脑外伤合并四肢骨折骨痂局部转化生长因子-β基因表达的实验研究

目的 通过对SD大鼠脑外伤合并四肢骨折不同时间段骨痂局部转化生长因子(TGF-β)的基因表达进行检测,了解TGF-β在增强骨折愈合过程中的作用。方法 纯种SD雄性大鼠80只,随机分为（每组40只）单纯四肢骨折组（对照组）与脑外伤合并四肢骨折组（实验组）（每组40只）,两组均利用逆转录PCR基因扩增技术对骨折后l、2、3、4周骨痂局部TGF-β基因表达的改变进行检测。结果 与对照组相比,实验组在骨折后1周TGF-β的基因表达明显升高,差异有统计学意义(P＜0.05);在骨折后3周其表达明显下降,差异有统计学意义(P＜0.05);两组TGF-β基因表达均在骨折后2周达到峰值,峰值与其他时间段比较差异均有统计学意义(P＜0.05),但两组间峰值比较差异无统计学意义(P＞0.05)。结论 脑外伤合并四肢骨折骨痂局部TGF-β基因表达模式的改变表明,TGF-β基因参与了增强的骨折愈合病理生理过程。     

Cyclooxygenase-2 Inhibition Delays the Attainment of Peak Woven Bone Formation following Four-Point Bending in the Rat

Fracture healing is retarded in the presence of cyclooxygenase-2 (COX-2) inhibitors, demonstrating an important role of COX-2 in trauma-induced woven bone adaptation. The aim of this experiment was to determine the influence of COX-2 inhibition on the remodeling and consolidation of nontraumatic woven bone produced by mechanical loading. A periosteal woven bone callus was initiated in the right tibia of female Wistar rats following a single bout of four-point bending, applied as a haversine wave for 300 cycles at a frequency of 2 Hz and a magnitude of 65 N. Daily injections of vehicle (VEH, polyethylene glycol) or the COX-2 inhibitor 5,5-dimethyl-3-3(3 fluorophenyl)-4-(4-methylsulfonal)phenyl-2(5H)-furanone (DFU, 2.0 mg · kg⁻¹ and 0.02 mg · kg⁻¹ i.p.), commenced 7 days postloading, and tibiae were examined 2, 3, 4, and 5 weeks postloading. Tibiae were dissected, embedded in polymethylmethacrylate, and sectioned for histomorphometric analysis of periosteal woven bone. No significant difference in peak woven bone area was observed between DFU-treated and VEH rats. However, treatment with DFU resulted in a temporal defect in woven bone formation, where the achievement of peak woven bone area was delayed by 1 week. Woven bone remodeling was observed in DFU-treated rats at 21 days postloading, demonstrating that remodeling of the periosteal callus is not prevented in the presence of a COX-2 inhibitor in the rat. We conclude that COX-2 inhibition does not significantly disrupt the mechanism of woven bone remodeling but alters its timing.     

Long-term effects of alendronate on fracture healing and bone remodeling of femoral shaft in ovariectomized rats

Aim:

To investigate the long-term effects of alendronate (Aln), a widely used oral bisphosphonate, on fracture healing and bone remodeling in ovariectomized rats.

Methods:

Adult female SD rats underwent ovariectomy, and then bilateral femoral osteotomy at 12 weeks post-ovariectomy. From d 2 post-ovariectomy, the animals were divided into 3 groups, and treated with Aln (3 mg·kg⁻¹·d⁻¹, po) for 28 weeks (Aln/Aln), Aln for 12 weeks and saline for 16 weeks (Aln/Saline) or saline for 28 weeks (Saline/Saline). At 6 and 16 weeks post-fracture, the fracture calluses were examined with X-ray radiography, and biomechanical testing and histological analysis were performed. The calluses were labeled with tetracycline and calcein to evaluate the mineral apposition rate (MAR).

Results:

The fracture line was less distinct in the 2 Aln-treated groups at 6 weeks post-fracture, and disappeared in all the 3 groups at 16 weeks post-fracture. The size of the callus and radiographic density of the femora in the Aln/Aln group were the highest among the 3 groups at 6 and 16 weeks post-fracture. Similar results were observed in the ultimate load at failure and energy absorption. However, the treatment with Aln delayed endochondral ossification of the callus, and significantly increased the total sagittal-sectional area, percentage callus area and callus thickness, and decreased the MAR at 6 and 16 weeks post-fracture.

Conclusion:

In the ovariectomized rat model, Aln is beneficial for the mechanical properties of the callus, but delays callus remodeling by suppressing the remodeling of woven bone into lamellar bone.     

茉莉酸甲酯诱导的白木香cDNA文库的构建及初步鉴定

目的 通过构建茉莉酸甲酯（MeJA）处理的白木香cDNA文库,为揭示伤害诱导沉香倍半萜合成的分子机制奠定基础。方法 以MeJA处理的白木香愈伤组织为材料,以改造的pGADT为载体,利用SMART技术构建白木香全长cDNA文库。结果 构建的文库原始滴度为8.99×10⁶ pfu/mL,阳性克隆子的比例为97%,插入片段的大小在500～3 000 bp,平均大于1 000 bp。结论 构建的文库无论从库容量、克隆效率,还是全长率方面评价,均达到了高质量文库的要求。     

不同激素配比对远志愈伤诱导及其黄酮积累的影响

目的:研究不同激素配比对远志根、茎、叶愈伤组织诱导的影响,并对远志根、茎、叶愈伤组织中的黄酮量进行测定分析。方法:以MS为基本培养基,分别以远志无菌苗的根、茎、叶为外植体,应用正交试验法确定2,4-二氯苯氧乙酸(2,4-D),萘乙酸(NAA),6-苄氨基嘌呤(6-BA)这3种激素对远志根、茎、叶不同部位愈伤组织诱导及其黄酮积累量的影响。结果:2,4-D,NAA,6-BA对远志根、茎、叶愈伤诱导率均有显著影响。叶的最佳愈伤诱导组合为MS+3. 0 mg·L~(-1)2,4-D+1. 0 mg·L~(-1)NAA+1. 5 mg·L~(-1)6-BA;茎的最佳愈伤诱导组合为MS+1. 0 mg·L~(-1)2,4-D+3. 0 mg·L~(-1)NAA+1. 5 mg·L~(-1)6-BA;根的最佳愈伤诱导组合为MS+1. 0 mg·L~(-1)2,4-D+1. 0 mg·L~(-1)NAA+1. 0 mg·L~(-1)6-BA。2,4-D,NAA,6-BA对远志茎愈伤组织中黄酮积累均影响显著,以MS+3. 0 mg·L~(-1)2,4-D+1. 0 mg·L~(-1)NAA+0. 5 mg·L~(-1)6-BA为最佳黄酮积累组合; NAA,6-BA对远志叶愈伤组织中黄酮积累影响均显著,而2,4-D则无明显影响,以MS+3. 0 mg·L~(-1)2,4-D+2. 0 mg·L~(-1)NAA+1. 0 mg·L~(-1)6-BA为最佳黄酮积累组合; 3种激素分别对远志根愈伤组织中黄酮积累无明显影响,以MS+2. 0 mg·L~(-1)2,4-D+1. 0 mg·L~(-1)NAA+1. 5 mg·L~(-1)6-BA为最佳黄酮积累组合。结论:在该条件下远志根、茎、叶愈伤组织诱导率均达100%,其中尤以远志叶愈伤组织长势最好,其次分别是远志茎和远志根。在该条件下远志根、茎、叶愈伤组织中黄酮量分别达到21. 31,24. 56,23. 61 mg·g-1。     

Noninvasive Quantitative Assessment of Bone Healing After Distraction Osteogenesis

One of the greatest challenges of limb lengthening and deformity correction is deciding when the bone has healed enough to remove the external fixator. Standard radiography is the most common imaging method used to assess bone healing after distraction osteogenesis because it is widely available, cheap, and relatively safe. However, other imaging technologies and methods are being investigated that will help quantify bone healing after distraction osteogenesis, providing an objective method for deciding when it is appropriate to remove an external fixator. This review will examine the latest techniques used to assess bone healing after distraction osteogenesis including dual-energy X-ray absorptiometry scans, ultrasound, quantitative computed tomography, and digital radiography (X-ray). Recommendations for clinical practice will be outlined.     

Anticancer activity of rice callus suspension culture

A multitude of natural products from plant extracts have been tested for their ability to inhibit the progression of several diseases including cancer. A novel approach of evaluating plant (rice) callus suspension cultures for anticancer activity is reported. The ability of different dilutions of rice callus suspension cultures to inhibit growth of two human cancer cell lines was tested employing varying cell numbers and different incubation times. A crystal violet assay was performed to assess cell viability of the cancer cell lines. Furthermore, microscopic analysis was carried out to determine the effect of the rice callus culture on the morphology of the cancer cells. Rice callus suspension cultures significantly inhibited the growth of human cancer and renal cell lines at densities of 5000 and 10000 cells/mL when incubated for 72 and 96 h. Rice callus suspension culture was more efficient than paclitaxel (Taxol®) and etoposide in selectively killing human colon and renal cancer cell lines compared with a control cell line (human lung fibroblasts). The use of plant callus suspension cultures is a novel approach for inhibiting the growth of cancer cells, which will lead to the development of new agents for selectively killing cancer cells. Copyright © 2011 John Wiley & Sons, Ltd.     

不同浓度的外源激素对黄芩愈伤组织的生物量和黄芩苷含量的影响

目的:探索有利于黄芩愈伤组织的诱导和次生代谢物积累的激素组合.方法:在培养基中添加不同浓度的外源激素,测定愈伤组织的生物量及采用高效液相色谱法测定黄芩愈伤组织中黄芩苷的含量,流动相为甲醇-水-磷酸(47∶ 53∶0.2),流速1 mL·min-1,检测波长280 nm,室温.结果:采用6-BA 1.5 mg· L-1 +NAA0.5 mg· L-1诱导培养的黄芩愈伤组织中黄芩苷含量较高,达到49.78 mg·g-1,生物量为29.34 g/瓶.结论:通过筛选适宜的外源激素组合可以促进黄芩愈伤组织生物量的提高和次生代谢物黄芩苷的积累.     

New alternative in treatment of callus

The pathological transformation of the skin into a thick and hard callus due to repetitive trauma or friction is commonly known as corn. Although a variety of medical and operative treatment choices have been proposed, an ideal treatment method is yet to be defined. Effectiveness of tangential excision together with topical cantharidin has been evaluated. We used Canthacur-PS as an adjunct to excision in an outpatient setting. Canthacur-PS is a commercially available topical solution that includes 1% cantharidin, 30% salicylic acid and 5% podophyllin. The treatment has been applied to 72 patients. We found that 65 patients (90.3%) had corn on their feet and seven patients (9.7%) on their hands. Thick, hard and hyperkeratotic skin area was scraped with the help of a no. 15 blade. The solution was applied on and around the periphery (up to 1–2 mm) of the lesion with a cotton swab, and kept closed for 5 days with an antibiotic dressing. All the patients had been followed up for at least 1 year and evaluated by clinical examination and patient satisfaction query. One session of treatment succeeded in 57 (79.2%) corn patients. Two sessions in nine corn patients (12.5%), three sessions in five corn patients (6.9%) and four sessions in one patient (1.4%) were needed. Only one recurrence (1.4%) was seen. No scar formation or other side-effects were seen. Our findings show that this treatment method is a simple, minimally invasive and reliable treatment for calluses.