Objective To investigate the regulatory effects of PTEN-PI3K/AKT pathway on the apoptosis of gastric cancer MKN28 cells and the possible mechanisms.Methods The specific sequence of PTEN was transfected to MKN28 cells by eukaryotic expression vector (transfection group),and then vacant vector (negative control group) and PBS (blank control group) were transfected to the MKN28 cells,respectively.The effects of PTEN-PI3K/AKT pathway on the apoptosis of MKN28 cells and the expressions of PI3K,AKT,Caspase-3 and Caspase-9 were investigated.The growth curve and apoptosis of the MKN28 cells were detected by MTT assay and TUNEL staining,respectively,and the protein expression was detected by the Western blot.MKN28 cells which did not transfected by the PTEN were processed by inhibitor of PI3K (LY294002) (treated group),and MKN28 cells in the control group were processed by PBS.The expressions of apoptosis protein and apoptosis-related protein were detected after inhibition of PI3 K.All data were analyzed using the t test.Results The model of over-expression of PTEN was obtained and transfected into MKN28 cells.The survival of MKN28 cells in the transfection group was significantly inhibited in a time-dependant manner ( r =0.938,P ＜ 0.05 ).The mean apoptotic rate of the transfection rate was 27.86％ ± 4.78％,which was significantly higher than 0.01％ ± 0.01％ of the negative control group ( t =9.527,P ＜ 0.05 ).The protein expression of PI3K in the transfection group was 0.25 ± 0.03,which was significantly lower than 0.93 ± 0.16 of the blank control group and 0.96 ± 0.15 of the negative control group (t =7.235,8.883,P ＜ 0.05 ).The protein expression of P-AKT in the transfection group was 0.21 ± 0.03,which was significantly lower than 0.93 ± 0.13 of the blank control group and 0.91 ± 0.12 of the negative control group (t =9.347,9.802,P ＜ 0.05 ).The expressions of Caspase-3 and Caspase-9 of the transfection group were 0.86 ± 0.11 and 0.87 ± 0.12,which were significantly higher than 0.16 ± 0.03 and 0.18 ± 0.04 of the negative control group,and 0.15 ± 0.02 and 0.16 ± 0.03 of the blank control group ( t =10.634,10.999,9.448,9.942,P ＜0.05).The apoptotic rate of the MKN28 cells of the treated group was 28.60％ ± 4.50％,which was significantly higher than 0.12％ ± 0.06％ of the control group (t =10.961,P ＜ 0.05 ).The protein expression of PI3K and P-AKT of the treated group were 0.18 ± 0.02 and 0.11 ± 0.01,which were significantly lower than 0.93 ± 0.14 and 0.90 ± 0.12 of the control group (t =9.186,11.363,P ＜ 0.05 ).The protein expression of PTEN of the treated group was 1.15 ± 0.15,which was significantly higher than 0.21 ± 0.08 of the control group (t =9.577,P ＜ 0.05 ).The relative expressions of Caspase-3 and Caspase-9 of the treated group were 0.86 ± 0.12 and 0.88 ± 0.11,which were significantly higher than 0.25 ± 0.02 and 0.21 ± 0.03 of the control group (t =8.685,10.178,P ＜ 0.05).Conclusions Over-expression of PTEN may enhance the apoptosis of gastric cancer cells through inhibition of PI3K/AKT pathway.Inhibition of PI3K can enhance the expression of PTEN.PI3K and PTEN regulate the apoptosis of gastric cancer cells.

Objective·To investigate and compare the clinical characteristic of non-organic dyspnea and asthma both with complaint of dyspnea.Methods·Seventy-four consecutive patients with non-organic dyspnea,and 74 age-,height-,weight and sex-matched patients with asthma were recruited for investigation in the study.The self-assessment surveys were conducted for the two groups by means of Hospital Anxiety and Depression Scale,Nijmegen Questionnaire and Athens Insomnia Scale.The words for describing dyspnea,clinical symptoms,effective sleep hours and items of pulmonary function test were collected and analyzed Results·Non-organic dyspnea patients tended to describe psychogenic aspects.Asthma patients tended to describe airflow limitation.Non-organic dyspnea group mainly performed psychogenic symptoms.The sleeping time in non-organic dyspnea group was significantly lower than that in asthma group(P＜0.05).The score of anxiety,depression,Nijmegen Questionnaire,Athens Insomnia Scale,FEV1,FEV1％Pred,FVC％Pred,FEV1/FVC in non-organic dyspnea group were significantly higher than those in asthma group (P＜0.05).There was no significant difference in FVC between two groups(P＞0.05).Conclusion·The non-organic group feel more anxiety,depressed and insomnic than the asthma group.Lung function test of asthma group is often abnormal.To discriminate non-organic dyspnea with asthma,clinicians should pay more attention to emotion,sleep,somatoform symptoms,medical history and so on,and do pulmonary function test,improve the understanding of the characteristics of the two diseases,decrease misdiagnosis and wrong diagnosis.

Objective To establish a human colon cancer multi-drug resistant cell line LS174T/5-fluorouracil(5-Fu) and to explore its biological characteristics. Methods A resistant human colon cancer line LS174T/5-Fu was established by stepwise increasing concentrations of fluorouracil selection from the parental cell line LS174T.Drug sensitivity was detected by MTT assay,and the changes of biological characteristics were determined by light microscopy,cell counting,flow cytometry and RT-PCR. Results LS174T/5-Fu cell line was developed after 6 months with stable resistance to 5-Fu and a resistance index of 40.24.The cells exhibited cross-resistance to cisplatin,etoposide,doxifluridine and hydeoxycamptothecin.LS174T/5-Fu cells grew more slowly than LS174T cells,which was longer than LS174T cell line.Cell cycle distribution of LS174T/5-Fu cells was changed compared with parental cells.The percentage of cells in G_(1) and S phase was increased,while in G_(2) phase was decreased.The level of MRP mRNA expression was increased according to the concentration of 5-Fu in resistant cell lines. Conclusion LS174T/5-Fu cells is a reliable multi-drug resistant cell subline of human colon cancer.The successful establishment of this cell subline has laid a solid foundation for further study of the multi-drug resistant mechanism of human colon cancer induced by 5-fluorouracil.

Objective To study the expression level of myostatin in the gastrocnemius muscle of cancer cachexia mice and to observe the improvement of the status and the influence on myostatin expression by use of meloxicam.(Methods The) tumor-bearing cachexia mice model was established by Lovo cell line subcutaneous inoculation.Twenty-four BALB/c mice were randomly divided into 3 groups(n=8 for each group):group A,control;group B,tumor-bearing mice plus saline;group C,tumor-bearing mice plus meloxicam(5 mg/kg).The food intake and body composition were documented.The expression of myostatin in the gastrocnemius muscle was investigated by RT-PCR and immunohistochemistry in all the animals,and the correlation analysis between serum TNF? and myostatin was conducted. Results The body weight of group B was about 60% of group A,and the average food intake was significantly declined(P

Objective To study the effects of granulocyte-macrophage colony stimulating factor (GM-CSF) on the expression of vascular endotllelial growth factor (VEGF) in human dermal fibroblast. Methods In vitro human dermal fibroblasts in good status were incubated with GM-CSF (GM-CSF group) or non-GM-CSF (control group) culture medium for different periods of time. The mRNA, protein expression of VEGF in derma fibroblast were determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively, and the secretion of VEGF in supernatant was measured by enzyme linked immunosorbant assay (ELISA). Results The expression of VEGF mRNA from dermal fibroblasts was increased significantly after l or more hours of incubation with GM-CSF comparing with the control (P＜0.05). 6 hours of stimulation by GM-CSF caused maximal expression of VEGF mRNA. The expression of VEGF protein in dermal fibroblasts was increased from 12 hours and was peaked at 24 hour after stimulation by GM-CSF. VEGF protein from the supernatant of the dermal fibroblasts was also raised persistently from 12 hour after stimulation by GM-CSF and was improved remarkably compared with the control. Conclusions GM-CSF can up-regulate directly the expression of VEGF in human derma fibroblast, which may be one of the mechanisms that GM-CSF accelerates neovascularization in wound healing.

Objective To investigate the proportion changes of the CD4 + CD25 + T cells and CD8 + CD28 - T cells in the peripheral blood of gastric cancer patients. Methods The proportions of CD4 + CD25 + T cells and CD8 + CD28 - T cells in the peripheral blood of gastric cancer patients were determined by flow cytometry. 30 patients with gastric cancer and 30 patients with chronic gastritis were enrolled in this study. Results Compared with the chronic gastritis controls, the proportion of the CD4 + CD25 + T cells in the peripheral blood of patients with gastric cancer (8.7%±1.3%) was not significantly changed. But the proportion of the CD8 + CD28 - T cell in the peripheral blood of patients with gastric cancer (30.3%±3.3%) was significantly higher than that of control group (20.3%±2.7%), P<0.05. There was no significant association between the proportions of T regulatory cells and the lymphomatic metastasis of gastric cancer. No relation was found between the proportions of T regulatory cells and pathological types of gastric cancer. Conclusion CD8 + CD28 - T ceils in peripheral blood may be of some value in the progression of gastric cancer.

Objective To evaluate clinical outcomes of treating displaced intra-articular calcaneal fractures with Orthofix external fixation (EF) in comparison with open reduction and internal fixation (ORIF).Methods Between February 2013 and December 2014,39 patients with 41 calcaneal fractures were treated operatively and fully followed up at our department.They were 27 men and 12 women,aged from 27 to 56 years(average,42.5 years).The left foot was affected in 15 cases and the right in 26.By the Sanders classification,there were 11 cases of type Ⅱ,22 ones of type Ⅲ and 8 ones of type Ⅳ.The 5 open fractures were Gustilo type Ⅰ (2 cases) and Gustilo type Ⅱ (3 cases).Of the 11 fractures complicated with soft tissue injury,4 were Tscherne-Oestern type Ⅰ and 7 Tscherne-Oestern type Ⅱ.The patients were divided into group EF (25 cases) and group ORIF (16 cases) according to the operation they had received.The 2 groups were compared in intraoperative indexes and follow-up radiological data.The American Orthopaedic Foot and Ankle Society (AOFAS) scores for ankle and hindfoot were recorded at the final follow-up for every patient.Results The mean duration of surgery,intraoperative bleeding,time for hospitalization and bone healing in group EF were significantly less than in group ORIF (P ＜ 0.05).There were no significant differences between the 2 groups in follow-up duration,AOFAS score,Bthler angle,Gissane angle,calcaneal axis,calcaneal horizontal length,tuberosity height,hillock calcaneal height,calcaneal width,or talar inclination angle (P ＞ 0.05).According to the AOFAS scores at the final follow-ups,15 cases were rated as excellent,8 as good and 2 as fair in group EF while 10 were rated as excellent,5 as good and one as fair in group ORIF.Conclusions All types of calcaneal fracture can be treated using Orthofix external fixation which may lead to advantages of earlier surgery regardless of soft tissue condition,minimal invasiveness with less devascularization to the calcaneus and decreased risk of complications,and good clinical outcomes.

Objective:To observe the effect of transdermal fentanyl combined with gabapentin for the treatment of malignant neu-ropathic pain (MNP). Methods:A total of 60 patients with MNP were randomly divided into two groups. A total of 30 cases in the con-trol group received transdermal fentanyl according to the dosages of opioid medicine that patients used. Such dosages were gradually in-creased until the pain relief visual analogue scale (VAS) fell below 3 or until the times of breakthrough pain became less than 3. For the combined group, gabapentin was co-administered with transdermal fentanyl, similar to the control group. Initially, 100 mg of gabapen-tin was administered thrice a day. This dosage was gradually increased until pain relief. However, gabapentin dosages were kept below 2,400 mg a day. VAS, quality of life (QOL), degree of pain relief, dosages of fentanyl and morphine, and side effects were evaluated be-fore treatment and at one, two, three, and four weeks after the treatment. Results:Both groups exhibited lower VAS after treatment (P<0.05), but the difference was observed to be more significant in the combined group than that in the control group (P<0.05). Both groups exhibited improved QOL (P<0.05), which was observed to be more significant in the combined group than in the control group (P<0.05). The effective rate was 96.7%in the combined group and 83.3%in the control group. The dosage of opioid medicine and the side effects in the combined group were less than those in the control group. Conclusion:Transdermal fentanyl combined with gabapen-tin is effective for the treatment of malignant neuropathic pain.

Objective To analyze the MRI features of central neurocytoma (CNC),and to improve the diagnostic accuracy of this lesion.Methods The MRI findings of 12 patients with CNC confirmed by surgery and pathology were evaluated retrospectively.All patients underwent contrast-enhanced and uncontrast-enhanced MR scan,and 3 patients also received plain CT scan.Results The lesions of 12 cases were all located in the lateral ventricles of septum pellucidum,which were close to the Monro's foramen.5 lesions located in the right lateral ventricle,5 lesions located in both lateral ventricles with 1 case reached the third ventricle,2 lesions located in the left lateral ventricle with 1 case reached the third ventricle.The mean diameter of the CNC lesions was 4.8 cm (ranged from 2.2 to 7.8 cm).11cases (92％) had vavious degrees of cystic change,8 cases (66.7％) had a spongy or soap bubble appearance and 1 case(8.3％) had completely cystic change.3 cases (25％) had a scalloping appearance on sagittal T1-weighted MRI.The solid part of the tumor was similar to the gray matter in signal intensity,with slightly high signal intensity on T2FLAIR.Among 9 cases with DWI,7 cases showed high signal intensity,1 case showed equal signal intensity,and 1 case showed slightly low signal intensity.On enhanced MRI,the solid part of tumors showed slight to medium (83％) or obvious (17％) enhancement.Conclusion CNC should be first considered when a young adult has a lesion located in the lateral ventricles of septum pellucidum,accompainied by varying degrees of cysts,showed slight to medium enhancement in the solid part of the tumor.

Objective To identify the risk factors for postoperative spinal cord injury in Stanford type A aortic dissection patients.Methods 210 Stanford type A aortic dissection(TAAD) patients underwent Sun's procedure in Beijing Aortic Disease Center during July 2014 to March 2015.14 patients had spinal cord injury after surgery.Clinical data and computed tomography angiography(CTA) imaging of aorta were retrospectively analyzed and multi-logistic regression analysis was performed to identify risk factors for spinal cord injury post operation.Results 14 out of 210(6.7％) patients had transient or permanent spinal cord injury after surgery.Univariate analysis showed only false lumen derived intercostal arteries at eighth thoracic vertebral level (T8) to first lumbar vertebral level (L1) was significantly associated with post-surgery spinal cord injury (P =0.000).Multi-logistic regression analysis showed that false lumen derived intercostal arteries (P =0.000) and age (P =0.016) were significantly associated with postoperative spinal cord injury.Conclusion Major intercostal arteries derived from false lumen and rapid thrombogenesis in false lumen are the major risk factors for postoperative spinal cord injury in Stanford type A aortic dissection patients.