Targeted mesenchymal stem cell and vascular endothelial growth factor strategies for repair of nerve defects with nerve tissue implanted autogenous vein graft conduits

Peripheral nerve gaps exceeding 1 cm require a bridging repair strategy. Clinical feasibility of autogenous nerve grafting is limited by donor site comorbidity. In this study we investigated neuroregenerative efficacy of autogenous vein grafts implanted with tissue fragments from distal nerve in combination with vascular endothelial growth factor (VEGF) or mesenchymal stem cells (MSCs) in repair of rat peripheral nerve defects. Six‐groups of Sprague‐Dawley rats (n = 8 each) were evaluated in the autogenous setting using a 1.6 cm long peroneal nerve defect: Empty vein graft (group 1), Nerve graft (group 2), Vein graft and nerve fragments (group 3), Vein graft and nerve fragments and blank microspheres (group 4), Vein graft and nerve fragments and VEGF microspheres (group 5), Vein graft and nerve fragments and MSCs (group 6). Nerve fragments were derived from distal segment. Walking track analysis, electrophysiology and nerve histomorphometry were performed for assessment. Peroneal function indices (PFI), electrophysiology (amplitude) and axon count results for group 2 were ?9.12 ± 3.07, 12.81 ± 2.46 mV, and 1697.88 ± 166.18, whereas the results for group 5 were ?9.35 ± 2.55, 12.68 ± 1.78, and 1566 ± 131.44, respectively. The assessment results did not reveal statistical difference between groups 2 and 5 (P > 0.05). The best outcomes were seen in group 2 and 5 followed by group 6. Compared to other groups, poorest outcomes were seen in group 1 (P ≤ 0.05). PFI, electrophysiology (amplitude) and axon count results for group 1 were ?208.82 ± 110.69, 0.86 ± 0.52, and 444.50 ± 274.03, respectively. Vein conduits implanted with distal nerve‐derived nerve fragments improved axonal regeneration. VEGF was superior to MSCs in facilitating nerve regeneration. © 2015 Wiley Periodicals, Inc. Microsurgery 36:578–585, 2016.     

Ultrasonographic evaluation of the femoral cartilage thickness in patients with chronic renal failure

Objective To investigate the effects of chronic renal failure (CRF) on the distal femoral cartilage thickness by using ultrasonography and to determine the relationship between cartilage thickness and certain disease-related parameters. Design Fifty-seven CRF patients (41 male and 16 female) (mean [SD] age, 44.7 [12.1] years) and 60 healthy controls (41 male and 19 female) (mean [SD] age, 43.5 [13.3] years) were enrolled in this study. Demographic and clinical characteristics were recorded. Cartilage thickness measurements were taken from the medial and lateral condyles, and intercondylar areas of both knees. Results Groups were similar in terms of age, weight, height, body mass index and gender (all p>0.05). The mean cartilage thickness was found to be less in CRF patients than in controls (statistically significant for medial condyles and intercondylar areas both in right and the left knees [all p<0.05]). Cartilage thickness showed no correlation with eGFR, and with the levels of serum urea, creatinine, calcium, magnesium, phosphor, hemoglobin, uric acid and as well as steroid use (all p>0.05) in CRF patients. Conclusion In the light of our findings, we imply that patients with CRF have thinner femoral cartilage than healthy controls. This result may support the view that patients with CRF are at increased risk for developing early knee osteoarthritis. Last but not least, clinicians should be aware of the importance of rehabilitation strategies aimed at decreasing onset and progression of knee osteoarthritis in patients with CRF.     

Two level pedicle substraction osteotomies for the treatment of severe fixed sagittal plane deformity: computer software-assisted preoperative planning and assessing

Purpose

To evaluate the efficacy of two level pedicle substraction osteotomies (PSOs) planned preoperatively with a computer software, in the patients with severe fixed sagittal plane deformities.

Methods

In the literature, there are studies indicating that two level PSOs may be required in severe cases. However, the results of two level PSOs preoperatively planned with computer software-assistance have not yet been reported in the English literature. Severe fixed sagittal plane deformities of 11 patients are described. Preoperative surgical planning was done with the aid of a computer software. Two level PSOs were indicated after the process. After the application of the indicated surgical technique, clinical and radiological results were evaluated in the preoperative, the early postoperative periods and during the last follow-up.

Results

The mean sagittal vertical axis was found as 190.5 (range 161–220) mm in the preoperative period, 23.5 (range ?27 to 61) mm in the early postoperative period (P < 0.001) (87.7 % correction) and 34.5 (range ?3 to 55) mm during the last follow-up (P < 0.001). The mean pelvic tilt (PT) significantly decreased from 38.3° (range 21°–63°) preoperatively to 23.8° (range 18°–42°) postoperatively (P = 0.008) and to 27.5° (range 17°–42°) during the last follow-up (P = 0.042). The mean lumbar lordosis (LL) was 2.8° (range ?29° to 20°) preoperatively, ?35.6° (range ?54° to 23°) early postoperatively (P < 0.001) and ?33.6° (range ?52° to 20°) during the last follow-up (P < 0.001). The average amount of bleeding was 5345 (range 2600–7415) ml.

Conclusion

Although a statistically significant correction was obtained, the mean PT and PI–LL value could not be restored in physiological limits during the last follow-up. Thus, two level PSOs performed after computer software (surgimap) assisted preoperative planning failed to correct severe fixed sagittal plane deformities. Besides, this procedure is of possible risks for major complications such as a result of excessive bleeding. We recommend that two level PSOs should be rarely indicated, but preferred as an alternative technique only in the most severe cases.

     

Perinatal trauma with and without loss experiences

Objective: The present study explored differences in mental health between women who experienced a trauma which involved a loss of fetal or infant life compared to women whose trauma did not involve a loss (difficult childbirth). Method: The sample consisted of 144 women (mean age = 31.13) from the UK, USA/Canada, Europe, Australia/New Zealand, who had experienced either stillbirth, neonatal loss, ectopic pregnancy, or traumatic birth with a living infant in the last 4 years. Results: The trauma without loss group reported significantly higher mental health problems than the trauma with loss group (F (1,117) = 4.807, p = .03). This difference was observed in the subtypes of OCD, panic, PTSD and GAD but not for major depression, agoraphobia and social phobia. However, once previous mental health diagnoses were taken into account, differences between trauma groups in terms of mental health scores disappeared, with the exception of PTSD symptoms. Trauma groups also differed in terms of perceived emotional support from significant others. Conclusion: The findings illustrate the need for a change in the focus of support for women’s birth experiences and highlighted previous mental health problems as a risk factor for mental health problems during the perinatal period.     

Determination of carnosic acid in Rosmarinus officinalis L. using square wave voltammetry and electrochemical behavior

A new, fast, sensitive and simple voltammetric method is established for the direct determination of carnosic acid (CA). And the electroreduction of carnosic acid (CA) was studied using electrochemical methods. The number of electrons transferred in electrode mechanisms were calculated for reversible and adsorption-controlled electrochemical reduction of CA at 17 mV versus Ag/AgCl at pH 7.0 in Britton–Robinson buffer (BR) on a hanging mercury drop electrode. Square-wave voltammetry was developed and validated for direct determination of CA. Square-wave parameters were optimized as accumulation potential = 0.0 mV, accumulation time = 5 s, frequency = 50 Hz, pulse amplitude = 50 mV, and staircase step potential = 5 mV. The developed method displays three linear responses from 2 to 9 μM, 10 to 30 and 40 to 90 μM for carnosic acid with a correlation coefficient of 0.996, 0.999 and 0.999. The detection limits were found to be 1.5 μM, 4.0 μM and 40.1 μM, respectively. The interference effect of most common organic and inorganic species was investigated. Proposed method was successfully applied for determination of CA in natural extract of rosemary and the average content was determined as 11.9 ± 1.0 (μg CA/1 g rosemary). The results were in agreement with that obtained by HPLC-UV comparison method. The developed method can be widely used in routine quality control of herbal materials as well as other in foods, medicinal, pharmaceutical and environmental analysis.     

过表达TrxR1重组HEK293细胞株的构建和鉴定

目的 构建稳定过表达硫氧还蛋白还原酶1（TrxR1）的HEK293细胞株,为TrxR1的功能研究以及靶向TrxR1药物筛选提供细胞模型。方法 通过PCR扩增,连接转化以及Sanger双脱氧测序构建并筛选出TrxR1的重组慢病毒表达载体pLVX-PuroTXNRD1,转染HEK293细胞,经嘌呤霉素筛选获得稳定转染细胞株。后续研究分为3组进行：①TrxR1过表达HEK293细胞：pLVX-Puro-TXNRD1载体稳定转染细胞;②对照HEK293细胞：pLVX-Puro空载病毒载体稳定转染细胞;③正常HEK293细胞;通过RT-qPCR、Western blot实验检测上述3组细胞中TrxR1的mRNA以及蛋白表达情况;通过胰岛素终点法以及TRFS-green探针成像检测上述3种细胞内TrxR1的酶活力;通过CCK8实验检测上述3种细胞对TrxR1特异性抑制剂auranofin的敏感性。结果 构建载体经DNA测序,成功获得插入TrxR1的重组慢病毒表达载体pLVX-Puro-TXNRD1。与HEK293以及HEK293-NC细胞相比,HEK293-TrxR1-OE细胞中TrxR1的mRNA和蛋白高表达,且酶活力也同样显著上升（P<0.005）;而与HEK293以及HEK293-NC细胞相比,auranofin对HEK293-TrxR1-OE细胞中TrxR1酶活力以及细胞增殖的抑制效率均显著下降（P<0.005）。结论 通过构建pLVX-Puro-TXNRD1慢病毒载体,成功获得过表达TrxR1酶的HEK293细胞株,该细胞对特异性靶向TrxR1的抑制剂的抗增殖作用具有抵抗,因而可用于靶向TrxR1药物的筛选。