[Mechanisms by which Mettl3 regulates pericyte-myofibroblast transdifferentiation through PI3K/AKT signaling pathway].

Objective: To investigate the role and underlying mechanisms of methyltransferase (Mettl) 3 in the process of angiotensin Ⅱ (Ang Ⅱ)-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis. Methods: C57BL/6J mice were used, in cell experiments, mouse renal pericytes were isolated and cultured using magnetic bead sorting. These pericytes were then induced to transdifferentiate into myofibroblasts with 1×106 mmol/L Ang Ⅱ, which was the Ang Ⅱ group, while pericytes cultured in normal conditions served as the control group. Successful transdifferentiation was verified by immunofluorescence staining, Western blotting, and real-time reverse transcription PCR (RT-qPCR) for α-smooth muscle actin (α-SMA). The levels of m6A modifications and related enzymes (Mettl3, Mettl14), Wilms tumor 1-associated protein (WTAP), fat mass and obesity protein (FTO), ALKBH5, YTHDF1, YTHDF2, YTHDC1, YTHDC2, YTHDC3 were assessed by Dot blot, RT-qPCR and Western blot. Mettl3 expression was inhibited in cells using lentivirus-mediated Mettl3-shRNA transfection, creating sh-Mettl3 and Ang Ⅱ+sh-Mettl3 groups, while lentivirus empty vector transfection served as the negative control (Ang Ⅱ+sh-NC group). The impact of Ang Ⅱ on pericyte transdifferentiation was observed, and the expression of downstream phosphatidylinositol 3-kinase (PI3K)/AKT signaling pathway proteins, including PI3K, AKT, phosphorylated AKT at serine 473 (p-AKT (S473)), and phosphorylated AKT at threonine 308 (p-AKT (T308)), were examined. PI3K gene transcription was inhibited by co-culturing cells with actinomycin D, and the half-life of PI3K mRNA was calculated by measuring residual PI3K mRNA expression over different co-culture time. The reversibility of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was assessed by adding the AKT activator SC79 to the Ang Ⅱ+sh-Mettl3 group. In animal experiments, mice were divided into these groups: sham group (administered 0.9% sterile saline), Ang Ⅱ group (infused with Ang Ⅱ solution), sh-Mettl3 group (injected with Mettl3 shRNA lentivirus solution), Ang Ⅱ+sh-Mettl3 group (infused with Ang Ⅱ solution and injected with Mettl3 shRNA lentivirus solution), and Ang Ⅱ+sh-Mettl3+SC79 group (administered Ang Ⅱ solution and Mettl3 shRNA lentivirus, with an additional injection of SC79). Each group consisted of six subject mice. Blood pressure was measured using the tail-cuff method before and after surgery, and serum creatinine, urea, and urinary albumin levels were determined 4 weeks post-surgery. Kidney tissues were collected at 28 days and stained using hematoxylin-eosin (HE) and Masson's trichrome to assess the extent of renal fibrosis. Results: Primary renal pericytes were successfully obtained by magnetic bead sorting, and intervened with 1×106 mmol/L Ang Ⅱ for 48 hours to induce pericyte-to-myofibroblast transdifferentiation. Dot blot results indicated higher m6A modification levels in the Ang Ⅱ group compared to the control group (P<0.05). RT-qPCR and Western blot results showed upregulation of Mettl3 mRNA and protein levels in the Ang Ⅱ group compared to the control group (both P<0.05). In the Ang Ⅱ+sh-Mettl3 group, Mettl3 protein expression was lower than that in the Ang Ⅱ group, with reduced expression levels of α-SMA, vimentin, desmin, fibroblast agonist protein (FAPa) and type Ⅰ collagen (all P<0.05). Compared to the control group, PI3K mRNA expression level was elevated in the Ang Ⅱ group, along with increased p-AKT (S473) and p-AKT (T308) expressions. In the Ang Ⅱ+sh-Mettl3 group, PI3K mRNA expression and p-AKT (S473) and p-AKT (T308) levels were decreased (all P<0.05). The half-life of PI3K mRNA was shorter in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ+sh-NC group (2.34 h vs. 3.42 h). The ameliorative effect of Mettl3 inhibition on Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation was reversible by SC79. Animal experiments showed higher blood pressure, serum creatinine, urea, and 24-hour urinary protein levels, and a larger fibrosis area in the Ang Ⅱ group compared to the sham group (all P<0.05). The fibrosis area was smaller in the Ang Ⅱ+sh-Mettl3 group than that in the Ang Ⅱ group (P<0.05), but increased again upon addition of SC79. Conclusion: Mettl3-mediated RNA m6A epigenetic regulation is involved in Ang Ⅱ-induced pericyte-to-myofibroblast transdifferentiation and renal fibrosis, potentially by affecting PI3K stability and regulating the PI3K/AKT signaling pathway.

[Expression and protective effect of chemerin in idiopathic pulmonary fibrosis].

Objective: To explore the expression and the role of chemerin in idiopathic pulmonary fibrosis (IPF). Methods: Quantitative PCR and Western blotting were used to determine the mRNA and protein levels of chemerin in lung tissues from IPF patients and the controls. Clinical serum level of chemerin was analyzed by enzyme-linked immunosorbent assay. The mouse lung fibroblasts isolated and cultured in vitro were divided into the control, TGF-ß, TGF-ß+chemerin and chemerin groups. Immunofluorescence staining was used to observe the expression of α-smooth muscle actin (α-SMA). C57BL/6 mice were randomly divided into the control, bleomycin, bleomycin+chemerin, and chemerin groups. Masson and immunohistochemical staining were performed to evaluate the severity of pulmonary fibrosis. Expression of epithelial to mesenchymal transition (EMT) markers was detected by quantitative PCR and immunohistochemical staining in the in vitro and in vivo models of pulmonary fibrosis, respectively. Results: Compared with the control group, the expression of chemerin was downregulated in both the lung tissue and the serum of IPF patients. Immunofluorescence showed that treatment of fibroblasts with TGF-ß alone resulted in a robust expression of α-SMA, whereas treatment with TGF-ß and chemerin together exhibited the similar expression levels of α-SMA as the control group. Masson staining indicated that the bleomycin-induced pulmonary fibrosis model was constructed successfully, while treatment of chemerin partially alleviated the damage of lung tissue. Immunohistochemical staining showed that the expression of chemerin in the lung tissue was significantly decreased in the bleomycin group. Quantitative PCR and immunohistochemistry showed that chemerin attenuated EMT induced by TGF-ß and bleomycin both in vitro and in vivo. Conclusions: The expression of chemerin was reduced in patients with IPF. Chemerin may play a protective role in the development of IPF by regulating EMT, providing a new idea for the clinical treatment of IPF.

[Advancement in endovascular therapy of aortoiliac occlusive disease].

Aortoiliac occlusive disease (AIOD) refers to the stenosis and occlusion of the distal abdominal aorta and(or) bifurcation of the aortoiliac artery,which is mainly caused by atherosclerosis,leading to pelvic and lower limb ischemia.Open surgery has always been the main treatment for complex AIOD.However,in recent years,with the development of endovascular surgery technologies and medical instruments,its treatment concept has been greatly changed.More and more clinical evidence has proved that the long-term efficacy of endovascular therapy is not inferior to that of traditional open surgery,so minimally invasive endovascular therapy has become the preferred treatment for AIOD.

[Establishment of a cytokine release syndrome associated with chimeric antigen receptor T cell treatment in SCID/Beige mice model].

Objective: To establish a cytokine release syndrome (CRS) mouse model related to CAR-T cell therapy and provide a research model for the clinical phenomena. Methods: CAR-T cells targeting human CD19 molecule were constructed by molecular cloning and lentiviral transfection. Flow cytometry (FACS) was used to detect the transfection efficiency of CAR-T cells. The tumor-killing efficiency of CAR-T cells was detected by ELISA and flow cytometry. The CAR-T cells were injected into the tumor-bearing SCID/Beige mice through tail vein, and divided into phosphate buffered solution (PBS) group, low-burden group (1×10(5) Raji-Luc2 cells) and high-burden group (5×10(5) Raji-Luc2 cells). The tumor treatment effect was detected by animal in vivo imaging. Serum levels of cytokines including human IFN-γ, human IL-2, mouse IL-6, and mouse GM-CSF were measured by ELISA. The health status of the mice was evaluated by pathological examination. Results: The health scores of T cell group and T cell+ OKT-3 group were (1.15±0.08) and (2.90±0.15), respectively, after the injection of human T cell and T cell + OKT-3 antibody through tail vein, and the difference was statistically significant (P<0.001). The serum levels of human IL-2, human IFN-γ, human IL-15, mouse IL-6 and mouse GM-CSF in T cell+ OKT-3 group were (1 064.00±50.14), (1 285.00±193.90), (202.4±18.76), (1 478.00±289.20) and (350.70±42.27) pg/ml, respectively, higher than (22.67±6.36), (23.67±3.71), (44.33±14.45), (147.30±36.20), (138.00±22.74) pg/ml in T cell group (P<0.05). OKT-3 combined with human T cells caused a rapid increase in serum levels of human IL-2, human IFN-γ, mouse IL-6 and mouse GM-CSF, accompanied by an increase in body temperature and weight loss. CD19-targeting CAR-T cells were successfully constructed, and the positive rate of CAR-T cells was >30% detected by flow cytometry. ELISA results showed that in the presence of CD19 antigen, IL-2 and IFN-γ secreted by CAR-T19 cells co-incubated with Raji and Nalm were (561.00±37.07), (680.30±71.27), (369±25.71) and (523.00±26.31) pg/ml, respectively, higher than (55.00±20.53) and (64.00±7.55) pg/ml in the co-incubated with K562 group (P<0.001). Activated CAR-T19 cells were reinjected through the tail vein on the seventh day after tumor formation. Imaging experiments in mice showed that on the thirteenth day after tumor formation, the fluorescence intensities of tumors in the low-burden and high-burden groups were lower than on the seventh day of tumor inoculation, and the fluorescence intensity of tumors in the high-burden group decreased from 144.00±24.69 to 5.02±2.35 (P=0.005). The fluorescence intensity of low burden group decreased from 58.47±9.36 to 3.48±1.67 (P=0.004). The serum levels of T cell activation related cytokines IL-2, IL-15 and IFN-γ increased rapidly, and the secretion of monocyte related cytokines IL-16 and GM-CSF increased, accompanied by the typical characteristics of CRS such as increased body temperature and weight loss at 72 hours after injection of CAR-T19 cells. Conclusions: CAR-T cells targeting CD19 molecule are successfully constructed, and CRS phenomenon is verified in tumor-bearing mice by CAR-T cell re-infusion, providing an animal model for the mechanism of CAR-T treatment-related CRS and CRS prevention strategies.

[Drug-coated balloons versus bare metal stent for treatment of femoropopliteal lesions:36 month follow-up results of single center].

Objective: To examine the therapeutic effects of drug-coated balloon (DCB) and bare metal stent (BMS) on primary femoropopliteal disease (FPAD) in the real world. Methods: This was a retrospective analysis of single-center follow-up results at 12,24,and 36 months of patients with FPAD lesions that were treated with DCB and BMS at Department of Aortic and Vascular Surgery, Fu Wai Hospital.One-to-one propensity score matching(PSM) was performed to balance the covariance between DCB group (137 cases) and BMS group (100 cases). Freedom from clinically driven target lesion reintervention rate(fCD-TLR) was determined by Kaplan-Meier curve.Log-rank test was used to compare the rates of fCD-TLR between DCB and BMS groups at 12,24,36 months post-operation. Results: After PSM, there were both 71 patients in each group,aged (68.0±9.6) years(range: 46 to 90 years) and (68.8±7.3) years(range: 48 to 87 years),lesion lengths were (119.6±14.2)mm(range:40 to 380 mm) and (110.8±13.1)mm(range:40 to 400 mm). The median follow-up period were 24.3 months (range:5.8 to 55.1 months).There was no death,amputation or reintervention within the 30 days after operation.The rates of fCD-TLR for DCB group at 12,24 and 36 months were 97.2%,85.9%,69.1%, and 95.8%,83.1%,59.2% for BMS group.There was no statistical difference between the two groups by Log-rank test (P=0.551). Conclusion: DCB and BMS can both maintain favorable clinical effects in FPAD patients at 12,24,36 months post-operation.

[Role and related mechanism of Mst-1 on regulating hypoxic reoxygenation induced autophagy and apoptosis in cardiomyocytes of mouse].

Objective: To explore the role and related mechanism of mammalian sterile 20-like kinase 1(Mst-1)in regulating hypoxia reoxygenation (HR) induced myocardial cell autophagy and apoptosis. Methods: Enzyme digestion method combined with differential adherent method was used to culture neonatal mouse myocardial cells. HR model was established by hypoxia for 24 hours and reoxygenation for 6 hours. The experimental groups including control group (normal cultured cardiomyocytes), Mst-1 empty virus group (cardiomyocytes transfected with recombinant lentiviral empty vector for 48 hours), Mst-1 knockdown group (recombinant lentivirus carrying Mst-1small interfering RNA (siRNA) was transfected into cardiomyocytes for 48 hours), Mst-1 overexpression group (cardiomyocytes were transfected with recombinant lentivirus carrying Mst-1 gene for 48 hours), HR group (cardiomyocytes exposed to HR), Mst-1 knockdown+HR group (HR model of cardiomyocyte was established 48 hours after transfection with recombinant lentivirus carrying Mst-1siRNA) and Mst-1 overexpression+HR group (HR model of cardiomyocyte was established 48 hours after transfection with recombinant lentivirus carrying Mst-1 gene). Real-time fluorescence quantitative RCR (qPCR) and Western blot were used to detect the relative expression of Mst-1 mRNA and protein in the cells, immunofluorescence staining was used to detect cardiomyocyte troponin T (cTnT), and autophagosomes and autophagy enzyme changes. TUNEL method was used to detect myocardial cell apoptosis, Western blot was adopted to detect autophagy-related protein microtubule-related protein 1 light chain 3 (LC3) Ⅱ/LC3 Ⅰ, P62 and apoptosis-related protein cleaved-caspase 9, pro-caspase 9, cleaved-caspase-3, pro-caspase-3, and myeloid leukemia 1 (MCL-1) expression. MCL-1 inhibitor A1210477 was used to validate the signaling pathway of Mst-1 on regulating cardiomyocyte apoptosis and autophagy. Results: Immunofluorescence detection revealed that the cultured cells expressed cardiomyocyte-specific marker cTnT. The expression of Mst-1 in cardiomyocytes increased in HR model. Lentiviral transfection could effectively inhibit or overexpress Mst-1 in treated cells. The levels of autophagosomes and autophagolysosomes in cardiomyocytes undergoing HR and in Mst-1 overexpression+HR group were lower than those of control group, while autophagosomes and autophagolysosomes in cardiomyocytes of Mst-1 knockdown+HR group was significantly higher than in the HR group (all P<0.05). The TUNEL results showed that the proportion of TUNEL positive cells was significantly increased in the HR group and Mst-1 overexpression+HR group than in the control group, while the proportion of TUNEL positive cells was significantly decreased in the Mst-1 knockdown group+HR group as compared to the HR group (all P<0.05). Western blot results showed that the LC3 Ⅱ/LC3 Ⅰ levels were significantly lower, while the expression levels of P62, cleaved-caspase-9 and cleaved-caspase-3 were significantly higher in the HR group and Mst-1 overexpression+HR group than in control group (all P<0.05). The LC3 Ⅱ/LC3 Ⅰ value was significantly higher, and the expression levels of P62, cleaved-caspase-9 and cleaved-caspase-3 were significantly lower in the Mst-1 knockdown+HR group than in the HR group (P both<0.05). The expression level of P-MCL-1 protein was significantly lower in cardiomyocytes of HR and Mst-1 overexpression+HR group than in control group, and the expression level of P-MCL-1 protein was higher in Mst-1 knockdown+HR group than in HR group (P both<0.05). The recovery experiment showed that inhibiting MCL-1 in cells can block the regulatory effect of Mst-1 siRNA on cell autophagy and apoptosis. Conclusion: Inhibiting Mst-1 expression in cardiomyocytes can promote the autophagy of cardiomyocytes induced by hypoxic reoxygenation and reduce the apoptosis of cardiomyocytes via activating McL-1.

[Surgical treatment of total subclavian artery occlusion: a single center experience of 67 cases].

Objective: To examine the outcomes of surgical repair for patients with total subclavian artery occlusion. Methods: A retrospective analysis was performed on 67 patients with subclavian artery occlusion disease admitted at Ward 1 of Aortic and Vascular Surgery Center, Fuwai Hospital from January 2016 to July 2019. The age was, and There were 51 male patients and 16 females with an age of (61.7±8.2) years (range: 37 to 79 years). The t-test, Mann-Whitney U-test, χ(2) test, and Fisher's exact test were used to analyze the factors related to the technique success. The Kaplan-Meier curve was used to calculate the cumulative patency rate and plot the corresponding survival curves, and the Log-rank test was used for comparison. The length from the subclavian artery ostial to the occlusion area was used as a variable to plot the receiver operating characteristic curve, and the optimal cut-off value was determined by the Youden index. Results: Eighteen patients received open surgery. Forty-nine patients with subclavian artery occlusion accepted endovascular repair, of which 38 patients succeeded (31 cases on left side and 3 cases on right side). Fifteen patients failed with endovascular therapy, of which 10 cases received elective surgery and 5 cases received conservative therapy. The success rate of endovascular repair was 69.4%(34/49). Among them, the success rate of left subclavian artery occlusion was 81.6%(31/38), while the right side was 3/11. Patients with the length from the subclavian artery ostial to the occlusion area ≥6 mm were more likely to get success (23/34 vs. 4/15, χ(2)=5.506, P=0.019). In the endo-group, one patient had hemorrhage in the left chest. In the open-group, one patient had lymphatic leakage. Follow-up period ranged from 3 to 46 months with a median of 22 months. The patency of endovascular repair group and the open surgery group was 92.6% and 90.8% at 12-month, while 82.9% and 84.3% at 24-month, respectively. The cumulative patency rates of smoking patients and non-smoking patients after endovascular treatment were 70.2% vs. 100% (P=0.048) at 24-month. No independent prognosis factors were identified through the Cox proportional risk model which significantly affected postoperative patency rates for patients with subclavian artery occlusion. Conclusions: Part of patients with subclavian artery occlusion can be treated by endovascular therapy. The success rate of left subclavian artery occlusions is higher than right sides. The length from the subclavian artery ostial to the occlusion area affected the success rate of repair.

Orally administered resveratrol enhances the therapeutic effect of autologous transplanted adipose-derived stem cells on rats with diabetic hepatopathy.

Stem cell therapy is a promising treatment for hepatopathy due to diabetes mellitus (DM); oral resveratrol treatment exhibits protective effects. We investigated whether protective effects could be produced in liver of diabetic rats receiving autologous adipose-derived stem cell transplantation (ADSC) plus oral resveratrol administration. Male rats were divided into four groups: sham group; streptozotocin induced DM group; DM + ADSC group, in which DM rats were treated with 106 stem cells/rat; and DM + R + ADSC group, in which DM rats were treated with ADSC and oral resveratrol. The DM group exhibited apoptosis, inflammation and fibrosis, whereas Sirt-1 and survival signaling were suppressed. Pathological conditions other than survival signaling were improved in the DM + ADSC group. All pathological conditions were improved in the DM + R + ADSC group. Also, the oxidative stress level in the blood was reduced in the DM + R + ADSC group compared to the sham group. Oral resveratrol administration appears to reduce oxidative damage and enhances survival signaling in diabetic liver. The therapeutic response in the DM + R + ADSC group was better than in the DM + ADSC group.

[First bite syndrome: clinical study of six cases and review of the literature].

Objective:The aim of this study is to explore the etiology, diagnosis and treatment of first bite syndrome（FBS）. Method:Clinical data of six patients with FBS were analyzed retrospectively. There were 4 cases of parapharyngeal space tumor resection complicated with FBS, 1 case of parotidectomy of deep lobe parotid gland tumor complicated with FBS, and 1 case of no definite cause. Analgesic treatments such as ibuprofen and tramadol capsules were given, in combination with carbamazepine （200 mg, 3 times a day） for four weeks. Result:The severity of symptoms decreased in five cases, 2-16 months after treatment. While another one had no amelioration during 10 months' follow-up. Conclusion:FBS is one of the possible complications of surgery of the upper cervical region, especially parotid deep lobe tumor resection, which should be distinguished from the usual postoperative pain. The cause of the disease may be related to intraoperative injury to the sympathetic plexus of the parotid gland. Currently, there is no standard treatment method, but the symptoms are usually relieved within a few months.

[Optical coherence tomographic analysis of in-stent neoatherosclerosis in lesions with restenosis after drug-eluting stent implantation].

Objective: To explore the imaging characteristics and related influencing factors of in-stent neoatherosclerosis (ISNA) in patients with restenosis after drug-eluting stent(DES) implantation with optical coherence tomography(OCT). Methods: A total of 25 cases of coronary heart disease patients(DES placement time ≥8 months) with coronary artery angiography showing DES in-stent restenosis (ISR) in Zunyi medical college affiliated hospital from July 2013 to December 2015 were included in this study and patient's data were retrospectively analyzed.In these patients with ISR, OCT images were acquired before percutaneous coronary intervention. Patients were divided into the ISNA group (12 patients and 12 lesions) and non-ISNA group(13 patients and 13 lesions) according to the result of OCT. ISNA on OCT was defined as neointima formation with the presence of lipids or calcification. Results: (1) The incidence of chronic kidney disease and increased low-density lipoprotein cholesterol level in ISNA group were significant higher than that in non-ISNA group(all P<0.05). The stent implantation time in ISNA group was longer than that in the non-ISNA group(53.0(14.0, 81.0) months vs. 15.0(8.5, 32.5) months, P<0.01). In addition, clinical manifestation of acute coronary syndrome was present in 8 out of 12 patientsin ISNA group, and stable angina pectoris was found in 10 out of 13 casesin non-ISNA group(P<0.01). (2) Quantitative analysis of OCT showed that the lumen area was less in ISNA group than in non-ISNA group((3.45±1.82)mm(2) vs. (4.17±1.68)mm(2), P<0.01), and neointimal area(3.89(2.26, 5.52)mm(2) vs. 2.96(1.99, 4.22)mm(2), P<0.01), neointimal load (53.15(40.18, 67.30)% vs. 41.54(32.08, 56.91)%, P<0.01), neointimal thickness(0.98(0.63, 1.36)µm vs. 0.72(0.51, 1.03)µm, P<0.01) were higher in ISNA group than in non-ISNA group.(3)Qualitative analysis of OCT showed that the prevalence of homogeneous intima was less in the ISNA group than in the non-ISNA group ((41.42±22.56)% vs.(72.06±18.68)%, P<0.05), on the contrary, the heterogeneous intima was more common in the ISNA group ((58.57±22.56)% vs. (27.94±18.68)%, P<0.05). There was no significant difference between two groups in the peri-stentmicrovessels (9/12 vs. 5/13,P>0.05), and prevalence of intraintimalmicrovessels was higher in the ISNA group than in non-ISNA group (7/12 vs. 2/13, P<0.05). In addition, thin cap fibrous plaque(7/12 vs. 0, P<0.01), disrupted intima with visible cavity (7/12 vs. 1/13, P<0.05),andintraluminal red thrombus(7/12 vs. 1/13, P<0.05) were significantly higher in ISNA group than in non-ISNA group. Conclusions: Results of OCT show that ISNA occurs frequently in patients with ISR after DES implantation. The stent implantation time, incidence of chronic kidney disease and higher low-density lipoprotein cholesterol level are associated with the formation of ISNA in these patients.

[Analysis of optical coherence tomography of early and very late stent restenosis after drug-eluting stent implantation].

Objective: Using optical coherence tomography (OCT) to analyze the morphological characteristics of lesions between early in-stent restenosis (E-ISR) and very late in-stent restenosis (VL-ISR) after drug-eluting stents (DES) implantation. Methods: In 25 patients (DES) with ISR, OCT images were acquired before percutaneous coronary intervention (PCI), the morphological characteristics of E-ISR (<1 years, n=14) and VL-ISR (>3 years, n=11) were compared.The quantitative and qualitative analysis of the entire stent and the minimum lumen area (MLA) site were carried out respectively. OCT quantitative restenotic tissue analysis included the assessment of mean lumen area, mean stent area, mean neointimal area and mean neointimal burden.OCT qualitative restenotic tissue analysis included the assessment of tissue structure [homogeneous or heterogeneous intima (lipid-rich neoitima, thin-cap fibroatheroma (TCFA)-like intima)], presence of microvessels, intraluminal material (red thrombus, pale thrombus), disrupted intima with cavity and tissue prolapse and was performed at every 1-mm slice of the entire stent. Results: Compared with the E-ISR group, the proportion of cross-sections with heterogeneous intima in the entire stent was significantly higher in the VL-ISR group (60.57% vs 32.93%, P=0.005), and both peristent and intraintimal microvessels were more frequently observed in the VL-ISR group (P<0.05). In addition, lipid-rich neointima (72.7% vs 21.4%, P=0.017), TCFA-like intima (54.5% vs 7.1%, P=0.021), disrupted intima with visible cavity (63.6% vs 7.1%, P=0.007) and red thrombus (63.6% vs 7.1%, P=0.007) were observed more frequently in the VL-ISR group compared with E-ISR group for the entire stent.The heterogeneous intima was observed more frequently in the VL-ISR group (90.9% vs 35.7%, P=0.012) at the MLA sites.Intraintimal microvessels and disrupted intima with visible cavity were observed only in the VL-ISR group. Conclusions: OCT imaging indicates that the morphological characteristics of restenosis lesions in VL-ISR are different from those in E-ISR.The atherosclerotic changes of neointima, such as lipid-rich neointima, disrupted intima with cavity and microvessels are more often observed in VL-ISR lesions after DES implantation compared with E-ISR.Progression of the atherosclerosis within neointima after DES implantation may be associated with VL-ISR.

[A control study of artificial oocyte activation in the same period].

Objective: To compare the fertility rate and embryo outcome between normal fertilization and the use of Calcium ionophore A23187 on the same period of the same cycle of human ovum for artificial activation. Methods: Patients who conducted the intracytoplasmic sperm injection (ICSI) cycle in vitro fertilization assisted reproductive from January 2015 to December in reproductive center of the Third Affiliated Hospital of Zhengzhou University were enrolled.The protocol of this study was approved by the ethics committee of The Third Affiliated Hospital of Zhengzhou University.The subjects must met at least one of the inclusion criteria: (1)the normal fertilization rate was less than 30% ICSI in the previous ICSI cycle; (2)no good quality embryos in a previous period of third days in ICSI; (3)patients with globozoospermia.The ovum were randomly divided into two groups, control group and artificial oocyte activation (AOA) group. In the control group, the eggs were treated with routine ICSI operation, and the AOA group was activated by A23187 after ICSI. Normal fertilization rate, cleavage rate, pregnancy and birth outcome of two groups were compared. Results: The 2PN fertilization rate in the AOA group 65.93% (60/91) was significantly higher than that in the control group 46.67% (41/89) (P<0.05). In addition, in patients who met the inclusion criteria of 1 categories (2PN≤30%), the 2PN fertilization rate was significantly higher (P<0.05) in the AOA group [79.59% (39/49)] compared with the control group [57.14% (28/49)]. In patients who met the inclusion criteria of 3 categories (globozoospermia), the 2PN fertilization rate was significantly higher (P<0.05) in the AOA group [75% (6/8)] compared with the control group [0% (0/5)]. Conclusions: The use of calcium ionophore A23187 assisted activation could be helpful to improve the normal fertilization rate of ICSI. But the effects of early embryonic development and the safety of generation need to be further studied.

[Control study of total percutaneous access with preclose technique versus open femoral artery exposure for endovascular aneurysm repair].

OBJECTIVE: To compare total percutaneous access using preclose technique with femoral artery cut-down in endovascular aneurysm repair (EVAR) and assess the safety and feasibility of preclose technique. METHODS: In the study, 81 cases undergoing EVAR from Dec. 2011 to Nov. 2014 in Peking University People's Hospital were retrospectively reviewed. Preoperative CT angiography (CTA) showed presence of infrarenal abdominal aortic aneurysm or descending aortic aneurysm in all the cases. The maximum diameter of aneurysm >4.5 cm met the indications for surgical treatment. The conditions of bilateral femoral artery and iliac artery CTA showed were good, and there was no moderate or severe stenosis, nor was there any severe calcification in anterior wall of femoral artery. Not only were the cases fit for percutaneous endovascular aortic aneurysm repair (PEVAR), but also feasible with open endovascular aneurysm repair (OEVAR). According to the intention of the patients about the surgical incision, the cases were divided into group PEVAR and group OEVAR. The data of the general situation, operation time, blood loss, technical success rate, length of hospital stay after procedure and wound complications were analyzed statistically. RESULTS: In the study, 44 cases (78 incisions) were enrolled in group PEVAR and 37 cases (65 incisions) in group OEVAR. There was no significant difference between the two groups in age, gender, body mass index (BMI), accompanying diseases, average number of stents and outer diameter of stent delivery system. Average operation time of group PEVAR was less than that of group OEVAR [(119.1±102.0) min vs. (163.6±61.9) min, P=0.025]. The blood loss in group PEVAR was less than that in group OEVAR [(64.7±97.0) mL vs. (98.6±88.3) mL], but there was no significant difference (P=0.106). There was no difference in the technical success rate (94.9% vs.95.4%, P=1.000). The average length of hospital stay after procedure was significantly shorter in group PEVAR [(7.8±2.8) d vs.(12.3±7.2) d, P<0.001]. There were 2 cases with subcutaneous hematoma of wound in group PEVAR and 7 cases of wound complications that occurred in group OEVAR including 3 cases with lymphatic leakage, 3 cases with lower limb ischemia and 1 case with subcutaneous hematoma. The analysis showed that PEVAR could reduce the wound complications (2.6%vs.10.8%), but there was no significant difference between the two groups (P=0.079). CONCLUSION: Using preclose technique in EVAR is safe and effective. It can shorten the operation time and length of hospital stay after procedure.

Impact of adherence to GOLD guidelines on 6-minute walk distance, MRC dyspnea scale score, lung function decline, quality of life, and quality-adjusted life years in a Shanghai suburb.

We performed a 1-year cluster-randomized field trial to assess the effect of standardized management of chronic obstructive pulmonary disease (COPD) on lung function and quality of life (QOL) measures in patients in China. We used the Global Initiative for Chronic Obstructive Lung Disease (GOLD) treatment guidelines and assessed indexes including pulmonary function, QOL, quality-adjusted life years (QALY), Medical Research Council (MRC) dyspnea scale, 6-min walk distance (6-MWD), number of emergency visits, and frequency of hospitalization. Of a total of 711 patients with chronic cough and asthma, 132 were diagnosed as having COPD and 102 participated in this study [intervention group (N = 47); control group (N = 55)]. We found that adherence to GOLD guidelines had a perceivable impact on 6-MWD, MRC dyspnea scale score, and QOL. The average QALY increased by 1.42/person/year in the intervention group, but declined by 0.95/person/year in the control group. We conclude that standardized management improves disease severity, QOL, and QALY in COPD patients when treatment protocols adhere to GOLD guidelines.

Predicting the Prognosis of Oral Tongue Carcinoma Using a Simple Quantitative Measurement Based on Preoperative MR Imaging: Tumor Thickness versus Tumor Volume.

BACKGROUND AND PURPOSE: Several studies indicated that tumor thickness or tumor volume might be helpful predictors for the prognosis of oral tongue squamous cell carcinoma. Our aim was to compare the value of tumor thickness versus tumor volume measurement based on preoperative MR imaging in predicting the prognosis of oral tongue squamous cell carcinoma, especially focusing on lymph node metastases and local recurrence. MATERIALS AND METHODS: Clinical, pathologic, and imaging data of patients with 46 oral tongue squamous cell carcinomas were retrospectively studied. Logistic regression analysis was used to evaluate the prognostic value of tumor thickness and tumor volume based on MR imaging. Receiver operating characteristic analysis was applied for the optimal cutoff value for the identified risk variable for prognosis. RESULTS: A higher intraclass correlation coefficient was achieved for the measurement of tumor thickness compared with tumor volume (0.990 versus 0.972). Multivariate analysis showed that tumor thickness was a significant predictor of lymph node metastases (P = .024), while tumor volume was not a significant predictor of either lymph node metastases or local recurrence (P > .05). Receiver operating characteristic results indicated that setting a tumor thickness of 8.5 mm as a cutoff value could achieve the optimal diagnostic efficiency for predicting lymph node metastases (area under the curve, 0.753; sensitivity, 0.889; specificity, 0.536). CONCLUSIONS: Tumor thickness based on preoperative MR imaging was useful in predicting the prognosis of oral tongue squamous cell carcinoma, especially lymph node metastases, in our patient population, while tumor volume was not.

DNA-PKcs activates the Chk2-Brca1 pathway during mitosis to ensure chromosomal stability.

The catalytic subunit of DNA-dependent protein kinase (DNA-PKcs) is known to have a critical role in DNA double-strand break repair. We have previously reported that DNA-PKcs is activated when cells enter mitosis and functions in mitotic spindle assembly and chromosome segregation. Here we report that DNA-PKcs is the upstream regulator of the Chk2-Brca1 pathway, which impacts microtubule dynamics, kinetochore attachment and chromosomal segregation in mitosis. Downstream from Chk2, Brca1 promotes monoubiquitination of γ-tubulin to inhibit microtubule nucleation and growth. We found that DNA-PKcs is essential for mitotic Chk2 phosphorylation at Thr68. As in Chk2- and Brca1-deficient cells, loss of DNA-PKcs resulted in chromosome misalignment and lagging during anaphase owing to elevation in microtubule dynamics. Importantly, these mitotic aberrations in DNA-PKcs-defective cells were alleviated by the overexpression of phosphomimetic Chk2 or Brca1 mutant proteins but not their wild-type counterparts. Taken together, these results demonstrate that DNA-PKcs regulates mitotic spindle organization and chromosomal instability via the Chk2-Brca1 signaling pathway.

Clinical feasibility of a novel biphasic osteochondral composite for matrix-associated autologous chondrocyte implantation.

OBJECTIVE: Matrix-associated autologous chondrocyte implantation has been used to treat cartilage defects. We developed a biphasic cylindrical osteochondral composite construct for such use, and conducted this study to determine its feasibility for treating osteochondral lesions in human knees. METHOD: Ten patients with symptomatic osteochondral lesions at femoral condyles were treated by replacing pathological tissue with the construct of dl-poly-lactide-co-glycolide, whose lower body was impregnated with ß-tricalcium phosphate and served as osseous phase. The construct had a chamber to load double-minced autologous cartilage, serving as source of chondrocytes. Osteochondral lesion was drill-fashioned a pit of identical dimension as the construct. Chondrocyte-laden construct was press-fit to fill the pit. Postoperative outcome was evaluated using Knee Injury and Osteoarthritis Outcome Score (KOOS) scale up to 24 months. Magnetic resonance image was taken, and sample tissue was collected with second-look arthroscopic needle biopsy at 12 months. Outcome parameters were primarily safety of surgery, and secondarily postoperative change in KOOS and regeneration of hyaline cartilage and cancellous bone. RESULTS: No patient experienced serious adverse events. Postoperative mean KOOS in "symptoms" subscale had not changed significantly from pre-operation until 24 months; whereas those in the other four subscales were significantly higher than pre-operation at 12 and 24 months. Second-look arthroscopy showed completely filled grafted sites, with regenerate cartilaginous surfaces flushed with surrounding native joint surface. Microscopically, regenerated cartilage appeared hyaline. CONCLUSION: This novel construct for chondrocyte implantation is safe for surgical application in knee. It repairs osteochondral lesions of femoral condyles by successful regeneration of hyaline cartilage.

Effect of Zn impurity in K0.8Fe(2-δ-x)Zn(x)Se2.

A series of K(0.8)Fe(2-δ-x)Zn(x)Se(2) single-crystal samples with nominal compositions 0 ≤ x ≤ 0.05 were grown and their physical properties were measured in order to study the effect of Zn impurity. It is found that the Zn impurity (x ≤ 0.02) does not affect the superconducting transition temperature T(c) significantly. Meanwhile the hump in resistivity which corresponds to the transition from the insulating to metallic phase quickly shifts towards low temperatures. The results imply that there should be a phase separation in this system and Zn impurity causes the enhancement of the insulating phase. The negligible effect of Zn impurity on T(c) suggests an s-wave pairing in the superconducting phase. Meanwhile there is a possibility that the Zn impurity may selectively enter into the insulting phase.

Comparison of spermatogenic damage induced at 6 months after ligation of the vas deferens at proximal and distal locations in the rabbit.

Recent study in rabbits demonstrated that vasectomy via the inguinal canal did not result in any spermatogenic damage 3 months postoperation; this study aimed to determine whether the damage would occur in a longer term. The left or right vas deferens was ligated near the epididymal head (unilateral proximal vasectomy, 12 animals) or via the inguinal canal (unilateral distal vasectomy, 11 animals) in adult male rabbits, with a sham operation being performed on the contralateral side. Six months postoperation, testes, epididymides and vasa deferentia were removed and methacrylate resin-embedded sections prepared to evaluate spermatogenesis by histological (qualitative) and stereological (quantitative) studies. The juxta-epididymal segment of the occluded vas deferens was severely distended (filled with sperm) in 10 of the 11 animals with distal vasectomy and moderately or slightly distended in nine of the 12 animals with proximal vasectomy. Severe spermatogenic damage occurred in seven animals with proximal vasectomy (the juxta-epididymal vas moderately or slightly distended), in only one animal with distal vasectomy (the vas not severely distended). In conclusion, spermatogenic damage occurred at 6 months postvasectomy in some animals, especially those with proximal vasectomy and therefore shorter occluded reproductive tract for sperm storage; the damage was probably intra-tract pressure mediated.