes of complex diseases. It was approved that it could complement the tranditional genome-wide association studies based on single nucleotide polymorphisms. Therefore, genomic structure variances are favorable for revealing the molecular mechanisms and genetic foundation of complex diseases.

Identification of tumor markers with SILAC proteomics approaches / 基础医学与临床

Mass spectrometry-based quantitative proteomics has become an indispensable tool for tumor marker discovery.Stable isotope labeling with amino acid in cell culture(SILAC),as a representative of in vivo metabolic labeling strategy,is a simple and relatively quantitative assay for comparative proteomics.Two kinds of cultured cells are grown in the medium containing either a light or heavy isotope labeled essential amino acids.After several cell doublings,the heavy amino acids are introduced into the nascent polypeptides in a sequence-specific fashion and the natural amino acid is completely replaced by its isotopically labeled analog.Accuracy quantification would be feasible by analyzing the peak intensities of every same peptide pairs labeled with light or heavy isotopes by the mass spectrometry.Compared with chemical labeling,SILAC requires much less amount of labeled proteins,and it is an efficient,straightforward,reproducible and accurate approach in cell-based systems.This method can not only investigate the dynamics of protein abundance,but also monitor the variation of posttranslational modifications and protein-protein interactions qualitatively and quantitatively.It is a powerful and important quantitative proteomics tool for tumor marker discovery.

Screening of human anti-ricin ScFv from large phage library / 中国免疫学杂志

Objective:To clone human anti-ricin antibodies from large phage antibody library.Methods:Panning for a large phage library against ricin toxin was conducted to select specific antibodies against ricin. The binding activities and specificities were tested by ELISA method. Soluble ScFvs were prepared through infecting E coli. HB2151 with the selected phage antibodies and induction with IPTG. Results:Forty positive clones were obtained after 5 rounds of panning, and 12 clones had specific binding ability to ricin toxin. DNA fingerprinting showed 7 different band patterns indicating 7 different positive clones. DNA sequencing showed that variable regions of these ScFvs belonged to different subgroups.Conclusion:Human anti-ricin antibodies were successfully obtained from large phage antibody library.

Glycosylation and localization of clusterin in esophageal squamous cell carcinoma / 第二军医大学学报

Objective:To study the glycosylation of different clusterin isoforms and the alterations of their intracellular localizations in esophageal squamous cell carcinoma(ESCC),so as to better understand the potential mechanisms of clusterin in tumorigenesis of ESCC.Methods: The glycosylation of clusterin proteins was studied after treated with PGNase-F by Western blot analysis in 23 ESCC specimens and their corresponding adjacent tissues.Immunohistochemical staining was used to visualize the intracellular localization of clusterin proteins.Results: Mature clusterin,a 40 000 glycosylated protein in SDS-PAGE gel,(usually) presents in the connective tissues of the lamina propria of esophageal epithelial mucosa,and is secreted by esophageal glands onto the esophageal mucosa and in plasm.The glycosylation of clusterin proteins was abnormal in the 23 ESCC specimens,with the premature clusterin and nuclear clusterin aggregated in the cells and the mature clusterin decreased or disappeared in the stromal layer of esophagus.Conclusion: The deglycosylation and localization alterations of clusterin in ESSC may be closely associated with the tumorigenesis of ESCC.

Construction of soluble anti-digoxin ScFv antibodies and diabody / 中国免疫学杂志

Objective:To clone soluble human anti-digoxin antibodies from large phage antibody library and construct a vector that expresses diabody.Methods:Soluble ScFvs were prepared through infecting E coli.HB2151 with the selected phage antibodies and induced with IPTG.The diabody vector was modified by enzyme digestion and checked by SDS-electrophoretogram.Results:It was showed by ELISA that soluble ScFv had specific binding ability to digoxin.The vector to express a soluble diabody was obtained by genetic modification,which was shown by Western blot.Conclusion:Soluble human anti-digoxin antibodies were successfully obtained from phage antibodies.The vector is efficient in creating diabody.