Objective:To provide chemical basis for processing of leech through investigating the variations of composition of leech scalded with talc powder. Methods: Amino acids were determined by HPLC post column derivatization and steroids by GC MS. Results: The results showed the total contents of amino acids in scalded leech were reduced and the steroids of it were changed. Conclusion: The processing method of leech used at the moment has a great influence in chemical components of leech.

PurposeThe restriction of strain ratio (SR) in the diagnosis of thyroid malignant tumor are caused mainly because of disadvantages including low sensitivity, in this research the authors used different calculation methods of strain ratio based on the size of tumors, to explore the clinical application value of them on the differentiation of benign and malignant thyroid nodules.Materials and Methods Data of 336 patients with thyroid nodule (340 nodules) who underwent elasticity imaging and ultrasound-guided fine-needle aspiration were retrospective analyzed, with the pathology combined follow up acted as golden standards, fixed surface area assay and true surface area assay of strain ratio were compared on the differential diagnosis of benign and malignant thyroid nodules with various size.Results Among 340 thyroid nodules, there were 91 small nodules and 249 big nodules. Pathological results showed that 36 of 91 small nodules and 104 of 249 big nodules were malignant, while the others were benign. The true surface area assay was significantly better for the diagnosis of smaller thyroid nodules with longitudinal diameter and transverse diameter<8 mm with higher consistency with pathology (χ2=20.89,P<0.01), and fixed surface area assay is more suitable of thyroid nodules with longitudinal diameter and transverse diameter>8mm (χ2=57.08,P<0.01). The sensitivity, specificity and consistency for the diagnosis of malignant nodules of true surface area assay were 57.10%, 83.00% and 72.30% (Kappa=0.413), which were 73.60%, 85.00% and 72.30% (Kappa=0.590) in the fixed surface area assay, and 85.00%, 85.50% and 85.30% (Kappa=0.699) in the selective usage of two methods according to the tumor size, respectively.Conclusion The sensitivity is highest when using the two methods selectively according to the nodule size, with a better consistence rate with pathological results as well.

Objective To assess the feasibility and effectiveness of percutaneous transluminal angioplasty (PTA)for the salvage of immature arteriovenous fistula (AVF) and to identify the incidence of arterial and venous puncture site spasm.Methods The medical records and radiological data of 88 patients with 112 interventional procedures for immature AVFs were retrospectively reviewed.Results The stenosis lesions were (2.0 ± 1.4) cm long.Technical success rate and clinical success rate were 80.4％ (78/97) and 92.8％ (90/97) for PTA via brachial artery,85.7％ (6/7) and 100％ (7/7) for PTA via vein,25％ (2/8) and 50％ (4/8) for PTA via both brachial artery and vein,respectively.Spasm of pure arterial PTA occurred in 2 patients (2.1％) and was mild and moderate.Spasm of pure venous PTA occurred in 2 patients (28.6％) and was both moderate.Spasm of combined arterial and venous PTA occurred in 3 patients (37.5％) and from being severe to completely occluded.By comparison,there were statistical differences of technical and clinical success rate (P =0.000,0.019 ; P =0.000,0.029),fistulas spasm rate was statistically significant different (P =0.000).Conclusions Endovascular therapy was effective in restoring the dysfunctional native AVFs,it was safer and more effective and with less sideeffects especially in selecting coronary balloon to treat patients without large phlebangioma and round fistulas.

Objective To explore the effect of epidermal growth factor (EGF) on tube formation of HUVEC induced by the secretion of angiogenesis factors of adipose-derived stem cells (ADSCs).Methods ADSCs were primarily cultured by enzyme digestion method.The flow cytomertry was performed to detect the expression of cell surface marker.ELISA was used to detect the expression of VEGF,HGF,and SDF-1 after given different doses of EGF.Tube formation assay was used to examine the effect of EGF on the tube formation induced by ADSCs.Results ADSCs were successfully isolated and cultured from human liposuction tissue and specific markers were expressed on ADSCs.EGF promoted the secretion of angiogenesis factors VEGF,HGF,and SDF-1,which were secreted by ADSCs.EGF pretreatment increased the ability of tube formation of HUVECs induced by ADSCs.Conclusions ADSCs induce the secretion of angiogenesis factors in vitro,and thus increase the ability of tube formation of HUVECs.EGF promotes the secretion ability of ADSCs,and the best concentration is 15 mg/L.

Objective To investigate the method of dermal-fat graft combined with secondary autologous fat transplantation in repairing severe facial depression and to evaluate the clinical effects.Methods Twelve cases of facial depression had been repaired by the transplantation of dermal-fat flap which was removed from the abdomen at the first stage.They were given fat granules injection 1 to 3 times postoperatively,and 3,6 and 3 patients were given fat granules injection three times,twice and once,respectively,with the interval period of 3 to 6 months.The result was based on comparison of the photos taken from preoperation and postoperation.Results All patients were healed primarily except one of which was formed hematoma after operation and scavenged thereafter.After 6 months to 2 years follow-up,all the patients had satisfactory facial contour.Conclusions Combined autologous fat granules with free dermal-fat graft to reconstruct severe facial depress is an easy,safe and effective technique and deserves to be recommended.

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 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.

Objective To study the locations, types and causes of different pseudoaneurysms in order to find out the optimal individualized treatment for different pseudoaneurysms. Methods Different methods were applied in treating 21 patients with pseudoaneurysm, which were located at limb (n = 11 ), spleen (n =3), kidney (n = 2) , common lilac artery (n = 1), internal iliac artery (n = 1), gallbladder (n = 1) and penis (n = 1 ). Different managements were employed in treating these pseudoaneurysms. Temporary obstruction of blood circulation with balloon together with arterial anastomosis or direct incision neoplasty was performed in 9 cases with pseudoaneurysms at limb arteries close to the larger joints. Endovascular stent graft was used to isolate the trunk type of pseudoaneurysm in 4 cases, in 2 of them branch arterial embolism and stent graft endovascular exclusion were applied as they had common iliae artery trunk type of pseudoaneurysm at the opening of internal iliac artery. Gelfoam together with metallic coils embolization was employed in 6 cases with terminal type of pseudoaneurysms. Results After different treatments, tumor cavities disappeared in the 21 cases with pseudoaneurysms. Distal arterial pulse returned to normal and no nerve damage occurred in 11 cases with limb pseudoaneurysms. No internal hemorrhage was observed and distal blood circulation returned to normal after graft endovascular exclusion in 2 eases with pseudoaneurysms at spleen artery trunk and in 2 cases with pseudoaneurysms at iliac artery trunk. In 6 cases with terminal type of pseudoaneurysms,the tumor cavity disappeared, hemorrhage stopped and no ischemic necrosis of organ occurred. But one of them with multiple traumatic pseudoaneurysms located at the second grade branch died one week after embolism due to a serious pelvic trauma accompanied with serious infection. Conclusion Based on the locations, types and causes of pseudoaneurysms, different individualized treatment should be adopted in order to obtain optimal results with least damages.

Objective. The goal of this paper is to investigate the relationship between the N-glycosylation of acetylglucosaminyltransferase V(Glc NAcT-V) and its activity and to know which site among the 6 N-glycosylation sites in the GlcNAcT-V gene is the most important.Methods. Wild type of GlcNAcT-V was transfected into COS-7 cells and its activity was measured 48 h later. The first site (Asn 110) was mutated with site-directed mutagenesis and transfected into COS-7 cells.Results. It was found that after the cells were added tunicamycin(TM, 1 μ g/ml), the activity was 11.7% of the wild type. The activity of the cells with mutating GlcNAcT-V was about 12.0% of the wild type. RT-PCR showed that there was no significant change in mRNA expression among the three groups.Conclusion. The N-glycosylation is important for its activity. Our results suggest that the N-linked carbohydrates on GlcNAcT-V are required for the posttranscriptional activity of the enzyme.

Objectives:To explore the impact of intensive lipid-lowering strategy on short-term prognosis of acute coronary syndrome(ACS)patients with multi-vessel disease. Methods:A total of 136 ACS patients with multi-vessel disease who received coronary stenting at Zhongnan Hospital of Wuhan University from August 2019 to November 2020 were enrolled in this study.Patients were divided into intensive lipid-lowering group(control low density lipoprotein cholesterol[LDL-C]below 1.0 mmol/L within 3 months,and continuously meet the standards within 12 months,n=69)or standard lipid-lowering group(gradually control LDL-C below 1.4 mmol/L within one year,n=67).The total cholesterol(TC),triglycerides(TG),LDL-C,high-density lipoprotein cholesterol(HDL-C),and lipoprotein(a)(Lp[a])data were collected.Incidence of major adverse cardiovascular events(MACE,including cardiac death,myocardial infarction,target vessel revascularization and stroke)were observed during 12 months of follow up. Results:The baseline data of the intensive lipid-lowering group and the standard lipid-lowering group were consistent before intervention.At the timeline of enrollment,there was no statistically significant difference in the blood lipid profiles(including TC,TG,LDL-C,HDL-C)between the two groups.After 3-months,patients in the intensive lipid-lowering group experienced significantly lower TC,TG,LDL-C and Lp(a)compared with baseline values(all P<0.05),while HDL-C remained unchanged(P>0.05).The standard lipid-lowering group showed a significant decrease in TC and LDL-C compared with baseline values(both P<0.05).The TC and LDL-C levels were significantly lower in the intensive lipid-lowering group than in the standard lipid-lowering group at 3/6/12 months follow up after discharge(all P<0.01).At 12 months follow-up,Kaplan-Meier survival analysis showed that the incidence of MACE was significantly lower in the intensive lipid-lowering group than in the standard lipid-lowering group(2.90%vs.14.93%,χ2=6.090,P=0.014).Multiple Cox regression analysis revealed that the intensive lipid-lowering strategy significantly reduced the risk of MACE compared with the standard lipid-lowering strategy(HR=0.177,95%CI:0.037-0.838,P=0.029). Conclusions:Our data show that intensive lipid-lowering strategy may probably reduce the incidence of short-term MACE in ASC patients with multi-vessel disease.Large-scale prospective multi-center studies are needed to further validate these results.