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Objective Blood trace and extraction is the important premising for forensic medicine appraisal reliability, this paper puts forward the blood trace and extraction method of using pyramidon. Methods Using pyramidon and benzidine to treat diluted fresh blood, and using Chelex-100 reagent treatment after the DNA in blood samples, and then applying the technology of PCR-STR and fluorescence detection test sample STR typing, observing the STR classification results and the detection sensitivity of DNA, comparing the different influence of two method on subsequent DNA - STR inspection. Results The pyramidon method can detect the minimum hemoglobin concentration is 25μg/mL,which was lower than 5μg/mL of the benzidine method. The results showed that the effective typing rate of the benzidine method was 18.09%, which was significantly lower than 96.67% of the method of the pyramidon method. The tail length of the detection group was 52.40±9.21, and the Oliva tail moment was 43.29±4.85%, and the tail intensity was 16.25±2.35, which was significantly higher than that of the pyramidon group (P < 0.01). Conclusions The sensitivity of the pyramidon method is moderate, and the influence of the following STR subtype and the nuclear DNA of the sample is significantly lower than that of benzidine, which can be used in the pre-experiment of blood stain in forensic medicine.
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Objective To explore the mechanism and inhibition of botulinum toxin type A (BTXA) on hypertrohic scar fibroblasts.Methods The cells were treated by 0 (control),0.2,0.4,0.8 U/ml BTXA for 48 h.Cell viability was detected by MTT assay.Cell apoptosis was detected by Hoechst staining.Cell cycle was detected by flow cytometry.The level of cell cycle related protein D1 (Cyclin D1),proliferation nuclear antigen (PCNA) and activation of phosphatidylinositol 3 kinase (PI3K) / protein kinase B (AKT) signaling pathway were assayed by western blot.Results Compared with control group(0.75±0.07),0.2,0.4,0.8 U/mL BTXA(0.59 ± 0.06,0.43 ± 0.04,0.34± 0.03) inhibited hypertrohic scar fibroblasts cell viability,increased cell apoptotic rate[control group(2.38±0.24)％;BTXA(15.79±1.54)％,(27.32±2.69)％,(38.46±3.90)％],down-regulated the expression of Cyclin D1(control group 1.57±0.18;BTXA 0.93±0.07,0.42±0.04,0.35±0.03) and PCNA(control group 1.46±0.16;BTXA 0.50±0.05,0.59±0.05,0.37±0.03),inhibited the expression of PI3K(control group 0.98±0.06;BTXA 0.49±0.04,0.50±0.04,0.39±0.03) and the phosphorylation of AKT(control group 1.38±0.08;BTXA 0.97±0.06,0.60±0.04,0.29± 0.02),made cell cycle arrested in G1 phase,The difference was statistically significant (P＜0.05).Conclusion These results suggested BTXA inhibit proliferation via blocking the activation of PI3K/AKT signal pathway and down-stream related cell cycle related protein.
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Objective To observe the effects of human embryonic fibroblasts ( FBs ) on the biological characteristics of diabetic FBs in vitro , and to explore the possible therapeutic mechanism on diabetic ulcers . Methods Diabetic FBs were isolated and cultured with embryonic FBs (experimental group), adult FBs(control group 1)and alone(control group 2)in transwell chambers.Diabetic FBs growth curves in the coculture systems were described.The cell shape was observed by microscopy with HE staining .On the 4th and 7th day of cocul-ture, diabetic FBs were isolated to measure the expression levels of hydroxyproline and transforming growth factor -beta 1(TGF-β1)in the supernatant of monoculture .The cell senescense was displayed by β-galactosidase histo-chemical staining on the 3th, 5th and 7th day of coculture.Eight patients with diabetic foot ulcers who were can-didates for embryonic FBs constructs after failed traditional treatment were included in the study .The pre and post self-control study was conducted by observing the wound healing process before and after embryonic FBs treat -ment.Results Diabetic FBs in the experimental coculture system displayed more typical pattern and faster grow -ing rate.The level of hydroxyproline and TGF-β1 in the supernatant of diabetic FBs monoculture was significantly higher in the experimental group than in the control group after treated 4 or 7 days ( hydroxyproline:4 d, 7 d :compared with control group 1:P=0.023,P=0.007;compared with control group 2: P=0.007, P=0.003;TGF-β1 4 d, 7 d:compared with control group 1:P=0.000, P=0.000;compared with control group 2: P=0.000, P=0.000).The expression of SA-β-gal tended to be more apparent in control group 1 and 2 than in the experimental group on the 3rd,5th,and 7th day.All of the 8 cases with diabetic ulcers were cured by using em-bryonic FBs constructs .Conclusion Embryonic FBs can promote the proliferation of diabetic FBs , correct cell senescence and increase the production of hydroxyproline and TGF-β1,which maybe enhance the process of dia-betic wound healing .
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Objective To study the factors relating to failure of vacuum assisted closure (VAC)therapy for pressure ulcers control in order to guide the clinical indications.Methods A retrospective cohort study of 75 patients with pressure ulcers to undergo VAC therapy was conducted.Correlations between 12 risk factors and VAC therapy failure were analyzed.The statistical prognostic factors were screened and the sensitivity,specificity,positive predictive value and negative predictive value of them were analyzed.Risk factors for VAC failure were determined by using Logistic regression analysis.Results Among the 75 patients,17 patients failed to respond to the VAC therapy.The failure rate of VAC therapy had correlations with positive blood cultures,bony exposure,hypoalbuminemia (alb ＜25 g/L) and wound depths≥ 4 cm (70.0％,50.0％,38.5％ and 40.9％,respectively),but the Logistic regression analysis found that the 4 factors were not the independent risk factors for failure of VAC therapy.Conclusions Pressure ulcer patients with positive blood cultures,bony exposure,hypoalbuminaemia (alb ＜25 g/L),and wound depth≥4 cm tend to have a higher failure rate in VAC therapy,which are the relative indications for VAC therapy.
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ObjectiveTo explore the application of the plastic principles in surgical treatment for diabetic foot ulcer and analyze the therapeutic efficacy.Methods51 cases of diabetic foot ulcer repaired with surgical treatment were analyzed.Results26 cases repaired with free skin graft,1 1 cases repaired with skin flap graft,3 cases repaired with amputation,the percentage of first stage healing rate was 91.4％.ConclusionApplication of the plastic principles in surgical treatment for diabetic foot ulcer,could improve the healing percentage and reduce the rate of amputation.