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Research on Screening Peptides Specifically Targeting Laryngeal Squamous Cell Carcinoma by Phage Display Technique

【摘要】　目的　篩選人源喉癌Hep-2細胞株特異結合的短肽，作為喉癌靶向治療的載體。　方法　體外培養Hep-2細胞株作為靶細胞，人正常喉黏膜上皮細胞為吸附細胞；用噬菌體展示十二肽庫進行3輪差減篩選，隨機挑取10個噬菌體克隆進行測序；采用酶聯免疫吸附（enzyme linked immunosorbent assay，ELISA）法鑒定噬菌體與Hep-2細胞的結合活性；通過免疫熒光鑒定喉癌細胞特異性結合肽（F2）噬菌體陽性克隆與喉癌細胞結合的特異性。　結果　經過3輪篩選后，噬菌體在靶細胞Hep-2上出現明顯富集；ELISA分析鑒定顯示5個陽性克隆能與Hep-2細胞特異結合，其中F2噬菌體克隆對喉癌細胞的結合靶向性明顯高于對照細胞（Plt;0.05）； 免疫熒光顯色顯示，F2能特異性地與喉癌細胞結合。　結論　利用噬菌體展示肽庫技術，可以成功篩選到F2，其可能成為喉癌靶向治療的載體。【Abstract】　Objective　To obtain the polypeptides specifically bound to laryngeal squamous cell carcinoma line (Hep-2) and use it as a potential therapeutic vector targeting laryngeal squamous cell carcinoma patients.　Methods　With the Hep-2 cells as the target cells and human normal laryngeal squamous epithelial cells (HNLE cells) as the absorber cells, 3 rounds of panning from a Ph.D.-12TM phage-display peptide library were carried out. Ten randomly selected phage clones were sent for sequence detection. The affinity of phage clones was detected by enzyme-linked immunosorbent assay (ELISA). The positive phage clones (F2) specifically bound to Hep-2 were identified by immunofluorescence detection.　Results　After 3 rounds of screening, 5 positive phage clones showed specific binding to Hep-2 cells and the affinity of positive phage clones (F2) was significantly higher than that of the control groups (Plt;0.05). The results of immunofluorescence detection indicated that F2 could be specifically bound to Hep-2.　Conclusions　Phage display peptide libraries technique can successfully screen the peptide specifically bound to Hep-2 cell line. Thus, it provides a potential vector for targeting therapy of laryngeal squamous cell carcinoma patients.

喉腫瘤切除會厭重建喉前壁22例

Effect of Down-regulated ATM Expression on Radiosensitivity for Laryngeal Squamous Cell Carcinoma in Nude Mice

目的　探討反義ATM多核苷酸（AS-Lipo）作用于裸鼠移植瘤對喉鱗狀細胞癌(Hep-2)放射治療敏感性的影響。 方法　用克隆生存率來檢測喉鱗狀細胞癌放射治療后的生存能力；用流式細胞儀來分析喉癌細胞的凋亡情況；用Tunel染色法檢測各組瘤體中細胞凋亡情況。 結果　在同等劑量的放射治療下，AS-Lipo組的生存分數最低。接受4 Gy放射治療后，行流式細胞儀檢測，結果發現AS-Lipo組的凋亡率為（30.7 ± 1.31）%，遠高于轉染Sen-Lipo、Mis-Lipo、Lipo組。ATM AS-Lipo+放療組的細胞凋亡數顯著高于單獨放射治療組；ATM AS-Lipo+放射治療組裸鼠瘤體內的凋亡指數為（17.12 ± 4.2）%，與其他組比較差異有統計學意義。 結論　反義ATM多核苷酸增強了裸鼠喉鱗狀細胞癌對放射治療的敏感性。

Comparison of Laryngeal Tube Suction II and LMAProseal for Airway Management in Elective Surgery: Asystematic Review

摘要:目的:評價II代引流型喉管(LTS II)和Proseal喉罩(PLMA)在擇期手術中應用效果。方法:檢索了Cochrane圖書館(2009年第3期)、Pubmed(1950～2009)、EMBase(1989～2009)、CNKI(1979～2009)、VIP(1989～2009)、CBM(1978～2009)中相關II代引流型喉管(LTS II)和Proseal喉罩在擇期手術中應用的隨機對照試驗(RCT),同時篩檢納入文獻的參考文獻。由2名研究者對文獻質量進行嚴格評價和資料提取,根據指標相應異質性進行描述性分析或Meta分析(RevMan 5.0)。結果:共納入3個RCT,共244例研究對象,文獻質量均為B級。3個RCT的結果顯示與PLMA相比,LTS II具有相似的首次置入成功率(P=0.45)、術畢即刻上呼吸道損傷發生率(P=1.00)、術后24 h咽痛發生率(P=0.81)、術后24 h吞咽困難發生率(P=0.12)。2個RCT的結果顯示兩組引流管置入均較容易。1個RCT的結果顯示兩組的操作者主觀評價相近(OR=1.86,95%CI 0.39～ 8.99)。氣道封閉效果由于采用方法學差異性較大,指標也不盡相同,尚不能得出準確結果。結論:LTS II在擇期手術中用于氣道管理具有較好的前景。但是現時仍不宜用于需在擇期術中進行控制通氣的病人。關于氣道封閉效果,尚需采用更合理規范的指標、更高質量的研究設計進一步研究。Abstract: Objective: To assess the efficacy of laryngeal tube suction II (LTS II) and LMAProseal (PLMA) for airway management in elective surgery. Methods:We searched Cochrane Library (2009),Pubmed (19502009)、EMBase (19892009),CNKI (19792009),VIP (19892009),CBM (19782009). The quality of the trials was assessed by two reviewers independently. RevMan 5.0 software provided by the Cochrane Collaboration was used for statistical analysis. Results:Three studies involving 244 participants were included. Same rates of fist successful attempt (P=0.45),upper airway trauma (P=1.00),sore throat (P=0.81) and dysphagia (P=0.12) were observed in LTS II and PLMA in all studies. Two studies indicated that the insertion of gastric tube was easy in both groups. The similarity of subjective maneuverability in two groups was reported in one study (OR=1.86, 95%CI 0.39 to 8.99). The correct result of effectiveness of airway seal could not be made because of various methods and measurements. Conclusion:LTS II have a good perspective in the airway management. Otherwise, it is not safe for patient required control ventilation because of lack of evidence on the effectiveness of airway seal. More RCTs of high quality need to be undertaken in the future.

The efficacy and safety of laryngeal mask versus endotracheal tubes for laparoscopic surgery: a meta-analysis

ObjectiveTo systematically review the efficacy and safety of laryngeal mask versus endotracheal tubes for laparoscopic surgery.MethodsPubMed, EMbase, The Cochrane Library, CNKI, WanFang Data and CBM databases were electronically searched to collect the randomized controlled trials (RCTs) about the efficacy and safety of laryngeal mask versus endotracheal tubes for laparoscopic surgery from inception to April, 2017. Two reviewers independently screened literature, extracted data and assessed the risk of bias of included studies. Then meta-analysis was performed by using RevMan 5.3 software.ResultsA total of 16 RCTs involving 1 593 patients were included. The results of meta-analysis showed that: there was no significant difference in the success rate of the first insertion (RR=0.99, 95%CI 0.96 to 1.02, P=0.55). The airway pressure of patients whose position were head higher than foot was significantly lower in the laryngeal mask group than in the tracheal intubation group (MD=–1.20, 95%CI –1.81 to –0.59, P=0.000 1), but there was no significant difference between two groups in reverse position patients (MD=0.48, 95%CI –0.90 to 1.87, P=0.49). The incidence of sore throat (RR=0.58, 95%CI 0.46 to 0.74, P<0.000 01), the incidence of blood stain (RR=0.48, 95%CI 0.30 to 0.77, P=0.002), the incidence of laryngeal spasm/bronchial spasm (OR=0.30, 95%CI 0.11 to 0.80, P=0.02) and the incidence of cough/hiccup (RR=0.10, 95%CI 0.07 to 0.15, P<0.000 01) in the laryngeal mask group were significantly lower than those in the tracheal intubation group.ConclusionThe current evidence shows that compared with tracheal intubation, laryngeal mask can effectively reduce airway pressure of patients whose position are head higher than foot. The risks of various complications are significant higher in tracheal intubation in laparoscopic surgery. Laryngeal mask can maintain patients' normal respiratory functions while reduce damage and do not increase the occurrence of reflux aspiration. Due to limited quantity and quality of the included studies, more high quality studies are needed to verify above conclusion.

The Indications and Safety Assessment of Total Thyroidectomy

Objective　To investigate the indications and prevention of complications of total thyroidectomy in the management of thyroid diseases. Methods　Eighty five patients who received total thyroidectomy between Jan. 2009 and Dec. 2011 were retrospectively analyzed with regard to the surgical procedures and postoperative complications. There were 46 thyroid cancers， 38 nodular goiters， and 1 Hashimoto thyroiditis. Results　The postoperative pathological exam-inations revealed that 9 （19.6%） of 46 thyroid cancers were bilateral， and all of nodular goiters were also bilateral multiple nodule. Bilateral recurrent laryngeal nerves were exposed in all of the patients in which 4 recurrent laryngeal nerves were invaded by cancer and 1 was sacrificed. There were 5 patients whose parathyroids were not identified and protected during the operation. Two patients developed postoperative bleeding and needed reoperation， 6 patients developed hoarseness of whom 5 patients recovered except for the one whose nerve was sacrificed. And in terms of hypoparathyroidism， 33 （38.8%） patients developed transient hypocalcemia related symptoms. The permanent hypoparathyroidism occurred only in 2 patients. Conclusions　Total thyroidectomy is a safe procedure in the management of thyroid cancer and bilateral nodular goiter. Exposing the recurrent laryngeal nerve and parathyroid is an effective method to prevent major complica-tions. Invasion of recurrent laryngeal nerve by thyroid cancer might not lead to hoarseness.

Lentivirus-mediated siRNA Targeting Cyclooxygenase-2 Gene Inhibits Human Laryngocarcinoma Cells Proliferation and Invasion

目的　構建沉默環氧化酶-2（COX-2）基因重組慢病毒，觀察其體外侵襲的抑制作用，從而探討干擾COX-2抑制喉癌細胞增殖的作用機理，為喉癌的治療提供新的思路。 方法　逆轉錄聚合酶鏈反應（RT-PCR）檢測COX-2基因在人表皮樣喉癌細胞（Hep-2）中的表達情況。利用上海吉凱公司RNA干擾（RNAi）慢病毒表達載體系統，構建針對COX-2基因慢病毒RNAi表達載體。轉染Hep-2細胞，干擾COX-2基因的表達，實時定量PCR檢測干擾前后基因表達變化。利用生長曲線測定干擾載體轉染前后細胞生長速度變化。流式細胞儀檢測細胞的生長周期。Boyden侵襲小室法測定體外侵襲力。 結果　成功構建了COX-2慢病毒RNAi表達載體，并建立了干擾COX-2基因的Hep-2細胞系。實時定量PCR檢測COX-2基因在Hep-2細胞系中過表達被顯著抑制。生長曲線測定，COX-2基因干擾后細胞增殖明顯變慢。流式細胞儀檢測細胞的生長周期可見干擾組誘導Hep-2細胞凋亡，轉染G0～G1期細胞數量明顯上升，S期細胞減少，表明siRNA干擾Hep-2細胞后，細胞由G0～G1期進入到S期受到阻滯，細胞增殖速度下降。體外侵襲實驗中，Hep-2-AS侵襲細胞數（31.0 ± 1.8）顯著低于Hep-2細胞（104.0 ± 2.6）及Hep-2-P細胞（99.0 ± 2.7），差異有統計學意義（P＜0.05）。 結論　喉癌中過表達的COX-2基因被干擾后表達明顯降低并顯著抑制細胞的生長速度和侵襲能力。同時驗證了COX-2基因RNA干擾在進行抗腫瘤的治療中潛在的應用前景。

SLIPATM 喉罩全身麻醉期間誤吸致肺不張一例

Treatment of Larynxderived Cough with Modified Zhisousan

目的：觀察古方止嗽散加味治療喉源性咳嗽的療效。方法：采用加味止嗽散治療喉源性咳嗽116例，并設西藥對照組113例。結果：治療組有效率87.3%，對照組有效率68.2%，兩組比較有顯著差異(Plt;0.01)。治療組不良反應例數為零；對照組不良反應例數約占10.2%。結論：加味止嗽散具有疏風宣肺、清熱解毒、利咽祛痰、益氣養陰、扶正祛邪、抗病毒、抗菌、抗炎和增強免疫功能等功效，為治療喉源性咳嗽的有效方藥。

自護技能指導前移對提高喉癌術后患者自護效果的影響

目的　討論自護技能指導前移對提高喉癌術后患者出院后自護能力的效果，尋求切實有效的健康教育方法。 方法　將2010年6月－2011年3月接受喉癌手術的113例患者按病床序號分為對照組54例和觀察組59例，對照組按常規在患者出院前2～3 d開始進行自護技能指導，觀察組則將指導時間提前至術后第5天，兩組自護技能指導內容相同。并于患者出院前1 d和出院后15 d對其自護技能及家庭護理效果進行評價。 結果　觀察組患者自護技能得分水平出院前后都明顯高于對照組（P＜0.05）；出院后在套管墊安置正確、能自行排痰、掌握保護氣道方法3個方面家庭護理效果優于對照組（P＜0.05）。 結論　自護技能指導前移更能提高喉癌術后患者出院后的自護能力水平，減輕患者及家屬對出院的焦慮感，促進家庭護理效果。