Immunogenicity Analysis of the Recombinant Plasmodium falciparum Surface-Related Antigen in Mice.

Plasmodium falciparum, mainly distributed in tropical and subtropical regions of the world, has received widespread attention owing to its severity. As a novel protein, P. falciparum surface-related antigen (PfSRA) has the structural and functional characteristics to be considered as a malaria vaccine candidate; however, limited information is available on its immunogenicity. Here, we expressed three fragments of recombinant PfSRA in an Escherichia coli system and further analyzed its immunogenicity. The results showed that rPfSRA-immunized mice produced specific antibodies with high endpoint titers (1:10,000 to 1:5,120,000) and affinity antibodies (i.e., rPfSRA-F1a (97.70%), rPfSRA-F2a (69.62%), and rPfSRA-F3a (91.87%)). In addition, the sera of immunized mice recognized both the native PfSRA and recombinant PfSRA, the rPfSRA antibodies inhibited the invasion of P. falciparum into the erythrocytes, and they were dose-dependent in vitro. This study confirmed PfSRA could be immunogenic, especially the F1a at the conserved region N-terminal and provided further support for it as a vaccine candidate against P.falciparum.

Interleukin-1ß Protection Against Experimental Sepsis in Mice.

The inflammatory response involving interleukin-1ß (IL-1ß) has been thought to play an important role in the development of late-phase sepsis. However, in this study, we wanted to explore the possibility of using IL-1ß to improve the prognosis of sepsis by triggering local differentiation of bone marrow cells (BMCs) into regulatory dendritic cells (DCs) in vivo, thereby reversing the immune paralysis in late-phase sepsis. Sepsis mouse models were induced by cecal ligation and puncture (CLP) and lethal Escherichia coli O18 infection. Mice were injected intraperitoneally with IL-1ß after CLP and after the lethal infection. Septic BMCs and liver immune cells were isolated at 0, 3, 6, 9, and 14 days post-CLP. BMCs and liver cells isolated from septic mice treated with IL-1ß were adoptively transferred into CLP mice. GFP+-C57BL/6 parabiosis models were established. Serum IL-1ß levels were determined by enzyme-linked immunosorbent assay (ELISA) kit, and the number, ratio, and phenotype of immune cells were observed by flow cytometry. IL-1ß treatment improved the survival of sepsis and increased the numbers of BMCs and liver immune cells in septic mice. Moreover, IL-1ß stimulation increased the number and the percentage of CD11c-CD45RBhigh DCs in septic BM and liver. Adoptive transfer of septic BMCs, liver immune cells, and CD11c-CD45RBhigh DCs treated with IL-1ß into CLP mice attenuated sepsis. IL-1ß triggered the redistribution of CD11c-CD45RBhigh DCs as well as BMCs in parabiosis models. IL-1ß protects against sepsis by stimulating local proliferation and differentiation of BMCs into CD11c-CD45RBhigh DCs at immune organs and non-immune organs.

High Mobility Group Protein 1 Reverses Immune System Paralysis in Late-Phase Sepsis.

High mobility group protein 1 (HMGB1) is considered to be the primary inflammatory factor triggering immune paralysis in late-phase sepsis. In this study, however, we wanted to explore the possibility of using HMGB1 to boost local differentiation of bone marrow cells (BMCs) into regulatory dendritic cells (DCs) in vivo, thereby inducing immune reversal in late-phase sepsis and improving the prognosis. For this purpose, sepsis was induced by cecal ligation and puncture (CLP). Mice were injected intraperitoneally with HMGB1 (10, 50, or 250 µg/kg of body weight) 7 days before CLP. BMCs and liver immune cells were isolated at 0, 3, 5, and 7 days post-CLP. Mice were intranasally infected with Pseudomonas aeruginosa 3 days post-CLP as a secondary pneumonia infection model. BMCs and liver cells isolated from septic mice pretreated with HMGB1 were adoptively transferred into CLP mice. GFP+-C57BL/6 and C3H/HeN-C3H/HeJ parabiosis models were established. We found that HMGB1 pretreatment improved the survival of sepsis and increased the numbers of BMCs and liver immune cells in CLP mice. Furthermore, HMGB1 stimulation improved survival in the secondary pneumonia infection model. HMGB1 increased the number as well as the percentage of CD11c- CD45RBhigh DCs in septic BM and liver. Adoptive transfer of septic cells pretreated with HMGB1 into CLP mice attenuated sepsis. HMGB1 enhanced the redistribution of CD11c- CD45RBhigh DCs through TLR4 signaling in parabiosis models. We conclude that HMGB1 triggers immune reversal through the mobilization, redistribution, and local immune differentiation of BMCs, thereby compensating for impaired immunity and leading to sufficient bacterial eradication.

Polysaccharides from Bupleurum Induce Immune Reversal in Late Sepsis.

BACKGROUND: Bupleurum chinense, a well-known Traditional Chinese Medicine, has been used for thousands of years in China. In this study, we would suggest that Bupleurum polysaccharides (BPS) could improve the prognosis of sepsis through its impact on redistribution of BMCs, which triggers immune reversal in late sepsis. METHODS: BALB/c mice were divided into five groups: sham burn group, burn plus P aeruginosa group, burn plus P aeruginosa with BPS (40 mg/kg, 100 mg/kg, and 250 mg/kg) treatment group, and they were sacrificed at post-burn day (PBD) 0, 3, 5, and 7. BMCs, liver cells, and dendritic cells (DCs) were harvested. Flow cytometry was used to determine the change of phenotypes of DCs and isolate these cells. Cytometric beads array was utilized to analyze the level of inflammatory factors. Cell therapy of BMCs, liver cells, and DCs was administrated to explore the protective role of regional organ immunity. RESULTS: BPS could decrease the lethality of burn sepsis in a dose-dependent fashion and increase both the percentage of CD11cCD45RB DCs in bone marrow (BM) and liver and the number of BMCs and liver cells significantly. Cell therapy of BMCs, liver cells, and CD11cCD45RB DCs at PBD7 could protect septic mice from sepsis. CONCLUSION: BPS has shown its potential in promoting the prognosis of post-burn sepsis through its effect on immune redistribution of BMCs, especially via differentiation of CD11cCD45RB DC cells in BM and nonimmune organs to induce immune reversal in late sepsis.

Lentinan ameliorates burn sepsis by attenuating CD4+ CD25+ Tregs.

AIM: The aim of our study was to investigate the effect of lentinan on regulatory T cells (Tregs) in sepsis, especially on the generation of interleukin (IL)-10 via regulation of Erk-FoxO1 signaling. METHODS: BalB/c mice were randomized into five groups: sham group, the group with burns plus Pseudomonas aeruginosa infection, and the groups with burns plus P. aeruginosa infection administered 40, 100, and 250mg/kg of lentinan. The mice were sacrificed on postburn days 0, 1, 2, 3, and 4, respectively, with eight animals per group at each time point. The peripheral blood CD4+ CD25+ Tregs and CD4+ T cells were isolated using magnetic microbeads. The phenotypes were analyzed by flow cytometry. The cytokine levels were determined with enzyme-linked immunosorbent assay (ELISA). Signal transduction was studied by Western blot, quantitative polymerase chain reaction (qPCR), and luciferase assay. RESULTS: The IL-10-producing capacity of CD4+ CD25+ Tregs was significantly enhanced in the group with burns plus P. aeruginosa infection, compared with the sham group. Administration of lentinan significantly decreased IL-10 production and FoxP3 expression of CD4+ CD25+ Tregs. The proliferative activities of CD4+ T cells, however, were restored. Lentinan decreased lipopolysaccharide (LPS)-induced IL-10 production in the Tregs isolated from burned mice. In addition, lentinan attenuated LPS-stimulated Erk-FoxO1 activation. CONCLUSIONS: Lentinan may improve the outcome of postburn sepsis by suppressing LPS-triggered Erk-FoxO1 activation. Consequently, the hyperfunction of CD4+ CD25+ Tregs is inhibited, leading to a shift in the inflammatory status from Th2 to Th1 in postburn sepsis.

Clinical patterns and characteristics of uveitis in a secondary hospital in southern China.

AIM: To investigate the characteristics of uveitis in a secondary hospital in southern China. METHODS: We reviewed all records of patients with uveitis at Hengli Hospital from January 2008 to December 2011. Demographic data, past history, ophthalmic examinations and other laboratory tests were analyzed. RESULTS: One hundred and ninety-nine uveitis patients were enrolled in this study, including 134 (67.3%) males and 65 females (32.7%) with an average age of 41.0±15.1y. The anatomical distribution included 103 (51.8%) cases of anterior uveitis, followed by panuveitis (65, 32.7%), posterior uveitis (29, 14.6%) and intermediate uveitis (2, 1.0%). Of the 98 (49.2%) non-idiopathic cases, there were 10.1% Behcet's disease, 9.5% Vogt-Koyanagi-Harada (VKH) syndrome, 7.5%infectious uveitis, 7.5% traumatic uveitis and 3.5% postoperative uveitis. CONCLUSION: Idiopathic anterior and posterior uveitis, Behcet's disease, VKH syndrome, infectious uveitis and traumatic uveitis are the most common uveitis entities in a secondary hospital in southern China. Additional measures should be taken to prevent infectious and traumatic uveitis.

[Chemical characteristics in airborne particulate matter (PM10) during a high pollution spring dust storm episode in Beijing, Tianjin and Zhangjiakou, China].

Atmospheric particulate matter (PM10) was collected at sampling locations of Beijing, Tianjin and Zhangjiakou from April 1st to May 24th, 2012. The mass concentration of PM10 and concentrations of ions, elemental carbon (EC) and organic carbon (OC) in PM10 were determined. The results showed that average mass concentration of PM10 were 233.82 microg x m(-3) for Beijing, 279.64 microg x (-3) for Tianjin and 238.13 microg x m(-3) for Zhangjiakou, respectively. Backward trajectories results confirmed dust storm events occurred from 27th to 29th April. The maximum daily mass concentrations of PM10 were 755.54 microg x m(-3) for Beijing, 831.32 microg x m(-3) for Tianjin and 582.82 microg x m(-3) for Zhangjiakou during the dust storm episodes, respectively. Water-soluble ions (Na+, NH4+, Ca2+, K+, F-, Cl-, NO3-, SO4(2-)), organic carbon (OC) and elemental carbon (EC) were major aerosol components during the dust storm episodes, and their concentrations were higher than non-dust storm days. In addition, dust storm caused increases in NO3-, SO4(2-) and enrichment of secondary organic carbon (SOC) concentration relative to OC, suggesting that chemical reaction processes involving gas-particle conversion occurred during the long-distance transport of aerosol particles.

Astragalus polysaccharides attenuate postburn sepsis via inhibiting negative immunoregulation of CD4+ CD25(high) T cells.

BACKGROUND: Astragalus polysaccharides (APS) isolated from one of the Chinese herbs, Astragalus mongholicus, are known to have a variety of immunomodulatory activities. However, it is not yet clear whether APS can exert an effect on the immune functions of regulatory T cells (Tregs). This study was carried out to investigate the effect of APS on the immune function of peripheral blood Tregs in postburn sepsis. METHODOLOGY/PRINCIPAL FINDINGS: BALB/C mice were randomly divided into six groups as follows: sham burn group, burn control (burn without infection animals) group, burn plus P. aeruginosa group, burn plus P. aeruginosa with APS (50 mg/kg) treatment group, burn plus P. aeruginosa with APS (100 mg/kg) treatment group, and burn plus P. aeruginosa with APS (200 mg/kg) treatment group, and they were sacrificed on postburn day 1, 3, 5, and 7, respectively, with seven animals at each time point. Magnetic microbeads were used to isolate peripheral blood Tregs and CD4(+) T cells. Phenotypes were analyzed by flow cytometry, and cytokine levels were determined with ELISA. In the burn plus P. aeruginosa group, forkhead/winged helix transcription factor p3 (Foxp3) expression on CD4(+)CD25(+)Tregs were strongly enhanced in comparison to the sham group, and the capacity of CD4(+)CD25(+)Tregs to produce interleukin (IL)-10 was markedly increased. Administration of APS to inhibit CD4(+)CD25(+)Tregs could significantly decrease expression of Foxp3 on CD4(+)CD25(+)Tregs, and IL-10 production in burned mice with P. aeruginosa infection. At the same time, proliferative activity and expression of IL-2 and IL-2Rα on CD4(+) T cells were restored. In contrast, anti-Toll-like receptor 4 (TLR4) antibody could block the effect of APS on Tregs immune function. CONCLUSION: APS might suppress CD4(+)CD25(+)Treg activity, at least in part, via binding TLR4 on Tregs and trigger a shift of Th2 to Th1 with activation of CD4(+) T cells in burned mice with P. aeruginosa infection.

[The regulatory effect and mechanism of Astragalus polysaccharides on CD11c(high)CD45RB(low) dendritic cell].

OBJECTIVE: To investigate immunomodulatory effect of Astragalus polysaccharides (APS) on IL-12-secreting dendritic cell (DC) subset CD11c(high)CD45RB(low) DC. METHODS: Spleen CD11c(high)CD45RB(low) DC and CD4(+)T lymphocytes in BALB/c mice were purified by magnetic beads sorting, and were treated with 0 (as control), 50, 100, 200 µg/mL APS. Immunofluorescence staining and flow cytometry were used to determine expressions of CD11c(high)CD45RB(low) DC surface molecules, including CD40, CD80, CD86, I-A/E, and Toll-like receptor (TLR) 4. IL-12 level in CD11c(high)CD45RB(low) DC culture supernatant was determined by ELISA. The CD4(+) T lymphocytes were divided into: normal control group, non-stimulation group (CD4(+)T lymphocytes cocultured with APS-unstimulated CD11c(high)CD45RB(low) DC), high-dose APS stimulation group (CD4(+)T lymphocytes cocultured with 200 µg/mL APS-stimulated CD11c(high)CD45RB(low) DC), high-dose APS stimulation+antibody 1 group (CD4(+)T lymphocytes cocultured with 200 µg/mL APS-stimulated CD11c(high)CD45RB(low) DC and IL-12 antibody), high-dose APS stimulation+ antibody 2 group (CD4(+)T lymphocytes cocultured with 200 µg/mL APS-stimulated CD11c(high)CD45RB(low) DC and IL-12 antibody isotype). Proliferation ability of CD4(+) T lymphocytes was determined with MTT method. IL-4 level as well as IFN-γ level in CD4(+)T lymphocyte culture supernatant was determined by flow cytometry. Data were processed with one-way analysis of variance. RESULTS: Compared with those in control, the expressions of CD11c(high)CD45RB(low) DC surface molecules (except for CD86) on CD11c(high)CD45RB(low) DC surface, as well as IL-12-secreting level with dose-dependence were increased in cells stimulated with 50, 100, 200 µg/mL APS. Proliferation ability of CD4(+)T lymphocytes in high-dose APS stimulation group was higher as compared with that in non-stimulation group (F = 13.438, P < 0.05). IFN-γ level in high-dose APS stimulation group \[(2784 ± 137) pg/mL\] was higher than that in non-stimulation group \[(1952 ± 101) pg/mL, F = 12.177, P < 0.05\]. IL-4 level in high-dose APS stimulation group was (172 ± 20) pg/mL, which was lower than that in non-stimulation group \[(193 ± 19) pg/mL, F = 11.963, P < 0.05\]. Proliferation ability of CD4(+) T lymphocytes, IFN-γ level, and IL-4 level in high-dose APS stimulation + antibody 1 group were all ameliorated when compared with those in non-stimulation group; while levels of the 3 indexes in high-dose APS stimulation + antibody 2 group were similar to those in high-dose APS stimulation group. CONCLUSIONS: APS can activate IL-12-producing CD11c(high)CD45RB(low) DC, and further induce the activation of immune function of T lymphocyte with shifting of Th2 to Th1 in vitro. APS can enhance the immune response via promoting the phenotypic and functional maturation of CD11c(high)CD45RB(low) DC.

High mobility group box 1 protein suppresses T cell-mediated immunity via CD11c(low)CD45RB(high) dendritic cell differentiation.

AIM: High mobility group box 1 protein (HMGB1) has been identified as a late proinflammatory cytokine and plays a key role in immune regulation. However, it is not yet clear whether HMGB1 can induce the activation and differentiation of dendritic cell (DC) subsets and subsequently modulate immune function of T cells. This study was performed to investigate the effect of HMGB1 on the differentiation of splenic DCs and its influence on T cell-mediated immunity in terms of DC subsets CD11c(low)CD45RB(high) DCs and CD11c(high)CD45RB(low) DCs in male BALB/c mice spleens in vitro. RESULTS: MACS microbeads were used to isolate splenic DCs, CD11c(low)CD45RB(high) DCs, CD11c(high)CD45RB(low) DCs and CD4(+) T cells. The percentage of CD11c(low)CD45RB(high) DCs was significantly increased after treatment with HMGB1 compared to their counterparts (CD11c(high)CD45RB(low) DCs). It was found that unlike the gradually increasing interleukin (IL)-12 secretion of CD11c(high)CD45RB(low) DCs induced by HMGB1, CD11c(low)CD45RB(high) DCs showed a obvious dose-dependent response between IL-10 production and HMGB1 stimulation. In order to verify whether the alteration of CD4(+) T cells was mainly associated with the differentiation of splenic DCs mediated by HMGB1 to CD11c(low)CD45RB(high) DCs, anti-IL-12 receptor (IL-12R) or anti-IL-10R monoclonal antibody was used to inhibit the effect of CD11c(high)CD45RB(low) DCs or CD11c(low)CD45RB(high) DCs in CD4(+) T cells mixed lymphocyte reaction culture. After treatment with anti-IL-12R or anti-IL-10 monoclonal antibody in CD4(+) T cells+CD11c(high)CD45RB(low) DCs or CD11c(low)CD45RB(high) DCs mixed lymphocyte reaction, the induction of these DCs on T cells was inhibited dramatically. CONCLUSION: These data demonstrated that HMGB1 might induce the differentiation of splenic DCs to CD11c(low)CD45RB(high) DCs followed by shifting of Th1 to Th2 with enhancement of T lymphocyte immune function in vitro. Also, the effect of HMGB1 on T cell differentiation to Th2 was not associated with the inhibition of IL-12 production in CD11c(high)CD45RB(low) DCs.

Naturally existing CD11c(low)CD45RB(high) dendritic cells protect mice from acute severe inflammatory response induced by thermal injury.

Despite a good understanding of the process that initiates and promotes host inflammation induced by acute injury, little is known about the host immune cells responsible for the inhibition of inflammatory response to thermal injury. The aim of this study was to investigate the potential effect of naturally existing CD11c(low)CD45RB(high) dendritic cells (CD11c(low)CD45RB(high) DCs) on acute severe inflammatory response and mortality rate in burned mice. Changes in the percentage of distinct subsets of splenic DCs and production of cytokines (IL-6, TNF-α, CCL-2) as well as CC chemokine (CCL)-2 were measured by FACS at various time points. The influence of CD11c(low)CD45RB(high) DCs on IL-6, TNF-α, CCL-2 as well as IL-10 levels and mortality rate was observed following a single intraperitoneal injection of DCs to scald mice. Levels of IL-6, CCL-2 and TNF-α were peaked at postburn hours (PBHs) 12, and percentages of CD11c(low)CD45RB(high) DCs were elevated at PBH 12-24. A single intraperitoneal injection of CD11c(low)CD45RB(high) DCs to 15% total body surface area in scald mice led to significant decrease in IL-6, TNF-α and CCL-2, and lethality, but up-regulation of IL-10 compared with untreated mice from PBH 0 to 48. CD11c(low)CD45RB(high) DCs can effectively down-regulate the production of inflammatory cytokines and reduce the mortality rate in 15% TBSA scald injury mice.

Astragalus polysaccharides regulate T cell-mediated immunity via CD11c(high)CD45RB(low) DCs in vitro.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragalus polysaccharides (APS) isolated from one of the Chinese herbs, Astragalus mongholicus, are known to have a variety of immunomodulatory activities. However, it is not yet clear whether APS can induce the activation and differentiation of dendritic cells (DCs) and subsequently activate T cells. AIM OF THE STUDY: This study was carried out to investigate the effect of APS on the differentiation of splenic DCs and its influence on T cell-mediated immunity through interleukin (IL)-12-producing CD11c(high)CD45RB(low) DCs in vitro. METHODOLOGY: MACS microbeads were used to isolate splenic DCs, CD11c(high)CD45RB(low) DCs, CD11c(low)CD45RB(high) DCs and CD4(+) T cells. Phenotypes were analyzed by flow cytometry, and cytokine levels were determined with cytometric bead array or ELISA. RESULT: The percentage of CD11c(high)CD45RB(low) DCs was significantly increased after treatment with APS compared to their counterparts. The cytokine secretion pattern of CD11c(high)CD45RB(low) DCs and CD11c(low)CD45RB(high) DCs was detected, and it was found that unlike the stable IL-10 secretion pattern of CD11c(low)CD45RB(high) DCs induced by APS, CD11c(high)CD45RB(low) DCs showed a dose-dependent relationship between IL-12 production and APS stimulation. In order to verify whether the activation of CD4(+) T was associated with the differentiation of splenic DCs mediated by APS to CD11c(high)CD45RB(low) DCs, anti-IL-12 receptor (IL-12R) as well as anti-IL-10R monoclonal antibody was used to inhibit the effect of CD11c(high)CD45RB(low) DCs and CD11c(low)CD45RB(high) DCs in CD4(+) T mixed lymphocyte reaction culture. After treatment with anti-IL-12R or anti-IL-10 monoclonal antibody in CD4(+) T+CD11c(high)CD45RB(low) DCs or CD11c(low)CD45RB(high) DCs mixed lymphocyte reaction, the inductions of these DCs on T cells were inhibited dramatically. CONCLUSION: APS might induce the differentiation of splenic DCs to CD11c(high)CD45RB(low) DCs followed by shifting of Th2 to Th1 with enhancement of T lymphocyte immune function in vitro. Also, the effect of APS on T-cell differentiation to Th1 was not associated with the inhibition of IL-10 production in CD11c(low)CD45RB(high) DCs.

Inflammatory response and immune regulation of high mobility group box-1 protein in treatment of sepsis.

Sepsis is an infection induced systemic inflammatory response syndrome and is a major cause of morbidity as well as mortality in intensive care units. A growing body of evidence suggests that the activation of a proinflammatory cascade is responsible for the development of immune dysfunction, susceptibility to severe sepsis and septic shock. The present theories of sepsis as a dysregulated inflammatory response and immune function, as manifested by excessive release of inflammatory mediators such as high mobility group box 1 protein (HMGB1), are supported by increasing studies employing animal models and clinical observations of sepsis. HMGB1, originally described as a DNA-binding protein and released passively by necrotic cells and actively by macrophages/monocytes, has been discovered to be one of essential cytokines that mediates the response to infection, injury and inflammation. A growing number of studies still focus on the inflammation-regulatory function and its contribution to infectious and inflammatory disorders, recent data suggest that HMGB1 formation can also markedly influence the host cell-mediated immunity, including T lymphocytes and macrophages. Here we review emerging evidence that support extracellular HMGB1 as a late mediator of septic complications, and discuss the therapeutic potential of several HMGB1-targeting agents in experimental sepsis. In addition, with the development of traditional Chinese medicine in recent years, it has been proven that traditional Chinese herbal materials and their extracts have remarkable effective in treating severe sepsis. In this review, we therefore provide some new concepts of HMGB1-targeted Chinese herbal therapies in sepsis.

Combined fascial flap and expanded skin flap for enveloping Medpor framework in microtia reconstruction.

The Medpor implant is another choice for a new auricular framework besides autogenous costal cartilage. However, its relatively frequent exposure and less-matching skin coverage discourage surgeons from using it. In this article, we present a new two-flap method, a combination of the temporoparietal fascial flap and the expanded skin flap, for wrapping the Medpor implant in microtia reconstruction. A staged surgical procedure was performed, including soft tissue expansion in the mastoid region, soft tissue expander removal, expanded skin flap and temporoparietal fascial flap formation, Medpor framework implantation, and the combined two-flap envelopment. Conventional lobule transposition and tragus reconstruction were accomplished for selected patients. In this study, a total of 22 microtias were reconstructed consecutively using this method. Eighteen patients were followed since the first surgery. The postoperative follow-up time ranged from 3 to 12 months. The draped soft tissue covering was thin enough to show the reconstructed ear with excellent, subtle contour when edema gradually vanished 3-6 months postoperatively. The new ear had a stable shape, and its skin color and texture matched the normal surrounding skin very well. No exposure or extrusion of the framework was observed in the series. The Medpor implant enveloped by both a temporoparietal fascial flap and an expanded cutaneous flap appears to be a promising alternative for the auricular framework in microtia reconstruction. Because of the wrapping tissues, auricular construction using a Medpor implant can be a safe, steady, and easily acceptable choice for both microtia patients and their physicians.

Glucose endothelial cytotoxicity and protection by Dan Gua-Fang, a Chinese herb prescription in huVEC in hyperglycemia medium.

BACKGROUND: Low success rate of blood glucose in diabetes is an international problem. The endothelia cytotoxicity of hyperglycemia has been widely accepted. However, it has not been seen in reports of the value of concentration of high glucose beginning to produce cytotoxicity and the relationship between hyperglycemia and cytotoxicity as well as how to effectively prevent and control hyperglycemia cytotoxicity. Dan Gua prescription is an effective Chinese herb prescription for diabetic vascular complications. METHODS: Dan Gua prescription was contained in Dan Gua liquor utilized in experiments. (1) The cytotoxicity experiment of Dan Gua was carried out with M199 medium whose glucose (Glu) was 5.55 mmol/l to seek for a suitable experimental concentration of Dan Gua. (2) The human vessel endotheliocyte was cultivated for 72 h with mediums containing glucose in different concentrations (Group G1 to Group G11, Glu: 5.5 to 99.9 mmol/l, respectively), and assayed an optical density (OD) value using the 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide method. (3) Experiment 2 was repeated. However, the medium of each group (Groups Y1 to Y11) contained Dan Gua liquor whose concentration was 1/300. RESULTS: There was a negative correlation between means of cell OD values and glucose concentrations (r=-.927, R(2)=.844), and it presented a notable linear correlation (y=0.681-0.002x). Based on the OD value of 5.5-mmol/l glucose concentration (group G1), when glucose concentration reached 22.2 mmol/l (G4), the difference in OD values has a statistical significance. OD values in Y1-Y11 were not less than that of G1. CONCLUSION: There is a notable linear correlation between the endothelial cytotoxicities of Glu and its concentrations. The spinodal point concentration of statistical significance of hyperglycemia cytotoxicity is 22.2 mmol/l; 1/300 Dan Gua can reverse the endothelia cytotoxicity in different concentrations of hyperglycemia.

Aesthetic rhinoplasty of the Asian nasal tip: a brief review.

Tip surgery, the most important part of the rhinoplasty procedure, has entered a new era in the past few decades. Various treatment protocols have been attempted. To date, however, opinions on the management of the Asian tip have not been solidified. To generalize and provide appropriate guidelines for the treatment of typical Asian tips, an English literature search from 1977 to March 2007 was conducted. Finally, a total of 26 papers were selected for review. The full text of each paper was read carefully, and data were extracted. Then all extracted information was imported into Microsoft Excel. Nine articles treating 11 groups of patients described the suitable techniques for Asian nasal tips, with 81.8% of the groups advocating that the protocol include a grafting technique, 64% reporting use of the grafting technique alone, and 9% applying cartilage reduction and a suturing technique. Of the 11 (18%) groups, 2 attempted more than one technique. Because of the Asian nasal tip's innate qualities, success with nasal tip plasty for Asians depends on the combined application of appropriate suturing, grafting, and defatting, with grafting techniques contributing the most.

[Application of both expanded cutaneous flap and temporoparietal fascia flap in ear reconstruction with Medpor framework].

OBJECTIVE: To investigate the feasibility and results of application of both expanded cutaneous flap and temporoparietal fascia flap in total ear reconstruction with Medpor framework. METHODS: The main procedure consists of two stages: Stage I-skin expansion; Stage II -auricle formation consists of orientation of Medpor implant and creation of coverage for the implant by both expanded skin flap and temporoparietal fascia flap. RESULTS: Twenty-two ears in 22 unilateral microtia patients were constructed using Medpor implants covered with both expanded cutaneous flap and temporoparietal fascia flap over the last three years, they were accepted as pleasing by the patients. CONCLUSIONS: Application of both expanded cutaneous flap and temporoparietal fascia flap can assure no extrusion of Medpor implant in ear reconstruction. Either more, the two layers of transferred tissues will not affect the profile details of the reconstructed ear. And because the skin covering the framework and fascia is derived from mastoid region, the appearance and profile of the reconstructed auricle is true to nature and close to that of the opposite one.

[Influence of the escharectomy during stock stage on the peripheral lymphocyte apoptosis and the antigen presentation function of monocytes in peripheral blood of scalded rats].

OBJECTIVE: To investigate the influence of escharectomy at different time-points after burn injury on the lymphocyte apoptosis and the antigen presentation function of monocytes in peripheral blood of scalded rats. METHODS: One hundred and thirty-six Wistar rats were randomly divided into normal control ( C,n = 8 ), scald ( S, n = 64,without treatment after scald) , A ( n = 40, with escharectomy at 36 post-burn hour( PSH) ) , B ( n = 24, with escharectomy at 72 PSH ) groups. The rats in A , B, S groups were inflicted with 30% TBSA full-thickness scald. The rats in S group were sacrificed on 6,12,24,72,120,168,216, 288 PSH, while those in A and B groups were sacrificed at 72 -288 PSH, 168 -288PSH, respectively. The rats in C group were also sacrificed as control. The apoptotic rate of peripheral lymphocytes, the positive expression rate of MHC- II in mononuclear cells, the changes in concentration of IL-4 and gamma-IFN were determined in each group. The correlation of above indices were also analyzed. RESULTS: (1) The apoptotic rate of peripheral lymphocyte in S group were increased dramatically at 6PSH, peaking at 24 PSH( 18. 19+/-1.42% ) , then decreasing gradually, reaching the lowest level at 72 PSH(8. 25+/-0.56% ) , then it increased gradually again, approaching almost the peak value at 288 PSH( 17.81 +/- 1.99% ). The values were all obviously higher than those in C group( P <0.05). The apoptotic rates of peripheral lymphocyte in A and B groups were evidently lower than that in S group ( P <0. 01). (2) The positive expression rate of MHC-II in monocyte was decreased sharply at 6 PSH, and it was 20% lower than that in C group (37. 2 +/- 2. 4% ) at 24 PSH. It then increased gradually, but it was significantly lower than that in A, B groups at 288 PSH (18. 8 +/-2. 8, P <0.01). (3) The plasma level of y-IFN in S group increased gradually from 6 PSH on, peaking at 24 PSH(440. 8 +/-25. 1 )ng/L,then decreasing gradually , and it reached the lowest level at 288 PSH (51.3 +/-37.0) ng/L. The IL-4 level in S group was increased gradually ,peaking at 288 PSH (78. 1+/-2. 8) ng/L. (4) There was negative correlation between the expression rate of MHC- II in S group and IL-4/gamma-IFN ratio in escharectomy groups during 72 - 288 PSH ( r = - 0. 96, P < 0. 05). CONCLUSION: Eacharectomy after scald can inhibit peripheral lymphocyte apoptosis, slow down the insertional tendency of IL-4/gamma-IFN , and ameliorate the antigen presentation function of monocytes. Moreover, escharectomy during shock stage can markedly promote the immune function of monocytes.