Application of modified no-touch technique in rabbit arteriovenous fistula model and its effect on venous neointimal hyperplasia.

BACKGROUND: To investigate the technical advantages of a modified no-touch technique (MNTT) in constructing arteriovenous fistulas (AVF) compared to the conventional technique (CT) and assess its potential to reduce neointimal hyperplasia in the outflow vein. METHODS: Forty-seven New Zealand rabbits were randomly divided into three groups: control, CT, and MNTT. Rabbits in control group were observed using ultrasound and then euthanized to obtain external jugular vein (EJV) for Hematoxylin-eosin (H-E). We established common carotid artery (CCA)-EJV AVF using MNTT in the MNTT group and the CT in the CT group. AVF patency and complications were compared between the CT and MNTT groups. Rabbits with patent AVF in both groups were observed using ultrasound 2 weeks after surgery to evaluate changes in the vessel diameter and blood flow spectrum of the AVFs. H-E staining measured the intima thickness of EJV adjacent to the anastomosis and histologic characteristics of the AVF at 2 and 4 weeks after surgery. RESULTS: Five rabbits died after surgery with common symptoms of sneezing, coughing, runny nose, anorexia, and diarrhea; two in the MNTT group and three in the CT group. There were significant differences in the diameter (p = 0.010) and peak systolic velocities (PSV) (p = 0.001) of EJV between the CT and MNTT groups 2 weeks after surgery. Spiral laminar flow (SLF) was observed in CCA and EJV adjacent to anastomosis in the MNTT group. Additionally, histological observations showed less venous neointimal hyperplasia in the MNTT group than in the CT group 4 weeks after surgery. CONCLUSION: The rabbit model of CCA-EJV AVF established using MNTT demonstrated fewer complications, larger vein diameters, and reduced venous neointimal hyperplasia, indicating that this maybe an ideal animal model to further investigate the application of MNTT in AVF surgery.

Mechanism of mitochondrial protease LONP1 in the biological behavior of prostate cancer / 肿瘤

Objective:To explore the mechanism of Ion peptidase 1,mitochondrial(LONP1)in the progression of castration-resistant prostate cancer(CRPC). Methods:The expression levels of LONP1,N-Myc downstream-regulated gene 1(NDRG1)and androgen receptor(AR)proteins in benign prostatic hyperplasia cell line BPH-1,androgen-dependent human prostate cancer cell line LNCaP and castration-resistant prostate cancer(CRPC)cell line PC3 were examined by Western blotting.Recombinant vector carrying full-length LONP1 gene was delivered into LNCaP cells via lentiviral infection to establish stable LONP1-overexpressing(OE-LONP1)cell line,and LNCaP cells transfected with empty vector was used as the corresponding negative control(OE-NC).shRNA specifically targeting LONP1 gene(shLONP1)was delivered into PC3 cells to establish stable LONP1-silencing cell line,and PC3 cells transfected with empty shRNA vector(shNC)was used as the negative control.The effect of LONP1-overexpression and silencing on cell proliferation and invasion was analyzed by CCK-8 assay,colony formation assay and Transwell invasion assay in OE-LONP1 and shLONP1 cell lines.The effect of LONP1 on the AR/DNRG1 signaling axis was analyzed by co-immunoprecipitation assay and chromatin immunoprecipitation assay.The effect of NDRG1-silencing on the proliferation and invasion of shLONP1-expressing PC3 cells was evaluated by CCk-8 assay and Transwell assay.Tumor xenograft model was established by subcutaneously inoculating shLONP1-expressing PC3 cells and the negative control cells(PC3-shNC cells)into BALB/c nude mice.The mice were treated with DMSO or AR antagonist enzalutamide(ENZ).Then,the expression level of Ki-67 in tumor tissues was examined by immunohistochemical staining,and the cell apoptosis in tumor tissues was evaluated by TUNEL assay. Results:Compared with BPH-1 cells,LNCaP cells had lower(P＜0.05)while PC3 cells had higher(P＜0.05)expression level of LONP1 protein.The expression of AR and NDRG1 were inhibited in LNCaP cells when LONP1 was overexpressed(P＜0.05),whereas the expression of AR and NDRG1 were increased in PC3 cells when LONP1 was silenced(P＜0.05).The proliferation and invasion of LNCaP cells were promoted when LONP1 was overexpressed(P＜0.05),whereas the proliferation and invasion of PC3 cells were suppressed when LONP1 was knocked-down(P＜0.05).LONP1 can directly bind with AR to recognize the androgen response element(ARE)of NDRG1,which further inhibits the AR/NRDG1 signaling pathway to promote the progression of prostate cancer.The treatment of xenograft tumors in mouse models showed that both LONP1-silencing and ENZ application could inhibit tumor growth,and the best inhibitory effect was observed in mice treated with LONP1-silencing in combination with ENZ.The results of immunohistochemical staining and TUNEL assay indicated that the tumor tissues in the shLONP1+ENZ group had lower level of Ki-67 expression and higher level of cell apoptosis(P＜0.001). Conclusion:LONP1 is highly expressed in CRPC cell line PC3,and promotes prostate cancer progression by inhibiting AR/NDRG1 signaling transduction.

Clinical efficacy of Nd:YAG laser adjunct to subgingival scaling and root planning for treating severe chronic periodontitis / 西安交通大学学报(医学版)

【Objective】 To evaluate the efficacy of Nd: YAG laser therapy adjunct to subgingival scaling and root planning (SRP) for treating severe chronic periodontitis. 【Methods】 We selected patients with severe chronic periodontitis whose teeth were distributed in 4 quadrants of the mouth, with probing depth (PD) of 5-8 mm, attachment loss (AL)≥5 mm and bleeding on probing (BOP). These teeth were randomly divided into three groups: SRP group, SRP+L group (Nd: YAG laser after SRP treatment), and L+SRP group (SRP after Nd: YAG laser treatment). We recorded parameters including BOP, PD and AL of the three groups at baseline and 8 weeks after treatment and made statistical analysis. 【Results】 At 8 weeks after treatment, BOP, PD and AL of the three groups were improved than those in the baseline (P<0.05). BOP positive percentage of SRP+L group and L+SRP group significantly reduced compared with SRP group (P<0.05). PD of SRP+L group significantly decreased compared with SRP group and L+SRP group (P<0.05), for sites with PD=7 mm, SRP+L group was significantly decreased compared with SRP and L+SRP groups (P<0.05). AL of SRP group significantly decreased compared with SRP+L group and L+SRP group (P<0.05). 【Conclusion】 Severe periodontal treatment with Nd:YAG laser adjunct to SRP is more effective in reducing BOP and PD, and for deeper pockets PD is significantly decreased in SRP+L group, but there is no advantage in the improvement of AL.

CT manifestations and pathologic characteristics of solitary fibrous tumor of the pleural / 实用放射学杂志

Objective To investigate the CT manifestations and pathology features of solitary fibrous tumors of the pleural (SFTP),to improve diagnostic accuracy of this disease.Methods CT manifestations of 12 cases with SFTP confirmed by surgery were analyzed retrospectively by comparison with the pathological findings.Results The CT images of the 12 cases showed intrathoracic soft-tissue shadow with well-defined margin which was widely connected with pleural.There were 4 cases with mound like and mushroom shape,3 cases with round or oval shape,2 cases with irregular shape growing into the pleural space,3 cases with large mass filling unilateral thoracic cavity,9 cases with pleural tail sign and 1 case with pleural pedicle sign.The plain CT showed homogeneous density in 7 cases,high-low mixed density in 4 cases and with internal calcification in 1 case.After contrast enhancement,5 cases were map like heterogeneous and progressive enhancement,and 4 cases have strip distorted vessel inside or on the edge of the lesion.Conclusion CT manifestations of SFTP have certain characteristics,such as broad base,shaping growth and pleural tail sign,which could be helpful to improve the diagnostic accuracy of SFTP by analyzing carefully.

Neovascularization aggravated hepatic fibrosis in combined hypoxia NASH rat model / 中华肝脏病杂志

Objective@#To optimize the construction of combined hypoxia NASH rat model on the basis of preliminary work, and to explore the role of neovascularization in the process of hepatic fibrosis.@*Methods@#32 rats were divided randomly to four groups that were null control group(A group ), hypoxia group(B group), high fat diet group(C group ) and high fat diet plus hypoxia group (D group ),treated with null , Intraperitoneal injection of NaNO2, high fat diet and high fat diet plus Intraperitoneal injection of NaNO2 respectively. Every group was observed for 16 weeks, B and D group was treated with Intraperitoneal injection of NaNO2 20 mg/kg.d at the laster 8 weeks. Liver histology NASH activity score(NAS) and Fibro score(FibroS), biochemical index were detected in this combined hypoxia NASH rat model(D group), meanwhile the changes of HIF1α, inflammatory factor and neovascularization were measured by ELISA, realtime PCR and immunohistochemistry.@*Results@#Liver tissue NAS > 4 was seen in C and D group. D group showed NASH characteristics, including significantly steatosis at liver acinar 3 area(mostly a microvesicular type fat droplets mixed with macrovesicular type), hepatocyte balloon degeneration, obvious lobular inflammation, while fibrosis score increased significantly, including visible hepatic sinusoid fibrosis, fibrosis around portal vein, and bridging fibrosis in a considerable portion of the rats. Compared with C group, biochemical indicators of aspartate aminotransferase (AST), HIF1α, neovascularization-related VEGFA, VEGFR2 mRNA level increased obviously and the expression of immunohistochemistry VEGFR2, CD34 enhanced markedly in D group(p < 0.05).@*Conclusion@#A combined hypoxia NASH rat model can be established throught feeding 16 weeks’ high-fat diet then intraperitoneal injection of NaNO2 20 mg / kg.d at the laster 8 weeks, meanwhile chronic hypoxia can accelerate this combined hypoxia NASH model liver fibrosis process. In this process neovascularization promoted the formation of hepatic fibrosis in this model.

Expression and clinical significance of DNA methyltransferases in hepatocellular carcinoma / 中华肝胆外科杂志

Objective To investigate the expression of DNA methyltransferase 1,3a,3b (DNMT1,DNMT3a,DNMT3b) in human hepatocellular carcinoma (HCC) and to determine their clinical significance.Methods The expression of DNMT1,3a,3b proteins was detected in 47 HCC tumor specimens and 42 HCC paracarcinoma liver tissues by immunohistochemistry.12 normal liver tissues were used as control.The results and clinicopathological parameters were analyzed.Results The positive expression rates of DNMT1,3a and 3b in HCC tissues were 80.9％,68.1％ and 78.7％ respectively.The positive expression rates of DNMT1,DNMT3a and DNMT3b in paracarcinoma tissues were 50.0％,52.4％ and 57.1％ respectively.The expression rates of DNMT1,3a and 3b in both HCC tissues and paracarcinoma tissues were significantly higher than normal liver tissues.The expression of DNMT1,DNMT3a,DNMT3b was correlated with tumor differentiation (P＜0.05) and hepatitis B surface antigen (HBsAg) positivity.Conclusions DNMT1,3a and 3b play important roles in carcinog(c)n(c)sis and development of HCC.

Overexpression of caveolin-1 inhibits the growth of human cervical squamous cell Hela cell line in vitro / 中国医师杂志

Objective To investigate the effects of caveolin-1 overexpressing on the growth of cervical squamous cell cancer Hela cell line. Methods Eukaryotic expression vector of human caveolin-1 gene was introduced into Hela cells by Lipofectamine. The clones stably overexpressed caveolin-1 were identiffed by RT-PCR, immunofluorescence cell staining techniques and Westernblotting. Cells proliferation viabihty was tested by MTT assay, and flow cytometry was used to assay the cell cycle and apoptosis, and the relative phosphorylation level of extracellular regulated protein kinases (Erk1/2) were detected by Westernblotting. Results The clones stably overexpressed caveolin-1 were obtained. Compared with the parental Hela cells, the tranfected cells exhibited a slower rate of growth. FAGS analysis results revealed that overexpression of caveolin-1 resulted in the cell cycle arrest in the G0/G1 [ ( 68. 04 ± 2. 57 ) % vs ( 53.41 ±1.01)%] phase and increased the apoptotic cell fraction[ (19. 18 ±2.20)% vs (5.63 ±0.55)%, P <0. 05 ]. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of Erk1/2(0.28 ±0.05 vs 0.81 ±0.07, P <0.05). Conclusions Overexpression of caveolin-1 suppressed the growth of Hela cells and induced apoptosis, down-regulation of Erk1/2 phosphorylation might be involved in its mechanism.