【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.
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.
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。
Objective To evaluate the expression and clinical significance of Survivin in the tissues of laryngeal carcinoma using meta-analysis. Methods The case-control studies published in China about the expression and association of clinical pathogenic features of Survivin in the tissues of laryngeal carcinoma were electronically retrieved in CBM (1994 to October 2012), CNKI (1994 to October 2012), VIP (1989 to October 2012) and WanFang Data (1996 to October 2012). The reviewers independently identified the literature according to inclusion and exclusion criteria, extracted data, and assessed the quality of the included studies. Then, meta-analysis was performed using RevMan 5.1 software. Results A total of 25 studies were included, involving 1 333 cases of laryngeal carcinoma and 528 cases of health laryngeal mucosa or polyp of vocal cord. The results of meta-analysis showed that, significant differences were found in groups of laryngeal carcinoma vs. health control, laryngeal carcinoma with vs. without lymphatic metastasis, clinical stages I-II vs. III-IV, cell differentiation G1 vs. G2-G3, T1 and T2 stages vs. T3 and T4 stages, and glottic carcinoma vs. non-glottic carcinoma (Plt;0.05). No significant difference was found in groups of age more than 60 vs. no less than 60, male vs. female, and smoke vs. non-smoke (Pgt;0.05). Conclusion Current domestic evidence shows that Survivin may be associated with the whole course of occurrence, advance and transfer of laryngeal carcinoma, and positively correlated to degree of tumor malignance, which may indicate poor prognosis.
【摘要】目的探討喉癌手術后患者對兩種不同霧化方式的耐受性,為選擇最佳霧化方式提供參考。方法將49例喉癌手術后患者隨機分為觀察組(25例)和對照組(24例),觀察組采用氧氣霧化吸入,對照組采用空氣壓縮泵霧化吸入。分別記錄兩組患者霧化吸入前及吸入15 min時脈搏血氧飽和度(SpO2)及心率;霧化過程中患者有無心慌、氣緊等不適以及霧化后痰液的性質及量。采用SPSS 13.0軟件進行統計分析。結果兩組患者霧化吸入15 min時的SpO2差異有統計學意義(Plt;001),觀察組高于對照組;而兩組患者霧化吸入前SpO2、心率、不適主訴及霧化后痰液的性質差異均無統計學意義(Pgt;005)。結論氧氣霧化吸入可以提高喉癌手術后患者霧化過程中的SpO2,使患者感覺更加舒適。【Abstract】Objective To investigate postoperative patients with laryngeal carcinoma atomization of two different forms of tolerance, in order to choose the best means of atomization. Methods Fifty postoperative patients with laryngeal carcinoma were divided into observation group using oxygen inhalation and control group using the air compression pump inhalation. Two groups of patients were recorded the value of SpO2 and heart rate before 15 minutes after the inhalation,as well as the discomforts such as flustered,gas tight during the atomization process and the nature and olume of sputum. Results The results of two groups of patients at the time of 15 minutes inhalation SpO2 statistically significant difference (Plt;001), the observation group than in the control group average SpO2 high; and two groups of patients with preinhalation SpO2 average, average heart rate, Discomfort chief complaint and the nature of sputum after aerosol compared no significant difference (Pgt;005). Conclusion Oxygen inhalation in patients with laryngeal cancer can improve the atomization process SpO2 value, so that patients feel more comfortable.
目的:探討改良氣管套管墊安置法對喉癌術后佩戴氣管套管、頸部傷口敷料加壓包扎期患者的適用性。方法:采用隨機分組的方法將38例喉癌術后佩戴氣管套管的患者分為傳統組20人和改良組18人, 傳統組采用“Y”型氣管套管墊,改良組采用“Y”型氣管套管墊,比較兩組患者在更換氣管套管墊時的SpO2值、SpO2降低值及刺激性咳嗽次數。結果:安置氣管套管墊的過程中,改良組SpO2值高于傳統組、SpO2降低值低于傳統組,且發生刺激性咳嗽的次數也低于傳統組,差異有統計學意義(Plt;0.01)。結論:與傳統氣管套管墊安置法相比,改良氣管套管墊安置法能減少患者換藥過程中刺激性咳嗽的次數,對SpO2值影響輕微,更適合于喉癌術后早期頸部傷口敷料加壓包扎患者的換藥。
ObjectiveTo explore the feasibility of reconstruction of phonatory function by using a tubular free flap from upper-lateral upper arm to repair the laryngotracheal circumferential defect after near-total laryngectomy for laryngeal cancer. Methods A retrospective study was conducted on 7 patients who underwent near-total laryngectomy between June 2021 and October 2023, aged from 48 to 70 years (median, 59 years), 6 males and 1 female. The disease duration ranged from 1 to 11 months, with a median of 6 months. Pathological diagnosis of preoperative biopsy was squamous cell carcinoma. Tumor classification: glottic type in 5 cases, supraglottic type in 1 case, transglottic type in 1 case; TNM staging: T4N0M0 in 6 cases, T4N2M0 in 1 case; American Joint Committee on Cancer (AJCC) staging in 2017 was stage Ⅳ. Preoperative MRI angiography of upper arm was performed to investigate the blood supply in the upper and lateral regions of the upper arm. After near-total laryngectomy and bilateral neck lymph node dissection, the area of the laryngotracheal defect was measured. A free flap measuring 7.0 cm×5.0 cm to 8.0 cm×7.0 cm was harvested from the upper-lateral upper arm, rolled into a tube shape, and connected between the stump of the cervical trachea in the neck root and that of the epiglottis at the tongue base. Four patients received adjuvant radiochemotherapy, 1 patient received radiochemotherapy and targeted therapy, 2 patients adopted no further adjuvant treatment. Results All 7 patients were followed-up 1-2 years (mean, 1 year and 3 months). Four patients had primary wound healing, 2 patients had minor pharyngeal fistulas that healed after dressing change, 1 patient experienced pharyngeal fistula because of flap necrosis and the wound still healed without secondary surgery. All patients took food orally within 1 month after operation, and the tracheal cannula was retained. Six patients with survived flap gradually adapted to their new pronunciation mode and obtained satisfactory phonatory function from 15 days to 2 months after operation. Four patients had slight aspiration after operation. Till the end of the follow-up, all patients survived and no local recurrence or distant metastasis had been observed. The motor function of the upper arm was not affected, only partial sensory loss occurred in the area near the incision. The scar of the incision could be covered by the short sleeve so as to obtain a better aesthetic effect. ConclusionUsing a tubular free flap from upper-lateral upper arm to repair the laryngotracheal circumferential defect after near-total laryngectomy for laryngeal cancer can achieve satisfactory phonatory restoration while preserve the motor function and aesthetics of the donor site.