Objective To establish HPLC method for assaying total flavonoids from the roots of Daphne genkwa(TFRD) in serum of mice and to elucidate the effect of mice serum containing TFRD on cell immunity in mice.Methods TFRD Concentration in serum was determined from the mice received single ig TFRD at certain time intervals using HPLC method.The effects of TFRD serum on lymphocyte proliferation,killing activities of NK and LAK cell,and phagocytic activity of macrophage were detected by MTT method.Results TFRD in serum reached its highest concentration in 20—30 min after ig admi-(nistration.) TFRD-containing serum significantly improved the proliferation of lymphocyte,enhanced the killing activities of NK and LAK cells,and enforced the phagocytic activity of macrophage.Conclusion(TFRD-)containing serum is an effective agents for enhancing cell immunity in mice.

Objective:To analyze imaging diagnosis and differential diagnosis of upper femur aneurysmal bone cyst.Methods: The pre-operation imaging appearances about digital radiography(DR), computerized tomography (CT) and magnetic resonance imaging (MRI) of 46 aneurysmal bone cyst cases confirmed by operation and pathology were retrospectively analyzed, and then these special imaging appearances were gained and analyzed.Results:①CT plain scans showed the irregular expansile lucency shadow in the upper femur with the uneven bone septum image; the complete or incomplete osteosclerosis edge can be found in some parts of lesions; some lesions extend to adjacent soft tissue, and the liquid-liquid level can be found in lesion of 8 cases, and their edge and separated intensity strengthening were visible after signals were enhanced.②MRI plain scans revealed that most lesions showed long T1, T2 signals, the periphery of lesions showed long T1, short T2 signal shadows, and septum image was visible within lesions, and the liquid-liquid level can be found in lesion of 8 cases; some lesions extend to adjacent soft tissue, and their edge and separated intensity strengthening were visible after signals were enhanced.Conclusion: Through combined examination of DR, CT and MRI, the specialty and diagnostic accuracy of aneurysmal bone cyst could be improved, and it was helpful in the differential diagnosis of other upper femur bone tumors.

Objective To investigate the needs of the main caregivers of children with cerebral palsy and probe into the nursing strategies. Method One hundred and fifteen main caregivers of children with cerebral palsy participated in the survey by a general state questionnaire and the family needs questionnaire. Results The score by the family needs questionnaire was (96.13 ± 19.47), with an average score of (3.00 ± 0.61). And the top needs of the main caregivers were information and econony need. Conclusions The needs of the main caregivers of children with cerebral palsy are high. More attention should be paid to community and family-based rehabilitation and supports to those care-givers.

Objective To investigate paclitaxel and ezrin-siRNA therapy for metastatic liver cancer.Methods Human metastatic liver cancer cells MHCC97-H were divided into four groups in vitro and in vivo,including control cells,exposure to paclitaxel alone,ezrin-siRNA alone,and the combination of paclitaxel and ezrin-siRNA.Nude mice lung lesions were formed by tail vein injection of tumor cells.The tumor cells'apoptosis rate,wound healing ability,the ability to cross an artificial membrane,and the numbers of metastatic lung lesions were measured.Results The apoptosis rate in normally cultured MHCC97-H tumor cells was 2.52％,compared to the other three groups at 11.66％,2.35％,7.38％ respectively.The wound healing ability was best in the negative control and weakest in cells treated with combination paclitaxel and ezrin-siRNA.The numbers of cells which could cross the artificial membrane in the negative,paclitaxel,ezrin-siRNA,and combination paclitaxel and ezrin-siRNA group were 52.5 ± 3.9,32.2 ± 3.2,26.6 ± 2.4,and 19.4 ± 1.1 respectively.The numbers of metastatic lung lesions in the nude mice (n =40) were 12.5 ±2.4,8.2 ± 1.5,7.0 ± 1.3,and 3.8 ± 0.3 respectively.Conclusions The ezrin-siRNA combined with paclitaxel therapy demonstrated a greater efficacy over single agent therapy for depressing liver cancer growth,motility,aggression,and metastasis in vitro and in vivo.

Aim To investigate the changes in the ex-pression of WNK1 in spinal cord of a rat model with bone cancer pain. Methods Female SD rats, weig-hing 170 ~200 g, were randomly divided into three groups:normal control group (group C, n=3), sham operation group ( group S, n =3 ) and bone cancer pain group ( group BCP, n =24 ) . Group C was not given any treatment, and group S was injected into the bone marrow of left tibia with 5 μl PBS solution while group BCP with 5 μl WALKER 256 mammary gland cancer cell suspension (approximately 1 × 105 cells). Mechanical paw withdrawal threshold ( MWT ) was measured at d1 before inoculation ( baseline) and d3, 6,9,10,11,12 after inoculation. Group S and C were sacrificed at d 12 while group BCP at d 3 ,6 ,9 ,12 after inoculation and spinal cord ( L4~6 ) were removed at different time points for detection of WNK1 mRNA ex-pression by qRT-PCR and WNK1 protein expression by Western blot. Results Compared with group C and S,group BCP’ s MWT started to decrease since d 3 ( P<0. 05 ) . The mRNA expression of WNK1 in spinal cord was up-regulated(P<0. 05) from d 3 showing an increasing trend over time until d 9 ( peak ) and then down-regulated to d 12 ( P >0. 05 ) while the protein expression upregulated since d6 and also showed an in-creasing trend to d 12 ( P<0. 01 ) . Conclusion The expression of WNK1 in spinal cord of a rat model with bone cancer pain increased abnormally, which may be involved in the occurrence and maintenance of a rat model with bone cancer pain.

Objective To investigate the effect of intrathecal CLP257 on the bone cancer pain in rats.Methods Forty adult female Sprague-Dawley rats,weighing 180-200 g,were divided into 4 groups (n=10 each) using a random number table:sham operation group (group S),bone cancer pain group (group BCP),dimethyl sulfoxide (DMSO) group,and CLP257 group.Bone cancer pain was induced by inoculating Walker 256 mammary gland carcinoma cell suspension (about 1× 105cells) 10 μl into the medullary cavity of the left tibia.On 7th-9th days after establishment of the model,5％ DMSO 10 μl was injected intrathecally once a day in group DMSO,and 10 μg/μl CLP257 10 μl was injected intrathecally once a day in group CLP257.The mechanical paw withdrawal threshold (MWT) was measured on 1 day before establishment of the model (T0),on 1st-6th days after establishment of the model (T1-6),and at 4 h after intrathecal administration on 7th-9th days after establishment of the model (T7-9).After the last intrathecal administration,the L4-6 segments of the spinal cord were removed for determination of the expression of potassium chloride cotransporter 2 (KCC2) protein and mRNA by Western blot and fluorescent quantitative real-time polymerase chain reaction,respectively.Results Compared with group S,the MWT was significantly decreased,and the expression of KCC2 protein and mRNA was down-regulated in BCP and DMSO groups,and the MWT was significantly decreased (P＜0.05),and no significant change was found in the expression of KCC2 protein and mRNA in group CLP257 (P＞0.05).Compared with group BCP,the MWT was significantly increased,and the expression of KCC2 protein and mRNA was up-regulated in group CLP257 (P＜0.05),and no significant change was found in the parameters mentioned above in group DMSO (P＞0.05).Conclusion Intrathecal CLP257 can attenuate the bone cancer pain in rats.

Objective: To investigate the effect of clopidogrel on antiplatelet therapy in patients with coronary artery disease (CAD) combining chronic kidney disease (CKD) in order to provide a medication reference in clinical practice. Methods: We retrospectively investigated 423 CAD patients with coronary angiography (CAG) conifrmed diagnosis in our hospital from 2014-01 to 2014-09. According to the value of eGFR, the patients were classiifed into 2 groups:CAD+ CKD- group,n=257 patients with eGFR ≥ 90 ml/(min?1.73 m2), including 182 male and 75 female at the mean age of (60.39 ± 11.09) years, and CAD+CKD+ group,n=166 patients with eGFR < 90 ml/(min?1.73 m2), including 107 male and 59 female at the mean age of (65.80 ± 10.84) years. The patients were treated either by aspirin 0.1 g/d with clopidogrel 75 mg/d for at least 7 days, or by PCI operation with the load of aspirin 0.3g and clopidogrel 300 mg. The thrombelastography was conducted to examine and compare the inhibitory rates of ADP receptor and arachidonic acid (AA) pathway in platelet between 2 groups. Results: The inhibitory rate of platelet ADP receptor in CAD+CKD- group (64.9 ± 27.2) % was higher than that in CAD+CKD+ group (56.6 ± 27.4) %,P=0.039. Based on clinical standard of platelet’s ADP and AA inhibitory rates, in CAD+CKD- group, there were 24/257 (9.4%) of patients only insensitive to clopidogrel, in comparison with 25 (9.7%) of patients only insensitive to aspirin,P=0.99. While in CAD+CKD+ group, there were 21/166 (12.7%) of patients only insensitive to clopidogrel, in comparison with 11 (6.6%) of patients only insensitive to aspirin,P= 0.045. Conclusion: Clopidogrel has decreased effect on anti-platelet therapy in CAD patients combining with CKD, such patients have reduced sensitivity to relevant medication.

OBJECTIVE To study the effect of epipolythiodioxopiperazine compound C87 on tumor cell proliferation and explore the potential mechanisms. METHODS Tumor cells were exposed to C87 0.05-1 μmol.L-1 for 24, 48 and 72 h, cell viability was determined by sulforhodamine B (SRB) assay and the half growth inhibition (Gl50 ) was calculated. After treatment with C87 0.1-2.5 μmol.L-1 for 6 h, or C87 2.5 μmol.L-1 for 0-6 h, the generation of reactive oxygen species (ROS) was measured using the compound 2′,7′-dichlorofluoresceindiacetate and flow cytometry analysis. After treatment with C87 2.5 μmol.L-1 , either alone or with antioxidant N-acetylcysteine (NAC), for 6 h, the generation of ROS was measured by flow cytometry analysis. Tumor cells were exposed to C87 0.05-1 μmol.L-1 , either alone or with NAC, for 24 and 48 h, while cell viability was determined by SRB assay. RESULTS The cell viability was significantly reduced following exposure to C87 0.05-1 μmol.L-1 for 24, 48 and 72 h in a concentration-dependent manner in A549, HCT116, HeLa and SMMC7721 cells(P<0.05). At 72 h, the value of r2 was 0.946, 0.989, 0.973 and 0.984(P<0.05), respectively. The cell viability was significantly reduced following exposure to C87 1 μmol.L-1 for 24 - 72 h in a time-dependent manner in A549, HCT116, HeLa and SMMC7721 cells(P<0.05). The value of r2 was 0.983, 0.956, 0.951 and 0.873(P<0.05), respectively. The generation of ROS was increased after exposure to C87 0.25-2.5 μmol.L-1 in a concentration-dependent manner in HCT116 and HeLa cells for 6 h (r2 = 0.760, P = 0.045: r2 = 0.987, P=0.001), and after exposure to C87 2.5 μmol.L-1 in a time-dependent manner in HCT116 and HeLa cells for 0.5-6 h (r2 = 0.886, P = 0.017: r2 = 0.994, P = 0.000).The C87-induced ROS generation could be blocked by NAC in HCT116 and HeLa cells(P<0.05). The C87 induced cell death could be blocked by NAC 5 and 10 mmol.L-1 , and the Gl50 value was 1.446 and 1.134 μmol.L-1 for 24 h (the Gl50 value of C87 group was 0.513 μmol.L-1 ), and 0.882 and 1.166 μmol.L-1 for 48 h (the Gl50 value of C87 group was 0.333 μmol.L-1 ). CONCLUSION The novel epipolythiodioxopiperazine derivative C87 exerts potent antitumor activity in vitro, possibly via triggering ROS production.

Tumor bone metastasis is one of the common complications of advanced tumor and one of the signs of poor prognosis.There are many targeted drugs for bone metastasis in the clinic,which play a very important role in the antitumor treatment and prevention of bone related events.Among them,zoledronic acid,a representative bisphosphonate drug,has been extensively studied.It has been found that zoledronic acid combined with other anti-tumor therapies can more effectively inhibit the occurrence of tumor and bone-related events.This article reviews the latest research on zoledronic acid combined with other antitumor methods to guide clinical use and provide research ideas.