BACKGROUND: Fibronectin serves not only as the supporter for cells,but also as an important intracellular linkage, possessing opsonic-like functions. It can promote the phagocytic function of mononucleophages and the repair in inflammation and trauma.OBJECTIVE: Type O plasma was freshly obtained from healthy males in search of the optimal preparation method for lower sampling, convenient and higher-yielding of fibronectin.DESIGN: Open experiment.SETTING: Institute of Traumatic Surgery, Fourth Affiliated Hospital of Guangzhou Red Cross Hospital, Jinan University.MATERIALS: This experiment was carried out in the Institute of Traumatic Surgery of Guangzhou Red Cross Hospital between July 2000 and July 2002. The major materials consisted of type O fresh plasma from healthy males, gelatin, sepharose 4B activated with cyanogen bromide,sephadex G-25, urea, and trishydroxymethylaminomethane.METHODS: Affinity column constituted by gelatin coupling with sepharose 4B activated with cyanogen bromide was used to purify fibronectin. ① Human plasma was added: 150 mL type O plasma was freshly obtained from healthy males and added into the column once by 25 mLat an interval of 20 minutes, the speed of flow was 3.5 mL/min. ② Removing mixed protein: the column was rinsed by Tris-sodium citrate equilibrium liquid (pH 7.5) until the absorbency of flow-out fluid decreased to ＜ 0.02 at 280 nm. Again lmol/L NaCl containing Tris-sodium citrate was used to wash off other mixed proteins. ③ Collecting fibronectin: the column was rinsed by urea-Trispurge Fluid (3 mol/L) to collect fibronectin.④ Removing urea from fibronectin collection: fibronectin collection liquid was filtrated by Sephade G-25 to remove urea. ⑤Disinfection: 0.22 μmgermtight filter was used for disinfection.MAIN OUTCOME MEASURES: ①Results of sodium dodecyl benzene sulfonate-polyacrylamide gel electrophoresis. ② The influence of retention time on the yield of purified fibronectin. ③ The influence of plasma quantity on the yield of purified fibronectin.RESULTS: ① Results of sodium dodecyl benzene sulfonate-polyacrylamide gel electrophoresis: The density of separation gel and condensed gel was 7% and 3%, respectively; fibronectin was electrophoresized into single protein lane. ② The influence of retention time on the yield of purified fibronectin: The yield of fibronectin was (65.24±3.45) %, (74.77±4.05) %,(86.99±4.10) %, and (80.47±3.75) %, respectively, when the loading amount of fibronectin was 150 mL with non-retention time and retention time of 10, 20 and 30 minutes. ③ The influence of plasma quantity on the yield of purified fibronectin: The yield of fibronectin was (72.56±3.63) %,(77.61±3.14) %, (86.99±4.12) %, and (74.67±3.05) % when the column retention time was controlled at minute 20 with the loading amount of 100,130, 150 and 180 mL, respectively.CONCLUSION: In a given column volume of gelatin, the quantity of purified fibronectin was closely related with the plasma retention time in column and the total loading amount of plasma. As a result, the optimal loading amount of plasma was 150 mL, and the retention time was 20 minutes.The preparation method, herein, has been proved to require small amounts of plasma and yield large amounts of fibronectin.

Objective To investigate the effect of siRNA targeted against Bcl-xl on cell proliferation of rheumatoid arthritis (RA) and the effect on expres-ion of apoptosis relevant factors Bcl-xl,Bax,Caspase-3.Methods Human RA synovial cells were cultured and passed by tissue block collagenase digestion method.The siRNA plasmid targeting Bcl-xl gene was constructed by Bcl-xl cDNA sequence provided by gene banks,while single missense sequences were used as negative controls.LipofectamineTM 2000 was used to transfect synovial cells.The effect of Bcl-xl silencing on proliferation of synovial cells was evaluated by MTT at 24,48,72 hours after transfection.The expressions of Bcl-xl,Bax,Caspase-3 protein,synovial apoptosisrelated factors were determined by Western blotting after transfected at different time points.The average of multiple-sample average was analyzed by single-factor x2 test or LSD-t and Tamhane's T2 test was used for twotwo comparison.Results MTT result s showed that RNA interference that specifically silent Bcl-xl could obviously suppress the proliferation of sliding film cells,which was most eveident at 48 hours.This inhibition was gradually weakened with prolonged time,but when compared was the control group,differences was significant (P＜0.05).Western blotting results displayed that when compared to the control group,Bcl-xl protein expression obviously declined after transfection P＜0.01,which was the least at 48 h time point.Bax,Caspase3 protein expression were obviously increased when compared to the coutrol group.Conclusion The expression of Bcl-xl,a RA synovial cell anti-apoptotic factor,is significantly reduced by specific RNA interference silencing Bcl-xl,which may play an important role in inhibiting the excessive proliferation of synovial cells.

Objective To evaluate the difference of clinical short-term effect and adverse reaction between paclitaxel liposome and paclitaxel in non-small cell lung cancer patients with lymph node metastasis after pulmonary resection.Methods Sixty-eight patients after pulmonary resection were divided into two groups by random digits table method,37 patients in experimental group with paclitaxel liposome (135mg/m2) combined with carboplatin (CBP) at 300 mg/m2 in chemotherapy,and 31 patients in control group with paclitaxel (135 mg/m2) combined with CBP at 300 mg/m2 in chemotherapy.Results All patients were evaluable.In experimental group,5 patients had complete remission,10 patients had partial remission,17patients were stable,5 patients' condition aggravated,the total effective rate was 40.5％(15/37),clinical control rate was 86.5％ (32/37).In control group,2 patients had complete remission,8 patients had partial remission,15 patients were stable,6 patients' condition aggravated,the total effective rote was 32.3％(10/31),clinical control rate was 80.6％(25/31).The treatment effectiveness in experimental group was significantly higher than that in control group (P ＜ 0.05).The main adverse reaction included marrow suppression,hair loss,muscle and joint pain and gastrointestinal symptom,there was no serious hypersensitivity.The rate of hypotension,face flushing,paresthesia,muscle and joint pain,erythra in experimental group was lower than that in control group [0 vs.9.7％ (3/31),5.4％ (2/37) vs.19.4％ (6/31),10.8％ (4/37) vs.22.6％ (7/31),13.5％ (5/37) vs.38.7％ (12/31),5.4％ (2/37) vs.25.8％ (8/31)] (P ＜0.0 1 or ＜0.05).Conclusion The curative effect rate of paclitaxel liposome is better than paclitaxel in patients with lymph node metastasis after pulmonary resection and with lower incidence of side effects.

BACKGROUND: Fibronectin(FN) plays the role of repair in the inflammation. There is no confirmed conclusion whether it can be applied to the refractory corneal epithelial defect.OBJECTIVE: To observe the repair effect of the human plasma FN for the refractory corneal epithelial defect.DESIGN: A controlled experimental study.SETTING: Guangzhou Red Cross Hospital, Guangzhou First People's Hospital, Guangzhou Children's Hospital, Guangzhou Hospital of Traditional Chinese Medicine and Guangzhou Second People's Hos pital.PARTICIPANTS: Totally 383 eyes with the refractory corneal epithelial defect were chosen, of which 309 were in the therapy group, and 74 in the control group.METHODS: The therapy group: Human plasma FN was administered by dropping it into the eyes once every two hours. The controlgroup: 10 g/L celacol M was administered by its dribbling into the eyes once every two hours. Weilesheng was taken orally in both groups, two pills once, three times per day. According to the state of illness, both groups received anti-bacterial or anti-viral treatment and reexamination was given every day or every other day after administration. 10 g/L fluorescein sodium was used to observe the changes of cornea.MAIN OUTCOSE MEASURES: The symptoms, results of staining using fluorescein sodium as well as the corneal epithelial healing of both groups.RESULTS: The symptoms, the results from staining using the fluorescein sodium and the corneal epithelial healing were used for the evaluation. In the therapy group, 309 eyes were followed up and the cure rate was 69.9%. The average therapeutic period was 6.5 days, while those of the control group were 58.1% and 8.7 days respectively. The difference in the curative effect between the two groups was significantly different( P＜0.01 ).CONCLUSION: The application of FN for the refractory corneal epithelial defect displays a more significant effect than conventional treatment.

Objectives To establish a cost-effective and reproducible procedure for induction of chronic left ventricular aneurysm (LVA) in rabbits. Methods Acute myocardial infarction (AMI) was induced in 35 rabbits via concomitant ligation of the left anterior descending (LAD) coronary artery and the circumflex (Cx) branch at the middle portion. Development of AMI was co n-firmed by ST segment elevation and akinesis of the occluded area. Echocardiography, pathological evaluation, and agar i n-tra-chamber casting were utilized to validate the formation of LVA four weeks after the surgery. Left ventricular end systolic pressure (LVESP) and diastolic pressure (LVEDP) were measured before, immediately after and four weeks after ligation. D i-mensions of the ventricular chamber, thickness of the interventricular septum (IVS) and the left ventricular posterior wall (LVPW) left ventricular end diastolic volume (LVEDV) and systolic volume (LVESV), and ejection fraction (EF) were recorded by echo-cardiography. Results Thirty one (88.6%) rabbits survived myocardial infarction and 26 of them developed aneurysm (83.9%). The mean area of aneurysm was 33.4% ± 2.4% of the left ventricle. LVEF markedly decreased after LVA formation, whereas LVEDV, LVESV and the thickness of IVS as well as the dimension of ventricular chamber from apex to mitral valve annulus significantly increased. LVESP immediately dropped after ligation and recovered to a small extent after LVA formation. LVEDP progressively increased after ligation till LVA formation. Areas in the left ventricle (LV) that underwent fibrosis included the apex, anterior wall and lateral wall but not IVS. Agar intra-chamber cast showed that the bulging of LV wall was prominent in the area of aneurysm. Conclusions Ligation of LAD and Cx at the middle portion could induce develo pment of LVA at a mean area ratio of 33.4%±2.4%which involves the apex, anterior wall and lateral wall of the LV.

In search of the optimal preparation method for large-scale purification of human plasma fibronectin, we adopted affinity chromatography with gelatin and the Sepharose 4B activated with cyanogen bromide to purify fibronectin from type "C" plasma of healthy males, and scanned the best method under the conditions of different amount of plasma loading and different residence time in column. In a given column volume of gelatin, the absorbent was related with the plasma residence time in column and the total amount of plasma loaded. As a result, the optimal loading amount of plasma is 150 ml, and the residence time is 20 minutes. The preparation method, herein, has been proved to require small amount of plasma and yield large amount of fibronectin.
Chromatography, Affinity ; methods ; Cyanogen Bromide ; chemistry ; Fibronectins ; blood ; isolation & purification ; Gelatin ; Humans ; Male ; Sepharose ; chemistry ; Wound Healing

Consensus ; Hirschsprung Disease ; diagnosis ; Humans

Objective: To investigate the expression of lncRNA LINC00886 in human esophageal squamous cell carcinoma (ESCC) tissues and cell lines, and its effects on proliferation, migration and invasion of Eca109 cells. Methods: The cancer tissues and corresponding para-cancerous tissues of 69 ESCC patients were collected in the biological specimen bank of the Fourth Hospital of Hebei Medical University from June 2014 to December 2016; the ESCC cell lines Eca109, TE13, TE1, Kyse150, Yes-2 and Kyse170 were also collected. LINC00886 gene expression in ESCC tissues and cell lines was detected by qPCR. Eca109 cells were transfected with pIRES2-LINC00886 and pIRES2-NC, respectively, and the overexpression efficiency of LINC00886 gene in Eca109 cells was detected by qPCR; MTS, clone formation assay, wound-healing assay and Transwell invasion assay were respectively used to detect the effect of LINC00886 over-expression on proliferation, migration and invasion ability of Eca109 cells. Results: The expression of LINC00886 gene in ESCC tissues was significantly lower than that in para-cancerous tissues (P<0.01), and its expression level was associated with tumor TNM stage and lymph node metastasis (both P<0.05). The expression level of LINC00886 gene in ESCC cell lines was also lower than that of the control group (all P<0.01). Compared with control group, the expression level of LINC00886 gene was significantly higher in Eca109 cells transfected with pIRES2-LINC00886 (both P<0.05). Compared with the control group, LINC00886 overexpression significantly inhibited the proliferation, migration and invasion abilities of Eca109 cells (all P<0.01). Conclusion: The decreased expression of LINC00886 gene may be related to the occurrence and development of ESCC. Over-expression of LINC00886 gene inhibits the proliferation, migration and invasion abilities of ESCC cells.

We proposed a new deep learning model by analyzing electroencephalogram signals to reduce the complexity of feature extraction and improve the accuracy of recognition of fatigue status of pilots. For one thing, we applied wavelet packet transform to decompose electroencephalogram signals of pilots to extract the δ wave (0.4-3 Hz), θ wave (4-7 Hz), α wave (8-13 Hz) and β wave (14-30 Hz), and the combination of them was used as de-nosing electroencephalogram signals. For another, we proposed a deep contractive auto-encoding network-Softmax model for identifying pilots' fatigue status. Its recognition results were also compared with other models. The experimental results showed that the proposed deep learning model had a nice recognition, and the accuracy of recognition was up to 91.67%. Therefore, recognition of fatigue status of pilots based on deep contractive auto-encoding network is of great significance.

Objective: To investigate the expression of long non-coding RNA NUP50-AS1 (lncRNA NUP50-AS1) in esophageal squamous cell carcinomas (ESCC) tissues and cell lines, and to explore its effect on proliferation, migration and invasion of human esophageal cancer Eca109 cells. Methods: 49 pairs of ESCC tissues and corresponding para-cancerous tissues obtained from the Biological Specimen Base of the Fourth Hospital of Hebei Medical University during Jan. 2015 to Jan. 2016 were used in this study. qRT-PCR method was applied to detect the expression of NUP50-AS1 in collected tissues samples and five esophageal cancer cell lines (TE1, TE13, Eca109, Kyse150 and Kyse170). ShRNAs were transiently transfected into Eca109 cells to interfere the expression of NUP50AS1 gene, and finally, sh2-NUP50-AS1 was used for the following experiments. The effect of NUP50-AS1 gene knockdown on the proliferation of Eca109 cells was detected by MTS and colony formation assay; the effect of NUP50-AS1 gene knockdown on the migration of Eca109 cells was detected by scratch test, and the effect on cell invasion was detected by Transwell assay. Results: The expression of NUP50-AS1 in ESCC was correlated with the lymphnode metastasis and TNM stage (all P<0.01). The expression of NUP50AS1 in ESCC tissues was significantly higher than that in corresponding normal tissues (2.003±0.870 vs 1.000±0.000, P<0.05). The expression of NUP50-AS1 in five esophageal cancer cell lines was significantly up-regulated (P<0.05), and it had the highest expression in Eca109 cell line. After transfection, sh2-NUP50-AS1 had the highest transfection efficiency, and knocking down NUP50-AS1 gene significantly inhibited the proliferation, invasion and migration of the Eca109 cells. Conclusion: The expression of lncRNA NUP50AS1 in ESCC tissues was significantly higher than that in the para-cancerous tissues, and correlated with the TNM stage and lymphnode metastasis. The down-regulation of NUP50-AS1 inhibited the proliferation, invasion and migration of esophageal cancer cells. The high expression of NUP50-AS1 gene may be closely related to the occurrence and development of ESCC.