Effects of synthetical methods in the treatment of keloid / 中华医学美学美容杂志

Objective To evaluate comprehensive therapy in the treatment of keloid effect and the patients' satisfaction.Methods From 2002 to 2015,a total of 523 patients with comprehensive treatment,according to the treatment the patients were divided into group A (surgery combined radiotherapy group) and group B (operation with corticosteroid hormone therapy group),group C (corticosteroids in combination with radiotherapy group),and the therapeutic effect of patients with satisfaction was evaluate.Results Total effective rate of three groups of patients were 85.7％,84.0％,64.9％;the efficiency in group A was higher than that of group B and group C;there was statistically significant difference between group A and C (P＜0.05),but there were no significant difference between the group A and group B (P＞0.05).All the patients were follow up for one years and the adverse reaction in the three groups was slight and happened in the 2 month after cure.And the adverse reaction was all self-cure in the final follow up.Difference was statistically significant between three groups of patients' satisfaction,group B better than in group A and group C;there was no significant difference between group A and group B (P＞0.05);the difference between group B and C group was statistically significant (P＜0.05).Conclusions Three kinds of comprehensive therapy effect are obvious,among which surgical adjuvant radiotherapy effect is best,being worth to recommend for clinical use.

Multicenter epidemiological investigation of hospitalized elderly, young and middle-aged patients with severe burn / 中华烧伤杂志

Objective@#To compare and analyze the epidemiological characteristics of hospitalized elderly, young and middle-aged patients with severe burn in recent years, so as to provide reference for the prevention and treatment of elderly patients with severe burn.@*Methods@#Relying on the entry system of epidemiological case data and biological sample of severe burn from multicenter in clinic, medical records of patients with severe burn, aged above 18, hospitalized in 8 burn wards from January 2012 to December 2015 were collected. Six hundred and fifteen patients who were more than 18 years old and less than or equal to 65 years old were included in young and middle-aged group (YM). Eighty-two patients aged more than 65 years old were included in elderly group (E). Data of age, gender, residence, education level, cause of injury, location of injury, season of injury, total burn area, occurrence and area of full-thickness burn injury, wound site, inhalation injury incidence and severity, post burn admission time, proportion of delayed resuscitation, proportion of escharectomy or tangential excision and skin grafting, preinjury systemic disease, system complication during hospitalization, length of hospital stay, outcome of treatment, and reason of abandoning treatment of patients were analyzed. Data were processed with chi-square test and Mann-Whitney U test. The odds ratios of preinjury systemic disease, system complication during hospitalization, and adverse outcome of patients in group YM were compared with those in group E.@*Results@#(1) The majority of patients in the two groups were male, but the proportion of male patients in group YM was higher. There was statistically significant difference in gender distribution of patients between the two groups (χ2=18.727, P<0.001). The majority of patients in the two groups were from rural areas, but the proportion of rural patients in group E was higher. There was statistically significant difference in residence distribution of patients between the two groups (χ2=9.306, P=0.002). Patients in group YM mainly had secondary education, while patients in group E mainly had primary education. There was statistically significant difference in distribution of education level of patients between the two groups (χ2=146.797, P<0.001). (2) The most common causes of injury of patients in the two groups were both flame, but the proportion of patients with flame burn injury in group E was higher. There was statistically significant difference in distribution of cause of injury of patients between the two groups (χ2=25.063, P<0.001). The main locations of injury of patients in groups YM and E were respectively public place and private residence. There was statistically significant difference in location distribution of injury of patients between the two groups (χ2=46.313, P<0.001). The main seasons of injury of patients in groups YM and E were respectively summer and winter. There was statistically significant difference in season distribution of patients between the two groups (χ2=23.143, P<0.001). There was statistically significant difference in distribution of total burn area of patients between the two groups (χ2=25.799, P=0.002). The occurrences of full-thickness burn injury of patients in the two groups were similar (χ2=2.685, P=0.101), while there was statistically significant difference in area of full-thickness burn injury of patients between the two groups (χ2=26.702, P=0.002). There was no statistically significant difference in distribution of wound site of patients between the two groups (χ2=3.954, P=0.785). There were no statistically significant differences in incidence and severity distribution of inhalation injury of patients between the two groups (with χ2 values respectively 0.425 and 0.672, P values above 0.05). (3) There was statistically significant difference in distribution of admission time of patients between the two groups (χ2=6.632, P=0.036), but there was no statistically significant difference in proportion of delayed resuscitation of patients between the two groups (χ2=1.261, P=0.261). The proportion of escharectomy or tangential excision and skin grafting of patients in group YM was 72.0% (443/615), which was significantly higher than 35.4% (29/82) of group E (χ2=44.498, P<0.001). The incidence of preinjury systemic disease of patients in group YM was 13.2% (81/615), which was significantly lower than 61.0% (50/82) of group E (χ2=108.337, P<0.001). The risk of preinjury systemic disease of patients in group E was 10.30 times of that of patients in group YM [with 95% confidence interval (CI) of 6.24-17.01, P<0.001]. During hospitalization, 59.8% (49/82) of patients in group E suffered from system complications, which was significantly higher than 36.6% (225/615) of group YM (χ2=16.282, P<0.001). The risk of system complication of patients in group E was 2.57 times of patients in group YM (with 95% CI of 1.61-4.12, P<0.001). The length of hospital stay of patients in group E was significantly shorter than that of group YM (U=36 735, P<0.001). There was statistically significant difference in treatment outcome of patients between the two groups (χ2=106.251, P<0.001). The risk of adverse outcome of patients in group E was 7.52 times of group YM (with 95% CI of 4.40-12.88, χ2=67.709, P<0.001). The proportion of abandoning treatment of patients in group E was significantly higher than that of group YM (χ2=150.670, P<0.001). The risk of abandoning treatment of patients in group E was 15.86 times of that of group YM (with 95% CI of 9.36-26.88, P<0.001). There was no statistically significant difference in distribution of reason of abandoning treatment of patients between the two groups (χ2=4.178, P=0.243).@*Conclusions@#There were significant differences in the epidemiological characteristics of patients in groups E and YM. In elderly burn patients, the proportion of rural population was higher and the education level was lower. Flame burn was common and burns mostly occurred in private residences and in winter. The total burn area was slightly lower but the area of full-thickness burn injury was larger. The length of hospital stay was shorter and the proportion of surgical treatment was lower. The incidences of preinjury systemic disease and system complication during hospitalization were higher, and therefore the risks of adverse outcome and abandoning treatment were higher.

Decreased intracellular TLR9 confers hyporesponsiveness of RAW264.7 cells to subsequent CpG ODN challenge.

Low-dose CpG ODN pretreatment is known to induce effective protective immunity against acute infectious diseases. In the present study, using primary murine peritoneal macrophages and the macrophage-like cell line, RAW264.7, we investigated whether low-dose CpG ODN pretreatment would induce hyporesponsiveness in response to a subsequent high-dose CpG ODN challenge and further investigated the molecular mechanisms underlying this event. Our results showed that pretreatment with a low dose of CpG ODN inhibits TNF-alpha production stimulated by later high-dose CpG ODN stimulation in a dose- and time-dependent manner. Interestingly, anti-mouse TLR9 blocking antibody added prior to CpG ODN pretreatment did not affect TNF-alpha release, but antibody added after CpG ODN pretreatment augmented the pretreatment effect of CpG ODN. This difference suggests that cell-surface TLR9 is indeed functional on activated cells. Flow cytometry revealed that low-dose CpG ODN pretreatment decreased cell-surface binding and internalization of a subsequent high-dose stimulation, suggesting that decreased internalization of succeeding CpG ODN is associated with reduced TNF-alpha release. Although both intracellular and cell-surface TLR9 expression are observed, low dose of CpG ODN pretreatment increased only cell-surface TLR9 levels. Importantly, low-dose CpG ODN pretreatment also significantly inhibited the activation of NF-kappaB, an important downstream regulator of various proinflammatory cytokines. In summary, our results demonstrate that suppression of TNF-alpha production by low dose of CpG ODN pretreatment correlates with decreased binding and internalization of subsequent CpG ODN, decreased intracellular content of TLR9, and inhibition of NF-kappaB activation.

A synthesized cationic tetradecapeptide from hornet venom kills bacteria and neutralizes lipopolysaccharide in vivo and in vitro.

Sepsis is a complex clinical syndrome that results from a harmful host response to infection, in which foreign bacteria and lipopolysaccharide (LPS) are potent activators of different immune cells, including monocytes and macrophages. To date, there are currently few effective adjuvant therapies in clinical use except activated protein C focusing on the coagulation system. Mastoparans (MPs) are wasp venom cationic amphiphilic tetradecapeptides; these are capable of modulating various cellular activities, including stimulation of GTP-binding protein, phospholipase C and can bind to a phospholipid bilayer. Masroparan-1 (MP-1, INLKAIAALAKKLL-NH2), a tetradecapeptide toxin isolated from hornet venom, was synthesized chemically. In this study, Escherichia coli 25922 (E. coli 25922) and LPS were used to induce sepsis in an animal model. We found that MP-1 treatment at 3 mg/kg protected mice from otherwise lethal bacteria and LPS challenges. MP-1 has antibacterial capabilities against Gram-negative and Gram-positive bacteria. Its antibacterial action against E. coli may result from the destruction of bacterial membrane structures. In addition, treatment of murine peritoneal macrophages with MP-1 potently inhibited the respiratory burst. This effect maybe related to an inhibition of NADPH oxidase in the membrane. Furthermore, MP-1, bound with high-affinity to LPS and lipid A with dissociation equilibrium constants of 484 and 456 nM, respectively, and neutralized LPS in a dose-dependent manner. MP-1 also significantly reduced the expression of TLR4, TNF-alpha and IL-6 mRNA and the release of cytokines in LPS-stimulated murine peritoneal macrophages. Our results shows that the MP-1-mediated protection of mice from lethal challenge by live bacteria and LPS was associated with its bactericidal action and inhibition of inflammatory responses by macrophages to both bacteria and LPS (the release of cytokines and reactive oxygen species).

The screening and isolation of an effective anti-endotoxin monomer from Radix Paeoniae Rubra using affinity biosensor technology.

Lipopolysaccharide (LPS) is a known trigger in the pathogenesis of sepsis, lipid A being the toxic component. One of several adjuvant therapeutic approaches for severe sepsis is currently focusing on the neutralization of LPS. In order to obtain the components from traditional Chinese herbs that can neutralize the endotoxin, aqueous extractions were tested using affinity biosensor technology. From amongst 42 herbs, eight were found to possess lipid A-binding abilities. Radix Paeoniae Rubras had the highest lipid A-binding ability; therefore an aqueous extraction from this plant was investigated further. After preparation using standard methods, including silica gel chromatography and HPLC, we obtained 1, 2, 3, 4, 6-beta-d-pentagalloylglucose (PGG), with lipid A-binding ability. It was found that in vitro, PGG directly bound to lipid A, with a Kd of 32 microM, and that it neutralized the endotoxin both in the Limulus Amebocyte Lysate (LAL) assay and in a TNF-alpha release experiment, in a dose-dependent manner. In in vivo experiments, PGG was found to protect mice from a lethal challenge by LPS, and significantly decreased the plasma endotoxin level both in endotoxemic mice and rats, the reduction of the endotoxin level in rats being tightly associated with the TNF-alpha level. In conclusion, we demonstrate the effectiveness of affinity biosensor technology in discovering useful agents amongst traditional Chinese herbs and using this approach we found a new anti-endotoxin agent.

Chloroquine protects mice from challenge with CpG ODN and LPS by decreasing proinflammatory cytokine release.

Bacterial DNA (bDNA) and lipopolysaccharide (LPS) are potent activators of immune cells such as monocytes and macrophages, which contribute to systemic inflammatory response syndrome (SIRS) and sepsis. To date, no effective anti-sepsis drugs have been developed for clinical use. Chloroquine (CQ), a diprotic weak base traditionally used for treating malaria, was recently shown to decrease cytokine release from macrophages induced by LPS and CpG oligonucleotide (CpG ODN). In the present study, Escherichia coli DNA (EC DNA), CpG ODN and LPS were used to induce SIRS/sepsis in animal models. We found that 30 mg/kg of CQ could protect mice from lethal challenge by CpG ODN and LPS, and 25 mg/kg of CQ could decrease serum TNF-alpha and IL-6 in rats injected with sublethal doses of CpG ODN and LPS. In addition, treatment of murine macrophage ANA-1 cells with 2 mM CQ potently inhibited the release of TNF-alpha, IL-6 and IL-12 induced by CpG ODN and LPS. In human peripheral blood mononuclear cells (hPBMC), 100-200 microM CQ almost completely abrogated release of both TNF-alpha and IL-6 induced by CpG ODN and LPS, whereas IL-6 release induced by EC DNA was not significantly affected by 50 microM CQ. Furthermore, CQ reduced the expression of TLR9 and TLR4 mRNA and the activation of NFkappaB and AP-1 stimulated by CpG ODN and LPS in ANA-1 cells. Flow cytometry and confocal microscopy revealed that CQ increased the accumulation of CpG ODN within ANA-1 cells without influence on its uptake, suggesting that the delayed degradation of CpG ODN was associated with the reduction of proinflammatory cytokine release from the cells. Our results demonstrated that CQ-mediated protection of lethal challenge by CpG ODN and LPS was associated with the reduction of proinflammatory cytokine release.

Comparison of sequence between adenoviral DNA and Escherichia coli DNA / 解放军医学杂志

Objective To identify the putative CpG-N ODNs in adenovisus 2 DNA (Adv2 DNA) and Adv5 DNA by comparing the sequence difference among Adv2, 5, 12 DNA and E.coli (EC) DNA. Methods Sequences of Adv2, 5, 12 DNA and EC DNA were obtained from the Entrez Nucleotides database at NCBI. The specific CpG motifs of Adv2 DNA and Adv5 DNA were identified after above sequences were analyzed and compared by softwares such as DNATools, BioEdit, and so on. All the 12-ODNs with specific CpG motif core were searched from Adv2 DNA and Adv5 DNA. Results 19 specific CpG motifs were ascertained and 504 12-ODNs were detected in Adv2 DNA and Adv5 DNA. Conclusion 504 12-ODNs were putative CpG-N ODNs in Adv2 DNA and Adv5 DNA.

Molecular mechanism of immune cells activated by bacterial DNA / 中国病理生理杂志

Bacterial DNA taken up by immune cells in a CpG motif-independent manner is translocated into endosome. Endosomal maturation is essential for subsequent bacterial DNA-mediated signal transduction. TLR9 is recruited into endosome to recognize bacterial DNA and initiate the TLR/IL-1R signal transduction pathway. As a result , transcription factors NF-?B and AP-1 are activated, which, in turn, leads to proinflammatory cytokine expression and induces a strong acute Th1-like inflammatory response.

Prokaryotic expression, purification and identification of anti-bacterial peptide gloverin / 第三军医大学学报

Objective To express anti-bacterial peptide gloverin in prokaryotic system and investigate its bioactivity. Methods E.coli BL21 was used to express gloverin. Western blot was used to identify the production and Tachypleus Amebocyte Lysate (TAL) was used to detect the bioactivity of the production. Results The production was identified as gloverin peptide by Western blotting, and the TAL data indicated that the production could neutralize LPS. Conclusion Anti-bacterial peptide gloverin is successfully expressed in E.coli BL21.

Screening of bacterial DNA antagonizing CpG oligodeoxynucleotides / 解放军医学杂志

Objective To obtain a potent bacterial DNA antagonizing CpG oligonucleotides (CpG-N ODN) from the structures of Adv2, 5 DNA. Methods Ten putative CpG-N ODNs were synthesized and investigated. Their abilities to inhibit the TNF-? release from hPBMC were observed. Based on the above results, the putative CpG-N ODN was redesigned according to the relationship between the structure and free energy using RNA structure software (version 3.71). Eleven putative CpG-N ODNs were synthesized and screened. Results Out of the ten initial CpG ODNs, ODN101 was the only CpG-N ODN with weak activity to inhibit TNF-? release from hPBMC induced by CpG-N ODN. After redesign, five CpG-N ODNs with strong activity were confirmed. Conclusion Six CpG-N ODNs were obtained with activity to inhibit TNF-? release from hPBMC induced by CpG-N ODN.