Objective To observe and explore the value of combined detection of serum BNP and TnI in patients with acute heart failure(AHF). Methods 60 AHF patients from February 2015 to April 2016 were selected as observation group, and divided into Level III group(n=32) and Level IV group(n=28) according to NYHA classification;60 healthy patients at the same period were selected as control group.BNP and TnI levels were detected by chemiluminescence in each group, and were compared.Results BNP and TnI of observation group were(2624.4 ±378.6) ng/mL and(0.87 ± 0.22) ng/mL, while in control group were(72.4 ±20.2) ng/mL and(0.13 ±0.02) ng/mL, the differences were significant(P<0.05);BNP and TnI in level III group before treatment were significantly lower than grade IV group(P<0.05), after treatment, the level of BNP and III in group IV and group TnI were significantly lower(P<0.05), and the levels of BNP and TnI were significantly lower than those in III group after treatment(P<0.05). Conclusion BNP combine with TnI levels monitoring play an important role in AHF diagnosis and prognosis evaluation, can help to guide the treatment of AHF.

Objective To explore the use of rhomboid skin flap in expanded skin flap transfer. Methods A rhomboid skin flap was designed if the top soft part could not be fully utilized after expanded in a rotation skin flap. The flap pedicels were designed near the incision side. It should be ensured that ra-tio of the length to the width of the composite flap, which was composed of the rhomboid skin flap and the rotation skin flap, was 2.5∶1.0. Results Among these 11 patients with re-designed rhomboid skin flaps in the rotation skin flaps, the ratio of the length to the width reached to 3∶1 in some cases, but 2. 5∶1.0 in most cases. All the skin flaps survived, except one patient with disturbance of blood circulation in a small area and one with mild congestion. Conclusion The expanded soft tissue can be fully and rationally utilized to repair the skin defect in this design. Attention should be paid to the ratio of the length to the width of the composited flap, and it is better to select axial flap as the composite flap for safety. This method is safe, and worthy of recommendation.

Objective To investigate the operative methods and merits of nasal reconstruction in terms of the aesthetics. Methods The noses of 12 patients were reconstructed with an expanded triangle-shaped forehead flap with unilateral supratrochlear artery, a skin expander was placed obliquely under galea aponeurotica of forehead. Liquid was injected with conventional expansion method. Using the skin and the scars on nasal dorsum and tip as lining, based on intercanthal distance and aesthetic standard, a triangle-shaped forehead myocutaneous flap was designed over the expanded forehead skin tissue and used for a nasal reconstruction. The triangle-shaped flap was trimmed to different layer and reshaped based on aesthetic subunit.Results In twelve post-operative patients with nasal defect, no flap necrosis was found and the appearance of reconstructed noses were almost normal and satisfactory after follow-up for 6 months to 2 years. Conclusion The modified forehead myocutaneous flap according to aesthetic standard is safe and ideal for major nasal reconstruction. Meticulous moulding of triangle-shaped flap, nose interior with good blood supply, and primary insertion of nose stretcher are the key to a satisfactory appearance.

Objective To explore the mutation situation of pancreatic cancer cell mitochondria DNA D-loop region. Method PCR and direct sequencing was used to analyze the mutational site of mitochondria DNA D-loop region in two pancreatic cancer cell lines SW1990 and JF-305 and normal primary cultured pancreas cell. Result The point mutations were found in two pancreatic cancer cell lines and normal primary cultured pancreas cell. Eight point mutations were found in SW1990 and 9 point mutations were found in JF-305. Three point mutations (73 site A-G,16223 site C-T, and 16358 site c-T) existed in all two pancreatic cancer cell lines and normal primary cultured pancreas cells, which can be considered as polymorphism. Other two point mutations (16211 site C-T and 16311 site T-C) were only found in two pancreatic cancer cell lines, which can be considered as special mutations. Conclusion The mitochondrial DNA D-loop region of pancreatic cancer cells existed polymorphism and special mutations, and the special mutations might be new molecule marker.

ObjectiveTo investigate the influence of connective tissue growth factor (CTGF) onthe collagen metabolism in human keloid fibroblasts with RNA interference (RNAi).Methods Human keloid fibroblasts (KFB) in vitro were transfected by 3 pairs of specific small interfering RNA (siRNA) CTGF plasmid synthesized for human CTGF,respectively.Reverse transcriptase-polymerase chain reaction (RT-PCR) contributed to the screening of the best siRNA in interfering of CTGF expression in human keloid fibroblasts to construct the plasmids,with the application of RNAi,to test the changes of expression level and collagen content of CTGF in transfected keloid fibroblasts through RT-PCR and Western blotting compared to its control groups.ResultsThe 3rd pair (C3) siRNA- CTGF expression of genes and proteins was remarkably inhibited after being interfered with human keloid fibroblasts,with inhibitory rates of 86.8 ％ and 65.6 ％.ConclusionsKeloid fibroblasts transfected by plasmid siRNA-CTGF effectively inhibite the expression of CTGF and deposition of collagen,and CTGF promotes the collagen synthesis in keloid development.

Objective To investigate the early diagnosis and disease evaluation value in patients with acute pancreatitis by various serum cytokines measurement.Methods Forty-eight acute pancreatitis patients were divided into two groups based on the results of computed tomography (CT) examination:mild acute pancreatitis group (30 cases) and severe acute pancreatitis group (18 cases).The other 30 normal persons were selected as control group.The various serum cytokines were measured by enzyme-linked immunosorbent assay (ELISA).Results The serum concentrations of interleukin(IL)-1,IL-10 and tumor necrosis factor (TNF)-α in mild acute pancreatitis group were significantly higher than those in control group [(25.00 ± 1.92) ng/L vs.(10.08 ± 2.65) ng/L,(59.78 ± 4.51) ng/L vs.(1.80 ± 0.66) ng/L,(55.31 ± 8.54) ng/L vs.(18.72 ± 7.84) ng/L,P ＜ 0.05].The serum concentrations of IL-1,IL-6,IL-8,TNF-α and platelet activating factor (PAF) in severe acute pancreatitis group were significantly higher than those in mild acute pancreatitis group [(93.27 ± 3.98) ng/L vs.(25.00 ± 1.92) ng/L,(397.84 ± 13.05) ng/L vs.(34.12 ± 4.96) ng/L,(93.32 ±3.40) ng/Lvs.(13.06± 1.86) ng/L,(181.94 ±7.54) ng/Lvs.(55.31 ±8.54) ng/L,(284.53 ±7.88) ng/L vs.(175.25 ±30.15) ng/L,P＜0.05].Conclusion The various serum cytokines measurement has great importance on the early diagnosis of acute pancreatitis and discrimination between the mild acute pancreatitis and severe acute pancreatitis.

Objective To explore the effects of recombinant plasmids of pGPU6/GFP/NeoshRNA-CTGF (shRNA-CTGF) on the type Ⅰ collagen (COL-Ⅰ) protein expression in keloid,through RNA interference on connective tissue growth factor (CTGF) in vivo and in vitro.Methods Recombinant plasmids were designed and constructed by specific shRNA-CTGF; After transfeced human keloid fibroblast with shRNA-CTGF in vitro,RT-PCR was used to detect the CTGF mRNA level,and Western blot to detect the secretion of COL-Ⅰ.After transfected the keloid of nude mice with shRNA-CTGF in vivo,RT-PCR was used to detect the CTGF and COL-Ⅰ mRNA level,and Western blot was used to detect the protein expression of COL-Ⅰ.Results Recombinant plasmids of CTGF were successfully constructed,and the CTGF gene expression was significantly decreased in vivo and in vitro by 86.8％ and 54.1 ％,respectively; Down-regulation of CTGF in vitro significantly inhibited the mRNA and protein level of COL-Ⅰ by 76.8％ and 65.6％,respectively; Down-regulation of CTGF in vivo significantly reduced the COL-Ⅰ mRNA and protein level by 52.7％ and 48.0％,respectively.Conclusions CTGF gene expression is successfully down-regulated by the recombinant plasmid of shRNA-CTGF in vivo and in vitro.shRNA-CTGF significantly reduces the COL-Ⅰ protein level in keloid.It implies that CTGF gene is a potential target in the therapy of pathological scar.

BACKGROUND:Early vascularization is crucial for wound healing. A high-porosity, macrovoid alogeneic skin leads to the rapid vascularization and celular infiltration.
OBJECTIVE: To obtain a new alogeneic skin product with high porosity, good cel permeability and good histocompatibility using an improved preparation method of human acelular dermal matrix.
METHODS: Cel components of healthy human skins were removed by the improved method and the traditional method, respectively. The improved method was to remove the subcutaneous fat, eliminate the epidermis (1 mol/L NaCl solution at 37℃ for 24 hours) folowed by shaking processing (2% NaOH at 45℃ for 4 hours), and then, the solution was neutralized with PBS rinsing, dried and stored at 4℃ for standby. We detected the porosity and degradation time in vitroof the acelular dermal matrices prepared by two methods and the cytotoxicity of the material infiltration liquid on the adipose-derived stem cels. Hematoxylin-eosin staining was used for the detection of the cel residual, the integrity of colagen and cel biocompatibility. Scanning electron microscopy was used to detect the pore size.
RESULTS AND CONCLUSION: Both the two methods could completely remove the cel components, and maintain the integrity of the colagen scaffold. The porosity of acelular dermal matrix with the improved method was (93.22±0.99)%, which was significantly higher than that with the traditional method [(74.28±2.06)%;P < 0.001]. However, there was no significant difference in in vitrodegradation time between the two kinds of acelular dermal matrices(P > 0.05). No obvious cytotoxicity of the acelular dermal matrix prepared with the improved method was detected. At 3-7 days of co-culture, the adipose-derived stem cels cultured on the acelular dermal matrix prepared with the improved method could penetrate the basement membrane to the deep dermis, while there was no obvious cel invasion and growth in the deep dermis prepared by the traditional method. Compared with the traditional method, the improved method is more suitable for cel infiltration and growth with higher porosity and larger pore size.

Objective To detect the characteristics and in vitro cell compatibility of human acellular dermal matrix (ADM) with the improved method.Methods Cell components of healthy human skins were removed by the improved method and the traditional method respectively.The porosity, degradation time in vitro of the ADM prepared by two methods and the cytotoxicity of the material infiltration liquid with the improved method on the adipose derived stem cells were detected.HE staining was used to detect the residual of the cells, the integrity of collagen and cell biocompatibility.Scanning electron microscopy (SEM) was used to detect the pore size.Results Both the two methods could completely remove the cells, and maintain the integrity of the collagen scaffold;The porosity of ADM with the improved method was higher (93.1±1.02)% than that of traditional method (74.27±2.04)% (P<0.05);There was no significant difference in the cytotoxicity and in vitro degradation time between the two kinds of ADM;While pore diameter of the improved method was significantly higher [(181.21±66.9) μm] than that [(102.38±15.63) μm] in dermal reticular surface with the traditonal method (P<0.05).Conclusions There is no obvious cytotoxicity of the ADM with the improved method, and therefore it is more suitable for cell adhesion growth with higher porosity and larger pore size.

Objective To study the significance of HSP47 gene in keloid formation after in vivo study with RNAi technology and the recombinant HSP4 7 siRNA against heat shock protein 47 to keloid in a nude mice model.Methods We injected RNAi mixture into the keloid of a nude mice model in experimental group and PBS water(0.25 ml)into control group at the 16th days after establishing the models.After interference we observed the specimens and harvested specimens at 7th days for biochemical and pathological analysis.Results The expression of HSP47 mRNA reduced obviously and the collagen content also reduced in the experimental group.The rusults had statistical significance.Conclusion We can suppress the expression of HSP47 gene and then reduce the production of collagen after in vivo interfering experiment with HSP4 7 siRNA in keloid nude mice models using RNAi technique.This study cornfirms the mechanism that HSP47 promotes the keloid formation,which provides a new target to treat keloid.