Objective To observe the changes of CD+8CD+28,CD+8CD-28 and CD+4CDhigh25CDlow127 regulate T (Treg)cellsin peripheral blood of lung cancer patients,and to analyze the correlation between CD+8CD-28 and CD+4CDhigh25CDlow127 Treg cells to reveal the role and clinical significance of them in lung cancer patients.Methods Flow cytometry was applied to evaluate the level of CD+8CD+28,CD+8CD-28 and CD+4CDhigh25CDlow127 Treg cells in peripheral blood of 60 untreated lung cancer patients and 60 healthy controls group.The association of each term with clinical features was analyzed.Results The percentage of CD+8CD-28 and CD+4CDhigh25CDlow127 Treg cells in lung cancer group[(58.430:15.749) ％,(7.365±2.025) ％]was significantly higher than those in healthy group [(41.057±15.436)％,(6.648±1.669)％,(t=6.102,P＜0.05;t=2.115,P＜0.05)],while the percentage of CD+8CD+28cells is lower(41.570±15.739)％ than that in healthy group[(58.700±15.298)％,(t=-6.043,P＜0.05)].No close associations were found between three index and gender,age,and biological characteristics.With the increase of TNM stage,The percentage of CD+4CDhigh25 CDlow127 Treg cells increased gradually,which was remarkably higher in patients rith stage Ⅳ than that with stage ⅢA(t=-3.898,P＜0.05).The percentage of CD+4CDhigh25 CDlow127 Tregcells was uncorrelated with CD+8CD-28 cells(r=-0.169,P＞0.05).Conclusion The higher percentage of CD+8CD-28 and CD+4CDhigh25 CDlow127 Treg cells,the lower percentage of CD+8CD+28 cells may be the important reasons of immune suppression in lung cancer patients.Though there is no correlation between CD+8CD-28 and CD+4CDhigh25 CDlow127 Treg cells,it is may be helpful to understand immunologic function and it may look for more specific therapy and provide a new reference in the prognosis of lung cancer.

Objective To detect the levels of CD4+CD25HiCD127Low regulatory T cells (Treg) in the peripheral blood and its clinical significance in patients with esophageal cancer. Methods The levels of Treg in the peripheral blood were detected by three-color flow cytometry (FCM) in 80 patients with esophageal cancer and 20 healthy controls. Among the 80 patients, 30 patients were also further studied for preoperative and postoperative comparison after operation. The clinical and pathological data of each patient were collected and analyzed for the correlation with the Treg levels. Results The level of Treg in the peripheral blood of the control group was lower than that of the esophageal cancer patients [(3.36±1.14) % and (5.70±1.96) %, respectively], with significant difference (P <0.01). The levels of Treg in the peripheral blood was higher in the patients with metastasis of lymph node (n=40) than that in the patients without metastasis of lymph node (5.96±1.36) % and (4.23±1.18) %, respectively] (n=30), with significant difference (P <0.01). The levels of Treg in the peripheral blood of the patients were negatively correlated with their TNM classification. As the TNM classification advanced, the level of the Treg in the peripheral blood increased. Conclusion The levels of Treg in the peripheral blood of the patients with esophageal cancer are significantly higher than that of the healthy subjects, which is correlated with the clinical and pathological conditions. The development of esophageal cancer may relate with suppression of immune function.

Objective To investigate the significance of resistant gene detection combined with adenosine triphosphate-tumor chemosensitivity assay (ATP-TCA) in the second-line chemotherapy of lung squamous cell cancer, and to provide a reference for clinical treatment. Methods 150 patients with lung squamous cell cancer diagnosed by histopathology or cytology and with the disease progressed after NP regime chemotherapy were enrolled. The mRNA expressions of excision repair cross complementation 1 (ERCC1) and ribonucleotide reductase M1 (RRM1) were detected by RT-PCR, and ATP-TCA was carried out. After detected by RT-PCR and ATP-TCA, the patients who were sensitive to gemcitabine plus cisplatin (GP) accepted the second-line systemic chemotherapy with GP regimen, and the others who were not sensitive to GP regimen or whose results of gene detection and ATP-TCA were on the contrary took the second-line chemotherapy regimens with docetaxel plus cisplatin (DP). Both groups accepted 2-4 cycles of systemic chemotherapy. The chest CT was followed up, and the response rate (RR), progression-free survival (PFS) and median survival time (MST) were investigated. Results The RR of GP group was 36.2 % (17/47), while the DP group was 28.1 % (26/92), and the difference was statistically significant (χ2= 4.274, P< 0.05). The media PFS of GP group and DP group were 4.2 months (95%CI 3.485-5.348 months) and 3.6 months (95 %CI 2.685-4.648 months), respectively, and the difference was statistically significant (P<0.05). The MST of GP group and DP group were 9.6 months (95 %CI 8.283-10.637 months) and 8.9 months (95 %CI 7.384-9.648 months), respectively, and there was no statistically significant difference (P> 0.05). Conclusion The resistance gene detection combined with ATP-TCA have certain guiding significance on the second-line chemotherapy for advanced lung squamous cell cancer.

Objective To evaluate the value and safety of transbronchial needle aspiration (TBNA) in the diagnosis of lung cancer. Methods The cytologic diagnosis of TBNA in 82 patients with enlarged hilar and/or mediastinal lymphnodes or lesions adjacent to the bronchial wall were analyzed retrospectively. All specimens were detected by the ThinPrep cytologic test. Results There were 43 positive cases in the 82 patients, and the positive rate was 52.4 %. There were 18 SCLCs,11squamous cancers, 9 adenocarcinomas and 5 undefinable cancers, respectively. There were 39 patients with local bronchial wall swelling accompanied with abnormal mucosae. TBNA, douche, brushing and forcep biopsy were applied, and the diagnostic rate was 64.1%, 7.7 %, 25.6 % and 48.7 %, respectively. The total positive rate was 76.9 %. 43 patients with normal mucous membrane only underwent TBNA. 18 cases were positive, and the positive rate was 41.9 %. There was no obvious complication in the 82 patients. Conclusion The technique of TBNA enlarged the inspection scope of bronchoscopy. It has significant meaning to the diagnosis of lung cancer. TBNA was an useful and safe method in clinical application and could be used widely.

BACKGROUNDTo evaluate the efficacy and toxicity of ifosfamide and cisplatin in the treatment of advanced non-small cell lung cancer.

METHODSFifty-six patients with advanced non-small cell lung cancer were treated by combination chemotherapy of ifosfamide and cisplatin for two cycles: ifosfamide 1.5-2.0g/m² iv drip on day 1-4, mesna 400mg iv at 0,4,8 hours after using ifosfamide; DDP 25-30mg/m² iv drip on day 5-7. The response, toxicity, relievable period and survival period were evaluated.

RESULTSThe total response rate was 50.0%. The response rate of patients in primary treatment was 52.8% and that of return cases was 45.0% (P > 0.05). The median relievable period was five months. The median duration of survival (MDS) was nine months. The major toxicity was inhibition of bone marrow, especially of leukocyte and platelet.

CONCLUSIONSCombination chemotherapy of ifosfamide and cisplatin is effective in the treatment of advanced non-small cell lung cancer including the return cases, and the toxicity is tolerable. If G-CSF is used as a complementary therapy, this regimen could be quite clinically valuable.

Objective To study the effect of lentiviral vector of MicroRNA-126 on survival differentiation of mesenchymal stem cells under ischemic condition.Methods Lentiviral vector of MicroRNA-126 was constructed and then transfected into mouse mesenchymal stem cells (MSCs).Flow detection was used to detect transfection ability and synthesis of collagen type Ⅰ.ELISA was used to detect osteocalcin,Insulin-like growth factor 1 (IGF-1) and transforming growth factor-β (TGF-β) in cell culture supernatant.Mandibular bone defect model was constructed and then injected the hydrogel mixed with MSCs.After 48h,the survival rate and proportion of differentiation into osteoblasts was determined.Results Flow results showed lentiviral vector of MicroRNA-126 could transfected into MSCs.ELISA results showed that MicroRNA-126 lentiviral vector transfected MSCs secreted more bone gla protein and IGF-1 and TGF-β,which increased by 1.43,1.87 and 2.63 times compared with the control virus treated group.Animal experiments showed the number of stem cells survived in the MicroRNA-126 group was 4.56 times that of the control group and the percentage of MSC differentiation into osteoblasts was 75％,which is 3.95 times more than that in the control group.Conclusion The research shows that lentiviral vector of MicroRNA-126 could maintain MSCs survival and differentiation under ischemic condition.

OBJECTIVE: To investigate the expression of breast cancer metastasis suppressor 1 (BRMS1) and CD44v6 protein in supraglottic cancer and to evaluate its clinical significance. METHOD: The expression of BRMS1 protein and CD44v6 protein were examined by using immunohistochemical method in 70 cases of paraffin-embedded supraglottic cancer tissues and their surrounding laryngeal normal mucosa tissues (LNT). RESULT: The expression of BRMS1 protein in LNT of supraglottic cancer was positive, and the positive rate was 85.7% (60/70); in tumor tissue was negative or lower expression, and the positive rate was 35.7% (25/70). The expression of CD44v6 protein in tumor tissue of supraglottic cancer was positive, the positive rate was 82.9% (58/70), in LNT was negative. There was a significant difference in BRMS1 and CD44v6 protein expression between the supraglottic cancer tissue and LNT (P<0.01). The expression of BRMS1 and CD44v6 protein had correlation with clinical stage and pathologic differentiation and cervical lymph node metastasis of supraglottic cancer (P<0.01). No correlation was found between the two proteins expression and sex and age (P>0.05). The expression of BRMS1 protein was related to the expression of CD44v6 protein (r = -0.9042, P<0.01). Calculated by Kaplan-Meier method, there is no survival difference at 3-year between the group with positive BRMS1 protein expression and the group with negative BRMS1 protein expression in tumor tissues (P>0.05), there is a significant survival difference at 3-year between the group with positive CD44v6 protein expression and the group with negative CD44v6 protein expression in tumor tissues (P<0.05). CONCLUSION The expression of BRMS1 protein in supraglottic cancer is significantly decreased and the expression of CD44v6 protein in supraglottic cancer is significantly increased. The expression of BRMS1 protein and CD44v6 protein has a close relationship with pathologic differentiation and clinical stage and cervical lymph node metastasis of supraglottic cancer. Combined detection of the expression of them in supraglottic cancer may provide a significant parameter to judge the cervical lymph node metastasis of supraglottic cancer.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Hyaluronan Receptors ; metabolism ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; Neoplasm Staging ; Prognosis ; Repressor Proteins

OBJECTIVE: To investigate the expression of breast cancer metastasis suppressor 1 (BRMS1) mRNA in supraglottic cancer and to evaluate its clinical significance. METHOD: The expression of BRMS1 mRNA was examined by using RT-PCR method which take beta-actin mRNA as reference template in 66 cases of supraglottic cancer tissues and their adjacent normal mucosa tissues (ANT). RESULT: The expression of BRMS1 mRNA in the tissues of supraglottic cancer is lower significantly than that in the tissues of ANT ( P<0.05). There is correlation between BRMS1 mRNA expression and the clinical stage, differentiation and cervical lymph node metastasis in the laryngeal supraglottic cancers (P<0.05). There is no correlation between BRMS1 mRNA expression and sex and age. CONCLUSION Expression of BRMS1 mRNA in supraglottic cancer is lower than that in adjacent normal mucosa. The decrease of BRMS1 mRNA expression may be related to clinical stage and low differentiation and lymph node metastasis of supraglottic laryngeal cancer.
Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Female ; Humans ; Laryngeal Mucosa ; metabolism ; pathology ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Proteins ; metabolism ; RNA, Messenger ; genetics ; Repressor Proteins

Objective: To investigate the effect of incisor retraction on three-dimensional morphology of upper airway and fluid dynamics in class Ⅰ adult patients with bimaxillary protrusion. Methods: Thirty class Ⅰ patients with bimaxillary protrusion that received fixed orthodontic treatment in Department of Stomatology, The First Affiliated Hospital of Wenzhou Medical University from January 2011 to September 2014 were selected using random number table. All the patients were treated with extraction of four first premolars and retraction of anterior teeth using implant anchorage. Cone-beam CT (CBCT) scans were performed before and after incisor retraction for all patients. The CBCT data of the upper airway were constructed using Mimics 16.0, and the flow field characteristics inside the upper airway were simulated using Ansys 14.0. The changes of volume (V), mean cross-sectional area (mCSA), maximum lateral diameters/maximum anteroposterior diameters (LP/AP) of cross section, the maximum pressure of airflow (P_max), the minimum pressure of airflow (P_min) and pressure drop (△P) of nasopharynx, oropharynx and hypopharynx before and after incisor retraction were measured and compared using paired t test. The correlation between the variation of △P in the most significant pharyngeal part and the morphological variables after incisor retraction was analyzed using Pearson correlation test. Results: No statistical differences were observed in the morphology and flow field in nasopharynx before and after incisor retraction (P>0.05). Before incisor retraction, the oropharyngeal volume and mCSA were (7 580±622) mm³ and (217±40) mm², respectively, and the hypopharyngeal volume and mCSA were (2 564±162) mm³, and (239±43) mm², respectively. After incisor retraction, the volumes of oropharynx and hypopharynx were (6 885±601) mm³ and (2 535±156) mm³, respectively, and mCSA of oropharynx and hypopharynx were (197±37) mm² and (236±42) mm², respectively. The volume and mCSA of oropharynx and hypopharynx were significantly decreased after incisor retraction (P<0.05). The greatest changes in pharyngeal volume and mCSA occurred in the oropharynx. In addition, the LP/AP of oropharynx after incisor retraction was changed from 1.9±0.6 to 2.1±0.7, which was significantly increased compared with the levels before incisor retraction (P<0.05). After simulation of pharyngeal airflow, the oropharyngeal P_min, hypopharyngeal P_max and P_min were (-13.7±4.3), (-8.3±3.8) and (-42.8±9.5) Pa, respectively, whereas the values turned to (-16.4±6.5), (-11.9±3.6) and (-46.0±11.0) Pa, respectively after incisor retraction, which was significantly reduced (P<0.05). △P of oropharynx was significantly increased from (42.7±10.1) Pa to (45.2±13.0) Pa after incisor retraction (P<0.05) and the variation of oropharyngeal △P was negatively correlated with the variation of V and mCSA in oropharynx before and after incisor retraction (r=-0.681, P=0.001; r=-0.844, P=0.000). Conclusions The oropharynx was constricted and the pharyngeal resistance was increased after incisor retraction in adult class Ⅰ patients with bimaxillary protrusion. A comprehensive and systematic evaluation of the pharyngeal morphology and ventilatory function were very important for making a scientific and rational clinical treatment plan.

OBJECTIVE To investigate the intervention effect and its potential mechanism of Yifei xuanfei jiangzhuo formula by inhibiting mitochondrial fission in a vascular dementia （VaD） model rats. METHODS VaD rat model was established by bilateral common carotid artery ligation. The experimental animals were randomly divided into sham operation group （SHAM）， model group （MOD），Yifei xuanfei jiangzhuo formula low-dose group （YFXF-L）， Yifei xuanfei jiangzhuo formula high-dose group （YFXF-H）， and Donepezil hydrochloride group （positive control）， with 9 animals in each group. After 30 days of intervention， the spatial learning memory ability was assessed by Morris water maze experiment； HE staining was used to observe histopathological changes in CA1 area of hippocampus； ELISA was used to detect the levels of serum inflammatory factors [interleukin-1β （IL-1β） and IL-4]； Western blot was used to detect the expressions of heat shock protein 90 （HSP90）/mixed lineage kinase domain-like protein （MLKL）/dynamin-related protein 1 （Drp1） pathway-related proteins， mitochondrial fusion proteins （MFN1， MFN2）， and adenosine triphosphate synthase 5A （ATP5A） in hippocampal tissues. The immunohistochemistry was used to detect the level of phosphorylated MLKL （p-MLKL）； real-time fluorescence quantitative PCR was adopted to detect mRNA expressions ofHSP90， MFN1， MFN2 and ATP5A. RESULTS Compared with SHAM group， the escape latency of rats in the MOD group was significantly prolonged， the number of crossing the platform was significantly reduced， and the hippocampal tissues showed typical neuronal damage characteristics， the positive expression level of p-MLKL and the serum level of IL-1β significantly increased， while the serum level of IL-4 significantly decreased， the protein and mRNA expression of HSP90， as well as the protein expressions of p-MLKL/MLKL and p-Drp1（Ser616）/Drp1 were all significantly increased in hippocampal tissue， the protein and mRNA expressions of MFN1， MFN2 and ATP5A， and protein expression of p-Drp1（Ser637）/Drp1 were all significantly decreased （P＜0.05）. After the intervention of Yifei xuanfei jiangzhuo formula， above indicators in each treatment group were all significantly reversed （P＜0.05）. CONCLUSIONS Yifei xuanfei jiangzhuo formula may alleviate neuronal damage and neuroinflammatory responses in VaD rats by regulating the HSP90/MLKL/Drp1 signaling pathway， inhibiting mitochondrial fission， thereby maintaining mitochondrial dynamic balance and improving mitochondrial function.