Objective To investigate the effects of applied electric fields on the migration of human retinal pigment epithelial (hRPE) cells, and probe the physiological effects and clinic significance of electric fields on the RPE cells during the wound healing process. Methods hRPE cells were clutured with Dubbeccolsquo;s modified Eagle medium (DMEM) (DMEM group ) or DMEM+10% fetal bovine serum (FBS) (DMEM+10% FBS group), and the cells unexposed to the electric fields were used as the control. After hRPE cells were exposed to direct current electric fields with the tension of 0,4, 6 and 8 V/cm, serial images were taken at 15 minuets intervals within 2 hours by photomicroscope to observe the migration of the cells, and then the migrating distances and angle between the cellular translocation direction and the field vector were measured. The results were analyzed by an image analyzer. Resutls The response of the hRPE cells cultured in DMEM group to the electric fields was not as obvious as that in DMEM+10% FBS group. The cells showed a tendency of cath odal migration when the field tension was added to 6 V/cm (the average cosine Phi; was compared with the normal control, P＜0.05.The effect of electric fields on cells clutured in DMEM+10%FBS group was ber than those in DMEM group. The long axes of the cells were perpendicular to the field line with the tension of 4 V/cm, and the cells continued migrating to the cathode, which was more obvious when the field tension increased (the average cosine Phi; was comparedwith the normal control, P＜0.05). After the polarity of the electric field reversed , the cells migrated to the opposite direction in accord with the direction of the cathode. Conclusion Applied electric fields can induce the cathode-directed migration of hRPE cells, which is positively correlated with the field tension and can be influenced by the serum. Physiological electric fields may be one of the inducing factors of cell migration at the early phase of healing process of wounded retina. (Chin J Ocul Fundus Dis, 2006, 22: 404-407)
ObjectiveTo investigate that the TGF- beta/Smad signaling pathway mediated epithelial mesenchymal transition (EMT) in trachea stenosis after transplantation. Methods180-220 g male rats (n=50) were randomly divided into a control group and an experimental group. no surgical operation rats were in the control group. tracheal transplantation rats (Wistar-SD rat) were in the experimental group. Graft specimens were obtained in rats on 3,7,10,14,35,90 days after operation. HE staining is used to explain the fibrosis degree of tracheal stenosis. The fibrosis degree of tracheal stenosis was detected by calculating the fibrosis rate. Immunohistochemical staining was used to detect transplanted tracheal, such as EMT related molecules E-cadherin, vimentin, alpha-SMA expression, p-Smad2/3 expression and transcription factor ZEB1, Snail1 expression in tracheal graft specimens. ResultsHE staining showed that the tracheal fibrosis rate of the control group was 0.171±0.020, fibrosis rate was 0.537±0.013 (P < 0.01) on the third day after transplantation. The result of immunohistochemical staining showed that vimentin positive epithelial cells increased significantly (P < 0.05). E-cadherin expression significantly reduced (P < 0.05). Compared with the control group, TGF- beta expression increased (P < 0.05) in the experiment group. Compared with the control group, the expression of p-Smad2/3, the transcription factor ZEB1 and Snail1 significantly increased (P < 0.05) in the experiment group. ConclusionMechanism of tracheal stenosis may be due to EMT. At the same time, TGF- beta/Smad signaling pathway and transcription factor ZEB1, Snail1 may regulate the EMT.
ObjectiveTo investagte the effects of single immunoglobin IL-1 receptor related protein (SIGIRR) on inflammation induced by cigarette smoke extract (CSE) in A549 cells derived from mouse alveolar epithelial cells. MethodsA549 cells were divided into a control group and an over-expressed SIGIRR group. Eukaryotic expression vectors pcDNA3.1(+) constructed with SIGIRR cDNA were transiently transfected into A549 cells, in which SIGIRR was forced to be over-expressed. The expression level of SIGIRR after transfection was detected with Western blot and RT-PCR method. After stimulated by CSE in both groups, the protein level of IL-6 was detected by ELISA, the transcriptional activity of cyclooxygenase-2(COX-2) was detected by dual-luciferase reporter assay system, and the release of reactive oxygen species (ROS) was measured by chemiluminescence method. ResultsThe expression level of SIGIRR increased significantly in A549 cells transfected with SIGIRR vectors. The COX-2 expression and the levels of ROS and IL-6 were significantly increased in the control group after CSE stimulation. Nevertheless, in the over-expressed SIGIRR group, the COX-2 expression and the release of ROS was reduced while the protein level of IL-6 was down-regulated compared with the control group(P < 0.05). ConclusionsUp-regulated SIGIRR expression can suppress the levels of ROS, COX-2 and IL-6 in A549 cells stimulated by CSE. It suggests that SIGIRR can inhibit airway inflammation caused by smoking.
Objective To explore the effectiveness of the transforming growth factor-β1(TGF-β1) and tumor necrosis factor-α(TNF-α) inducing human bronchial epithelial(HBE) cells to optimize epithelia-mesenchymal transformation(EMT) model. Methods Blank control, TGF-β1 10 ng/ml, TNF-α 10 ng/ml, TGF-β1 10 ng/ml+TNF-α 10 ng/ml induced human epithelial cells for 24 hours. Then the change of morphological alteration were observed by applying CCK8, cells migration assay and Western blot technique. Results When TGF-β1 plus TNF-α induced human epithelial cells for 24 hours, most of HBE cells traits changed including morphological alteration from cobblestone to fusiform, connection between cells vanishing, intercellular space broadening. In the experiments of checking cell migration capacity by the vitro scratch test, the group spacing was 420.06±10.38 μm in the blank control group, 499.86±34.00 μm in the TGF-β1 10 ng/ml group, 514.93±10.56 μm in the TNF-α 10 ng/ml group, 569.68±33.58 μm in the TGF-β1 10 ng/ml+TNF-α10 ng/ml group. TGF-β1 cooperated with TNF-α led to scratch spacing narrowing significantly. Western blot analysis showed that expression of E-cadherin and Vimentin varied significantly in the TGF-β1+TNF-α group. Conclusion Inducing human bronchial epithelial cell by TGF-β1 cooperated with TNF-α optimizes EMT model.
ObjectiveTo analyze the surgical outcome of patients with lung cancer using double micro-portal video-assisted thoracic surgery (VATS) technique.MethodsWe retrospectively analyzed the perioperative data of 200 patients with primary lung cancer who underwent successful two micro-portal VATS lobectomy between September 2016 and June 2018 at our unit. There were 125 males and 75 females, aged 61.01±8.71 years. The length of the main operating hole was about 2.0–2.5 cm, the size of the secondary operating hole and the observation hole was 0.5 cm individually. Thus, the total length of the three incisions was 3.0–3.5 cm.ResultsThe mean operating time was 99.18±21.77 min, blood loss was 170.35±105.12 ml, and the mean number of dissected lymph node was 15.82±3.33. The mean volume and duration of chest tube were 446.90±195.32 ml and 3.67±1.85 days. The postoperative hospital stay was 5.54±2.41 days. Only one patient died of pulmonary embolism after surgery. There were 7 patients who were converted to thoracotomy. Postoperative pulmonary infection after lobectomy was found in 8 patients. Postoperative air leak over 5 days was developed in 7 patients.ConclusionThe double micro-portal VATS procedure is a safe and effective strategy for patients with lung cancer, which is associated with decreased surgical trauma and less postoperative pain. This emerging technology may benefit patients by enhancing comfort during their postoperative hospitalization.
ObjectiveTo summarize the clinical experience of Da Vinci robotic-assisted left upper lobectomy for treating lung cancer.MethodsWe retrospectively analyzed the perioperative data of 33 patients with primary lung cancer who underwent Da Vinci robotic-assisted left upper lobectomy between December 2016 and December 2018 in our hospital. Meanwhile, the perioperative data of 41 patients with lung cancer who underwent video-assisted thoracoscopic left upper lobectomy during the same period by the same surgeon were studied as a control group. The resection was followed by the principle of "from back down to front up" way. Systemic lymph node dissection including No.4-9 was performed for all patients.ResultsAll patients received successful surgery with no case of conversion to thoracotomy and perioperative death. Comparing to video-assisted thoracoscopic surgery, the Da Vinci robotic-assisted left upper lobectomy had longer operating time (191.21±61.77 min vs. 154.51±38.81 min, P=0.003), more cost (82 307.75±11 859.03 yuan vs. 58 966.57±5 640.07 yuan, P=0.000), shorter chest tube duration (4.58±1.77 d vs. 5.41±1.52 d, P=0.031) and postoperative hospital stay (6.48±1.82 d vs. 7.66±2.12 d, P=0.014). However, there was no significant difference between the two groups regarding to blood loss, lymph node dissection, postoperative pain score, total chest drainage volume, chest drainage volume per day and the rate of pulmonary complications.ConclusionThe Da Vinci robotic-assisted left upper lobectomy for treating lung cancer is safe and more minimally invasive, but more expensive.
ObjectiveTo reveal the true value of plasma detection of epidermal growth factor receptor (EGFR) mutation for early-stage non-small cell lung cancer (NSCLC) gene diagnosis and to predict survival prognosis. MethodsTissue samples of positive EGFR mutations by using amplification refractory mutation system (ARMS) method were surgically resected from 198 patients with stage I-IV NSCLC between February 2014 and June 2015 in Tangdu hospital. Paired blood samples were collected before surgery. And the cellfree DNA (cfDNA) in plasma was extracted, plasma EGFR mutations were detected by real-time polymerase chain reaction (PCR). Concentration of cfDNA was measured by ultraviolet spectrophotometry. Follow-up observation for stage ⅢA patients was put into force after surgery. Kaplan-Meire was used in survival analysis. ResultsThe sensitivity of EGFR mutation for the 198 paired tissues and plasma samples was 17.2%.The sensitivity was positively correlated with TNM stage and negatively correlated with tumor differentiation. The sensitivity of sage ⅢA was 33.3%, significantly higher than that of the patients at stage ⅠA (1.6%, P=0.000) and stage ⅠB (7.9%, P=0.004). The sensitivity of poor differentiation was 36.8%, significantly higher than that of high differentiation (0.0%, P=0.000) and moderate differentiation (15.7%, P=0.010). There was no correlation between plasma cfDNA concentration and patient characteristics. Survival analysis showed that plasma detection was a vital factor for predicting postoperative survival prognosis of stage ⅢA patients (P=0.014). ConclusionTissue samples cannot be replaced by plasma samples for epidermal growth factor receptor (EGFR) mutation test in early-stage NSCLC patients, currently. When the sensitivity increases dramatically in the plasma samples of stage ⅢA NSCLC and poor differentiation tumor, we recommend using plasma detection for gene diagnosis, dynamic monitoring of EGFR mutations in stage ⅢA or poorly differentiated tumors, especially in NSCLC patients whose tissue samples cannot be obtained by surgery. And plasma EGFR detection is a valuable method of forecasting survival prognosis for locally advanced NSCLC patients.
ObjectiveTo evaluate the clinical outcomes of larynx-preserving limited resection with total thoracic esophagectomy and gastric pull-up reconstruction for the treatment of cervical esophageal squamous cell carcinoma (ESCC) without tumor involvement of the larynx and hypopharynx compared with the upper thoracic ESCC.MethodsRetrospective and comparative analysis of consecutive patients with cervical and upper thoracic ESCC who underwent R0 surgical resection from 2006 to 2011 in our center was performed. Kaplan-Meier method was used to calculate the patients’ survival.ResultsIn total, 44 pairs of patients, including 71 males and 17 females with an average age of 60.66±8.49 years were enrolled in the study after propensity score matching. The baseline characteristics of the two groups of patients were well balanced. There was no statistical difference in the operation time (P=0.100), blood loss (P=0.685), mortality rate in 30 days (P=1.000), total complication rate (P=0.829), cervical anastomosis leakage (P=0.816), mechanical ventilation (P=1.000), normal oral diet within 15 days (P=0.822) and anastomosis recurrence rate (P=0.676) between the two groups. Survival analysis showed that there was no statistical difference in survival time between the cervical group [31.83 (95%CI 8.65-55.02) months] and upper thoracic group [37.73 (95%CI 25.29-50.18) months, P=0.533]. The 5-year survival rates were 32.6% and 42.1%, respectively.ConclusionLarynx-preserving limited resection with total thoracic esophagectomy and gastric pull-up reconstruction for the treatment of cervical ESCC without involvement of the larynx and hypopharynx may result in a similar clinical outcome to upper thoracic ESCC.
ObjectiveTo explore whether surgery combined with adjuvant chemotherapy can bring survival benefits to patients with cervical and upper thoracic esophageal squamous cell carcinoma (ESCC).MethodsThe clinical data of patients with cervical and upper thoracic ESCC who underwent R0 resection and neck anastomosis in our department from 2006 to 2010 were retrospectively analyzed. Patients received neoadjuvant therapy or adjuvant radiotherapy were excluded. The adjuvant chemotherapy group was given a combination of taxanes and platinum based chemotherapy after surgery; the surgery alone group did not receive adjuvant chemotherapy. The Kaplan-Meier method was used to analyze the survival difference between the adjuvant chemotherapy group and the surgery alone group. ResultsA total of 181 patients were enrolled, including 141 (77.9%) males and 40 (22.1%) females, with an average age of 61.0±8.2 years (80 patients aged≤61 years, 101 patients aged>61 years). There were 70 (38.7%) patients of cervical ESCC, and 111 (61.3%) patients of upper thoracic ESCC. Eighty-seven (48.1%) patients underwent postoperative adjuvant chemotherapy, and 94 (51.9%) patients underwent surgery alone, and the basic clinical characteristics were well balanced between the two groups (P>0.05). The median survival time of patients in the adjuvant chemotherapy group and the surgery alone group was 31.93 months and 26.07 months, and the 5-year survival rate was 35.0% and 32.0%, respectively (P=0.227). There was no statistical difference in median survival time between the cervical ESCC and upper thoracic ESCC group (31.83 months vs. 29.76 months, P=0.763). For cervical ESCC patients, the median survival time was 45.07 months in the adjuvant chemotherapy group and 14.70 months in the surgery alone group (P=0.074). Further analysis showed that the median survival time of lymph node negative group was 32.53 months, and the lymph node positive group was 24.57 months (P=0.356). The median survival time was 30.43 months in the lymph-node positive group with adjuvant chemotherapy and 17.77 months in the lymph-node positive group with surgery alone. The survival curve showed a trend of difference, but the difference was not statistically significant (P=0.557).ConclusionThere is no statistical difference in the long-term survival of cervical and upper thoracic ESCC patients after R0 resection. Postoperative adjuvant chemotherapy may have survival benefits for patients with cervical ESCC and upper ESCC with postoperative positive lymph nodes, but the differences are not statistically significant in this setting.