【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.
ObjectiveTo evaluate the safety and necessity of recurrent laryngeal lymph node resection by comparing the complications and prognosis of patients with recurrent laryngeal nerve injury receiving different recurrent laryngeal lymph node resections.MethodsWe reviewed the clinical data of 153 patients with stage T1N0M0 esophageal squamous cell carcinoma who underwent radical esophageal cancer surgery at the Department of Thoracic Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology from June 2014 to May 2016. Among them, 125 were male and 28 were female, at an average age of 62 years. All patients underwent bilateral recurrent laryngeal nodes sampling. They were divided into 3 groups according to the dissection situation: patients with only one recurrent laryngeal lymph node resection on both sides during the operation were treated as a sampling group (n=49); patients with only one recurrent laryngeal lymph node resection on one side and more than one recurrent laryngeal lymph nodes resection on the other side were treated as a unilateral dissection group (n=49); patients with more than one recurrent laryngeal lymph nodes resection on both sides were treated as a bilateral dissection group (n=55). Follow-up was performed to compare the prognostic differences among the three groups. Seven days after the operation, the vocal cords of the patients were examined with an electronic laryngoscope and classified using the Clavien-Dindo system. The differences in complications related to recurrent laryngeal nerve injury among the three groups were compared.ResultsThe 5-year overall survival (OS) rate of the patients in the sampling group, unilateral dissection group and bilateral dissection group was 66.8%, 88.5%, 93.8%, respectively. There was statistical difference between the sampling group and the unilateral dissection group or the bilateral dissection group (P<0.05), and no statistical difference between the unilateral dissection group and the bilateral dissection group (P>0.05). The incidence of complications among the three groups was not statistically different (P>0.05).ConclusionFor patients with esophageal squamous cell carcinoma of stage T1N0M0, the lymph nodes of the bilateral recurrent laryngeal nerves should be removed during the operation as many as possible, which will help improve the 5-year survival rate of the patients.
摘要:目的: 探討選擇性內皮素A受體拮抗劑BQ123對人喉癌Hep2細胞裸鼠種植瘤的生長及血管形成的影響。 方法 :將實驗動物裸鼠隨機分為3組:BQ123［n =8,2mg/(kg·day)］、氟尿嘧啶組［n =8,2mg/(kg·day)］、生理鹽水組(n =8),比較各組裸鼠成瘤體積、微血管密度(MVD)。 結果 :BQ123組腫瘤體積為(162±053)cm3,明顯小于生理鹽水組及氟尿嘧啶組,差異具有統計學意義;BQ123組的腫瘤組織中MVD高倍鏡下為232,明顯低于生理鹽水組(586)及氟尿嘧啶組(395),差異具有統計學意義。 結論 :BQ123對人喉癌Hep2細胞在裸鼠體內有明顯抑瘤作用,腫瘤的體積、腫瘤組織MVD顯著低于對照組,表明BQ123可通過抑制腫瘤血管生成而顯著抑制腫瘤生長。Abstract: Objective: To study the effects of endothelin A receptor blockade BQ123 on the implanted human laryngeal carcinoma angiogenesis of nude mouse. Methods : From March 2008 to July 2009, 24 Balb/c nude mice were randomly divided into three groups: BQ123 group ［〖WTBX〗n =8, BQ123 at 2mg/(kg·day)］, 5Fu group ［〖WTBX〗n =8, fluorouracil at 2mg/(kg·day)］ and the control group (〖WTBX〗n =8, normal saline). The carcinoma volume and microvascular density of each group were compared. Results : The tumor size of BQ123 group, which was (162±053)cm3 in average, was significant smaller than the tumor sizes of the other two group s. The average microvascular density score of the tumors in BQ123 group was 232 per hyper power len (HP), which was also significantly less than the average scores of control groups (586 and 395 respectively). Conclusion : Nude mouse experiments show that the carcinoma volume and microvascular density of BQ123 group are significantly lower than those of the control groups. BQ123 inhibits the growth of carcinoma by its inhibition of carcinoma angiogenesis.
Objective To explore the clinical significance on protection of parathyroid and recurrent laryngeal nerve （RLN） by meticulous capsular dissection and exposure of RLN in thyroid lobectomy. Methods Clinical data of 452 patients who underwent thyroid lobectomy by meticulous capsular dissection and exposure of RLN in our hospital from Jan. 2010 to Dec. 2012 were retrospectively analyzed. Results All of the 452 cases underwent thyroid lebectomy successfully without death， and the operative time was 45-110min （average 60 min）， the blood loss was 5-100mL （average 20mL）. The pathological results showed that there were thyroid adenoma in 193 cases， nodular goiter in 175 cases， Hashimoto thyroiditis in 38 cases， thyroid cancer in 46 cases. After operation， 4 cases suffered RLN injury， 1 of the 4 cases recovered after removal of drainage tube， and other 3 cases recovered during 0.5 to 3.0 months. In addition， 2 cases suffered laryngeal nerve injury whose symptoms disappearred within 1 week， 5 cases suffered parathyroid founctional damage without permanent hypocalcemia whose symptom had kept 1-5 days after treatment. Three cases were reoperated because of bleeding， including branch of anterior venous bleeding in 1 case， thyroid side arterial tube bleeding in 1 case， and thyroid stump bleeding in 1 case. Twenty one cases suffered hypothyroidism in 1 month after operation， and no recurrence happened during the followed up period. Conclusions Meticulous capsular dissection can effectively protect function of parathyroid and reduce the injury probability of RLN. Exposure of RLN is safe and feasible， which plays an important role in avoiding serious RLN injury.
目的 研究人喉表皮癌細胞系Hep-2中乳腺癌易感基因1（BRCAl）、P53結合蛋白1（53BP1）和DNA損傷檢測點介質1（MDC1）的表達及臨床意義。 方法 采用逆轉錄聚合酶鏈式反應檢測BRCA1、53BP1、MDC1在喉癌細胞系Hep-2中mRNA的表達，同時用免疫印跡法檢測蛋白的表達。 結果 在所檢測的人喉癌細胞系Hep-2中BACR1、53BP1、MDC1在基因與蛋白兩個水平均有表達。 結論 BRCA1、53BP1、MDC1可能在喉癌的發生發展中有一定作用。
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。
目的 探討雙管喉罩與氣管插管用于全身麻醉婦科腹腔鏡手術的安全性和可行性。 方法 2009年1月-5月擇期婦科腹腔鏡手術患者60例，ASAⅠ～Ⅱ級，隨機分為喉罩組（P組）和氣管插管組（T組）。記錄入室基礎值（T0），置罩（管）前（T1），置罩（管）后即刻（T2），置罩（管）后5 min（T3 ），拔除罩（管）即刻（T4），拔除罩（管）后5 min（T5）的收縮壓（systolic pressure, SBP），舒張壓（diastolic pressure, DBP），心率（heart rate, HR）和脈搏血氧飽和度（pulse oxygen saturation, SpO2），喉罩和氣管插管控制呼吸時氣腹前后不同時段的氣道峰壓（airway. maximum pressure, Pmax），潮氣量（vital volume, VT）和呼氣末二氧化碳分壓（end tidal CO2, PETCO2）。記錄插罩（管）成功率，及相關并發癥。 結果 T2時T組SBP，DBP和HR顯著高于P組（P＜0.05），兩組術中通氣均滿意；Pmax，VT和PETCO2組間比較各時點無差異（P＞0.05）。氣腹后Pmax和PETCO2組內比較均高于氣腹前，差異有統計學意義（P＜0.05）。置罩（管）成功率組間比較差異無統計學意義，拔罩（管）期及術后24 h并發癥，喉罩組明顯低于氣管導管組，差異顯著（P＜0.05）。 結論 雙管喉罩用于全麻婦科腹腔鏡手術通氣效果滿意，安全可行。