ObjectiveTo observe the therapeutic effect of early comprehensive rehabilitation and the value of intensity time (I/t) curve examine for peripheral facial paralysis.Methods89 patients with peripheral facial paralysis were treated by early comprehensive rehabilitation program in 1 week and were examined by I/t curve in 10～20 days.ResultsThe rate of the cure and excellent was 93.26%.Normal nerve detected by I/t curve were 62.92% and partial denervation were 37.08%.ConclusionPatients with peripheral facial paralysis who were treated by early comprehensive rehabilitation will have good prognosis. As a kind of examine ways, I/t curve can be used in clinic.

AIM: To develop the methods for the quantitative analysis of gallic acid and total phenolic acid in Sedum Aizoon L. METHODS: Gallic acid was analyzed by HPLC on a Hypersil BDS C_(18) column and detected at 271 nm.The mobile phase was methol-water(adjusted to pH=3.0 with H_3PO_4)(90∶10)at a flow rate of 1.0 mL/min.Total phenolic acid was analyzed by spectrophotometry at 720 nm.The colour-developing agent was the mixture solution of 0.6%FeCl_3—0.9%K_3[Fe(CN)_6](1∶0.9). RESULTS: Calibration graphs were constructed in the range of 0.343 0-1.200 ?g(r=0.999 7) for gallic acid and 0.4640-2.320 ?g/mL(r=(0.999 3)) for total phenolic acid.The average recoveries were 97.91%(n=6) for gallic acid and 99.21%(n=6) for total phenolic acid.The RSD of the methods were 1.8% and 2.0%,respectively. CONCLUSION: The methods were fast,reliable,accurate and suitable for analysis of gallic acid and total phenolic acid and quality control in Sedum Aizoon L.

AIM:To investigate the effect of AG 490 on the expression of VEGF and HIF-1α, and the capacity of invasion in human erythroleukemia (HEL) cells.METHODS:The HEL cells were treated with AG490 at different con-centrations .The cell viability was detected by CCK-8 assay.The apoptosis was detected by Hoechst staining .The apoptosis and the cell cycle were analyzed by flow cytometry .The capacity of migration was evaluated by Transwell assay .The mRNA expression level of JAK2 was measured by RT-PCR.The protein levels of p-JAK2, VEGF and HIF-1αwere determined by Western blot.RESULTS:The HEL cell viabilities were 88%, 75%, 48%, 10%and 0.12%after treated with AG490 at 20, 40, 60, 80 and 100 μmol/L for 48 h, respectively.The results of Hoechst staining showed that brilliant blue cells in 80 μmol/L AG490 group was significantly increased compared with control group for 48 h.The apoptosis rate of 80μmol/L AG490 group was significantly increased compared with control group at 48 h after AG490 treatment.The number of membrane-permeating HEL cells in 20μmol/L AG490 group at 24 h after AG490 treatment was significantly lower than that in control group (P<0.05).The mRNA level of JAK2 decreased in a concentration-dependent manner after the HEL cells were treated with different concentrations of AG 490 for 48 h.The protein levels of p-JAK2, VEGF and HIF-1αwere lower in AG490 treatment groups than those in control group (P<0.05).CONCLUSION: AG490 inhibits the expression of VEGF and HIF-1αin HEL cells by inhibiting JAK2 pathway.

Objective To investigate the isokinetic strength of knee extensors and flexors in patients with knee osteoarthritis (KOA), and to establish the correlation between the isokinetic strength and function in patients with KOA. Methods 23 patients with bilateral KOA and 14 matched normal controls finished the isokinetic test of knee extensors and flexors, the Five Times Sit-to-Stand Test (FTSST), Gait analysis, and Balance test. The KOA patients were evaluated with Visual Analog Scale (VAS) for pain and Western Ontario and McMaster University Osteoarthritis Index (WOMAC). Results It was less of the peak torque, average peak torque, average power, max rep total work, total work of knee extensors and flexors in the mainly involved limbs than the contralateral limbs (P<0.05), and more of the hamstring/quadriceps peak torque (H/Q) percentage (P<0.05) in the patients. It was less of all the observed indicators of knee extensors and average power of knee flexors in the patients than in the normal controls (P<0.05), and was more of the H/Q percentage (P<0.05). The peak torque of knee extensors correlated with the results of FTSST, walking speed, walking distance, falling index, VAS, and WOMAC-pain (P<0.05), but the peak torque of knee flexors did not (P>0.05). Conclusion It is weak of the isokinetic strength of knee extensors and flexors in patients with KOA in the mainly involved limb, as well as the isokinetic strength of knee extensors compared with the normal controls. The changes in the extensors and flexors are not equivalent. The peak torque of knee extensors significantly correlated with the pain and functions of the knee.

Objective: To investigate the regulation of JAK2 tyrosine kinase inhibitor ruxolitinib on extracellular matrix metalloproteinase (MMP in JAK2V617F positive myeloproliferative neoplasms (MPN) cells. Methods: ①Forty cases of newly diagnosed JAK2V617F positive MPN patients and 15 healthy volunteers as control in Baoding No.1 Hospital between January 2012 and December 2015 were enrolled in this study. JAK2V617F/JAK2 ratio was detected by real-time-PCR; the expression levels of phosphorylation protein tyrosine kinase 2 (p-JAK2) , MMP-2 and MMP-9 in pathological tissues of bone marrow were detected by immunohistochemistry. The bone marrow cells of JAK2V617F positive MPN patients were treated with ruxolitinib, then the migration ability and MMP-2, MMP-9 gene and protein expression levels were detected. ②The human erythroleukemia cell line HEL cells were treated with different concentrations of ruxolitinib (0, 50, 100, 250, 500, 1 000 nmol/L) . The cell viability was detected by CCK-8 test; cell migration ability was tested by transwell chambers. The mRNA expression levels of JAK2, MMP-2 and MMP-9 were detected by real-time-PCR. The protein expression levels of p-JAK2, MMP-2 and MMP-9 were detected by Western blot. Results: ①The expression levels of p-JAK2, MMP-2 and MMP-9 in the newly diagnosed group were significantly higher than control group respectively [ (78.56±24.55) % vs (41.59±17.29) %, P<0.05; (48.25±18.74) % vs (22.79±13.89) %, P<0.05; (53.29±19.28) % vs (15.56±14.96) %, P<0.05]. Spearman correlation analysis showed the positive correlation of MMP-2 and MMP-9 protein expression levels with JAK2V617F mutation (r=0.526, P=0.001; r=0.543, P=0.001) . ②The proliferation of HEL cells was inhibited by different concentrations of ruxolitinib in time and dose dependent manner. ③Cell migration test showed the number of cells leaked to the low chamber in MPN patients bone marrow cells and HEL cells treated with 5 nmol/L of ruxolitinib group were significantly lower than that without ruxolitinib treatment after 24 h [ (154.7±27.5) vs (320.3±67.3) , t=13.47, P<0.05; (70.7±10.5) vs (135.3±16.7) , t=13.89, P<0.05]. The mRNA and protein expression levels of JAK2, MMP-2 and MMP-9 decreased with the increased concentration of ruxolitinib. Conclusion Ruxolitinib inhibits MPN cell migration and expression of MMP-2 and MMP-9 via JAK2 signal pathway.

Objective To determine the effect of rapamycin(Rapa) on JAK2, ABCA3, and the immune checkpoint PD-1/PD-L1 in exosomes derived from JAK2 V617F positive HEL cells. Methods Human erythroleukemia HEL cells (JAK2 V617F mutation-positive) were cultured in vitro, and rapamycin was added at concentrations of 10, 50, and 100 nmol/L. The control group was established and cell proliferation inhibition rate was detected by CCK-8. Based on the inhibition rate of cell proliferation, cells intervened with 10 and 50 nmol/L Rapa were selected. Exosomes were extracted using a kit and identified by Western blot and flow cytometry. JAK2, ABCA3, and PD-1/PD-L1 mRNA changes in exosomes were detected by fluorescent quantitative PCR. The expression of exosome PD-1/PD-L1 protein was determined by flow cytometry. Results The exosomes extracted with the exosome kit all expressed characteristic CD9, CD63, and CD81 proteins, which were consistent with the general characteristics of exosomes. JAK2 mRNA can be detected in exosomes from HEL cells; Rapa reduced exosome production by HEL cells, and dose-dependently decreased gene expression of JAK2, ABCA3, and PD-L1 in exosomes. In addition, Rapa inhibited exosomal PD-L1 protein expression and had no significant effect on PD-1 protein expression. Conclusion HEL cells may transmit JAK2 mutant gene signals via exosomes. Rapa can reduce the production of exosomes and inhibits JAK2 mutated signals delivered by exosomes and suppresses PD-L1 protein expression.