中华长城椎弓根螺钉系统对胸腰椎骨折后脊髓功能的恢复作用

目的观察中华长城椎弓根螺钉系统(ChinaGreat-WallSpinaSystem)在胸腰椎爆裂性骨折中的应用效果,以探讨其对骨折后脊髓功能的恢复作用。方法采用中华长城椎弓根螺钉系统治疗胸腰椎爆裂性骨折18例,并用X线检查评估Cobb角、椎体成角、上下终板成角椎体前缘高度。结果术后及随访期间拍X片测定Cobb角(由术前平均7.8°到术后平均1.3°)、椎体成角(由术前平均-17.2°到术后平均-3.2°)、上下终板成角(由术前平均-15.6°到术后平均3.9°)、椎体前缘高度与正常高度的比值均明显改善,随访期间测量以上结果与术后相比无明显变化。最后一次随访按Frankel分级评估,4例A级Frankel分级无改变,其余均改善1～2级。3例C级和4例D级恢复到E级,2例B级和3例C级恢复到D级,1例B级恢复到C级,1例A级恢复到级,4例A级Frankel分级无改变,但感觉平面有所下降。结论中华长城椎弓根螺钉系统(CGWS)能有效减轻胸腰骨折后的脊椎移位,促进脊髓功能的恢复。     

定量RT-PCR方法检测衰老大鼠肾脏PAI-1基因的表达

目的采用SYBR 绿色荧光染料Ⅰ(SYBR GREENⅠ)定量PCR法,对来源珍贵的微量组织中的基因进行检测,并对其可行性及适用性进行研究. 方法取24月龄的老年大鼠、3月龄青年大鼠各6只,提取其肾脏组织的RNA, 采用SYBR GREEN I实时检测的逆转录-聚合酶链反应(RT-PCR)检测纤溶酶系活化剂抑制物-1(PAI-1)基因的表达,并用磷酸甘油醛脱氢酶(G3PDH)作为外参. 结果标准曲线有良好的相关性(R2>0.99),PCR产物特异,将PAI-1的拷贝数与G3PDH的拷贝数相除,青年大鼠肾脏PAI-1表达水平为(1.27±0.90)×10-3,老年大鼠肾脏PAI-1表达水平为(3.84±2.00)×10-3,两者比较,差异有显著性(P<0.05). 结论只要严格控制PCR反应条件,SYBR GREENⅠ定量PCR法可以作为一种良好的定量PCR方法对来源珍贵的微量组织的基因表达进行检测,并证实了老年大鼠肾脏PAI-1表达水平明显增加.     

Clinical outcomes and ergonomics analysis of three laparoscopic techniques for Hirschsprung's disease

AIM: To report the clinical outcomes and ergonomics analysis of three laparoscopic approaches in the management of Hirschsprung's disease(HD).METHODS: There were 90 pediatric patients(63 boys, 27 girls; mean age: 3.6 ± 2.7 mo; range: 1.0-90.2 mo) who underwent laparoscopic endorectal pull-through Soave procedures for short- and long-segment HD in our hospital. Three laparoscopic approaches were used: conventional laparoscopic pull-through(CLP) in 30 patients between 2009 and 2013, single-incision laparoscopic pull-through(SILP) in 28 patients between 2010 and 2013, and hybrid single-incision laparoscopic pull-through(H-SILP) in 32 patients between 2011 and 2013. We applied the hybrid version of the single-incision approach in 2011 to preserve the cosmetic advantage of SILP and the ergonomic advantage of CLP. We retrospectively analyzed the clinical data, cosmetic results, and ergonomics of these three approaches to have a better understanding of the selection of one approach over another. RESULTS: The CLP, SILP, and H-SILP groups were similar in regard to age, sex, transition zone, blood loss, hospital stay, and intraoperative complications. Early and late postoperative results were not different, with equal daily defecation frequency and postoperative complications. No conversion to open technique was needed and none of the patients had recurrent constipation. With proper training, the ergonomics challenges were overcome and similar operative times were registered for the general operative time in the patients 1 year of age and the short-segment HD patients. However, significantly shorter operative times were registered compared to SILP for patients 1 year of age(CLP and H-SILP: 120 ± 15 min and 119 ± 12 min, respectively, vs 140 ± 7 min; P 0.05) and for long-segment HD patients(152 ± 3.5 min and 154 ± 3.6 min, respectively, vs 176 ± 2.3 min; P 0.05). The best cosmetic result was registered with the SILP(scarless), followed by the H-SILP(near scarless appearance) and the CLP(visible scars) procedures. CONCLUSION: Based on the results, we believed that the laparoscopic approach should be selected according to the age, transition zone, and desired cosmetic result.     

Ataxia telangiectasia-mutated-Rad3-related DNA damage checkpoint signaling pathway triggered by hepatitis B virus infection

AIM： To explore whether acute cellular DNA damage response is induced upon hepatitis B virus （HBV） infection and the effects of the HBV infection. METHODS： We incubated HL7702 hepatocytes with HBV-positive serum, mimicking a natural HBV infection process. We used immunoblotting to evaluate protein expression levels in HBV-infected cells or in non-infected cells; immunofluorescence to show ATR foci ands Chkl phosphorylation foci formation; flow cytometry to analyze the cell cycle and apoptosis; ultraviolet （UV） radiation and ionizing radiation （IR）-treated cells to mimic DNA damage; and Trypan blue staining to count the viable cells. RESULTS： We found that HBV infection induced an increased steady state of ATR protein and increased phosphorylation of multiple downstream targets including Chkl, p53 and H2AX. In contrast to ATR and its target, the phosphorylated form of ATM at Ser-1981 and its downstream substrate Chk2 phosphorylation at Thr-68 did not visibly increase upon infection. However, the level of Mre11 and p21 were reduced beginning at 0.5 h aEer HBV-positive serum addition. Also, HBV infection led to transient cell cycle arrest in the S and the G2 phases without accompanying increasedapoptosis. Research on cell survival changes upon radiation following HBV infection showed that survival of UV-treated host cells was greatly increased by HBV infection, owing to the reduced apoptosis. Meanwhile, survival of IR-treated host cells was reduced by HBV infection. CONCLUSION： HBV infection activates ATR DNA damage response to replication stress and abrogates the checkpoint signaling controlled by DNA damage response.     

损伤控制性手术救治严重胸部创伤患者的护理

目的探讨损伤控制性手术救治严重胸部创伤患者的护理方法和效果。方法对98例严重胸部创伤患者采取损伤控制性手术救治,术前严密动态观察病情并评估伤情,遵循严重创伤VIPCO(通气、输液、搏动、控制出血、手术)救护流程行重症监护治疗;术后进一步复温,加强器官功能监护,重视胸肺物理治疗,加强围术期气道管理,积极预防肺部并发症。结果治愈出院89例,家属放弃治疗自动出院2例,死亡7例,救治成功率为90.82%。结论有效的护理措施对改善严重胸部创伤患者的预后,提高救治成功率发挥了至关重要的作用。     

利用单纯形法对氯霉素及其分解产物GLC分离条件的优化

本文以色谱响应函数CRF_(RT)作为色谱分离性能的参考尺度,利用单纯形法对氯霉素及其分解产物的GLC分析条件进行了优化。通过9次试验,找到了以2%SE-30 Chromosrb W Aw-Dmcs 80-100目为固定相时满意的分离条件:柱温为245℃,载气流速为56ml/min。在保证峰的良好分离情况下使分析时间由原来的13.5分缩短至5.09分。本实验表明单纯形法是药物色谱分离最优化中一种简单有效的方法。     

椎间盘置换术治疗腰椎间盘突出症

目的 探讨人工椎间盘置换术治疗腰椎间盘突出症的临床效果。方法 对6例腰椎间盘突出症患者用Link SB Charite Ⅲ型人工腰椎间盘行椎间盘置换术,共置换假体6例。手术节段为L_(4、5)4例,L_5S_12例。结果 随访3～9个月,平均6个月,术中和术后未出现并发症,术后X线片显示人工椎间盘位置正确,椎间隙高度恢复正常,5例患者腰腿痛症状完全消失,直腿抬高试验阴性,1例劳累后仍感腰痛,6例均恢复工作和日常生活能力。临床评价:优5例,良1例。结论 人工椎间盘置换术是治疗椎间盘突出症的有效方法,近期效果满意,但应严格掌握手术适应证。     

血清胱抑素C在糖尿病肾病早期诊断中的临床意义

目的:探讨血清胱抑素C(Cys C)在早期糖尿病肾病患者中水平变化及其诊断价值。方法:选择130例糖尿病(DM)患者,其中单纯糖尿病组68例,早期糖尿病肾病(DN)组62例,并选择65例健康体检者作为对照。检测3组血清肌酐(Scr)、尿素氮(BUN)及Cys C水平并进行统计学分析。结果:早期DN组中Cys C水平明显高于单纯DM组及健康对照组(P<0.01),但单纯DM组和早期DN组之间Scr、BUN水平差异无统计学意义(P>0.05);早期DN组中Cys C阳性检出率为58.04%,明显高于Scr的22.58%(P<0.01)和BUN的19.35%(P<0.01);早期DN组中CysC与Scr、BUN均呈高度正相关(P<0.01)。结论:血清Cys C是诊断早期糖尿病肾病的敏感指标。     

准分子激光原位角膜磨镶术与准分子激光角膜上皮瓣下磨镶术比较治疗近视效果的系统评价

目的 系统评价准分子激光原位角膜磨镶术(LASIK)与准分子激光角膜上皮瓣下磨镶术(LASEK)比较治疗近视的疗效与安全性.方法 计算机检索PubMed、Embase、Cochrane图书馆(2010年2期)、CNKI、VIP 及万方数据库(1990.01～2010.10),检索语种限制为中、英文,纳入LASIK对比L...     

VEGF受体单抗与Luffin双靶融合毒素的构建及其对非小细胞肺癌细胞的抑制作用

目的构建含血管内皮生长因子(vascular endothelial growth factor,VEGF)受体单链抗体(KDRscFv)、尿激酶纤溶酶原激活剂(urokinase plasminogen activator,uPA)裂解位点(uPAcs)、丝瓜毒素(luffin-β)与KDEL(Lys-Asp-Glu-Leu)内质网驻留信号序列的双靶向融合毒素,并探讨其对非小细胞肺癌(non-small-cell lung carcinoma,NSCLC)细胞的抑制作用。方法全基因合成KDRscFv基因,RT-PCR法克隆luffin-β,重叠PCR法将两基因融合,其连接部位与C端分别引入uPA裂解位点uPAcs与KDEL,形成融合基因KDRscFv-uPAcs-Luffin-β-KDEL。将该融合基因克隆至原核表达载体pET-32a(+)中,转入大肠杆菌后,诱导其表达融合毒素蛋白Trx-EK-KDRscFv-uPAcs-Luffin-β-KDEL(TEKPLK)并纯化TEKPLK。用肠激酶(enterokinase,EK)切割TEKPLK后,纯化与回收靶向融合毒素KDRscFv-uPAcs-Luffin-β-KDEL(KPLK)。采用CCK-8(cell counting kit-8)、RT-PCR、Western blot等方法,体外检测靶向融合毒素KPLK经uPA酶裂解后释放Luffin-β对NSCLC细胞的抑制作用。结果成功诱导重组载体pET-32a(+)/KDRscFv-uPAcs-Luffin-β-KDEL表达相对分子质量约7.5×104含载体表达标签(Trx)的融合毒素蛋白TEKPLK,EK酶切该蛋白获得相对分子质量约5.8×104的靶向融合免疫毒素KPLK。CCK-8法检测表明,KPLK毒素的杀瘤活性成剂量-效应关系,其IC50约为35 ng/mL;RT-PCR和Western blot检测结果显示,KPLK经uPA酶体外裂解后能释放细胞毒素小分子Luffin-β,上调瘤细胞促凋亡基因caspase-3及其蛋白的表达。结论成功构建了KDRscFv-uPAcs-Luffin-β-KDEL融合基因及其原核表达载体,并获得相对分子质量约5.8×104的靶向融合毒素KPLK,该毒素经uPA酶体外裂解后能释放具杀瘤活性的Luffin-β毒素小分子。