Objective To investigate the features of lymph node metastasis and its effects on the prognosis of patients after radical operation for thoracic esophageal squamous cell cancer, and investigate the reasonable postoperative adjuvant protocol. Methods Multivariate analysis of the clinical data of 204 patients was carried out by Spearman correlation analysis, Cox model and Kaplan-Meier method. Results The lymph node metastasis rate was 40.2% (82/204), and 166 out of 2193 dissected lymph nodes had metastasis with the rate of 7.57%. The analysis of related factors revealed that the invasion depth, tumor length and differentiation grade were significantly associated with the postoperative lymph node metastasis (χ2 = 17.466, 11.494, 6.767, P <0.05), while age, tumor site were not significantly correlated with the postoperative lymph node metastasis (χ2=1.086, 3.897, P > 0.05). Kaplan-Meier analysis showed that the 1-, 3-, 5-year survival rates of patients with < 4 lymph nodes metastasis were significantly higher than those with ≥4 lymph nodes metastasis (χ2=4.493, 4.494, 4.450, P < 0.05). The recurrence and metastasis were more often occurred in patients with lymph node metastasis compared with those without lymph node metastasis (r=-2.060, -4.296, P <0.05). Multivariate analysis confirmed that the pathological stage, tumor differentiation grade, and the postoperative adjuvant treatment were the independent prognostic factors. Conclusions The invasion depth, tumor length and differentiation grade are significantly associated with the postoperative lymph node metastasis. The lymph node metastasis state and the number of involved lymph nodes affect the prognosis of patients. Oral administration of 5-FU is benefit to the patients without lymph node metastasis.

Objective To observe the short-term efficacy and toxicity of HEPP regimen in treatment of refractory Non-Hodgkin's Lymphoma. Methods HEPP regimen: HCPT 8 mg/m2 iv gtt d1~ d5, VP16 100 mg/d iv gtt d1~d5, PDD 20 mg/d iv gtt d1~d5, PDN 60 mg/m2 po d1~d14. The chemotherapy was repeated every 4 weeks as a cycle.The clinical effect was evaluated after 2 cycles and toxicity was observed during every cycle. Results 25 patients were eligible for toxicity evaluation and 22 patients for clinical response evaluation. The objective response rate was 60.0 %, including three cases complete remission and ten cases partial remission. Six cases achieved stable disease and three cases progressive disease. The major toxicity was bone marrow suppression, including 24.0 % grade Ⅲ/Ⅳ leukopenia and 12.0 % grade Ⅲ/Ⅳ thrombocytopenia. The incidence of nausea/vomiting, mucositis and hepatic toxicity was low. Conclusion HEPP regimen can achieve a satisfy result in the treatment of refractory Non-Hodgkin's Lymphoma. It is low toxic and well tolerated.

OBJECTIVETo guide doctors in precisely positioning surgical operation, a new production method of minimally invasive implant guide template was presented.

METHODSThe mandible of patient was scanned by CT scanner, and three-dimensional jaw bone model was constructed based on CT images data The professional dental implant software Simplant was used to simulate the plant based on the three-dimensional CT model to determine the location and depth of implants. In the same time, the dental plaster models were scanned by stereo vision system to build the oral mucosa model. Next, curvature registration technology was used to fuse the oral mucosa model and the CT model, then the designed position of implant in the oral mucosa could be determined. The minimally invasive implant guide template was designed in 3-Matic software according to the design position of implant and the oral mucosa model. Finally, the template was produced by rapid prototyping.

RESULTSThe three-dimensional registration technology was useful to fuse the CT data and the dental plaster data, and the template was accurate that could provide the doctors a guidance in the actual planting without cut-off mucosa.

CONCLUSIONThe guide template which fabricated by comprehensive utilization of three-dimensional registration, Simplant simulation and rapid prototyping positioning are accurate and can achieve the minimally invasive and accuracy implant surgery, this technique is worthy of clinical use.

Computer-Aided Design ; Dental Implantation, Endosseous ; Dental Implants ; Dental Models ; Humans ; Jaw, Edentulous ; Mandible ; Patient Care Planning ; Surgery, Computer-Assisted

Adult ; Aged ; Antineoplastic Agents ; adverse effects ; therapeutic use ; Arsenicals ; adverse effects ; therapeutic use ; Chemical and Drug Induced Liver Injury ; Female ; Humans ; Liver Neoplasms ; drug therapy ; Male ; Middle Aged ; Oxides ; adverse effects ; therapeutic use ; Remission Induction ; Treatment Outcome ; Vomiting ; chemically induced

Objective:To investigate the expression of microRNA-296 (miR-296) in rabbit hypertrophic scars and its role in human fibroblasts (HFbs).Methods:The experimental method was used. Twelve healthy adult New Zealand long-eared rabbits regardless gender were randomly divided into normal control group and scar group, with 6 rabbits in each group. The rabbit ear hypertrophic scar model was created in scar group according to the literature, and the rabbits in normal control group did not receive any treatment. On 60 days after setting up the models in scar group, hematoxylin-eosin staining was performed to observe the growth and arrangement of fibroblasts (Fbs) in the ear scars and skin tissue of rabbits in the two groups. The mRNA expressions of miR-296 and transforming growth factor-β 1 (TGF-β 1) in ear scars and skin tissue of rabbits in the two groups were detected by real-time fluorescent quantitative reverse transcription polymerase chain reaction, and the correlation of mRNA between miR-296 and TGF-β 1 was performed with Pearson regression analysis. Two batches of HFbs were used and transfected respectively with corresponding sequences, with the 1st batch being divided into TGF-β 1 wild type+miR-296 negative control group and TGF-β 1 wild type+miR-296 mimic group and the 2nd batch being divided into TGF-β 1 mutant type+miR-296 negative control group and TGF-β 1 mutant type+miR-296 mimic group. At 48 h after transfection, luciferase reporter gene detection kit was used to detect the luciferase and renal luciferase expression of TGF-β 1 in the cells of each group, with their ratio being used to reflect the gene expression level. Two batches of HFbs were used, and each batch of cells were divided into miR-296 negative control group and miR-296 mimic group, being transfected with the corresponding sequences. At 0 (immediately), 12, 24, 36, and 48 h after transfecting the first batch of cells, the cell proliferation was detected by thiazolyl blue method. At 24 h after transfecting the second batch of cells, the expression of TGF-β 1 and collagen type Ⅰ was detected by Western blotting. The number of samples in cell experiments was 3. Data were statistically analyzed with analysis of variance for factorial design, independent sample t test. Results:On 60 days after setting up the models in scar group, the Fbs of rabbit ear scar tissue in scar group proliferated and arranged disorderly, while the growth and arrangement of Fbs in rabbit ear skin tissue in normal control group were normal. The mRNA expression of miR-296 of rabbit scar tissue in scar group (0.65±0.11) was significantly lower than 1.19±0.12 of rabbit ear skin tissue in normal control group ( t=5.175, P<0.01). The mRNA expression of TGF-β 1 of rabbit ear scar tissue in scar group (1.47±0.06) was significantly higher than 1.10±0.03 of rabbit ear skin tissue in normal control group ( t=12.410, P<0.01). Pearson regression analysis showed that there was a negative correlation between the mRNA expression of miR-296 and TGF-β 1 in the ear scars and skin tissue of 12 rabbits ( F=7.278, P<0.05). At 48 h after transfection, the gene expression of TGF-β 1 of cells in TGF-β 1 wild type+miR-296 mimic group was significantly lower than that in TGF-β 1 wild type+miR-296 negative control group ( t=35.190, P<0.01), while the gene expression of TGF-β 1 of cells in the two TGF-β 1 mutant type groups were close ( P>0.05). The HFbs proliferation ability in miR-296 mimic group was significantly lower than that in miR-296 negative control group at 12, 24, 36, and 48 h after transfection( t=3.275, 11.980, 10.460, 17.260, P<0.05 or P<0.01). At 24 h after transfection, the protein expressions of TGF-β 1 and type Ⅰ collagen of cells in miR-296 negative control group were significantly higher than those in miR-296 mimic group ( t=3.758, 29.390, P<0.05 or P<0.01). Conclusions:The miR-296 expression in rabbit hypertrophic scars is down-regulated; miR-296 can inhibit the proliferation of HFbs and the expression of type Ⅰ collagen by down regulating the expression of TGF-β 1.

Objective:To explore the effect of chronic intermittent hypoxia(CIH)on learning and memory dysfunc-tion in rats,as well as the expression of high mobility group box-1(HMGB1)and nuclear transcription factor-KB(NF-κB)in the hippocampus region.Methods:The CIH rat model was established,and forty SD rats were randomly divid-ed into four groups:normoxia group,hypoxia for 4 weeks group(CIH4 group),hypoxia for 8 weeks group(CIH8 group),and hypoxia for 12 weeks group(CIH12 group).Morris water maze was used to assess the learning memory ability of rats,and immunohistochemistry and ELISA were used to detect the expression of HMGB1 and NF-κB in the hippocampus of rats.Results:Compared with the normoxia group,the CIH12 and CIH8 groups had longer escape la-tency,the number of crossing the platform and the residence time in the quadrant of the platform were significantly shortened,but there was no significant difference in the CIH4 group.Additionally,there was no significant expression of HMGB1 and NF-κB in the hippocampal region of the normoxia group,but little expression was observed in CIH4 group,and significantly expressed in CIH8 group and CIH12 group,and the expression of CIH12 group was significantly higher than that of CIH8 group.Conclusion:CIH can lead to a decline in learning and memory function in rats,and the longer time of intermittent hypoxia led to the more significant effect on their learning and memory function.In addi-tion,CIH also leads to increased expression levels of HMGB1 and NF-κB in the hippocampus region,and the expres-sion increased more significantly after hypoxia for 12 weeks,comparing to hypoxia for 8 weeks.

ObjectiveTo investigate the effects of berbamine hydrochloride on sorafenib resistance in hepatocellular carcinoma cells and the underlying mechanisms. MethodThe sorafenib-resistant cell line SMMC-7721/S was selected by the concentration increment method starting at 1.25 μmol·L^-1 sorafenib. Both SMMC-7721 and SMMC-7721/S cells were treated with 0， 2.5， 5， 10， 15， 20 μmol·L^-1 sorafenib， and the cell counting kit-8 （CCK-8） assay was employed to determine the half maximal inhibitory concentration （IC₅₀） and calculate the resistance index （RI）. Western blot was conducted to compare the expression of proteins involved in autophagy and phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway between SMMC-7721 and SMMC-7721/S cells. Furthermore， SMMC-7721/S cells were treated with 5 μmol·L^-1 berbamine hydrochloride alone or in combination with 2.5， 5， 10 μmol·L^-1 sorafenib， and the cell growth was assessed by the CCK-8 assay. In addition， SMMC-7721 and SMMC-7721/S cells were treated with 5 μmol·L^-1 berbamine hydrochloride alone or in combination with 5 μmol·L^-1 sorafenib， and the cell proliferation was examined by the colony formation assay. The immunofluorescence assays with Microtubule-associated protein 1 light chain 3 （LC3） and LysoTracker as probes were employed to assess the lysosomal acidification in SMMC-7721 cells treated with 5 μmol·L^-1 berbamine hydrochloride or 0.1 μmol·L^-1 autophagy inhibitor bafilomycin A1 （Baf）. Further， the expression of proteins involved in autophagy and PI3K/Akt/mTOR signaling pathway was determined by Western blot and compared between groups. ResultSorafenib showed the IC₅₀ of 9.56 mol·L^-1 （P<0.01） and 7.99 mol·L^-1 for SMMC-7721/S and SMMC-7721 cells， respectively， at 24 h. The resistance index （RI） of SMMC-7721/S for sorafenib was 1.20 （P<0.01）， which indicated mild resistance. Compared with SMMC-7721 cells， SMMC-7721/S cells exhibited up-regulated expression of p-mTOR， p-Akt， and LC3Ⅱ， down-regulated expression of p62 protein （P<0.01）， and unchanged Akt protein level. CCK-8 and colony formation assays demonstrated that the combination of berbamine hydrochloride and sorafenib exhibited a synergistic effect （Q>1.15）， with berbamine hydrochloride partially reversing the resistance of liver cancer cells to sorafenib. The immunofluorescence detection of LC3 revealed that berbamine hydrochloride and Baf significantly increased LC3 in SMMC-7721 cells. The detection with LysoTracker as the probe showed that berbamine hydrochloride inhibited the acidity of lysosomes in SMMC-7721 cells （P<0.01）， indicating the suppression of autophagy. Berbamine hydrochloride further enhanced the downregulation of p-mTOR and p-Akt protein levels and did not change the Akt protein level in SMMC-7721 cells exposed to sorafenib. Berbamine hydrochloride inhibited the increase in p-mTOR expression， down-regulated the p-Akt protein level， and did not change the total Akt protein level in the SMMC-7721/S cells exposed to sorafenib. ConclusionBerbamine hydrochloride can ameliorate the resistance of liver cancer cells to sorafenib by inhibiting cellular autophagy and the PI3K/Akt/mTOR signaling pathway.