Objective To study the effectiveness of comprehensive rehabilitative therapy with paraplegic spinal cord injury (SCI) patients. Methods Forty patients with SCI-caused paraplegia were divided randomly into a rehabilitation group and a control group. The routine treatment was administered to both groups, and comprehensive rehabilitative therapy was also administered to patients in the rehabilitation group as an addition. Functional assessments and somatosensory evoked potential (SEP) tests were performed with the two groups pre-treatment and 30 days post-treatment. Results The rehabilitation group achieved, on average, greater improvement in their physical functions , as demonstrated by their much higher scores in terms of the Barthel index than those of the control group ( P

Objective: To study the influence of pre-operative disease course on the operation and post-operation quality of life of adolescents with idiopathic scoliosis (AIS). Methods: A total of 110 AIS patients who were treated with classic posterior correction, pedicle internal fixation were divided into two groups according to their disease courses (n=55): short course group with a pre-operation course <2 years (S group), long course group with a pre-operation course ≥2 years (L group). The gender, Lenke type, and major curve Cobb angle were matchable between the two groups. Various radiographic measurements and indices like fusion level, intraoperative blood loss and scores of SRS-22 scale were compared between the two groups before operation, immediately, and 2 years after operation. Results: The pre-operative Cobb angles of the major curve were similar between the two groups, but the flexibility of the major curve averaged (55.7±18.77)% in the S group and (48.1±18.24)% in the L group (P = 0.034). Pre-operative Cobb angles of the minor curve were larger in L group than those in S group ([30.1±12.10]° in the S group and [34.8±10.85]° in the L group, P = 0.035). The post-operative radiographic measurements and the blood loss/infusion were similar between the two groups. The number of fused vertebrae in the L group was significantly more than that in the S group (P = 0.027). The parameters in the SRS-22 scale, including function/activity, pain, self-image/appearance, and mental health were similar between the two groups during follow-up. And the L group had a significantly lower satisfaction rate of treatment compared with the S group ([4.0±0.70] vs [3. 7±0.78], P = 0.037). Conclusion: The flexibility of the curve in AIS decreases with the increase of disease course, and the disease course might be a risk factor for the scoliosis progression of the minor side. Patients with a disease course ≥2 years have more fused vertebrae than those with a disease course <2 years. The length of pre-operative disease course has influence on the quality of life of AIS patients after operation.

AIM:To observe whether modified epitopes from osteosarcoma high-expressing antigen papilloma-virus-binding factor (PBF) have HLA-A2 restricted antitumor ability,and to develop peptide-based immunotherapy for os-teosarcoma. METHODS:RT-PCR and Western blot were used to determine the expression of PBF in the osteosarcoma cell lines U2OS and Saos-2. HLA-A2 epitopes from PBF protein were predicted by NetCTL 1.2, SYFPEITHI and IEDB. The modified peptides from PBF containing HLA-A2 binding anchor motifs were designed by replacing the anchor residues. The peptides were synthesized by standard solid-phase methods,and the binding affinity of the peptides to HLA-A?0201 was evaluated by T2A2 cell binding assay. ELISPOT assay was used to investigate the seretion of interferon-γ(IFN-γ) from the peptide-induced specific cytotoxic T-lymphocytes (CTLs). The ability of inducing T-cell response was analyzed by lactate dehydrogenase (LDH) release assay and carboxyfluorescein succinimidyl ester (CFSE) cytotoxicity assay in vitro. RE-SULTS:The expression of PBF was observed in the U2OS and Saos-2 cells. The candidate peptides P75-1Y2L,P412-1Y, P416-1Y2L9V, P107-1Y and P435-1Y2L showed moderate affinity toward HLA-A2 molecule. The modified peptides showed significantly higher affinity with HLA-A2 than the native peptide. ELISPOT assay showed that P412, P412-1Y, P416,P416-1Y2L9V and P435-1Y2L induced specific CTLs to secrete IFN-γ, and P412-1Y and P416-1Y2L9V induced more secretion of IFN-γ than the native peptide. The CTLs induced by P412, P412-1Y, P416 and P416-1Y2L9V lysed U2OS cells. P412-1Y and P416-1Y2L9V peptide-specific CTLs showed higher cytotoxicity against U2OS cells than the native peptide-specific CTLs. CONCLUSION:Compared with the native peptide,modified epitopes P412-1Y and P416-1Y2L9V have higher binding affinity with HLA-A?0201 and retain immunogenecity. In addition,the anti-tumor immunity effects of modified epitopes P412-1Y and P416-1Y2L9V are stronger than the native peptide. The peptides P412-1Y and P416-1Y2L9V is excellent HLA-A?0201 restricted CTL epitopes from tumor antigen PBF,which could serve as new can-didates towards antitumor peptide vaccines.

0.05);the body weight of 170 and 357 mg/kg 3,4-DCA exposure groups decreased significantly(P

Tacrolimus is a novel immunosuppressant used in the treatment of a variety of autoimmune diseases.More and more studies have shown that tacrolimus has a certain therapeutic effect on myasthenia gravis (MG).This article reviews the mechanism,clinical researches,adverse reactions,dosage and clinical evaluation of tacrolimus in the treatment of MG.

OBJECTIVETo analyze the influence of retinoic acid (RA) on the undifferentiated state and EB formation abilities of human embryonic stem cells.

METHODSThe biological characteristics of H9 ESCs after RA treatment were characterized by real-time PCR, MTS proliferation assay and immunofluorescence staining. The expression of three germ layers markers, osteogenic differentiation markers and adipogenic differentiation markers in H9-differentiated embryoid bodies (EBs) with RA treatment were quantified by real time PCR.

RESULTSThe proliferation of H9 ESCs in the early logarithmic growth phase was accelerated by RA treatment. In addition, RA induced differentiation of H9 ESC coupled with morphology changes, decreased expression of undifferentiated markers Oct4, Nanog, Sox2 and OCT4 mRNA binding protein Lin28 at mRNA level, and reduced expression of Oct4 at protein level. RA induced formation of cavities in EBs. Real time PCR results showed that the expressions of ectodermal markers: NeuroD1, Noggin; mesodermal markers: Brachyury, Twist and endodermal markers: AFP, GATA-4 were significantly increased (P < 0.05), especially for AFP (P < 0.01), by RA treatment in a dose-dependent manner. In addition, the expression of adipogenic differentiation marker C/EBPalpha was increased while the osteogenic differentiation marker OPN was decreased in EBs after RA treatment for 5 days.

CONCLUSIONSHigh concentrations of RA induced the loss of stemness in H9 ESCs and excessive differentiation in EBs, and damaged the balance between osteogenic and adipogenic differentiation during early EB differentiation, which may be relevant to the congenital malformations.

Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Embryonic Stem Cells ; cytology ; drug effects ; Humans ; Tretinoin ; pharmacology

OBJECTIVE:To establish the method for the simultaneous determination of anticancer activity components of flavonoids from Hedyotis diffusa,i.e. quercetin,kaempferol. METHODS:HPLC was applied to determine the contents and performed on Alltima C18(250 mm?4.6 mm,5 ?m) column. Mobile phase consisted of methanol(A)-0.5% glacial acetic acid,(gradient elution). The detection wavelength was aet at 350 nm. RESULTS:The linear range of quercetin was 0.006 2～0.244 0 ?g(r=0.999 8)and that of kaempferol 0.007 8～0.310 6 ?g(r=0.999 9). The average recovery of quercetin was 101.84%(RSD=1.79%,n=6) and that of kaempferol 99.04%(RSD=2.90%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible for the quality control of H. diffusa.

Objective To investigated the role of antithrombin Ⅲ (AT-Ⅲ) levels in the early diagnosis of disseminated intravascular coagulation (DIC) in patients with sepsis and the predictive effect of AT-Ⅲ on the development of DIC.Methods A retrospective study was conducted. Patients admitted to intensive care unit (ICU) of the First Affiliated Hospital of China Medical University from January to December in 2015 were enrolled. The patients were divided into sepsis group and non-sepsis group according to the diagnostic criteria of sepsis. In addition, sepsis patients were divided into 3 subgroups according to the international society on thrombosis and haemostasis (ISTH) scores on the first day: overt DIC (ISTH ≥ 5), non-overt DIC (ISTH 1-4) and none DIC group (ISTH = 0). Blood routine test, prothrombin time (PT), fibrinogen (Fib), D-dimer, fibrin degradation products (FDP), acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) scores, sequential organ failure assessment (SOFA) scores and ISTH scores were recorded on the first ICU day. AT-Ⅲ was recorded during 7 days. The differences were compared among these 3 groups. Correlations of AT-Ⅲ with various parameters were calculated by using Pearson correlation analysis in sepsis group and overt DIC group. Receiver operating characteristic (ROC) curves for diagnosis of DIC with AT-Ⅲ, AT-Ⅲ+PT were drawn to evaluate the diagnostic efficiency. The AT-Ⅲ levels of DIC patients were compared between early-onset DIC and late-onset DIC during their ICU stay. The change of AT-Ⅲ levels with time and prognosis in patients with early-onset DIC was compared between groups.Results Totally 445 patients were recruited, with 138 patients in sepsis group, and 307 in non-sepsis group. There were 20 patents diagnosed with overt DIC on the first ICU day, 115 patients non-overt DIC and 3 patients of none DIC. Twenty-five sepsis patients were diagnosed overt DIC during the ICU days. AT-Ⅲ level in sepsis patients on the first ICU day were lower than that in non-sepsis patients [(55.29±13.92)% vs. (76.54±12.31)%,P < 0.01]. Patients with overt DIC had a lower AT-Ⅲ level than non-overt DIC or none DIC patients [(43.85±13.00)% vs. (56.95±13.03)%, (68.00±16.52)%, bothP < 0.05]. It was shown by Pearson correlation analysis that AT-Ⅲ level of sepsis patients on the first ICU day was negatively correlated to ISTH score and PT (r value were -0.467, -0.654, bothP < 0.01). AT-Ⅲ level of overt DIC patient on the first ICU day was negatively correlated with PT (r = -0.675,P = 0.001). It was shown by ROC curve that area under ROC curve (AUC) of AT-Ⅲ combined with PT for diagnosis overt DIC in sepsis patients was higher than that of AT-Ⅲ or PT alone (0.843 vs. 0.763, 0.834), the sensitivity 90.0%, specificity 73.7%. The cut-off value for overt DIC diagnosis in sepsis patients of AT-Ⅲ level alone was 48.5%, sensitivity was 78.0%, specificity was 70.0%. On the first ICU day, AT-Ⅲ level was risen when ISTH score improved in patients with sepsis. There was similar change of AT-Ⅲ level between patients with early-onset DIC and late-onset DIC. AT-Ⅲ level increased with DIC improvement.Conclusion AT-Ⅲ level can be used for diagnosing sepsis-associated overt DIC independently or with a combination of PT. When ISTH score improved, AT-Ⅲ level was risen in patients with sepsis associated DIC.

Objective To evaluate the value of fecal high mobility group box-1 (HMGB 1 ) in early diagnosis and prediction of the severity of neonatal necrotizing enterocolitis (NEC). Methods From July 2013 to June 2015 , the neonates who had distention, vomit, or gross blood in stool and were suspected of NEC were recruited as NEC group while hospitalized children without abdominal distension, vomiting, bloody diarrhea, or other gastrointestinal symptoms were recruited as the control group. Stool samples were collected on day 1 , 3 , 5 and 7 after admission. The level of HMGB 1 was measured by enzyme linked immunosorbent assay (ELISA). Results In the end, there were 46 cases in NEC group and 15 cases in control group. In NEC group, 29 cases were conifrmed of stageⅠof NEC by abdominal radiograph within 24 h after hospitalized, all of them were deteriorated to stageⅡphase in 4 days, and 10 cases were deteriorated to stageⅢ. Seventeen cases were conifrmed of stageⅡby abdominal radiograph within 24 h after hospitalized, 7 cases were deteriorated to stageⅢ. In 17 stageⅢcases, 11 cases received surgical treatment and 6 cases gave up. Eight cases survived and 3 died after surgery. The levels of HMGB 1 in NEC group on day 1 , 3 , 5 and 7 after hospitalized were higher than those in control group (P

Objective To study the effects of diagnostic ultrasound combined with microbubble contrast agent on augmentation thrombolysis in vivo and to discuss the synergism and mechanism of combination of urokinase. Methods Thirty-two rabbits of femoral vein thrombus were divided into four groups: simple urokinase group, ultrasound and contrast microbubble group, ultrasound and contrast microbubble combined with urokinase group, control group. Then according to the best parameter combination which was definited in previous experiment, the experiments combination of urokinase were performed. Color Doppler was used to evaluate the recanalization of different group in 15 rain,30 min and 60 min. Results After the treatment of 15 rain and 30 min,the recanalization rate in ultrasound and contrast microbubble combined with urokinase group improved significantly compared with other groups (P<0.05);after 60 min, the recanalization grade of ultrasound and contrast microbubble and urokinase group was evidently better than other two group. The result of immunohistochemistry showed that disaggregation of clot and fibrous in this group at 60 min was better than the results at 15 min, 30 min. Conclusions Diagnostic ultrasound with microbubble contrast agent showed superior effects of thrombolysis and the synergism of combination of urokinase in vivo.