Objective To investigate the correlation of diabetic skeletal muscle disease with macroangiopathy, and to explore the related genes of Shenqi Compound Recipe (SCR) in preventing and treating diabetic skeletal muscle disease by using gene chip technique, thus to reveal the molecular mechanism. Methods KKAy mice were fed with water containing nitri oxide synthase inhibitor of Nω-nitro-L-arginine methyl ester ( L-NAME) and high fat diet to induce the macroangiopathy complicated with type 2 diabetes. The experimental animals were divided into normal c57BL/GJ group, KKAy group, model group, SCR group (in the dosage of 14.4 g·kg-1·d-1) and rosiglitazone group ( in the dosage of 1.33 mg·kg-1·d-1) , 15 in each group. The medication groups were administered the corresponding agents for 8 consecutive weeks just as the modeling began. During the experiment period, blood glucose was monitored. At the end of the experiment, the abdominal aorta and skeletal muscle of mice were taken out for the observation of morphological changes, and differentially expressed genes of skeletal muscle between SCR group and model group, and between model group and KKAy group were detected by gene chip technique. Results SCR had an effect on relieving the atrophy, edema, fracture, and inflammatory changes in the skeletal muscle. There were 198 genes differentially expressed between model group and KKAy group, including 119 up-regulated genes and 79 down-regulated genes. There were 70 genes differentially expressed between SCR group and model group, including 33 up-regulated genes and 37 down-regulated genes. In the two comparison groups, 7 genes ( Celsr2, Rilpl1, Dlx6as, 2010004M13Rik, Anapc13, Gm6097, Ddx39b) showed reversed differential expression. Conclusion Diabetic skeletal muscle disease is associated with macroangiopathy. SCR has preventive effect on diabetic skeletal muscle lesion, and the mechanism may be related to the regulation of Celsr2, Rilpl1, Dlx6as, 2010004M13Rik, Anapc13, Gm6097, Ddx39b gene expression.

Objective To investigate whether the inhibition of caveolin-1 (Cav-1) phosphorylation will regulate effectively nuclear factor-erythroid 2-related factor (Nrf2) signal pathway and downstream effector molecules and protest against ventilation induced lung injury (VILI) in an animal model in vivo. Methods Ninety male Sprague-Dawley (SD) rats were randomly divided into nine groups (each n = 10): sham group in which rats did not receive ventilation but received tracheotomy; lung protective ventilation (PV) for 1 hour or 2 hours group; mechanical ventilation (MV) at high volume tidal (VT, 40 mL/kg) for 1 hour or 2 hours group; protein tyrosine kinase inhibitor PP2 or rosiglitazone (Rsg) pretreatment + high VT ventilation for 1 hour or 2 hours groups. The two pretreatment groups were given intraperitoneal injection PP2 15 mg/kg or intragastric administration of Rsg 5 mg/kg 1 hour before ventilation respectively. The rats were sacrificed after model reproduction, and bronchoalveolar lavage fluid (BALF) was collected. Pulmonary vascular permeability was measured by Evans blue (EB). The levels of tumor necrosis factor-α (TNF-α), activator protein-1 (AP-1), nuclear factor-κB (NF-κB), and interleukin-8 (IL-8) in BALF were determined by enzyme linked immunosorbent assay (ELISA). Then the lung tissues were collected, the lung wet/dry ratio (W/D) was calculated, the changes in pathology was observed with light microscope, and myeloperoxidase (MPO) activity was determined by colorimetric analysis. Nrf2 mRNA was determined by reverse transcription-polymerase chain reaction (RT-PCR). The expressions of Cav-1 tyrosine residues 14 phosphorylation (pCav-1-Y14), Cav-1, peroxisome proliferators-activated receptor γ (PPARγ) and claudin-5 as well as Nrf2 in cytoplasm and nucleus were determined by Western Blot. The positive expressions of PPARγ and claudin-5 in lung tissues were assayed with immunohistochemistry staining. Results There were no obvious pathological changes in the lung tissue in sham group and PV groups, and there were no significant differences in all the parameters between the two groups either. However, the injury in lung tissue was severe in the high VT groups in which W/D ratio, EB contents, MPO activity, and TNF-α, AP-1, IL-8, NF-κB levels in BALF as well as the protein expressions of Cav-1 and pCav-1-Y14 were significantly higher than those of sham group and PV groups, and the protein expressions of PPARγ and claudin-5 were significant lower than those of sham group and PV groups with a dose-dependent manner; but Nrf2 expressions in cytoplasm and nucleus did not show a statistical increase. After pretreatment of PP2 or Rsg, W/D ratio, MPO activity, EB contents, TNF-α, AP-1, IL-8, and NF-κB in BALF were significantly decreased as compared with those of high VT group, and RT-PCR showed significant up-regulation of Nrf2 mRNA in lung tissues too. Moreover, there was a statistically significant increase in expressed Nrf2 proteins in nucleus in PP2 or Rsg groups as compared with those of high VT groups [Nrf2 in nucleus (gray value): 0.61±0.06, 0.56±0.06 vs. 0.31±0.02 at 1 hour, 0.38±0.06, 0.43±0.07 vs. 0.22±0.03 at 2 hours; all P < 0.05], but no significant difference was found in the expression of Nrf2 protein in the cytoplasm among all groups. The protein expressions of pCav-1-Y14 in PP2 pretreatment groups were significantly lower than those of high VT groups (gray value: 0.89±0.04 vs. 1.48±0.02 at 1 hour, 0.86±0.02 vs. 1.31±0.01 at 2 hours; both P < 0.05); but expressed PPARγ proteins and expressed claudin-5 proteins in PP2 or Rsg pretreatment groups were significantly higher than those of high VT groups [PPARγ (gray value): 0.34±0.07, 0.42±0.13 vs. 0.17±0.07 at 1 hour, 0.38±0.09, 0.33±0.07 vs. 0.16±0.03 at 2 hours; claudin-5 (gray value): 0.33±0.05, 0.38±0.07 vs. 0.14±0.03 at 1 hour; 0.30±0.06, 0.31±0.04 vs. 0.17±0.04 at 2 hours; all P < 0.05]. Conclusions The inhibition of Cav-1-Y14 phosphorylation can increase the expression of Nrf2 in the nucleus, then result in an increase in the protein expressions of PPARγ and claudin-5 of its effector molecules. This effect can reduce the inflammation and capillary permeability of lung tissue in the model of VILI.

ObjectiveTo determine whether the inhibition of caveolin-1 tyrosine residues 14 (Cav-1-Y14) phosphorylation with protein tyrosine kinase inhibitors (PP2) will upregulate heme oxygenase-1 (HO-1) activity to protect against ventilation induced lung injury in vivo of an animal model.Methods Fifty-four male Sprague-Dawley (SD) rats were randomly divided into nine groups (eachn = 6). Group A served as normal control group, in which rats did not receive ventilation but tracheotomy. Groups B1 and B2 received lung protective ventilation respectively for 1 hour or 2 hours. Groups C1 and C2 received high tidal volume (40 mL/kg) ventilation for 1 hour or 2 hours, respectively. The group D1 or D2 also received high tidal volume ventilation for 1 hour or 2 hour respectively, but they were given PP2 1 hour before high tidal volume ventilation. The groups E1 and E2 also received high tidal volume ventilation respectively for 1 hour or 2 hours, but tyrosine kinase inhibitor PP2 and HO-1 inhibitor zinc protoporphyrinⅨ(ZnPPⅨ) were given to animals 18 hours before high tidal volume ventilation. All the animals were sacrificed after ventilation, and the specimens of lung tissues and bronchoalveolar lavage fluid (BALF) were harvested. Then the changes in pathology of lung tissue was observed, and diffuse alveolar damage scores (DAD) were calculated, myeloperoxidase (MPO) activity was measured by colorimetric analysis, lung wet/dry ratio (W/D) was estimated. The expressions of phosphorylated caveolin-1 (P-Cav-1-Y14), caveolin-1 (Cav-1) and HO-1 were determined by Western Blot. The expressions of high mobility group B1 (HMGB1) and advanced glycation end product receptor (RAGE) in lung tissues were assayed with immunohistochemistry staining. The levels of tumor necrosis factor-α(TNF-α) in BALF were measured by enzyme linked immunosorbent assay (ELISA).Results There was no significant difference in all the parameters between group A and groups B. Compared with group B1, DAD score, W/D ratio, the activity of MPO and the concentration of TNF-α in BALF in group C1 were significantly increased [DAD score:7.97±0.59 vs. 0.55±0.13, W/D ratio: 5.70±1.61 vs. 5.04±0.63, MPO (U/g): 1.82±0.14 vs. 0.77±0.26, TNF-α(ng/L): 370.10±29.61 vs. 54.38±8.18, allP< 0.05], and the injury in ventilation 2 hours group was more serious than that in ventilation 1 hour group. Compared with groups C, all the parameters in groups D were significantly decreased. The parameters in groups E were significantly higher than those in groups A, B, and D, but no significant difference was found as compared with groups C. Compared with groups B, the protein expressions of Cav-1 and P-Cav-1-Y14 (gray value) in groups C were significantly increased (1 hour: 1.49±0.02 vs. 1.26±0.13, 1.34±0.02 vs. 0.87±0.04;2 hours: 1.58±0.02 vs. 1.27±0.27, 1.31±0.01 vs. 0.95±0.02, allP< 0.05), and the expression of HO-1 protein (gray value) was significantly decreased (1 hour: 0.59±0.02 vs. 1.10±0.01, 2 hours: 0.49±0.01 vs. 1.20±0.02, both P< 0.05). No significant difference in Cav-1 protein expression between groups D as well as groups E and groups C. The protein expression of P-Cav-1-Y14 in groups D and E was significantly lower than that in groups C. The protein expression of HO-1 in groups D was significantly higher than that in groups C, but the phenomenon was not found in groups E as compared with groups C. Compared with group A, the positive expression of HMGB1 and RAGE in lung tissue in groups C and E was significantly increased, but no significant difference was found between groups B as well as groups D and group A.Conclusion Cav-1-Y14 phosphorylation is the key factor for ventilator induced lung injury, which can not only lead to a decrease in vascular barrier function, but also inhibit the activity of HO-1 enzyme, thus further aggravates inflammatory injury of the lung as induced by mechanical ventilation.

Objective To investigate the characteristics of motor development in children with mental retardation. Methods Motor development was assessed with the Peabody Developmental Motor Scale 2 (PDMS-2) and mental development with adaptive developmental quotient (ADQ) of Gesell developmental schedules (GDS) in 430 infants (6~36 months old) with mental retardation. Results The gross (GDQ), fine (FDQ) and total motor developmental quotient were poor in all the children, and significantly different among children with slight, mild, and serious retardation (P<0.01). ADQ correlated with each sub-score of PDMS-2 (P<0.01). For PDMS-2, FDQ correlated with sub-scores of gross motor, and GDQ with sub-scores of fine motor (P<0.01) in all the mental retardate children. Conclusion The development of both gross and fine motor is poor in children with mental retardation. Mental development correlate with motor development, and gross with fine motor development.

Objective To describe the clinical manifestation of the chronic atlantoaxial rotatory dislocation (AARD) with C1,2 lateral facets locked,and explore the effect of the operation combined with anterior retropharyngeal approach and posterior approach.Methods Data of 8 patients with chronic AARD with C1,2 lateral facet locked who had undergone open reduction with the anterior retropharyngeal approach and C 1,2 transpedicular screw fixation with autologous iliac bone graft from Oct.2010 to Jun.2013 were retrospectively analyzed.There were 4 males and 4 females with an average age of 31 years old (range,11-57 years old).The intervals from onset to diagnosis were from 29 to 180 days and the mean time was 70.6 days.5 cases were chronic AARD with right C1,2 lateral facets locked and 3 with left C1,2 lateral facets locked.Reduction was failed to obtain by traction for two to four weeks.Subsequently,after open reduction with the anterior retropharyngeal approach,the patients were performed C 1,2 transpedicular screw fixation with autologous iliac bone graft for one stage or two.Results 1 patient underwent open reduction with the anterior retropharyngeal approach in stage one and C1,2 transpedicular screw fixation in stage two because of an overall severe condition,and the other patients did anterior and posterior operation in one stage.The average operation time was 205 min (range,160-260 min).The mean blood loss was 210 ml (range,100-300 ml).There were no operation complications except one young girl reflected pain in autologous iliac donated area,and pain relieved when taking non-steroidal anti-inflammatory by oral for one week.All patients had been followed up for a mean period of 14.8 months (range,5-37 months).Three-dimensional computed tomography revealed C1,2 arthrodesis bone graft fusion from two to four months and the average was 3.1 months.Neither recurrence of symptoms nor dislocation was observed at the latest follow-up.Conclusion Open reduction through the anterior retropharyngeal approach with C1,2 transpedicular screw fixation is an effective treatment strategy for chronical AARD with C1,2 lateral facets locked,which reduces surgical complications with less operation difficulty.

Objective To observe the effect of combining virtual reality technology with occupational thera-py in treating children with spastic hemiplegia resulting from cerebral palsy. Methods Thirty-eight spastic and hemiplegic children with cerebral palsy were randomly divided into a treatment group ( n=18) and a control group ( n=20) . Both groups received occupational therapy, while the treatment group was additionally provided with virtual reality-based treatment. The sessions lasted 30 min, 6 times a week for 3 months. Before and after the treatment the upper extremity motor function of both groups was assessed using the fine motor quotients of the Peabody developmen-tal motor scale, the Caroll hand function scale and a activities of daily life scale. Results Before the treatment there was no significant difference between the two groups in terms of any of the measurements. After the 3 months, significant improvement was observed in both groups, but the improvement of the treatment group was significantly greater than that of the control group. Conclusion Virtual reality technology can further improve the motor function of the upper ex-tremities and ability in the activities of daily life beyond that achievable through occupational therapy alone.

Objective To explore the characteristics of visual P300 event-related potentials (ERP) in children with mental retardation and evaluate the correlations between visual P300 in event-related potentials and intelligent quotient.Methods Thirty children with mental retardation were selected as the test group who were inpatients or saw a doctor in the Department of Rehabilitation of Hunan Children Hospital.At the same time,thirty normal children were selected as the control group who saw a doctor in the Department of Child Healthcare of Hunan Children Hospital.Two groups had no statistically significant difference in sex and age (P ＞ 0.05) and were comparable.All children were administered with the Wechsler intelligence test in the Department of Child Healthcare and the visual event-related potential test in the Department of Rehabilitation.The results of the visual P300 latency and amplitude were analyzed.Another,the relevance between intelligent quotient (IQ) and P300 latency and amplitude were also compared by statistics.Results Compared to the control group,the visual P300 latency in children with mental retardation (MR) was apparently longer and the amplitude decreased significantly in children with MR,with a statistically significant difference between two groups(P ＜0.05).P300 latency in MR expected negative correlation with intelligent quotient and the amplitude in MR expected no correlation with intelligent quotient.Conclusions P300 latency and amplitude of ERP could objectively reflect cognitive function in children with MR,could be regarded as one of the objective measurement in the evaluation of cognitive function in children with MR.It can be used as an objective electrophysiological index for assisting in the diagnosis of mental retardation of children in clinic.

A new kind of Cage made of poly-DL-lactic acid was designed and an in-vitro study was conducted to evaluate the biomechanical effect of PDLLA Cage on a goat cervical spine model. 27 goat cervical spines were divided into four groups randomly: intact group, PDLLA Cage group, titanium Cage group, and autologous tricortical iliac crest bone group. Different implants were implanted after complete discectomy (C3-4) was performed, then they were tested in flexion, extension, axial rotation, and lateral bending with a nondestructive stiffness method using a nonconstrained testing apparatus, and three-dimensional displacement was measured. The range of motion (ROM) and the mean stiffness values were calculated and compared between groups. The result showed that the ROM values between PDLLA Cage group and the titanium Cage group were not significantly different in extensional and rotational movement (P > 0.05), but they were lower than that of rest groups (P < 0.05). In flexional loading mode, the ROM values showed statistically significant difference between the four groups (P < 0.05). And in lateral bending loading mode, no significant difference was found between PDLLA Cage group and iliac crest bone group (P > 0.05), but significant difference was found among the other groups (P < 0.05). The stiffness of cervical spine was raised after Cage was implanted. In flexional and rotational loading mode, significant difference in stiffness was found between PDLLA Cage group and control group or iliac crest bone group (P < 0.05). So PDLLA Cage can provide enough primary stability for cervical intervertebral fusion.
Animals ; Biomechanical Phenomena ; Bone Transplantation ; Cervical Vertebrae ; physiology ; surgery ; Equipment Design ; Female ; Goats ; Joint Instability ; prevention & control ; Lactic Acid ; Orthopedic Fixation Devices ; Polyglycolic Acid ; Prostheses and Implants ; Spinal Fusion ; instrumentation

OBJECTIVE: To study the mTOR expression of cancer stem cells(CSCs) in nasopharyngeal carcinoma and preliminarily explore the mechanism of inhibiting its proliferation with rapamycin. METHOD: Nasopharyngeal carcinoma spherical cells were gathered by using serum-free suspension culture method, CCK8 assay was used to detect cell proliferation, Western blot assay was used to detect the expression of CD44, OCT4, SOX2 and mTOR signaling. The spherical cells and CNE2 were treated with rapamycin in concentrations of 0, 0.1, 1.0, 10.0, 100.0, 1000.0 nmol/L, CCK8 assay was used to detect cell inhibition ratio, Western blot assay was used to detect the expression of mTOR signaling of nasopharyngeal carcinoma spherical cells. RESULT: Compared with CNE2, the spherical cells exhibited a high proliferation rate in RPMI 1640 medium supplemented with fetal bovine serum, and overexpressed in OCT4, SOX2 (P < 0.05), but not that of CD44 (P > 0.05). Although the expression levels of mTOR, P70S6, 4EBP1 were not significantly different between the two kinds of cells (P > 0.05) the proteins of phosphorylation activation form of them (P-mTOR, P-P70S6, P-4EBP1) were highly expressed in spherical cells (P < 0.05). The spherical cells and CNE2 were treated with rapamycin in different concentrations, the concentrations for 50% of maximal effect of spherical cells and CNE2 were 2.59 nmol/L and 78.12 nmol/L respectively, rapamycin inhibited the spherical cells more strongly compared with CNEZ. The expression levels of P-mTOR, P-70S6, P-4EBP1 in spherical cells were gradually decreased with increasing of the concentrations of rapamycin, but the difference of the expression levels of mTOR, P70S6, 4EBP1 were not significant. CONCLUSION The proteins of mTOR signaling pathway of CSCs in nasopharyngeal carcinoma are overexpressed, and rapamycin can effectively inhibit cell proliferation of CSCs in nasopharyngeal carcinoma by blocking mTOR signaling pathway.
Adaptor Proteins, Signal Transducing ; metabolism ; Carcinoma ; Cell Proliferation ; Humans ; Nasopharyngeal Carcinoma ; Nasopharyngeal Neoplasms ; metabolism ; pathology ; Neoplastic Stem Cells ; drug effects ; metabolism ; Phosphoproteins ; metabolism ; Phosphorylation ; Signal Transduction ; Sirolimus ; pharmacology ; TOR Serine-Threonine Kinases ; metabolism ; Tumor Cells, Cultured

Objective:To evaluate salvage surgery in patients with early gastric cancer after noncurative endoscopic resection .Method:A total of 56 cases with early gastric cancer receiving salvage surgery after noncurative endoscopic resection were enrolled and the clinicopathological and follow-up information were analyzed to evaluate the necessity and safety of salvage surgery.Results:Among the 44(79%)patients with submucosal invasion, 38 (68%) were with SM2 (invasion submucosal invasion≥500 μm) according to the pathological results after endoscopic resection. 33 (59%)cases had positive margin. The rate of lymph node metastasis and positive residual tumor as found by salvage gastrectomy were 11% (6/56) and 25% (14/56) . In the multivariate analysis, deeper submucosal invasion resulted as independent risk factor for residual tumor( OR=1.001, 95% CI=1.000-1.002, P=0.036). Among the 12(21%)cases with postoperative complications, 3 (5%)underwent unplanned reoperations because of anastomotic or intra-abdominal bleeding. There was no difference in the number of retrieved lymph nodes and rate of postoperative complications between laparoscopic and open surgery(all P>0.05). Conclusion:For patients with the risk factors of lymph node metastasis after noncurative endoscopic resection, salvage surgery was necessary and laparoscopic approach was safe and feasible.