Child ; Child, Preschool ; Clinical Laboratory Techniques ; DNA, Fungal ; genetics ; Evidence-Based Medicine ; Fungi ; genetics ; isolation & purification ; Humans ; Infant ; Mycological Typing Techniques ; Mycoses ; diagnosis ; microbiology ; Polymerase Chain Reaction ; Serologic Tests ; Specimen Handling

Trypsin was purified from crude material of pig pancreas by AOT/isooctane reversed micellar system.The influence of the main operating parameters such as ethanol concentration in forward and backward extraction,pH,KCl and AOT concentration,temperature were investigated.Forward and backward extraction recovery of trypsin reached almost 90% and neared 100%,respectively.Finally,about 88% of total yield was obtained,and the specific activity of trypsin was increased to over 1800U/mg with purification factor of 5 folds more.In AOT-isooctane reverse micellar extraction system,denaturation or precipitation of proteins always occured due to strong electrostatic interaction between AOT-proteins molecules.It had been resolved by adding ethanol into reverse micellar system,and no denaturation was observed.Otherwise,the phase separation time was shortened significantly because of ethanol added.It was only 10 minutes or less to reach phases separation after forward and backward extraction.If this method can be applied in industry,efficiency will be greatly improved.

Objective To study the clinical characteristics by using CT, MRI and biochemistry test on the X linked adrenoleukodystrophy (X ALD) Methods The clinical,neuro imaging and determinations of plasma very long chain fatty acids (VLCFAs) of 12 cases with X ALD were analysed Results The main clinical features consisted of the childhood onset,progressive auditory, visual and/or intelligent impairment, behavioural changes,epileptic seizures and melanodermia. CT or MRI scans demonstrated that the demyelinating lesions were located on the bilateral white matter in occipital,posterior parietal and temporal lobes.The levels of hexacosanoic acid (C 26:0 ) and the ratio of C 26:0 to docosanoic acid (C 22:0 ) were increased. Conclusions ALD is an inherited disease as an X linked trait.The childhood X ALD is characteristiced by progressive auditory, visual and intelligent deterioration.The abnormalities of CT or MRI scan and the levels of the plasma VLCFAs are crucial to the diagnosis of X ALD.

Objective To discuss the blood substitutes during the perioperative period for hemophilic children with surgical disease as well as the coping strategies to its postoperative complications.Methods A retrospective study between 2003 and 2013 from one our centre identified a total of 41 perations performed in haemophiliac patients.33 patients were diagnosed with haemophilia A,among whom 10 were severe cases,13 moderate cases and 8 mild cases.8 patients were diagnosed with haemophilia B,among whom 2 were severe cases and 6 were mild cases.Different kinds of operation were required for each case.Results According to the monitoring of the coagulation tests PT and APTT before and after the operation respectively,after the use of the blood substitutes such as FV Ⅲ,PPSB and fresh frozen plasma,38 patients underwent surgical treatment successfully and had full recovery without any surgical complications,1 patient was dead,1 suffered intraperitoneal hemorrhage and 1 had delayed wound healing.Conclusion Full preparation and thorough plan before the operations,combined with appropriate blood substitutes can effectively reduce postoperative bleeding for hemophilic children.

Acute Disease ; Adult ; Humans ; Male ; Occupational Diseases ; diagnosis ; therapy ; Phosphines ; poisoning ; Poisoning ; diagnosis ; therapy

The morphological indexes of the Scuophularia ningpoensis seedlings including the longth, diameter and weight were measured, clustering analysis was used to set up the standard quality grading of seedlings of S. Ningpoensis by SPSS. Field experiment was carried out to measure the indicators of plants growth and development, the yield and the quality. The results showed that the growth and yield of class I seedlings were better than those of class II and III. The content of main active ingredients was affected barely by seedlings classification. To ensure the quality, class II seedlings or above should be used for plantation. The established quality classification standard of S. ningpoensis seedlings was scientific and feasible, and provides the basis for the standardized cultivation of S. ningpoensis.
Drugs, Chinese Herbal ; analysis ; standards ; Quality Control ; Scrophularia ; chemistry ; classification ; growth & development ; Seedlings ; chemistry ; growth & development

Objective To investigate the distribution and antimicrobial resistance of major pathogens causing bloodstream infection in a tertiary hospital,so as to guide the clinical rational use of antimicrobial agents.Methods Blood specimens from patients with suspected bloodstream infection in this hospital between January 2012 and January 2015 were cultured,identified and performed antimicrobial susceptibility testing,the major pathogens and antimi-crobial susceptibility testing results were analyzed statistically.Results A total of 1 658 strains of pathogens were isolated from 19 326 blood culture specimens,isolation rate was 8.58%,in which gram-negative bacilli,gram-posi-tive cocci,and fungi accounted for 58.57% (n=971 ),39.38% (n=653),and 2.05% (n=34)respectively,the major gram-negative bacillus,gram-positive coccus,and fungus were Escherichia coli ,coagulase-negative staphylo-cocci (CNS),and Candida albicans respectively.The isolated pathogens mainly distributed in departments of hema-tology,gastroenterology,and cardiovascular surgery.Detection rates of extended-spectrumβ-lactamases (ESBLs)-producing Escherichia coli and Klebsiella pneumoniae were 56.35% and 36.14% respectively,but were highly sen-sitive to cefoperazone/sulbactam and carbapenems (resistance rates <3%).The percentage of methicillin-resistant coagulase-negative staphylococcus (MRCNS ) and methicillin-resistant Staphylococcus aureus (MRSA ) were 78.60% and 32.28%respectively,but were highly sensitive to vancomycin and linezolid(resistance rates < 1 %). Conclusion Escherichia coli and CNS are predominant pathogens causing bloodstream infection in this hospital,antimicro-bial resistance rates are high;regular analysis on distribution and antimicrobial resistance of pathogens causing bloodstream infection is helpful for the prevention and control of infection caused by multidrug-resistant organisms.

Objective To construct the prokaryotic vector and to express a novel candidate gene IDD01 associated with hepatocellular carcinoma (HCC) for its functional study. Methods The open reading frame (ORF) of IDD01, amplified from HCC tissue by RT PCR, was inserted into the expression vector pMAL c2X. The recombinant vector was transformed into E. coli TB1, and the expression products were analyzed by SDS PAGE. Results The length of PCR product was about 770 bp. The restriction enzyme digestion and sequencing conformed that the gene segment was correctly inserted into the vector. SDS PAGE and density scanning indicated that the protein was expressed as a fusion protein with 28 KD of molecular weight and the fusion protein possessed 30.4% of the total bacterial protein. Conclusion The recombinant vector is constructed successfully and IDD01 can be expressed at a high level, which lays a foundation for the further research on the functions of IDD01 gene.

Aim To investigate the inhibitive effects of trichostatin A(TSA) on telomerase activity of HL-60 cells and expression of subunit hTERT during apoptosis in vitro and its mechanism.Methods The proliferative activity of HL-60 cells was assessed using morphology and MTT assay.Cell apoptosis was confirmed using Flow Cytometer.Telomerase activity was examined using TRAP-ELISA.The expression status of telomerase subunits was analyzed using RT-PCR.Results A time-and dose-dependent inhibition was detected in HL-60 cells treated with TSA.After 48 h TSA(300 nmol?L~(-1)) treatment,the apoptotic rate detected using cytometric assay(Annexin V/PI double staining)of HL-60 cells was 42.6%.Telomerase activity and expression level of hTERT and the key subunit of telomerase decreased at 24-hour after TSA treatment.No significant changes were observed in the expression of hTR,hTP and the other two subunits of telomerase.Conclusion TSA inhibits telomerase activity and induces apoptosis in HL-60 cells.The underlying mechanism might be related to the down regulation of hTERT transcription.

Exosomes are nanosized membrane microvesicles secreted by various living cells.They contain proteins,lipids,RNA,and a variety of other biological macromolecules.Exosomes play an important role in many pathological and physiological processes,such as antigen presentation in the immune system,repair of damaged tissues,and growth and migration of tumors.Tumor-derived or tumor-associated exosomes play a vital role in regulating the occurrence and development of tumors.The analysis and detection of exosomes in tumors is helpful for the early diagnosis of tumors and provide new treatment methods.This article reviews exosomes' origin,composition,and functions in the development,migration,diagnosis,and treatment of tumors and provides new ideas for the treatment of tumors.