Objective To investigate midkine (MK) level in serum of patients with esophageal squamous cell carcinoma (ESCC) before and after operation,detect the expression of MK protein in tissues and analyze its relationship with the malignant biological behavior of ESCC.Methods The MK levels in serum were detected by enzyme-linked immunosorbent assay in 30 healthy cases and 55 patients with ESCC when the day before operation and the tenth-day after operation.Then the expression of MK protein in 55 patients surgically removed ESCC were detected by immunohistochemistry using monoclonal antibodies against human MK.Results The average optical density of MK in serum of ESCC patients before operation was 0.1006±0.0624,0.0455±0.0155 in normal control group,0.0752±0.0267 in postoperative control group (t =6.203,P ＜ 0.001).The MK levels in serum of ESCC patients before operation were significantly higher than those after operation (t =4.357,P ＜ 0.001) and those in healthy controls (P ＜ 0.05).The expression of MK protein was no statistically significant correlation with tumor size (x2 =0.131),TNM stage (x2 =0.315) and lymph node metastasis (x2 =0.282) in ESCC (all P ＞ 0.05).But significant correlation was noted between the expression of MK protein and the vasculature involving of the cutting edge (x2 =4.223),degree of cellular differentiation (x2 =10.326),invasive extent of the carcinoma (x2 =20.556) (all P ＜ 0.05).Conclusion MK is overexpressed in ESCC patient' s serum and tissue.The high level of preoperative serum MK is caused by the tumor.

Objective To compare the sensitivities of the stimulation effect of human cord blood mesenchymal stem cells(CB-MSCs) and CB-MSCs of neuronal differentiation to lymphocytes(LCs) detected with carboxyfluorescein didcetate(CFSE) and MTT. Methods To prepare LCs from SD rat and divided into four group stimulating cells: 1. CB-MSCs;2. Dif-CB-HSCs;3. SH-SY5Y(positive control);4. Auto-LC(negative control).Stimulating cells were respectively Co-cultured with LCs. The proliferation of LCs was detected with MTT and CFSE ( n =3). Results CB-MSCs and Dif-CB-HSCs stimulated LCs to proliferate more weakly than positive control detected with MTT and CFSE. The quantity of proliferation of lymphocytes Co-cultured with CB-MSCs and Dif-CB-HSCs were higher than that of Auto-LC detected with CFSE. But MTT OD value of CB-MSCs and Dif-CB-HSCs was a little lower than that of Auto-LC. Statistical analysis results showed no significant difference.Conclusion CFSE can reflect proliferation status of lymphocytes better than MTT. CFSE shows more advantages in practical use.

Objective To study the effects of amniotic epithelial cells conditioned medium on the differentiation of bone marrow stromal cells into neural cells. Methods Bone marrow stromal cells and amniotic epithelial cells were isolated and cultured in vitro,then the cell surface antigen was detected by flow cytometry and the expressions of nestin and ki67 were detected by immunofluorescence staining method.When the cells were co-cultured with amniotic epithelial cells conditioned medium,the morphological character of cells was observed by inverse phase-contrast microscope,and the expressions of NSE(neurone specific enolase),TH(tyrosine hydroxylase) and DAT(dopamine transporter) were detected by immunofluorescence staining method. Results Amniotic epithelial cells conditioned medium had obvious inductive effect on bone marrow stromal cell's neural differentiation.Conclusion The amniotic epithelial cells conditioned medium may have inductive effect neuron-like cell's differentiation and dopaminergic neuron-like cell's differentiation of bone marrow stromal cells in vitro.

Objective To investigate the function of amnion endothelial cell in the differentiation of human umbilical cord blood stem cells into dopaminergic neurons.Methods Primary human amnion endothelial cells were separated and cultured in vitro;the conditioned medium(CM) was prepared through high speed centrifugation.The cord blood mesenchymal stem cells of P_1 passage were induced by the conditioned medium,and the mophology of cells was observed under the inverted phase contrast microscope.The expression of tyrosine hydroxylase(TH)and dopamine transportor(DAT) of the induced cord blood mesenchymal stem cells were detected by immunocytochemistry staining method and immunoblotting(Western blotting).Results The masculine rate of TH and DAT of the cord blood mesenchymal stem cells of P_1 passage which were induced by amnion endothelial cells conditioned medium was higher than that of the control group,with a significant difference(P

Objective To study the proliferation inhibition effect by silencing PLCε gene expression with RNA interference in BIU-87 cells. Methods The specific short hairpin RNA recombinant plasmids were constructed by gene clone technology.The expression level of PLCε protein and mRNA were detected by Western blot and RT-PCR respectively after transfected recombinant plasmids into BIU-87 cells.The influence on proliferation was check by MTT.The changes of proliferating cell nuclear antigen(PCNA)were analyzed by immunocytochemical method,and the distribution of cell cycle was analyzed using flow cytometry. Results After transfected with the specific recombinant plasmids,PCNA expression was decreased 33.08%,and the analysis of cell cycle indicated that cells of G0/G1 phase were increased comparision with(40.75±2.30)%and(40.00±1.76)0A,and its G2/M phase cells(8.16±0.51)%were decreased strikingly compared with group control(31.20±1.76)%and group NP(35.94±1.58)%.Cells were blocked at G0/G1 phase,the cell proliferation was inhibited obviously. Conclusion PLCε may play an important role in proliferation of bladder cancer cells,which could be a potential target of biological treatment on bladder cancer in the future.

GFAP.Conclusion RA is the best factor for neurons and astroglia,and RA+EGF+bFGF are the best for oligodendrocytes.

Objective:To explore the efficacy of a flat ground exoskeleton robot in improving the walking ability of stroke survivors.Methods:Fifty-eight stroke survivors with mobility difficulties were randomly divided into a robot group ( n=29) and a control group ( n=29). In addition to routine rehabilitation, the control group received conventional walking training, while the robot group underwent exoskeleton robot-assisted gait training. The 30-minute training sessions were held twice a day, 5 days per week for 5 weeks. Before as well as after 2 and 4 weeks of treatment, everyone′s walking ability was tested using the 6-minute walk test (6MWT) and functional ambulation scale (FAC). General lower limb motor function was quantified using the Fugl-Meyer Lower Extremity assessment (FMA-LE). Moreover, gait analysis was conducted before and after 4 weeks of treatment. Results:After 2 and 4 weeks of treatment, the average 6MWT times of both groups were significantly better than before the treatment, with the improvement of the robot group significantly greater than that of the control group after 2 weeks. After 2 and 4 weeks the average FMA-LE and FAC scores of both groups had improved significantly compared with before treatment. After 4 weeks the stride frequency and gait cycle of both groups had improved significantly.Conclusions:Exoskeleton robot-assisted gait training can improve walking ability and lower limb motor function of stroke survivors about as well as conventional walking training.

Objective To explore the effectiveness of the Five Elements Music Therapy(FEMT)in relieving stress in participants with different cultural backgrounds,and to compare the differences between the FEMT and the Western Art Music Therapy(WAMT)in stress relief.Methods This was a comparative pilot randomized pre-post repeated measures study,37 subjects were allocated with 2 dropped out,with an inclusion-ended sample of 35 subjects,23 from Canada and 12 from China.After informed consent,all subjects were randomly assigned to listening to either Five Elements Music or Western Art Music at home for 30 minutes,twice a week for four weeks.Participants were asked to use headphones,measure their pulse rate before and after each session,and fill out five questionnaires,including a background and demographic survey(reporting age,gender,education,cultural background,listening experience).Self-assessment of stress(pre-post after each session),General Hospital Anxiety/Depression Scale(HADS,weekly after 2nd session),Perceived Stress Survey(PSS,weekly after 2nd session),the Music Therapy Intervention Survey(MTIS,pre-post each session).Results ①There was a significant decrease in self-assessed stress scores after the second session in the FEMT compared with the WAMT group(t=-2.057,P=0.046).②In both groups,there was a significant decrease in stress scores pre-post treatment in each group(WAMT t=5.026;FEMT T=7.645,P=0.000).③There was no significant difference between the two groups in post-intervention HADS scores(P>0.05);In the Chinese sample,there was a significant difference in HADS scores after the eighth session in both FEMT and WAMT groups(t=-3.862,P=0.003),and a statistically significant difference in HADS pre-post intervention in the FEMT((t=5.117,P=0.004).There was a significant difference in MTIS pre-post treatment in the WAMT(t=-2.572,P=0.023),but in not the FEMT group(t= 1.331,P=0.207).Conclusion This pilot trial explores a safe and feasible self-administered music therapy approach for stress in two distinct cultural groups,and for the first time provides preliminary comparison and evidence of effectiveness of FEMT and WAMT in alleviating stress and anxiety.Further investigation with bigger randomized samples is needed to elucidate the effects of different kinds of music and cultural groups on stress and anxiety levels.

Objective To report our efficacy of one-stage definite treatment of Gustilo type Ⅲ open fractures of long bone by plate osteosynthesis.Methods A retrospective case series study was conducted of the 69 cases who had undergone plate osteosynthesis for one-stage definite treatment of Gustilo type Ⅲ open fractures of long bone from January 2006 to June 2016 at Microsurgery and War Trauma Center of Chengdu Military Command,59 Hospital of Chinese PLA.They were 47 males and 22 females with an average age of 34.2 years (from 2 to 62 years).There were 27 shaft fractures of tibia or fibula (13 cases of type ⅢA,12 cases of type Ⅲ B and 2 cases of type Ⅲ C),4 fractures of distal tibia (2 cases of type Ⅲ A and 2 cases of type ⅢB),14 shaft fractures of ulna or radius (9 cases of type ⅢA,3 cases of type ⅢB and 2 cases of type Ⅲ C),12 factures of humeral shaft (7 cases of type Ⅲ A,3 cases of type Ⅲ B and 3 cases of type Ⅲ C),3 fractures of distal humerus (all type ⅢC),6 fractures of femoral shaft (5 cases of type ⅢA and one type Ⅲ C),and 3 fractures of distal femur (2 cases of type ⅢA and one type ⅢC).The intervals between injury and operation ranged from 4 to 17 hours,averaging 9.6 hours.After thorough debridement,osteosynthesis was performed with locking compression plate,limited contact dynamic compression plate or/and reconstruction locking plate,or 1/3 tubular plate.Direct closure with decreased tension or without tension was used for type Ⅲ A injury;deep open defects were repaired with perforator flaps,neurovascular axis flaps,traditional axis flaps and muscular flaps,or local flaps;limb reconstructions included neurovascular repair in 12 cases,tendon and ligament repair in 5 cases,and muscle reconstruction in 3 cases.Superficial defects were covered by skin grafts simultaneously or secondarily.Results The duration of hospitalization averaged 19 days (from 5 to 37 days).Partial necrosis occurred in one case of sural neurovascular axis flap.Superficial infection with multiple antibiotic-resistant bacteria occurred in 2 cases.Follow-up for the 69 patients ranged from 12 to 27 months (average,19.2 months).No deep bone infection occurred.Implant breaking occurred in 4 cases and implant loosening in one.The implant failures were corrected by change into intramedullary nails or plate refixation (respectively in 2 cases) in addition to bone graft.Bone union was achieved after 5 to 15 months (average,7.7 month)with satisfactory aesthetic and functional outcomes.Conclusion For patients with Gustilo type Ⅲ open fracture of long bone,especially those with metaphyseal,intraarticular or upper limb fracture and pediatric ones,plate osteosynthesis can be a satisfactory one-stage definite treatment besides intramedullar nailing and external fixation,providing that through debridement and satisfactory soft-tissue coverage can be achieved.

Purpose: Mixed-lineage leukemia protein 4 (MLL4/KMT2D) is a histone methyltransferase, and its mutation has been reported to be associated with a poor prognosis in many cancers, including lung cancer. We investigated the function of MLL4 in lung carcinogenesis. Materials and Methods: RNA sequencing (RNA-seq) in A549 cells transfected with control siRNA or MLL4 siRNA was performed. Also, we used EdU incorporation assay, colony formation assays, growth curve analysis, transwell invasion assays, immunohistochemical staining, and in vivo bioluminescence assay to investigate the function of MLL4 in lung carcinogenesis. Results: We found that MLL4 expression was downregulated in non–small cell lung cancer (NSCLC) tissues compared to adjacent normal tissues and tended to decrease with disease stage progression. We analyzed the transcriptomes in control and MLL4- deficient cells using high-throughput RNA deep sequencing (RNA-seq) and identified a cohort of target genes, such as SOX2, ATF1, FOXP4, PIK3IP1, SIRT4, TENT5B, and LFNG, some of which are related to proliferation and metastasis. Our results showed that low expression of MLL4 promotes NSCLC cell proliferation and metastasis and is required for the maintenance of NSCLC stem cell properties. Conclusion Our findings identify an important role of MLL4 in lung carcinogenesis through transcriptional regulation of PIK3IP1, affecting the PI3K/AKT/SOX2 axis, and suggest that MLL4 could be a potential prognostic indicator and target for NSCLC therapy.