OBJECTIVETo explore the RCP protein expression and its clinicopathological significance in laryngeal squamous cell carcinoma (LSCC).

METHODSRCP protein expression in human head and neck squamous cell carcinoma cell lines (NP-69, Tu686, Tu212, M2 and M4) was analyzed by Western blotting. Besides, its expression in 87 cases of LSCC, 18 cases of adjacent epithelial mucosa and 16 cases of vocal cord leukoplakia was detected by immunohistochemistry, and their correlation with clinicopathological parameters and patients' outcome was analyzed.

RESULTSThe NP-69, Tu212 Tu686, M2 and M4 cells showed a gradual increase in the expression of RCP protein. The average relative expression levels of RCP protein in the NP-69, Tu212, Tu686, M2 and M4 cells were 0.05±0.01, 0.38±0.05, 0.63±0.02, 0.84±0.06 and 0.96±0.04, respectively. The same situation occurred in the adjacent mucosa, vocal cord leukoplakia and LSCC. Specifically, only 3 of 18 adjacent mucosa showed a low RCP expression (scored 0-2). Although the 16 cases of vocal cord leukoplakia had a low RCP expression, all their scores ranged from 0 to 3. While in the LSCC specimens, 59 (67.8%) cases demonstrated a high RCP expression (scored 8-15), 18 cases showed a lower RCP expression (scored 4-7), and only 10 cases were scored 2-3. Among the 87 LSCC cases, there were 28 cases (32.2%) of low RCP expression and 59 cases of high RCP expression. All the 18 cases of cancer-adjacent tissues and 16 cases of vocal cord leukoplakia were of low RCP expression. RCP overexpression was significantly associated with T classification, clinical staging, lymph node metastasis and recurrence (P<0.05 for all). Survival analysis revealed that the 5-year survival rate was 40.0% in the patients with high RCP expression and 75.0% in the patients with low RCP expression, the tumor-free 5-year survival rate was 30.7% and 64.0%, respectively, both showing a significant difference between the two subgroups (P<0.05). Univariate analysis revealed that alcohol history; smoking, T classification, clinical staging, lymph node metastasis and RCP expression were significantly associated with a poor prognosis (P<0.05 for all). The multivariate analysis showed that only recurrence and RCP expression were independent prognostic factors affecting the prognosis for patients with LSCC (P<0.05 for both).

CONCLUSIONSExpression of RCP protein may contribute to the malignant progression of LSCC, and may become a novel marker predicting tumor recurrence, cervical lymph node metastasis and prognosis for patients with laryngeal squamous cell carcinoma.

Aged ; Biomarkers ; Biomarkers, Tumor ; Carcinoma, Squamous Cell ; diagnosis ; metabolism ; Disease Progression ; Female ; Head and Neck Neoplasms ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Laryngeal Neoplasms ; diagnosis ; metabolism ; Lymphatic Metastasis ; Male ; Middle Aged ; Neck ; Neoplasm Recurrence, Local ; Prognosis ; Survival Rate

Objective To explore the RCP protein expression and its clinicopathological significance in laryngeal squamous cell carcinoma ( LSCC ) . Methods RCP protein expression in human head and neck squamous cell carcinoma cell lines (NP?69, Tu686, Tu212, M2 and M4) was analyzed by Western blotting. Besides, its expression in 87 cases of LSCC, 18 cases of adjacent epithelial mucosa and 16 cases of vocal cord leukoplakia was detected by immunohistochemistry, and their correlation with clinicopathological parameters and patients’ outcome was analyzed. Results The NP?69, Tu212 Tu686, M2 and M4 cells showed a gradual increase in the expression of RCP protein. The average relative expression levels of RCP protein in the NP?69, Tu212, Tu686, M2 and M4 cells were 0. 05 ± 0. 01, 0. 38 ± 0. 05, 0. 63 ± 0. 02, 0. 84 ± 0. 06 and 0. 96 ± 0. 04, respectively. The same situation occurred in the adjacent mucosa, vocal cord leukoplakia and LSCC. Specifically, only 3 of 18 adjacent mucosa showed a low RCP expression (scored 0?2). Although the 16 cases of vocal cord leukoplakia had a low RCP expression, all their scores ranged from 0 to 3. While in the LSCC specimens, 59 (67. 8%) cases demonstrated a high RCP expression (scored 8?15), 18 cases showed a lower RCP expression (scored 4?7), and only 10 cases were scored 2?3. Among the 87 LSCC cases, there were 28 cases (32. 2%) of low RCP expression and 59 cases of high RCP expression. All the 18 cases of cancer?adjacent tissues and 16 cases of vocal cord leukoplakia were of low RCP expression. RCP overexpression was significantly associated with T classification, clinical staging, lymph node metastasis and recurrence (P<0. 05 for all). Survival analysis revealed that the 5?year survival rate was 40. 0% in the patients with high RCP expression and 75. 0% in the patients with low RCP expression, the tumor?free 5?year survival rate was 30. 7% and 64. 0%, respectively, both showing a significant difference between the two subgroups (P<0. 05). Univariate analysis revealed that alcohol history;smoking, T classification, clinical staging, lymph node metastasis and RCP expression were significantly associated with a poor prognosis (P<0. 05 for all). The multivariate analysis showed that only recurrence and RCP expression were independent prognostic factors affecting the prognosis for patients with LSCC (P<0. 05 for both). Conclusions Expression of RCP protein may contribute to the malignant progression of LSCC, and may become a novel marker predicting tumor recurrence, cervical lymph node metastasis and prognosis for patients with laryngeal squamous cell carcinoma.

Objective To explore the RCP protein expression and its clinicopathological significance in laryngeal squamous cell carcinoma ( LSCC ) . Methods RCP protein expression in human head and neck squamous cell carcinoma cell lines (NP?69, Tu686, Tu212, M2 and M4) was analyzed by Western blotting. Besides, its expression in 87 cases of LSCC, 18 cases of adjacent epithelial mucosa and 16 cases of vocal cord leukoplakia was detected by immunohistochemistry, and their correlation with clinicopathological parameters and patients’ outcome was analyzed. Results The NP?69, Tu212 Tu686, M2 and M4 cells showed a gradual increase in the expression of RCP protein. The average relative expression levels of RCP protein in the NP?69, Tu212, Tu686, M2 and M4 cells were 0. 05 ± 0. 01, 0. 38 ± 0. 05, 0. 63 ± 0. 02, 0. 84 ± 0. 06 and 0. 96 ± 0. 04, respectively. The same situation occurred in the adjacent mucosa, vocal cord leukoplakia and LSCC. Specifically, only 3 of 18 adjacent mucosa showed a low RCP expression (scored 0?2). Although the 16 cases of vocal cord leukoplakia had a low RCP expression, all their scores ranged from 0 to 3. While in the LSCC specimens, 59 (67. 8%) cases demonstrated a high RCP expression (scored 8?15), 18 cases showed a lower RCP expression (scored 4?7), and only 10 cases were scored 2?3. Among the 87 LSCC cases, there were 28 cases (32. 2%) of low RCP expression and 59 cases of high RCP expression. All the 18 cases of cancer?adjacent tissues and 16 cases of vocal cord leukoplakia were of low RCP expression. RCP overexpression was significantly associated with T classification, clinical staging, lymph node metastasis and recurrence (P<0. 05 for all). Survival analysis revealed that the 5?year survival rate was 40. 0% in the patients with high RCP expression and 75. 0% in the patients with low RCP expression, the tumor?free 5?year survival rate was 30. 7% and 64. 0%, respectively, both showing a significant difference between the two subgroups (P<0. 05). Univariate analysis revealed that alcohol history;smoking, T classification, clinical staging, lymph node metastasis and RCP expression were significantly associated with a poor prognosis (P<0. 05 for all). The multivariate analysis showed that only recurrence and RCP expression were independent prognostic factors affecting the prognosis for patients with LSCC (P<0. 05 for both). Conclusions Expression of RCP protein may contribute to the malignant progression of LSCC, and may become a novel marker predicting tumor recurrence, cervical lymph node metastasis and prognosis for patients with laryngeal squamous cell carcinoma.

BACKGROUND:As a dominant breed pig in southwest China,the southern Yunnan small-ear pig has been widely used as an experimental animal in the basic research of other disciplines,but there are still no reports on its application in anterior cruciate ligament reconstruction. OBJECTIVE:To establish a large animal model of the southern Yunnan small-ear pig with anterior cruciate ligament with autologous Achilles tendon was established. METHODS:Twenty adult female Yunnan small-ear pigs were equally randomized into two groups.In the autologous Achilles tendon group,the right knee anterior cruciate ligament was reconstructed with autologous Achilles tendon as a graft,while in the sham-operated group,a similar operation was performed on the right knee without any treatment of the anterior cruciate ligament.General conditions of each pig were observed and recorded before and 12 months after surgery.Ligaments and grafts were taken for gross observation and MAS scoring.Hematoxylin-eosin staining was performed to observe morphological characteristics of ligaments.The staining and arrangement of type I and type Ⅲ collagen were evaluated by immunohistochemistry.Transmission electron microscopy was used to observe the type,size,diameter,ratio,and distribution of collagen fibers in ligaments. RESULTS AND CONCLUSION:All animals had normal diet and activity,good wound healing,no obvious inflammatory reaction,no local purulent infection,and no significant changes in mental and urinary conditions compared with those before surgery.The reconstructed cruciate ligament of the knee was intact,with no stiffness and normal range of motion.Both the anterior drawer and Lachman tests were negative.Gross observation of the graft:12 months after surgery,the grafts was in good position,with good integrity,obvious tension,ligament color close to the original anterior cruciate ligament,and complete surface synovial coverage.Most of the intraarticular ligaments in the autologous Achilles tendon group were defined as MAS I type and a few were defined as MAS Ⅱ type.In the sham-operated group,the intraarticular ligament was defined as MAS I type.Hematoxylin-eosin staining indicated that,12 months after surgery,collagen fibers in the autologous Achilles tendon group began to appear bundled,isotropic,and uniformly arranged,with more obvious isotropic corrugations,and the nuclei were mainly linear or spindle-shaped,which were similar to those in normal anterior cruciate ligament tissue of the sham-operated group.Immunohistochemistry results indicated that,12 months after surgery,there was a higher expression of type I collagen and significantly less expression of type Ⅲ collagen in the reconstructed anterior cruciate ligament in the autologous Achilles tendon group.The degree of type I and type Ⅲ staining was similar in the two groups.Under the transmission electron microscope,the diameter,arrangement and density of collagen fibers in the reconstructed anterior cruciate ligament of the autologous Achilles tendon group were similar to those of the original anterior cruciate ligament at 12 months after surgery,indicating that the ligament remodeling process had been basically completed in the autologous Achilles tendon group at 12 months after surgery.Through a comprehensive evaluation of animal general conditions,ligament general view,MAS score,hematoxylin-eosin staining,immunohistochemistry,and transmission electron microscopy observation,we successfully established a large animal model of anterior cruciate ligament reconstruction using autogenous Achilles tendon in southern Yunnan small-ear pigs,with good morphological,histological and ultrastructural results.

OBJECTIVE: To review the studies about the tibial-graft fixation methods on anterior cruciate ligament (ACL) reconstruction, in order to provide clinical reference. METHODS: The literature about the tibial-graft fixation methods on ACL reconstruction at home and abroad was extensively reviewed, and the factors that affect the selection of fixation methods were summarized. RESULTS: The knee flexion angle, graft tension, and graft fixation device are mainly considered when the tibial-graft was fixed on ACL reconstruction. At present, the graft is mainly fixed at 0°/30° of knee flexion. The study shows that the knee joint is more stable after fixed at 30°, while the incidence of knee extension limitation decrease after fixed at 0°. In terms of graft tension, a good effectiveness can be obtained when the tension level is close to 90 N or the knee flexion is 30° to recover the affected knee over-restrained 2 mm relative to the healthy knee. In terms of the graft device, the interference screw is still the most commonly used method of tibial-graft fixation, with the development of all-inside ACL reconstruction in recent years, the cortical button fixation may become the mainstream. CONCLUSION Arthroscopic reconstruction is the main treatment of ACL rupture at present. However, there is no optimal fixation method for the tibial-graft, the advantages and disadvantages of each fixation methods need to be further studied.
Humans ; Tibia/surgery* ; Anterior Cruciate Ligament Reconstruction ; Knee Joint/surgery* ; Anterior Cruciate Ligament Injuries/surgery* ; Bone Screws