Objective To use gene engineering technique to express the humanin.Methods Humanin segment was synthesized and cloned into the prokaryocyte expression vector pGEMEX-1 and pet44a. The plasmid was transformed to E. coli BL21(DE3) by screening and identifying to obtain the high expressing positive clone.Results The humanin segment was cloned into vector successfully,and expressed efficiently with the inducement of IPTG. The molecular weight of the expressed recombinant protein of pGEMEX-1-humanin was about 28 kd while it was 63 kd in pet44a-humanin,and the expressed quantity of fusion protein was 40 percentage of the total bacterial protein while it was 20 percent in pet44a-humanin.Conclusion The humanin has been cloned and expressed successfully and the high efficiently expressed recombinant protein might contain the intact important structure domain of the humanin.

Objective To observe the interactions between a anti-interleukin-8 monoclonal antibody (anti-IL-8 mAb) carried targeted ultrasound contrast agents and the injured vascular endothelial cells,as well as to explore the role of IL-8 in the formation of atherosclerotic plaques and a new assessing method of vascular endothelial functions.Methods The targeted ultrasound contrast agent was prepared by using a erosslinking agent to couple a anti-IL-8 mAb with SonoVue microbubbles.The interactions between SonoVue microbubbles/the targeted microbubbles and normal/injured endothelial cells were observed under an inverted microscope,respectively.The numbers of endothelial cells and adhered microbubbles were counted under high power magnification.The ratio of microbubbles to endothelial cells was calculated for the quantitative analysis of the interactions.Results In the control group,only a slight amount of original SonoVue microbubbles were bound to normal/injured endothelial cells.In contrast,it was visible under the microscope that the anti-IL-8 mAb carried SonoVue could be bound to endothelial cells,and the number of microbubbles bound to the surface of injured endothelial cells was significantly higher than that bound to the normal endothelial cells.Conclusions The anti-IL-8 mAb carried targeted ultrasound contrast agent could be readily bound to the surface of the injured cells specifically,and thus suggesting a new direction for ultrasonic detection of vascular endothelial injury and the ultrasonic assessment of vascular endothelial functions.

OBJECTIVETo develop the knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for occupational groups, and to provide a convenient and effective tool for the survey of knowledge, attitude, and behavior on the prevention and control of occupational diseases in occupational groups and the evaluation of intervention effect.

METHODSThe initial questionnaire which was evaluated by the experts was used to carry out a pre-survey in Guangzhou, China. The survey results were statistically analyzed by t test, identification index method, correlation analysis, and Cronbach's a coefficient method. And then the questionnaire was further modified, and the content of the questionnaire was determined finally.

RESULTSAfter modification, there were 18 items on knowledge, 16 items on attitude, and 12 items on behavior in the "Knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for enterprise managers"; there were 19 items on knowledge, 10 items on attitude, and 11 items on behavior in the "Knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for workers".

CONCLUSIONThe knowledge, attitude and practice questionnaire on the prevention and control of occupational diseases for occupational groups is developed successfully, and it is a convenient and effective tool for the survey of knowledge, attitude, and behavior on the prevention and control of occupational diseases in occupational groups and the evaluation of intervention effect.

China ; Health Knowledge, Attitudes, Practice ; Humans ; Occupational Diseases ; prevention & control ; Surveys and Questionnaires

Objective To construct the bp26 deletion mutant of Brucella vaccine strain M5-90 (M5-90Δbp26),to compare its immunogenicity with parental strain(M5-90),and to develope a new candidate vaccine strain of Brucella melitensis with reduced virulence that can be used to distinguish vaccinated livestock from infected animals.Methods Mutant vaccine strain of Brucella melitensis was constructed by conventional molecular biology techniques then the genetic stability of mutant M5-90Δbp26 was tested and its conventional bacteriological nature was identified; 1.0 × 109 CFU/2 ml doses of M5-90Δbp26 strain and the parental strain were used to vaccinate 3 sheep; sera were analyzed for reactivity against BP26 by Western blotting and for agglutination activity; to analyze the virulence of mutant and parental strain,mice were injected with 1.0 × 106,6.0 × 106 and 2.0 × 107 CFU/0.2 ml doses of M5-90 and M5-90Δbp26,respectively,and clinical symptoms were monitored and the death of mice was recorded.Results The M5-90Δbp26 was successfully generated and reversion was not observed in 15 generations.The size of PCR products was 629 bp while the parental strain was 1279 bp.The sequence analysis showed a 650 bp missing in M5-90Δbp26.The conventional bacteria identification tests confirmed that the mutant was depth variant strain,including mono-specific antiserum M type transformed into R,and the BK2 phage based splitting assay converted from the positive to the negative.Western blotting showed the purified BP26 protein was recognized by the serum against the parental strain while not by the serum against M5-90Δbp26 strain.Agglutination test showed the level of the serum antibody induced by M5-90Δbp26 strain(1:50) was significantly lower than that of serum induced by parental strain(＞ 1:800).Virulence test showed that M5-90Δbp26 strain was less virulent than parental strain.Conclusions M5-90Δbp26 has been successfully constructed.M5-90Δbp26 of Brucella melitensis has the characteristic of reduced virulence and has a potential as brucellosis candidate vaccine strain permitting serological discrimination between diseased and vaccinated livestock.

Female ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Infant, Newborn ; Lung ; metabolism ; pathology ; Macrophage Migration-Inhibitory Factors ; analysis ; genetics ; Male ; Respiratory Distress Syndrome, Adult ; blood ; genetics ; pathology

OBJECTIVETo investigate the expression of aquaporin 3, 4, and 8 in the colonic mucosa of rat models with slow transit constipation (STC).

METHODSSTC rat model was established by giving the rats the compound solution of diphenoxylate. Real time polymerase chain reaction (RT-PCR) was used to measure the expression of aquaporin mRNA in colonic mucosa of STC rat models (study group,n=16) and normal rats (control group,n=16). Gray scale ratio of aquaporin to β-action (internal reference) was used for quantification.

RESULTSRT-PCR revealed that the mean gray scale ratios of aquaporin 3 in the proximal colon of the study group and control group were 0.344 and 0.602 (P<0.05), and were 0.419 and 0.509 in the distal colon (P>0.05), respectively. The mean gray scale ratios of aquaporin 4 in the proximal and the distal colon were 0.764 and 0.759 in the study group (P>0.05), and were 0.776 and 0.736 in the control group (P>0.05), respectively. However, there was no expression of aquaporin 8 in the proximal and the distal colon in either the study group or the control groups.

CONCLUSIONSExpression of aquaporin 3 in the proximal colon of STC rat models is down-regulated, which regulates water absorption. There are no significant changes in the expressions of aquaporin 4 and 8.

Animals ; Aquaporin 3 ; metabolism ; Aquaporin 4 ; metabolism ; Aquaporins ; metabolism ; Colon ; metabolism ; Constipation ; metabolism ; Disease Models, Animal ; Female ; Intestinal Mucosa ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

To improve the efficiency of halophilic actinobacteria screening and carry out the rapid selection of targeted strains, we tested 34 strains of Streptomonospora by PCR-single strand conformation polymorphism analysis (PCR-SSCP) based on genus-specific primers for the PCR identification. This approach employs PCR with two pairs of primers located in the 16S rRNA sequence flanking two variable region, then build clustering tree according as SSCP data. Synchronously, we sequenced all the 16S rRNA partial sequences for these strains to verify them. The results showed that the PCR-SSCP analysis was an efficient, easy-to-handle and economic method for rapid selection of halophilic actinobacteria resources.

Objective To observe the normal structures of arteria circumflexa femoris lateralis (ACFL) and to establish digitized visible models of anterolateral thigh(ALT) flap,which can be used in clinical training and operation design.Methods The cross-sectional images from the VCH Male 3 dataset were reviewed to study ACFL structures on a section-by-section basis.Next,one adult fresh cadaver specimen was perfused with lead ox- ide-gelatine mixture to be subject to radiographic CT scanning on its lower limbs.Likewise,the cross-sectional images from the CT images were reviewed to study ACFL structures on a section-by-section basis.Three-dimensional computerized reconstructions of ACFL structures and their adjacent struetures were conducted from the two sets of data using Amira 3.1 (TGS) software respectively.Results The three-dimensional reconstructed visible models established by the above two methods perfectly displayed the anatomic relationships of ACFL structures and their adjacent structures.Conclusions Since the digitized images of ACFL structures can offer section by section in- sights into the ACFL anatomy,their 3D reconstructive models ean be applied in clinical training,pre-operative designing and virtual operation procedures.

Cancer is a group of various diseases, all of which involve unregulated cell growth. Many currently used chemotherapeutic drugs are derived from botanicals. Thus, searching botanical sources for novel oncology medications, including identifying the lead compounds and their derivatives for chemoprevention, is an essential step in advancing cancer therapeutics. This article mainly focuses on the data from our previous American ginseng anti-colon cancer studies. In addition to the potential role of American ginseng on cancer, the herb as an adjuvant for cancer treatment is presented, including describing the attenuation of adverse events induced by chemotherapeutic agents and increasing of quality of cancer patient life. Since heat-treated American ginseng and ginsenoside gut microbiome metabolites showed significant increases in cancer chemopreventive effects, active constituents of the steamed herb and their gut metabolites should be clearly identified, and the structure-activity relationship should be further explored. Data obtained from herbal medicine studies and clinical trials will help develop useful anticancer agents.
Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Colorectal Neoplasms ; drug therapy ; pathology ; Ginsenosides ; isolation & purification ; metabolism ; pharmacology ; therapeutic use ; Hot Temperature ; Humans ; Panax ; chemistry ; Phytotherapy ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Structure-Activity Relationship

Adolescent ; Child ; Child, Preschool ; Clubfoot ; physiopathology ; Electromyography ; Female ; Humans ; Infant ; Male