Objective:To compare the effectiveness of two types of robotic training in improving the lower extremity motor functioning of children with spastic cerebral palsy (SCP).Methods:Twenty-eight children with SCP were randomly divided into a control group and an experimental group, each of 14. Both groups received conventional exercise therapy, paraffin therapy, neuromuscular electrical stimulation, and massage. Both also performed 30 minutes of gait training five days a week for eight weeks assisted by either a Lokomat or a Relink lower limb rehabilitation robot. Before and after the treatment, both groups were evaluated using the Gross Motor Function Measure (GMFM), the Pediatric Balance Scale (PBS), the Modified Tardieu Scale (MTS), the six-minute walk test (6MWT), the Physiological Cost Index (PCI) and their self-selected walking speed (SWS).Results:Significant improvement in all of the measurements were observed in both groups. After the treatment, there were no significant differences between the two group in the average GMFM (section D and E) or PBS scores. The average MTS R1 and R2, SWS, 6MWT and PCI results of the experimental group were, however, significantly better than those of the control group.Conclusion:Applying either the Lokomat or Relink robot in lower extremity rehabilitation improves the lower extremity motor function of children with grade II-III SCP. The Relink robot is the more effective in improving triceps surae spasm and walking ability.

Objective:To provide the risk stratification method of hepatoblastoma (HB) suitable for implementation in China and explore the new treatment method for high-risk HB patients.Methods:A total of 100 cases of children and adolescents under 18 years old with newly diagnosed HB in Sun Yat-sen University Cancer Center and Sun Yat-sen University First Affiliated Hospital from September 2014 to September 2018 were included. According to the clinical stage, AFP level, pathological subtype and other factors, patients were stratified into four groups: extremely low-, low-, intermediate- and high-risk. The patients at very low risk were treated with surgery only and followed-up. The patients at very low risk were treated with C5V(Cisplatin+ 5-Fluroracil+ Vincristine) regimen for 4 courses. The patients at intermediate risk were treated with C5VD(Cisplatin+ 5-Fluroracil+ Vincristine+ Doxorubicin)regimen before and after surgery for 6-8 courses. The patients at high risk were treated with C5VD and IIV (ifoshamide+ irinotecan+ vincristine) alternately before and after surgery for 8 courses.Results:One hundred patients were stratified into extremely low-risk, low-risk, medium-risk and high-risk groups for 2, 10, 51 and 37 cases, respectively. Eighty three cases had evaluable lesions before chemotherapy. Among them, 65 patients achieved partial remission, stable disease and progressive disease were observed in 10, and 8 cases, respectively, with a response rate of 78.3%. During a median follow-up of 20 months, 30 patients experienced tumor relapse or progression, and 27 of them died. The 2-years progression-free survival (PFS) and overall survival (OS) rates were 69.2% and 72.0%, respectively. The 2-years PFS rates of patients with extremely low risk, low risk, medium risk and high risk were 100%, 88.9%, 75.3% and 43.2%, respectively. The 2-years OS rates were 100%, 100%, 81.0% and 44.8%, respectively.Conclusions:The novel HB risk classification is simple and feasible. With active comprehensive treatment, patients at extremely low-, low- and medium-risk have excellent outcomes. The survival rate of high-risk HB patients remains to be improved, and new treatment strategies need to be explored.

Objective:To provide the risk stratification method of hepatoblastoma (HB) suitable for implementation in China and explore the new treatment method for high-risk HB patients.Methods:A total of 100 cases of children and adolescents under 18 years old with newly diagnosed HB in Sun Yat-sen University Cancer Center and Sun Yat-sen University First Affiliated Hospital from September 2014 to September 2018 were included. According to the clinical stage, AFP level, pathological subtype and other factors, patients were stratified into four groups: extremely low-, low-, intermediate- and high-risk. The patients at very low risk were treated with surgery only and followed-up. The patients at very low risk were treated with C5V(Cisplatin+ 5-Fluroracil+ Vincristine) regimen for 4 courses. The patients at intermediate risk were treated with C5VD(Cisplatin+ 5-Fluroracil+ Vincristine+ Doxorubicin)regimen before and after surgery for 6-8 courses. The patients at high risk were treated with C5VD and IIV (ifoshamide+ irinotecan+ vincristine) alternately before and after surgery for 8 courses.Results:One hundred patients were stratified into extremely low-risk, low-risk, medium-risk and high-risk groups for 2, 10, 51 and 37 cases, respectively. Eighty three cases had evaluable lesions before chemotherapy. Among them, 65 patients achieved partial remission, stable disease and progressive disease were observed in 10, and 8 cases, respectively, with a response rate of 78.3%. During a median follow-up of 20 months, 30 patients experienced tumor relapse or progression, and 27 of them died. The 2-years progression-free survival (PFS) and overall survival (OS) rates were 69.2% and 72.0%, respectively. The 2-years PFS rates of patients with extremely low risk, low risk, medium risk and high risk were 100%, 88.9%, 75.3% and 43.2%, respectively. The 2-years OS rates were 100%, 100%, 81.0% and 44.8%, respectively.Conclusions:The novel HB risk classification is simple and feasible. With active comprehensive treatment, patients at extremely low-, low- and medium-risk have excellent outcomes. The survival rate of high-risk HB patients remains to be improved, and new treatment strategies need to be explored.

Objective To measure the tear film lipid layer thickness (LLT) in dry eye patients and investigate the correlations of LLT with ocular surface signs.Methods One hundred and thirty dry eye patients (130 eyes),including 64 meibomian gland dysfunction (MGD) patients and 66 non-MGD patients,were included in this study.LLT,break-up time (BUT),fluorescein staining (FL),Marx's line (ML) score and Schirmer I test were performed and examined.The distribution of LLT in different age groups and the correlations between LLT and other examinations were analyzed.Results There was significant difference in LLT among different age groups (P =0.007),while LLT was not significantly different between male and female in each age group (P ＞ 0.05).LLT was positively correlated with age (r =0.334,P ＜ 0.001) and was not correlated with sex (r =0.107,P =0.226).LLT was positively correlated with upper eyelid ML score (r =0.295,P =0.001) and lower eyelid ML score (r =0.233,P =0.008).There was no significant correlation of LLT with BUT,FL or Schirmer Ⅰ test (all P ＞0.05).In the MGD group,there were positive correlations of LLT with upper eyelid ML score and lower eyelid ML score (all r =0.306,P =0.014),and no correlation of LLT with other examinations (all P ＞ 0.05).In the non-MGD group,there was no correlation of LLT with other examinations (all P ＞ 0.05).In a multivariate linear regression analysis,age and upper eyelid ML score were significantly related to LLT (β =0.254,P =0.005 for age and β =0.207,P =0.022 for upper eyelid ML score) in all dry eye patients.Age was the only factor related to LLT (β =0.382,P =0.002) in the MGD group.Upper eyelid ML score and lower eyelid ML score were higher in the MGD group than the non-MGD subgroup (all P ＜ 0.001).Conclusion LLT is positively correlated with age and ML score in dry eye patients.The measurement of tear film LLT,as an auxiliary examination in the diagnosis of dry eye disease,should be analyzed with the influential factors including age.

Objective To observe what changes the brown adipose tissue (BAT) of T2DM rat models would have,including morphology,function and specially expressed uncoupling protein (UCP1) after the gastric bypass (Roux-en-Y,RYGB) and to explore the effects of RYGB on BAT of T2DM rat models and its related mechanism in order to provide a theoretical and experimental basis for treatment of T2DM patients with RYGB.Methods SD rats were given a high-fat and high-sugar diet for two weeks,by injecting streptozotocin (STZ) 30 mg/kg intraperitoneally to build models.Blood glucose was measured after 72 h and 1 week by the fast blood glucose meter.The models were built successfully if blood glucose at both times were ≥ 16.7 mmol/L.Feeding environment:individually caged,standard rat feed,natural circadian cycle,indoor temperature (18±2)℃,indoor humidity (50±2)％.50 rats were randomly selected and dividing into four groups according to intervention methods:diabetes operation group (group A,n=10),undergoing RYGB surgery with the whole stomach kept;diabetes sham operation group (group B,n=10),the same anesthesia and incision as the previous RYGB group.The operation mode was anterior gastric wall incision and suture,jejunum transection in corresponding position and in situ anastomosis with the same suture method as group A;diabetes control group (group C,n=10),normally feeding after building models;and the last one was the healthy control group (group D,n=10):no special treatment,adequate water feeding ensured.The rest of rats remained to be used.The body mass (BM),fasting blood glucose (FBG),fasting serum insulin(Fins)before and at the 1st,2nd,4th and 8th week after surgery were measured.The number of transversal ceils was calculated by IPP6.0 image software and the average radius of fat cells was calculated.UCP1 expression was tested with western blot.Results ① The fasting blood glucose,fasting serum insulin level and the body weight of dia betic rats were higher than those of the control group,but the insulin sensitivity index was significantly lower.② HE Staining showed:diabetes operation group (group A) rats,compared with diabetes control group and diabetes sham operation group(group B),had obviously higher brown fat cell counts transversally and average radius,and the difference was statistically significant (P＜0.01).Diabetes operation group (group A) rats had no significant difference from the healthy control group(group D) rats,and the diabetes control group (group C) rats had no significant difference from sham operation group (group B) rats as well.③ Western blot showed that after the gastric bypass surgery,compared with the diabetes sham operation group (group B) and the diabetes control group (group C),UCP1 expression of brown adipose tissue of the diabetes operation group (group A) increased significantly (P＜0.05).The diabetes sham operation group (group B) had no significant difference from the diabetes control group (group C),and the diabetes operation group(Group A) had no significant difference from the healthy control group (Group D) as well (P＞0.05).Conclusion RYGB can reduce the body mass and insulin resistance (IR) of diabetic rats and,at the same time,promote the expression of UCP1 of brown adipose tissue.RYGB might increase the activity of brown adipose tissue by regulating the UCP1 signaling pathway to improve body's insulin resistance.

Background Corneal transplantation faces a great challenge because of the shortage of corneal donors and difficulty of human corneal endothelial cells (HCECs) regeneration in vitro.So the study on tissue engineering cornea is still a main topic.Previous research showed that mouse embryonic stem cell conditioned medium (ESC-CM) improved the proliferative capacity of HCECs in vitro,and acellular porcine corneal stroma (APCS) was a good saffold material.However,whether HECEs cultured by mouse ESC-CM can form cell sheet in vitro were rarely studied.Objective This study was to investigate the potential that HCECs cultured by mouse ESC-CM form a monolayer cell sheet.Methods The supernatant of ESC-CM was collected after mouse ES-E14 cells were cultured,and the cultured medium was centrifuged and mixed with 75％ human corneal endothelium medium (CEM)at a proportion of 1 ∶ 3 to prepare the 25％ ESC-CM system.Primary cultures of HCECs were established from explants of corneal limbal with Descemet's membrane,and the cells were identified by using reverse-transcription PCR to determine the expressions of collagen Ⅷ (Col Ⅷ) mRNA and neuron-specific enolase (NSE) mRNA in the cells.APCS was prepared by decellularization with phospholipase A2 and bicarbonate solution,and the second generation of HCECs were inoculated on the sterilized APCS at a 800/mm2 density.The morphology of the cells was observed by hematoxylin-eosin staining under the phase-contrast microscope.The expressions of zona occludens protein-1 (ZO-1)and Na+-K+-ATPase in the cell sheet were detected by immunofluorescence staining.Results The second generation of HCECs cultured with 25％ ESC-CM in vitro showed the hexagon in shape with positive expressions for Col Ⅷ mRNA and NSE mRNA.Decellularization APCS was transparent,and no corneal cells were seen,the structures of corneal collagenous fibres were regular.HCECs attached closely to APCS and formed monolayer sheet 7 days after culture on the APCS with the cell density of (2 694±143)/mm2.ZO-1 and Na+-K+-ATPase were positively expressed on the HCECs sheet.Conclusions Twenty-five percent ESC-CM promotes the proliferation and maintains the normal morphology of HCECs.APCS provide a good scoffold and microenvironment for the formation of HCEC sheet.The HCEC sheet Possesses the pump function of HCECs and is a good corneal donor for transplantation.

Background The fate of adult stem cells is associated with its surrounding microenviroment.Our previous work found that embryonic stem cells (ESCs) micro-environment enhance the stemness of human limbal stem cells (LSCs),but its mechanism has not been elucidated.Objective This study was to explore the molecular mechanism of ESC micro-environment enhancing the stemness and inhibiting the apoptosis of LSCs.Methods Human LSCs were cultured by explant culture method with CnT-20 medium and CnT-20+20％ ES culture supernatant (ESC-CM),respectively.Colony formation assay was used to analyze the proliferation ability of cells.Telomerase reverse transcriptase (TERT) siRNA (19-25nt siRNA) or siRNA (sc-37007) was transfected into the cells of ESCCM group.Apoptosis and mitochondrial membrane potential were assayed by flow cytometry,and the expressions of telomerase and reactive oxygen species (ROS) in TERT siRNA-or siRNA-F-transfected cells by immunofluorescence and flow cytomery.RT-PCR,immunofluorescence staining and Western blot were employed to determine the expressions of p63,ATP-binding cassette transporer G2 (ABCG2),integrin β1 mRNA and proteins and cytokeratin 3 (C K3) in the cells.The levels of focal adhesion kinase (FAK),Akt,glycogen synthase kinase 3β (GSK3β) and p21 protein and phosphorylation proteins in the cells were detected by Western blot.Results The LSCs presented an increased proliferative capacity and passaged to the eighth generation with the colony-forming efficiency (CFE) of (7.6±0.6) ％ in ESC-CM group,but the cells to the sixth generation with the CFE of (5.6±0.6)％,showing a significant difference between them (t =4.454,P =0.011).The apoptotic rates of the cells from 2 through 6 generations were lower in the ESC-CM group than those in the CnT-20 group (all at P＜0.05).The apoptotic rate of the cells was (7.67± 1.31)％ in the siRNA-F transfected group,which was significantly lower than (32.33 ±3.13)％in the siRNA-TERT transfected group (t =-12.588,P =0.000).No significant differences were seen in the expression levels of p63,ABCG2,integrin β1 mRNA and proteins and TERT protein in the primary cells between the ESC-CM group and the CnT-20 group (all at P＞0.05),but significantly declined expressions of CK3 mRNA and protein were found in the ESC-CM group compared with the CnT-20 group (all at P＜0.01).However,the expressions of p63,ABCG2,integrin β1 mRNA and proteins and TERT protein in the second generation of the cells were significantly higher in the ESC-CM group compared with the CnT-20 group (all at P＜0.01).The telomerase activity was (4.83±0.67) ％ in the siRNA-TERT transfected group,which was significantly lower than (46.71±1.22) ％ of the siRNA-F transfected group (t =52.116,P =0.000).The expression of pFAK,pAkt,pGSK3β proteins were weakened,but the expression of p21 was increased in the ESC-CM group after addition of FAK inhibitor,GSK3β inhibitor and TERT-siRNA transfected group.Mitochondrial membrane potential in the second generation of cells was elevated in the ESC-CM group in comparison with the CnT-20 group and the siRNA-TERT transfected group (all at P＜0.01),and the rates of ROS positively reaction was lower in the ESC-CM group and the siRNA-F transfected group than those of the CnT-20 group and siRNA-TERT transfected group (all at P＜0.01).Conclusions ESC-CM culture system can effectively keep the stemness of LSCs and inhibit apoptosis.ESC-CM culture system plays functions probably via telomerase-p21-mitochondrial axis and the activation of the FAK/Wnt signaling pathways.

Objective To investigate the risk factors of pancreatic fistula after pancreaticoduodenectomy.Methods The clinical data of 310 patients who received pancreaticoduodenectomy at the Ruijin Hospital of Shanghai Jiaotong University from January 2005 to May 2013 were retrospectively analyzed.The risk factors associated with the interoperative pancreatic fistula were analyzed.The univariate and multivariate analysis were done using the Pearson chi-square test and non-conditional Logistic regression model.Results A total of 134 patients had postoperative complications,including 103 (33.23％) with pancreatic fistula,among them 40 patients developed additional complications.The results of univariate analysis showed that preoperative levels of hemoglobin,total bilirubin,diameter of the pancreatic duct and postoperative level of albumin were risk factors of pancreatic fistula after pancreaticoduodenectomy (x2 ＝ 4.543,6.087,6.265,5.311,P ＜ 0.05).The results of multivariate analysis showed that preoperative level of total bilirubin equal to or above 34.2 μmol/L,the diameter of the pancreatic duct under 3 mm and the level of postoperative albumin under 28 g/L were the independent risk factors of pancreatic fistula (OR =1.806,1.936,1.780; 95％ confidence interval:1.107-2.948,1.170-3.206,1.002-3.165,P ＜ 0.05).Conclusion Preoperative jaundice (the level of total bilirubin ≥ 34.2 umol/L),pancreatic duct diameter ＜ 3 mm and postoperative malnutrition (albumin ＜ 28 g/L) indicate a higher incidence of postoperative pancreatic fistula.

Discrimination of abnormal images from the numerous wireless capsule endoscope (WCE) video sequence images is laborious and time-consuming, so that a computer-based automatic image recognition system is desired for this task. We propose an algorithm to allow feature extraction from each image channel and decision fusion using multiple BP neural networks. The algorithm was tested and the results demonstrated its high efficiency and accuracy in identification of abnormalities in the WCE images.
Algorithms ; Capsule Endoscopy ; methods ; Image Interpretation, Computer-Assisted ; methods ; Pattern Recognition, Automated ; methods