The Angiotensin II/transforming growth factor-β1 (AngII/TGF-β1) signal axis is an important regulatory pathway for atrial fibrosis, which can contribute to atrial fibrillation (AF). Fused in sarcoma (FUS) was recently found to regulate cardiac diseases. This study aimed to investigate whether FUS could regulate AngII induced fibrosis and uncover the possible mechanisms. The expression of FUS in AF patients and AngII-induced cardiac fibroblasts was measured by RT-qPCR and western blot assays. Fus was silenced in cells using short hairpin RNA (shRNA), then cell proliferation, migration, collagen synthesis and TGF-β1/Smad signaling were detected by CCK-8, wound healing and western blot assays, respectively. The possible target for Fus was predicted by searching Starbase database and verified by RNA-binding protein immunoprecipitation (RIP) and RNA pull down. Cells were overexpressed with Pax3 in the presence of Fus silence and AngII stimulation, then the above cellular processes were further evaluated. Results showed that FUS was upregulated in AF patients and AngII-induced cardiac fibroblasts. Fus knockdown inhibited AngII-enhanced cell proliferation, migration, collagen synthesis and TGF-β1/Smad signaling activation. Furthermore, Fus functions as an RNA-binding protein to bind to Pax3 mRNA and positively regulate its expression. Further studies demonstrated that Pax3 overexpression canceled the above effects of Fus knockdown on cell proliferation, migration, collagen synthesis, and TGF-β1/Smad signaling activation in AngII-induced cells. In conclusion, Fus could target Pax3 to increase the pro-fibrotic effect of AngII in cardiac fibroblasts via activating TGF-β1/Smad signaling. Knockdown of Fus/Pax3 axis may provide a potential therapy for relieving AF.

The aim of the present study was to investigate the protective effects of ischemic preconditioning on rats with renal ischemia-reperfusion injury and the effects on the expression of Bcl-2 and Bax. Thirty-six SD rats were randomly divided into three groups (n=12) including sham operation (S) group, ischemia-reperfusion group (I/R) group and ischemic preconditioning (IP) group. After anesthesia with intraperitoneal injection of chloral hydrate, bilateral renal pedicles were clipped for 45 min, followed by perfusion for 6 h to establish the I/R model. Both kidneys in rats of S group were separated and exposed for 45 min, but renal pedicles were not clipped. In IP group, bilateral renal pedicles were clipped for 5 min, followed by perfusion for 5 min, this procedure was repeated 3 times. Then bilateral renal pedicles were clipped for 45 min, followed by perfusion for 6 h. Blood samples were collected and rats were sacrificed to collect renal tissue. Levels of serum creatinine (Cr) and blood urea nitrogen (BUN) were measured. Activity of superoxide dismutase (SOD) was measured by xanthine oxidase assay. Degree of renal injury was evaluated by H&E staining. TUNEL kit was used to detect the number of apoptotic cells in renal tissue. Expression levels of Bcl-2 and Bax were detected by semi-quantitative PCR and western blot analysis at mRNA and protein levels, respectively. Results showed that levels of Cr and BUN in I/R and IP groups were significantly higher than those in S group, and levels of Cr and BUN in I/R group were significantly higher than that in IP group (P<0.05). Activity of SOD in I/R group and IP group were significantly lower than those in S group, and activity of SOD in I/R group were significantly lower than those in IP group (P<0.05). H&E staining showed that, compared with S group, renal injury in the I/R and IP groups was more serious than that in the S group, and I/R group was more serious than the IP group (P<0.05). TUNEL apoptosis assay showed that number of apoptotic cells in IP and I/R groups were significantly higher than that in the S group (P<0.01). Semi-quantitative PCR and western blot analysis showed that, compared with the S group, expression levels of Bcl-2 mRNA and protein were significantly decreased, expression levels of Bax mRNA and protein were significantly increased, and the ratio of Bcl-2/Bax was significantly decreased in the IP and I/R groups (P<0.01). Compared with the I/R group, expression level of Bcl-2 was significantly increased, the level of Bax was significantly deceased, and the ratio of Bcl-2/Bax was significantly increased in the IP group (P<0.01). As a result, ischemic preconditioning can protect rats with renal ischemia-reperfusion injury possibly by increasing the expression level of Bcl-2 and decreasing the expression level of Bax.

目的:探讨肾移植术后中远期出血的特点和诊治方法。方法:回顾6例术后并发血管疾病的肾移植患者临床资料,对其发病特点和诊治方法进行分析总结。结果:6例患者中,并发移植肾破裂2例,移植肾动脉破裂出血3例,移植肾动脉破裂伴瘤样动静脉内瘘形成1例。6例均经彩色多普勒血流显像(CDFI)作出诊断,其中2例经CT明确诊断,3例进一步行磁共振血管成像明确诊断,1例经血管造影进一步明确诊断。所有患者经手术治疗,2例移植肾破裂,1例修补成功,1例行移植肾切除;3例移植肾动脉破裂均行移植肾切除;1例移植肾动脉破裂伴瘤样动静脉内瘘形成患者经血管修补整形成功。结论:肾移植术后的出血发展迅速,应根据具体情况及时采取相应治疗手段,处理不及时往往导致严重后果,因此早期诊断非常重要,CDFI可作为首选筛查手段。

BACKGROUND:The vascular anastomosis technique is directly related to the success or failure of the renal transplantation. OBJECTIVE:To improve the techniques of variable vascular anastomosis in renal transplantation. METHODS:878 cases received renal transplantation in the Department of Organ Transplantation, Guangdong No.2 Provincial People’s Hospital from September 1999 to December 2010 were selected, and the donor renal vascular variation including renal artery and renal vein were treated with trim, merge, extend and other reconstruction processes for renal transplantation, and the accessory renal arteries were anastomosed to inferior epigastric arteries by a modified stereotaxic method. RESULTS AND CONCLUSION:Among the 878 renal transplantation cases, 55 donor kidneys had renal&nbsp;artery variation and 22 donor kidneys had renal vein variation. Al the variable vessels were anastomosed to the internal iliac or external iliac vessels successful y after vascular reconstruction. Twenty-three cases of accessory renal arteries with the diameter of more than 1 mm were anastomosed to inferior epigastric arteries successful y. Color Doppler ultrasound image showed that blood stream was smooth in al anastomosed vessels. Transplant renal vascular variation and accessory renal arteries can be transplanted safely and effectively by correct vascular rebuilding or vascular anastomosis.

BACKGROUND: It is a hot investigation to many scholars that how to cure and prevent renal ischemic reperfusion injury (IRI) in a utility way, but the mechanism is unclear at present. The investigation indicates that hypoxia inducible factor (HIF) plays an important role during this process. OBJECTIVE: To investigate the effects and possible mechanism of HIFs on renal ischemia reperfusion injury in mice. METHODS: Two kinds of mice were selected: one kind was aryl hydrocarbon receptor nuclear translocator (ARNT) gene inactivation, and the other kind was used as controls. Each kind of mice were randomly divided into sham operation group, ischemic reperfusion group, ischemic reperfusion+ recombinant human erythropoietin (rhEPO) group, ischemic reperfusion+0.9% normal saline (NS) group. Mice were established for renal IRI models. Serum erythropoietin (EPO) levels after 1 hour and 24-hour serum creatinine (Cr) values were examined, 24-hour renal tubular injury score following PAS staining was recorded, and 24-hour renal tubular cell apoptosis was counted by using TUNEL method. RESULTS AND CONCLUSION: One hour after reperfusion ARNT knockout serum EPO levels were significantly lower than the control group (P < 0.01). At 24 hours, Cr levels of ARNT knockout mice were significantly higher than the control group (P < 0.01). Renal Tubular injury scores of ARNT knockout group were significantly higher than the control group (P < 0.01). The number of TUNEL positive cells of ARNT knockout group was significantly more than the control group (P < 0.01). After rhEPO injection, there were no significant difference between ARNT knockout mice and control mice (P > 0.05). The results showed that HIF system on renal ischemia reperfusion injury has important protective role. It may be the maximum protective effect mediated by EPO.

Background: Mitochysis of mouse embryonic fibroblasts is inhibited by mitomycin C treatment or γ irradiation. Cells keep the activity, but dose not proliferate, which can produce factors enhancing the growth of embryonic stem cells and inhibiting the differentiation of embryonic stem cells, but the life span is limited. Objective: To investigate the isolation proper age of mouse embryonic fibroblasts and the preparation of embryonic fibroblast feeder layer. Design, time and setting: The cytology in vitro experiment was performed at the Laboratory of Guangdong Family Planning Special Hospital from November 2005 to July 2006. Materials: Clean pregnant Kunming rats (7.5, 10.5, 13.5, 16.5, 19.5 days of pregnancy) (five each) were used in this study. Methods: Mouse primary embryonic fibroblasts were sterilely prepared from mouse fetus of different embryonic ages. Cells at densities of 1 × 10 7 L -1, 1 × 10 9 L -1, 1 × 10 11 L -1 were digested in trypsin. When embryonic fibroblasts grew and contacted each other, and then treated with mitomycin C for 2-4 hours, trypsin solution supplemented with Ca 2+-free and Mg 2+-free phosphate buffered saline for 2-5 minutes. Cells were blown and hit with a pipette to isolate cells, Subsequently, cells were inoculated in DMEM containing 10% fetal bovine serum into a density of 5 × 10 8 L -1. This suspension was inoculated in gelatin-treated culture dish to prepare fibroblast feeder layer. Main outcome measures: Effects of embryonic age, cell concentration and passage times on the proliferation of mouse embryonic fibroblasts. Effects of feeder layer preparation of mouse embryonic fibroblasts at different embryonic ages. Results: Fibroblasts were harvested from pregnant mice at embryonic days 7.5-19.5. Number of fibroblasts were few and with short life span in mice at embryonic day 7.5 and day 10.5, and with many other ypes of cells. Number of fibroblasts was more and with rapid proliferation in mice at embryonic day 13.5, with few other types of cells. Fibroblasts grew poorly, and proliferated slowly, with many other types of cells in mice at embryonic days 16.5-19.5. Under the same condition, fibroblasts at a density of 1 × 10 9 L -1 grew well, with suitable monolayer confluence and long life span in mice (P < 0.05). Mouse embryonic fibroblasts at embryonic day 13.5 were cultured at fourth passage. Fibroblast strain was successfully established. Under the same condition, the morphology, volume and growth condition of 1st-4th passages of cells had no significant difference. The feeder layer of mouse embryonic fibroblasts at embryonic day 7.5-19.5 had similar monolayer confluence (P > 0.05), and the cell life span could maintain for 1-2 weeks. Conclusion: The optimal embryonic age and cell concentration are respectively 13.5 days and 1 × 10 9 L -1 in mouse embryonic fibroblasts. Passage time within the fourth passage cannot affect the proliferation of mouse embryonic fibroblasts. No significant difference is detected in cells in feeder layer at different embryonic days.

目的研究大剂量静脉滴注免疫球蛋白(IVIG)对降低肾移植患者群体反应抗体(PRA)的临床治疗效果。方法31例等待肾脏移植的PRA阳性患者,年龄24~67岁,PRA值为18%~91%,平均为40.7%。采用静脉滴注的方式,1次/d,5 g/次,两周为1个疗程。两个疗程之间,间隔1周时间。用药开始时检测一次PRA,疗程结束立即再检测一次PRA。结果31例患者中有19例患者用药1个疗程,11例患者用药2个疗程,1例患者用药3个疗程。31例患者静脉应用免疫球蛋白后PRA降为阴性13例(41.9),完全未降4例(12.9),有不同程度下降14例(45.2)。31例患者治疗后2~3周内,因有合适的供肾均进行了肾脏移植手术。术中均未出现超排。2例患者术后1周左右因急性排斥并发移植肾破裂切除移植肾,切除后病理检查报告为急性血管性排斥。1例急性排斥OKT35 mg/d连续冲击14 d后,急性排斥逆转,肾功能逐渐恢复正常。结论大剂量IVIG能有效的降低肾移植患者PRA,提高肾移植的成功率;IVIG的替代作用和免疫调节作用对移植术后患者的顺利康复也有较大帮助。

OBJECTIVETo explore the prevention and management of human cytomegalovirus (HCMV) infection accompanied with acute pancreatitis after kidney transplantation.METHODS AND RESULTSA retrospective analysis of 5 patients with acute pancreatitis after kidney transplantation was conducted. The incidence of acute pancreatitis after kidney transplantation was 2.3% (5/217). All the 5 cases were complicated by active HCMV infection, 3 of which had hyperlipemia and 2 had liver dysfunction. Three cases were finally cured while the other 2 died, one of which was due to respiration failure arising from HCMV interstitial pneumonia accompanied with hemorrhagic necrotizing pancreatitis, and the other due to fulminating liver function failure because of active HCMV infection accompanied with hemorrhagic necrotizing pancreatitis.CONCLUSIONActive HCMV infection is the most important factor responsible for acute pancreatitis, and early diagnosis and treatment are crucial to lower mortality rate.