Objective:To map dominant antigenic determinants on SAK by gene-targeted fragmemt library . Methods:①The PcAbs specificly against SAK were produced by immunning BALB/C mice. Pu rified the antisera through a SAK-Sepharose 4B affinity chromatography column. The purified PcAbs were biotinylated for next step. ② After constructing SAK r andom epitope library we sequenced 12 isolated clones randomly to ensure its int egrity ,capability and randomness.③The library was screening by situ-clone hy bridization.④Constructed mSAK ,the A1 region deleted mutant of SAK ,and used Western-blot assay to identified its immunoreactivity. Results:①Got a dominant epitope at amino acid 71-89,called A1 region; ②Western-bl ot assay suggested that mSAK, a mutant SAK without A1 region., didn't combine to the anti-SAK PcAbs.Conclusion:A dominant epitope of SAK was mapped successfully with a simple,effective method . [

Progesterone has nongenomic effects on inducible nitric oxide synthase (iNOS), which is mediated by mitogen activated protein kinase (MAPK) pathways. This effect is supposed to have some potential association with asymptomatic gonococcal infections in women by immunological depression. In this study, polymorphonuclear leukocytes (PMNs) challenged by gonococci were used to study the nongenomic effects of progesterone. The activation of iNOS was assessed by measuring [(3)H] L-arginine converses to [(3)H] L-citrulline, and the activity of MAPK was detected by Western blot. It was found that the activity of iNOS and the yields of NO were enhanced significantly in gonococci-challenged PMNs compared with the controls (P<0.01). Progesterone could repress the activation of iNOS through P38MAPK pathway within PMNs (P<0.05), which could be blocked by SB203580 (P<0.01), but not by actinomycin D (P>0.05). It was also found subsequently that in the serum specimens collected from gonococci-infected but asymptomatic women, the progesterone level was higher than that in women with severe symptoms (P<0.01). Moreover, the yield of NO had an inverse correlation with progesterone. With these results it suggested that the rapid nongenomic effects of progesterone may inhibit iNOS activation and NO yields mediated by P38MAPK pathways, which were supposed to be concerned with asymptomatic women infected with gonococci.

Both interleukin-1 and IgE are important in the pathogenic mechanism of the allergy asthma. cDNA of interleukin-1receptor antagonist (IL-1ra) and IgE were cloned and a prokaryotic expression vector IL-1ra-Fcepsilon/pBV220 was constructed. The vector was transformed into Escherichia coli BL21(DE3). The fusion protein was expressed successfully in the form of inclusion body. The recombination protein of IL-1ra-Fcepsilon was highly purified by chromatography of gel filtration and ion exchange, which was identifited by Western blotting. The cell assay showed that the activity of IL-1ra-Fcepsilon was as high as IL-1ra in vitro after refolding. The pharmacokenetic profile of IL-1ra-Fcepsilon and L-1ra was analyzed, and the half time of IL-1ra-Fcepsilon is 4.78 times than that of IL-1ra.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Gene Fusion ; Immunoglobulin E ; genetics ; metabolism ; Immunoglobulin Fc Fragments ; genetics ; metabolism ; Interleukin 1 Receptor Antagonist Protein ; genetics ; metabolism ; Recombinant Fusion Proteins ; genetics ; metabolism

The cDNA of Insulin-like growth factor binding protein 3 was subcloned into a eukaryotic secretory expression vector pSectagA to construct pSectag-IGFBP3. Human renal cell carcinoma (RCC) 786-0 cells were transfected with pSectag-IGFBP3 using lipofectamine 2000. After 48 h, the secretory IGFBP-3 was tested and identified by western blotting. Meanwhile, Annexin V-EGFP stain was used to analyze the apoptosis of 786-0 cells induced by IGFBP-3. Secretory IGFBP-3 protein could express successfully in the 786-0 cells and the expressed IGFBP-3 directly displayed an apoptotic effect on the host cells. This work provides a basis for further study on the apoptosis-inducing mechanism of IGFBP-3 and the development of a new anti-tumor drug.
Apoptosis ; drug effects ; Base Sequence ; Carcinoma, Renal Cell ; metabolism ; pathology ; Humans ; Insulin-Like Growth Factor Binding Protein 3 ; genetics ; secretion ; Kidney Neoplasms ; metabolism ; pathology ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; physiology ; Transfection ; Tumor Cells, Cultured

The different concentration of specific ion species and the electrodiffusion of the ions down their electrochemical gradient generate transmembrane potential. The regulation of membrane potential for the function of numerous membrane proteins such as ion channels, transporters, pumps and enzymes plays primary role in the conversion of extracellular electric stimulation into a sequence of intracellular biochemical signals. Some ion channels regulated by membrane potential are well known, and the membrane non-ion channels protein is also modulated physiologically by membrane potential.
Humans ; Ion Channel Gating ; physiology ; Ion Channels ; metabolism ; Membrane Potentials ; physiology ; Phosphoric Monoester Hydrolases ; metabolism ; Receptors, G-Protein-Coupled ; metabolism

OBJECTIVETo study the regulation trend of resveratrol on TNF-alpha, IL-1beta, IL-6 expressions in bronchoalveolar layage fluid (BALF) of RSV-infected BALB/c mice at different time points.

METHODRSV-induced BALB/c mice were orally administered with resveratrol. Their BALFs were collected at 24, 72 and 144 h after the first nasal drip with RSV to detect the level of TNF-alpha, IL-1P3, IL-6 by EILSA.

RESULTThe expression of TNF-alpha, IL-1Pf and IL-6 in BALF increased significantly compared with the normal group (P <0. 01) after 24 hours of RSV infection, while the expression of TNF-alpha (P < 0.01), IL-1beta (P < 0.05), IL-6 (P < 0.01) in the resveratrol group decreased notably compared with the model group. After 72 hours of infection with RSV, although the expression of TNF-alpha (P < 0.05), IL-1beta (P < 0.01) and IL-6 (P < 0.01) in BALF in model group were higher than those in the normal group, they were much more lower than at 24 h. The expression of IL-1beta and IL-6 (P < 0.05) in the resveratrol groups were down-regulated significantly, but no difference had been shown in TNF-alpha expression compared with the RSV infection group. After infection with RSV for 144 h, the expression of IL-1beta (P < 0.01) and IL-6 (P < 0.05) in BALF in the model group were higher than those in the normal group, but there was no difference in the secretion of TNF-alpha. The expression of TNF-alpha, IL-1beta and IL-6 showed also no remarkable difference between the resveratrol groups and the RSV infection group.

CONCLUSIONResveratrol can inhibit the over expression of inflammatory factors TNF-alpha, IL-1beta, IL-6 in bronchoalveolar lavage fluid of RSV-induced BALB/c mice and keep them at a low level with the passing of infection time.

Animals ; Bronchoalveolar Lavage Fluid ; immunology ; Female ; Interleukin-1beta ; analysis ; Interleukin-6 ; analysis ; Mice ; Mice, Inbred BALB C ; Respiratory Syncytial Virus Infections ; drug therapy ; immunology ; Stilbenes ; pharmacology ; therapeutic use ; Tumor Necrosis Factor-alpha ; analysis

OBJECTIVETo analyze the epidemiological characteristics of hospitalized children with burn injuries in Fujian Medical University Union Hospital, so as to provide evidence to complete an adequate, timely, and effective prevention and treatment system of children with burn injuries.

METHODSMedical records of children with burn injuries, aged 14 and under, hospitalized in the Department of Burns from July 2012 to June 2015 were collected. Data of gender and age, location and cause of injury, time of injury, state of injury, admission time after injury, first aid, length of hospital stay, and treatment and so on were recorded. They were divided into 4 age brackets: less than or equal to 1 year old, more than 1 year old and less than or equal to 3 years old, more than 3 years old and less than or equal to 7 years old, more than 7 years old and less than or equal to 14 years old, then gender and cause of injury of children in the 4 age brackets were analyzed. Admission months of the children were divided into spring (March to May), summer (June to August), autumn (September to November) and winter (December to February of the following year), and then the cause of injury of children in each season was analyzed. Severities of male and female children, length of hospital stay of children with different causes of injury were analyzed. Data were processed with chi-square test, Wilcoxon rank-sum test.

RESULTSOut of 2 608 inpatients with burn injuries, 1 407 children with burn injuries, aged 14 and under, accounting for 53.9%, were admitted in the recent 3 years. The ratio of male to female was 1.6 ∶1.0. Children more than 1 year old and less than or equal to 3 years old ranked the largest number (68.3%, 961/1 407) in the 4 age brackets. There was statistically significant difference in constituent ratios of gender of children among the 4 age brackets (χ(2)=11.00, P=0.012). One thousand three hundred and seventy-two children were burned indoors (97.5%), while 35 children were burned outdoors (2.5%). Scalding with hot fluids was the most common cause of burn (95.0%, 1 337/1 407). There was statistically significant difference in constituent ratios of injury cause of children among the 4 age brackets (χ(2)=107.23, P<0.01). There was statistically significant difference in constituent ratios of injury cause of children more than 7 years old and less than or equal to 14 years old compared with those of the other 3 age brackets (with χ(2) values from 12.88 to 119.85, P values below 0.01). Most burn accidents occurred between 17: 00-20: 59 (33.5%, 472/1 407). Burns were more likely to happen in April to October. July (10.4%, 146/1 407) and August (10.5%, 148/1 407) were the crest-time. Most of the children were burned in summer (35.3%, 496/1 407). There was statistically significant difference in the injury cause of children among each season (χ(2)=14.61, P=0.024). The burn degrees of male and female children were mainly mild or moderate, and there was no statistically significant difference in the severity (Z=-0.39, P>0.05). The trunk was the most involved anatomic site (61.1%, 859/1 407). Most of children were admitted to hospital within 2 hours post burn (79.7%, 1 121/1 407). Majority of children were taken off clothes as first aid on spot or did not receive any treatment. Most of the children were discharged within 2 weeks after admission (80.0%, 1 126/1 407). There was statistically significant difference in length of hospital stay of children with causes of hot liquid scald, flame burn, electric burn, high temperature solid burn, chemical burn (χ(2) =17.33, P=0.002). Most of the children were treated with non-surgical methods, and the majority of the children got better condition or totally recovered and then discharged.

CONCLUSIONSThe majority of hospitalized children with burn injuries in our unit are young boys in preschool period, who were burnt by hot fluid at the time of dinner and bathing at home during summer. So we should make more effort on popularization of prevention about burn.

Adolescent ; Burns ; classification ; epidemiology ; Burns, Chemical ; Burns, Electric ; Child ; Child, Hospitalized ; statistics & numerical data ; Child, Preschool ; Female ; Hospitalization ; Humans ; Infant ; Inpatients ; Length of Stay ; Male

Objective: To explore the pain experiences of adult burn patients so as to lay foundation for practical analgesic measures. Methods: Using phenomenological method in qualitative research, semi-structured interviews were conducted on 12 adult burn patients hospitalized in our burn units from May to November 2015, aiming at pain experiences from immediately after burns to 3 to 7 months after being discharged from hospital. Then the Colaizzi′s analysis method was applied to analyze, induce, and refine themes of interview data. Results: After analysis, pain experiences of adult burn patients were generalized into 6 themes: deep pain experiences, heavy psychological burden, limited daily life, poor assessment and treatment of pain, different attributions of pain, and different ways of coping of pain. Conclusions Burn pain brings harm to the patients′ physiology, mentality, and daily life. Nevertheless, pain processing modes of medical staff and patients themselves are the key factors affecting patients′ pain experiences. Therefore, according to the deficiency of current situation of pain management, the targeted analgesic intervention measures should be carried out from the perspectives of medical staff and patients.

Objective:To investigate the effects of sodium butyrate(NaB)on pathological processes such as autophagy and inflammation in an Alzheimer's disease(AD)model and related mechanisms.Methods:Aβ 25-35's action on PC12 cells was used to establish an in vitro AD model, in which microglial BV2 were treated with NaB.Based on treatments, there were a PC12 cells group, a PC12 cells+ Aβ 25-35 group, a PC12 cells+ Aβ 25-35+ NaB group, a BV2 cells group, a BV2 cells+ Aβ 25-35 group, and a BV2 cells+ Aβ 25-35+ NaB group.The expression of markers of the autophagy pathway and inflammatory proteins was evaluated by Western blot, immunofluorescence and EdU staining.Results:NaB was able to promote cell proliferation in the AD model and reduce the level of Tau protein hyperphosphorylation.NaB also inhibited the inflammatory response of microglia and reduce the expression of inflammatory response marker NLRP3(148.313±0.973 vs.113.291±1.218, t=38.91, P<0.001). At the same time, NaB promoted the expression of autophagy pathway proteins(including Beclin-1, LAMP-1 and LC3 Ⅱ/Ⅰ)in microglia. Conclusions:NaB can mitigate pathological changes in an AD model by promoting autophagy and reducing the inflammatory response.

Pentraxin 3 (PTX3) was identified as a marker of the inflammatory response and overexpressed in various tissues and cells related to cardiovascular disease. Honokiol, an active component isolated from the Chinese medicinal herb Magnolia officinalis, was shown to have a variety of pharmacological activities. In the present study, we aimed to investigate the effects of honokiol on palmitic acid (PA)-induced dysfunction of human umbilical vein endothelial cells (HUVECs) and to elucidate potential regulatory mechanisms in this atherosclerotic cell model. Our results showed that PA significantly accelerated the expression of PTX3 in HUVECs through the IkappaB kinase (IKK)/IkappaB/nuclear factor-kappaB (NF-kappaB) pathway, reduced cell viability, induced cell apoptosis and triggered the inflammatory response. Knockdown of PTX3 supported cell growth and prevented apoptosis by blocking PA-inducted nitric oxide (NO) overproduction. Honokiol significantly suppressed the overexpression of PTX3 in PA-inducted HUVECs by inhibiting IkappaB phosphorylation and the expression of two NF-kappaB subunits (p50 and p65) in the IKK/IkappaB/NF-kappaB signaling pathway. Furthermore, honokiol reduced endothelial cell injury and apoptosis by regulating the expression of inducible NO synthase and endothelial NO synthase, as well as the generation of NO. Honokiol showed an anti-inflammatory effect in PA-inducted HUVECs by significantly inhibiting the generation of interleukin-6 (IL-6), IL-8 and monocyte chemoattractant protein-1. In summary, honokiol repaired endothelial dysfunction by suppressing PTX3 overexpression in an atherosclerotic cell model. PTX3 may be a potential therapeutic target for atherosclerosis.
Apoptosis/drug effects ; Atherosclerosis/chemically induced/*drug therapy/immunology/pathology ; Biphenyl Compounds/chemistry/isolation & purification/*pharmacology ; C-Reactive Protein/*genetics/immunology ; Down-Regulation/drug effects ; Drugs, Chinese Herbal/chemistry/isolation & purification/*pharmacology ; Human Umbilical Vein Endothelial Cells ; Humans ; I-kappa B Kinase/*immunology ; Lignans/chemistry/isolation & purification/*pharmacology ; Magnolia/chemistry ; Palmitic Acid ; Protein-Serine-Threonine Kinases/*immunology ; Serum Amyloid P-Component/*genetics/immunology ; Signal Transduction/drug effects