Role of C3a and C5a in focal segmental glomerulosclerosis

Objective To study the role of C3a and C5a in focal segmental glomerulosclerosis (FSGS) patients. Methods (1) A total of 66 patients with FSGS confirmed by renal biopsy were selected, including 18 cases of tip lesion, 11 cases of perihilar, 22 cases of not otherwise specified (NOS), 10 cases of cellular, and 5 cases of collapsing FSGS. The normal renal tissue resected from patients with kidney tumor was taken as a negative control. The expression of C3a and C5a in renal tissues was detected by immunohistochemistry. (2) Serum and urine samples from these 66 FSGS patients were collected, and serum and urine samples from 10 healthy adult selected from the same physical examination center in the same term were used as normal controls. The levels of C3a and C5a in serum and urine were detected by enzyme-linked immunosorbent assay (ELISA). Results (1) Immunohistochemical results showed that C3a and C5a were deposited in glomerulus of FSGS patients, and no deposition in normal renal tissues. The semi-quantitative score showed that kidney C3a score was significantly correlated with serum creatinine (r=0.547, P＜0.001) and 24 h urine protein (r=0.329, P=0.007) in FSGS patients, and kidney C5a score was also significantly correlated with serum creatinine (r=0.415, P＜0.001) and 24 h urine protein (r=0.414, P＜0.001) in FSGS patients. (2) The levels of serum C3a and C5a in FSGS patients were higher than those in healthy adults (both P＜0.05), but there was no significant difference among the five pathological types (P＞0.05). The levels of urinary C3a/urinary creatinine, urinary C5a/urinary creatinine were higher in FSGS patients than those in healthy adults (all P＜0.05). The levels of urine C3a/urinary creatinine and urinary C5a/urinary creatinine in collapsing FSGS were higher than other FSGS types (all P＜0.01), but there was no significant difference among the tip lesion, the perihilar, the not otherwise specified and the cellular (P＞0.05). (3) Urinary C3a/urinary creatinine levels were significantly correlated with serum creatinine (r=0.774, P＜0.001) and 24 h urine protein (r=0.430, P＜0.001) in FSGS patients, and urinary C5a/urinary creatinine levels were also significantly correlated with serum creatinine (r=0.677, P＜0.001) and 24 h urine protein (r=0.333, P=0.007) in FSGS patients. Conclusion Complement C3a and C5a may be involved in the pathogenesis of FSGS and may be related to the severity of FSGS.     

Comparison of main molecular markers involved in autophagy and apoptosis pathways between spermatozoa of infertile men with varicocele and fertile individuals

Abnormal dilatation and tortuosity of the pampiniform plexus within the spermatic cord are termed varicocele which leads to impaired spermatogenesis due to heat‐related oxidative stress and cell death. Previously, it was shown that both apoptosis and autophagy pathways were activated by heat in germ cells of mouse in vivo and in vitro. But, status of these pathways is not clear in chronic state of heat stress such as varicocele. Therefore, we aimed to access sperm apoptotic markers (active caspases 3/7 and DNA fragmentation), and autophagic markers (Atg7 and LC3 proteins) as primary outcomes, and also sperm parameters and protamine deficiency as secondary outcomes between 23 infertile men with varicocele and 16 fertile individuals. Sperm parameters were assessed according to World Health Organization 2010 protocol. Apoptotic markers (active caspases 3/7 and DNA fragmentation), autophagic markers (Atg7 and LC3 proteins), and protamine deficiency were evaluated by flow cytometry, fluorescence microscope, and western blotting techniques. Mean of autophagy and apoptosis markers, and also protamine deficiency have significantly increased in infertile men with varicocele compared to fertile individuals, but autophagy and apoptosis markers did not significantly correlate with each other. In conclusion, it seems that both apoptosis and autophagy pathways are independently active in spermatozoa of infertile men with varicocele.     

14‐3‐3 epsilon plays an important role in testicular germ cell apoptosis: A functional proteomic study of experimental varicocele

The latest perspective indicates that apoptotic dysregulation is an important mechanism in male infertility induced by varicocele. To elucidate the molecular mechanism of apoptosis caused by varicocele, we used proteomics (2D‐MALDI‐TOF MS) to identify the altered proteins in the testes of experimental varicocele rats compared with the control. Here, 21 significantly different protein spots were detected by proteomics technology. 14‐3‐3 epsilon (14‐3‐3ε) was our subsequent research target because of its function in apoptosis. The expression of 14‐3‐3ε in rat testes was confirmed by Western blot and immunohistochemistry, and the terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL) method was used to analyse the apoptosis of germ cells. GC‐1 spg cells transfected with small interfering RNA were used to confirm the function of 14‐3‐3ε in vitro. 14‐3‐3ε protein expression decreased, accompanied by a higher apoptosis index in rat testes of the varicocele group. Furthermore, 14‐3‐3ε siRNA‐treated GC‐1 spg cells caused the upregulation of the apoptotic rate detected by flow cytometry. The expression of Bax and Bcl‐2 was found to be regulated by 14‐3‐3ε in vitro. Our investigation demonstrated the pro‐apoptotic function of the downregulation of 14‐3‐3ε, which may play an important role in germ cell apoptosis induced by varicocele.     

The role of PI3K/Akt signaling pathway in non‐physiological shear stress‐induced platelet activation

The PI3K/Akt signaling pathway has been implicated in playing an important role in platelet activation during hemostasis and thrombosis involving platelet‐matrix interaction and platelet aggregation. Its role in non‐physiological shear stress (NPSS)‐induced platelet activation relevant to high‐shear blood contacting medical devices (BCMDs) is unclear. In the context of blood cells flowing in BCMDs, platelets are subjected to NPSS (>100 Pa) with very short exposure time (<1 s). In this study, we investigated whether NPSS with short exposure time induces platelet activation through the PI3K/Akt signaling pathway. Healthy donor blood treated with or without PI3K inhibitor was subjected to NPSS (150 Pa) with short exposure time (0.5 s). Platelet activation indicated by the surface P‐selectin expression and activated glycoprotein (GP) IIb/IIIa was quantified using flow cytometry. The phosphorylation of Akt, activation of the PI3K signaling, was characterized by western blotting. Changes in adhesion behavior of NPSS‐sheared platelets on fibrinogen, collagen, and von Willebrand factor (vWF) were quantified with fluorescent microscopy by perfusing the NPSS‐sheared and PI3K inhibitor‐treated blood through fibrinogen, collagen, and vWF‐coated microcapillary tubes. The results showed that the PI3K/Akt signaling was involved with both NPSS‐induced platelet activation and platelet‐matrix interaction. NPSS‐sheared platelets exhibited exacerbated platelet adhesion on fibrinogen, but had diminished platelet adhesion on collagen and vWF. The inhibition of PI3K signaling reduced P‐selectin expression and GPIIb/IIIa activation with suppressed Akt phosphorylation and abolished NPSS‐enhanced platelet adhesion on fibrinogen in NPSS‐sheared blood. The inhibition of PI3K signaling can attenuate the adhesion of unsheared platelets (baseline) on collagen and vWF, while had no impact on adhesion of NPSS‐sheared platelets on collagen and vWF. This study confirmed the important role of PI3K/Akt signaling pathway in NPSS‐induced platelet activation. The finding of this study suggests that blocking PI3K/Akt signaling pathway could be a potential method to treat thrombosis in patients implanted with BCMDs.     

Safety of mucous fistula refeeding in neonates with functional short bowel syndrome: A retrospective review

PurposeMucous fistula (MF) refeeding of proximal stoma effluent in neonates after small bowel resection can promote nutrient absorption and prevent atrophy of the unused distal bowel. This study aimed to assess the safety of this practice in neonates.MethodsA retrospective chart review of all patients admitted to the neonatal intensive care unit (NICU) between 2009 and 2015 who underwent a laparotomy with creation of an enterostomy and mucous fistula was performed. Patients were included if they were refed proximal stoma effluent into the MF.ResultsThirty-one patients were identified that were refed. There were no major complications (perforation, stricture, death) related to refeeding. Patients were refed for an average of 41 days (± 22), with patients gaining an average of 25.7 g/day (± 10.1) while being refed. Total parental nutrition (TPN) was administered for an average of 55 days (± 31.4) between resection and reanastomosis, with only 7 (23%) developing cholestasis and 15 (48%) reaching full feeds in this time. Mean time to full feeds after reanastomosis was 36 days (± 58.6) with two patients having anastomotic leaks.ConclusionMF refeeding is a safe technique that has the potential to contribute to significant weight gain and a decreased dependence on total parenteral nutrition.Level of evidenceII     

Effectiveness of Antithymocyte Globulin Induction Dosing Regimens in Kidney Transplantation Patients: A Network Meta-analysis

BackgroundAntithymocyte globulin (ATG) is an induction therapy in kidney transplantation, but our knowledge about the relation between outcomes and ATG regimens is limited. We compared ATG effectiveness in kidney transplantation according to dosage and administration schedule.MethodsReports from 1970 until May 2018 in CENTRAL, MEDLINE, EMBASE, and Science Citation Index Expanded were searched. We performed direct and indirect network meta-analysis using Bayesian models and generated rankings for ATG dosage and injection number variations by generation mixed treatment comparison.We compared ATG dose and schedule in kidney transplantation in relation to all-cause death, graft failure, antibody-mediated rejection, T-cell mediated rejection, biopsy-proven acute rejection, and bacterial and viral infection.ResultsTen studies (N = 1065) were analyzed by forming 6 groups: ATG alternate doses, 9 mg/kg, 6 mg/kg, and 4.5 mg/kg; single dose, 6 mg/kg, and 4.5 mg/kg; and control. Compared to placebo, ATG regimen variations were not associated with significant differences in survival, viral infection, renal function, or graft survival. ATG regimens 9 and 4.5 mg alternate dosing tended to reduce biopsy-proven acute rejection but without statistical significance. According to the highest rank probability, the 9 mg alternate dosing group had the highest tendency for cytomegalovirus and bacterial infections but without statistical significance.ConclusionsThe rejection frequency tended to be lower for the 9 and 4.5 mg alternate dosing groups. Infections occurred at a higher rate in the 9 mg alternate dosing group, but the differences in the risk of infection among the groups with different ATG regimens were not statistically significant.     

3-D 打印技术结合经皮椎弓根螺钉内固定术治疗中老年胸腰椎爆裂骨折的疗效分析

目的：探究 3-D 打印技术在经皮椎弓根螺钉内固定术治疗中老年单纯性胸腰椎骨折手术中应用的可行性。方法：回顾分析我院 2014 年 1 月—2016 年 6 月收治的中老年胸腰椎爆裂性骨折且符合手术条件的患者 28 例,其中男性 11 例,女性 17 例;年龄 52 ～ 56 岁,平均 56.4 岁。所有患者术前均收集 CT 数据,在 Mimics 软件中进行骨折三维重建,打印出 1:1实体模型,并在 3-D 模型上行模拟手术设计内固定方案,并行经皮椎弓根螺钉内固定术。观察患者围手术期指标 ( 术中失血量、手术时间、引流量、住院时间 ) 、术前术后视觉模拟评分 ( VAS) 、Cobb 角及椎管占位率改变、术后并发症发生情况。结果：手术时间平均 (58.37±10.43) min,术中失血量平均 (51.93±7.65) mL,术后切口引流量平均 (32.41±12.19) mL,住院时间平均 (9.53±2.48) d,所有患者术后随访 14~19 个月,平均 15.7 个月。术前平均 Cobb 角 (16.25±4.29)°,术后2 天 (5.00±1.64) °,术后 12 个月 (5.00±1.55) °。术前平均椎管占位率 (28.18±3.60) %,术后 2 天 (9.71±1.80) %,术后 12 个月 (8.39±1.73) %。Cobb 角及椎管占位率术前术后比较差异有统计学意义 (P<0.05)。术前 VAS (8.07±1.05) 分,术后 1 周 (2.75±0.84) 分,术后 12 个月 (1.50±0.88) 分。术后 VAS 评分较术前显著降低,差异有统计学意义 (P<0.05)。术后 1 周 VAS 评分与术后 12 个月比较差异存在统计学意义（P<0.05）。所有患者均无断钉及内固定物松动、伤椎再骨折、感染、下肢深静脉血栓等并发症。1 例术后 12 个月出现腰背部慢性疼痛,给与口服非甾体类止疼药,症状明显缓解。结论：3-D 打印技术结合经皮椎弓根螺钉内固定术治疗中老年单纯性下胸腰椎骨折可获得满意的早期复位效果,明显改善临床症状,增加伤椎的稳定性,降低并发症发生风险,有很强的临床指导作用。