OBJECTIVETo investigate the ability of invasion and migration of breast cancer MDA-MB-231 cells under serum starvation and hypoxia, and the effect of antiangiogenic drugs, rh-endostatin and bevacizumab, on the ability of invasion and migration of breast cancer cells under serum starvation and/or hypoxia, in order to explore the potential risk of antiangiogenic therapy in clinics.

METHODSThe cells were randomized into 4 groups, i.e., group A: 10% fetal bovine serum (FBS) group; group B: hypoxia + 10% FBS group; group C: serum starvation group; group D: hypoxia + serum starvation group; each group was further divided into three subgroups as blank control, treated with rh-endostatin and bevacizumab, respectively. Cell counting kit-8 (CCK-8) was used to assess the inhibition rate of cell growth induced by endostatin and bevacizumab, in order to determine the proper working concentration and time of the two drugs. Transwell assay was conducted to detect the cell invasion and migration in vitro. The expressions of c-Met and MMP-9 were detected by Western blot. The cells treated with rh-endostatin or bevacizumab under serum starvation were tested by hybridization using Exiqon miBase 18.0 microarray. The miRNAs which exibited significant differences (P < 0.05) in miRNA hybridization were verified by real-time PCR assay.

RESULTSCCK-8 assay showed that the inhibition rates of MDA-MB-231 cells cultured with 800 mg/L rh-endostatin for 48 h and 24 h were (32.2 ± 2.5)% and (27.0 ± 1.3)%, respectively, showing a significant difference (P = 0.023). The inhibition rates of MDA-MB-231 cells cultured with 80 mg/L bevacizumab for 48 h and 24 h were (30.5 ± 1.4) % and (26.1 ± 2.4) %, respectively, showing also a significant difference (P = 0.015). The Transwell assay showed that in the starvation blank group, the number of invaded and penetrated cells were 28.8 ± 2.2 and 31.4 ± 1.5, respectively, significantly different from that in the rh-endostatin and bevacizumab groups (P < 0.05). The relative expressions of c-Met and MMP-9 were 0.213 ± 0.017 and 0.542 ± 0.048, respectively, with a significant difference from those of the groups treated with each drug (P < 0.05 for both). The numbers of penetrated cells in the Transwell assay treated with rh-endostatin in hypoxia were 17.5 ± 2.1 and 16.5 ± 2.8, respectively, and the numbers of penetrated cells in the Transwell assay treated with bevacizumab were 16.3 ± 3.5 and 17.5 ± 2.4, respectively, showing no significant difference among them (P > 0.05 for both). The ability of migration and invasion of MDA-MB-231 cells and the expression of c-Met and MMP-9 were not impacted by hypoxia (P > 0.05). Real-time PCR assay showed that only the levels of miR-2355 and miR375 were significantly and stably decreased in the cells which had increased ability of invasion and migration. The relative expression levels of miR375 and miR-2355 in the serum starvation blank group were 0.550 ± 0.036 and 0.852 ± 0.121, respectively, significantly lower than that in the groups treated with rh-endostatin or bevacizumab (P<0.05). In the serum starvation group, the expression levels of miR375 and miR-2355 of cells treated with rh-endostatin were 0.295 ± 0.012 and 0.253 ± 0.011, and the expression levels of cells treated with bevacizumab were 0.234 ± 0.020 and 0.309 ± 0.022, respectively, (P > 0.05 for all). Compared with the serum starvation blank group, the expression levels of miR2355 and miR375 were significantly decreased when cells were treated with rh-endostatin/bevacizumab under serum starvation, but no significant difference was found between the two drugs (P > 0.05). However, hypoxia did not affect the expressions of miR2355 and miR375 (P > 0.05).

CONCLUSIONSThe results of this study suggest that serum starvation can increase the ability of invasion and migration of breast cancer cells. Furthermore, both rh-endostatin and bevacizumab may enhance their invasion and penetration ability under serum starvation condition.

Angiogenesis Inhibitors ; adverse effects ; Bevacizumab ; adverse effects ; Breast Neoplasms ; pathology ; Cell Hypoxia ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Culture Media, Serum-Free ; Endostatins ; adverse effects ; Female ; Humans ; Matrix Metalloproteinase 9 ; metabolism ; MicroRNAs ; analysis ; Neoplasm Invasiveness ; Proto-Oncogene Proteins c-met ; metabolism ; Random Allocation ; Real-Time Polymerase Chain Reaction ; Time Factors

Objective\ To further clarify the biological effects of TGF?1 on blastocyst implantation. Methods We used female adult Kunming mice to mate with male for observing the distribution of TGF?1 and its receptor at different stages of implantation with immunohistochemical method. Results\ Compared with nonpregnancy,TGF?1 and its receptor immunostaining markedly increased in epithelial and glandual cells on day 4 of pregnancy.The moderate extracellular staining of TGF?1 was present around the fibroblasts.On day 5\|6 of pregnancy,strong immunostaining in intensity of TGF?1 and its receptor was noted in primary decidual zone.With blastocyst invasion,TGF?1 and its receptor immunostaining diffused in decidual cells. Conclusion\ TGF?1 might play an important role via its receptor during blastocyst implantation.\;

Objective To explore the correlation of adiponectin (ADP) and type 2 diabetes mellitus (T2DM) depressive symptoms.Methods Random sample of hospitalized confirmed patients with T2DM were 76 cases,who were divided in to T2DM group (DM group,30 cases) and T2DM with depressive symptoms group (DD group,40 cases) and randomly selected 36 cases in the medical center as control group.The level of height,weight,waist circumference,hip circumference,plasma ADP,fasting plasma glucose (FPG),total cholesterol (TC),triglyceride (TG),high density lipoprotein cholesterol (HDL-C),low-density lipoprotein protein cholesterol (LDL-C),C-reactive protein (CRP),fasting plasma cortisol (COR),fasting insulin (FINS) were measured.Body mass index (BMI),waist to hip ratio (WHR) and insulin resistance index (HOMA-IR) was calculated.Results The plasma ADP level in DD group was lower than that in DM group and control group [(3.05 ± 1.07) g/L vs.(4.06 ± 1.12),(6.97 ± 1.64) g/L],there was significant difference (P ＜ 0.01).The plasma ADP level was negatively correlated with CRP,HOMA-IR,TC,FPG and SDS,but positively correlated with HDL-C.HDL-C and SDS was the independent risk factor influencing the ADP level in DD group.Conclusions T2DM patients with depressive symptoms exist the greater insulin resistance,the level of plasma ADP is specific drop; the lower level of plasma ADP is an independent risk factor for T2DM patients with depressive symptoms,there is independent correlation between them.

OBJECTIVE:To synthesize the aryl-pyridazinone acid and curcumin ester. METHODS:With the raw material of 5-methyl-2-(3-chloro-4-fluorophenyl)-2-oxo-pyridazine acid (compound 1) and curcumin,there was ester-forming in the two sides of a phenolic hydroxyl and pyridazin-6-one carboxyl in curcumin by the catalysis of N,N-dicyclohexyl carbodiimide(DCC)/4- dimethylaminopyridine (DMAP). The targeted product,MS and NMR characterization of the structure were obtained through column chromatographic separation. Single factor was adopted to detect the effect of mixture ratio of curcumin and compound 1,re-action temperature and time and catalyst on the reactions. RESULTS:The productivity of targeted product (aryl-pyridazinone acid and curcumin ester)was 56.3%(in curcumin),the content by HPLC was 98.1%. The optimum conditions were as follows as the mixture ratio of curcumin and compound 1 was 1∶3,reaction temperature was 50 ℃,reaction time was 10 h and the catalyst was DCC/DMAP. CONCLUSIONS:The aryl-pyridazinone acid and curcumin ester is successfully synthesized with stable process.

BACKGROUND:Discectomy is an important therapy for lumbar disc herniation, but a smal number of patients undergoing discectomy wil relapse.
OBJECTIVE:To investigate the spinal stability fol owing posterior pedicle screw fixation combined with interbody fusion cage for treatment of recurrent lumbar disc herniation.
METHODS:Twenty-six patients with recurrent lumbar disc herniation from January 2007 to December 2011 were enrol ed and subjected to posterior pedicle screw fixation combined with interbody fusion cage. Pain relief and lumbar stability were observed postoperatively. We analyzed the spinal stability in recurrent lumbar disc herniation patients after posterior pedicle screw fixation combined with interbody fusion cage depending on literature search.
RESULTS AND CONCLUSION:Al the 26 patients were fol owed up for 12-36 months. After treatment, al patients effectively al eviated the symptoms of low back pain, and lumbar interbody fusion was good, with a good rate of 96.2%. There was no pedicle screw loosening, broken, non-fusion phenomenon. Posterior decompression and interbody fusion cage combined with posterior pedicle screw fixation for recurrent lumbar disc herniation, characterized as fast symptom relief, strong fixation, exact interbody fusion exact, is an ideal treatment for recurrent lumbar disc herniation.

OBJECTIVE:To provide reference for further promoting the implementation ofseparation of prescribing and dis-pensingpolicy which fit China's national conditions. METHODS:Based on literature review,informant interview,and filed in-vestigation the development ofseparation of prescribing and dispensingwere compared between typical eastern and western coun-tries. Constraints ofseparation of prescribing and dispensingin China were explored,international successful experiences were summarized to inspire appropriate implementation of this policy in China. RESULTS & CONCLUSIONS:There are four con-straints in the implementation ofseparation of prescribing and dispensingin China. Firstly,the legal foundation is yet established for the professional development of pharmacists. Secondly,the medicine pricing and hospital financing systems are unreasonable. Thirdly,patients'ownership of prescriptions and right of dispensing options are monopolized by hospitals. Fourthly,safety of medi-cines use is yet ensured.Separation of prescribing and dispensingshould follow the rules of social and economic development and should be adjusted to adapt local conditions.Separation of prescribing and dispensingpolicy itself may not help to achieve the objectives of cost containment and rational use of medicines. To achieve the intended objectives,legislation of Pharmacist Law should also be accelerated to clearly define the role of pharmacist and guide the behavior of prescribers with economic levers.

Objective To analyze the efficacy and safety of different chemotherapy regimens for treatment of progressive patients with small cell lung cancer (SCLC) brain metastasis after radiotherapy. Methods 96 SCLC brain metastasis patients with progressive intracranial lesions after radiotherapy were divided into four groups: carmustine group (Group A, 28 cases), temozolomide group (Group B, 19 cases), topotecan group (Group C, 24 cases) and no chemotherapy group (Group D, 25 cases). Results In terms of brain metastases, there were no complete response cases in the whole groups. The rates of partial remission (PR), stable disease (SD) and progression of disease (PD) in Group A were 17.8%(5/28), 53.6%(15/28) and 28.6 % (8/28), respectively, the response rate (RR) of intracranial lesions was 17.9 % (5/28), and disease control (CR+PR+SD) rate was 71.4%(20/28). The rates of PR, SD and PD in Group B were 15.8%(3/19), 63.2 % (12/19) and 21.1 % (4/19), respectively, the RR of intracranial lesions was 15.8 % (3/19), and disease control rate was 78.9 % (15/19). The rates of PR, SD and PD in Group D were 8.3 % (2/24), 54.2 %(13/24) and 37.5 % (9/24), respectively, the RR rate of intracranial lesions was 8.3 % (2/24), and disease control rate was 62.5 % (15/24). In Group D, there was no response case, and 20 patients with PD (80.0 %) were found. The median progression-free survivals (PFSs) were (3.64 ±0.43) months, (4.68 ±0.49) months,(3.58 ±0.50) months, (2.60 ±0.31) months in Group A, B, C and D, respectively, and the median overall survivals (OSs) were (18.80±1.74) months, (18.76±1.85) months, (19.10±1.64) months and (9.64±0.84) months, respectively. The median OS of Group A, B or C was longer than that of Group D (P=0.002). The differences of grade Ⅲ-Ⅳhematologic toxicities among the four subgroups were not statistically different. Patients in Group B had better tolerance to nausea and vomit. In Group D, the central nervous system symptoms such as fatigue and headache occurred frequently. Conclusions The response rate and OS of SCLC brain metastasis patients with progressive intracranial lesions after radiotherapy are improved after chemotherapy, however, PFS is not significantly prolonged. The efficacies of carmustine, temozolomide and topotecan are similar in short and long term, besides, temozolomide shows less adverse events and a higher disease control rate. The application of chemotherapy that could penetrate the blood-brain barrier can improve the efficacy on SCLC brain metastasis patients with progressive intracranial lesions after radiotherapy with well tolerance.

Small cell lung cancer is the most common primary neuroendocrine malignancy of the lung and is characterized by high malignant degree,rapid doubling time,easy metastasis in early stage and poor prognosis.Accurate staging of small cell lung cancer can formulate personalized therapeutic plans and improve the prognosis of patients.PET/CT can obtain metabolism and anatomical images of the whole body in one scan and improve the diagnostic accuracy and integrity.PET/CT has been widely applied to clinical practice now.PET/CT will play a more and more important role in diagnosis,staging,treatment and prognosis assessment of patients with small cell lung cancer.The value of PET/CT in staging and treatment of small cell lung cancer was reviewed in this article.

Objective To investigate the effect of electroacupuncture on the expressions of in-terleukin(IL)-33 and ST2 in spinal cord and dorsal root ganglion in rats with neuropathic pain. Methods Forty adult female SD rats were randomly divided into sham operation group (group Sham),model group (group MC),ipsilateral electroacupuncture group (group SE)and contralateral electroacupuncture group (group VE).Each group had 10.The sciatic nerve injury models were es-tablished.The rats in group SE and group VE were respectively accepted electrical acupuncture treat-ment in Yanglingquan and Zusanli in the right and left limbs.Respectively,before treatment (T0 ), treated for 7 d (T1 ),3 d (T2 )and 7 d (T3 )after treatment,the MWT and TWL of rats in the four groups were measured.At T3 ,the expressions of IL-33 in spinal cord and dorsal root ganglion in rats were detected by using quantitative PCR.Results Compared with T0 ,MWT and TWL in group MC, group SE and group VE at T1-T3 were decreased significantly (P < 0.05).Compared with sham group,MWT in group SE and group VE,TWL in group MC,group SE and group VE at T1-T3 were decreased significantly (P <0.05).Compared with group MC,MWT and TWL in group SE and group VE at T2 ,T3 were increased significantly (P <0.05).Compared with sham group,the relative expression levels of IL-33 mRNA and ST2 mRNA in spinal cord and dorsal root ganglion in the group MC,group SE and group VE were elevated,the differences were statistically significant (P <0.05). Compared with group MC,the relative expression levels of IL-33 mRNA and ST2 mRNA in spinal cord and dorsal root ganglion group SE and group VE were decreased,the differences were statistical-ly significant (P <0.05).Conclusion The electroacupuncture treatment could significantly reduce the pain symptoms of neuropathic pain in rats,possibly via inhibiting the expressions of IL-33 and ST2 in spinal cord and dorsal root ganglion and blocking the IL-33/ST2 mediated inflammatory to play role.

Objective:To compare the therapeutic and adverse effects of chemotherapeutic regimen based on three drugs (taxol+carboplatin/cisplatin+etoposide) and two drugs (carboplatin/cisplatin+etoposide) on the combined small cell lung cancer (CSCLC). Methods:A retrospective study was conducted based on the data of 62 CSCLC patients who were admitted to and treated at Tianjin Medical University Cancer Institute and Hospital between July 2000 and April 2013. Of the 62 patients, 19 received the three-drug regi-men and 43 received the two-drug regimen. All patients received at least two cycles of chemotherapy and completed follow-up proce-dures. For each patient, the therapeutic efficacy was evaluated every two cycles, and toxicity was evaluated every cycle. Results:The response rates between the three-drug and two-drug groups were statistically significant (90%vs. 53%, P=0.033). However, no statisti-cal differences were observed in the disease control rate between the two groups (100% vs. 86%, P=0.212). The three-drug regimen could induce a better median progression-free survival compared with the two-drug regimen, but with no statistical significance (10.5%vs. 9.8%, P=0.484). Similarly, no statistical differences were noted in the median overall survival between the three-drug and two-drug groups (24.0%vs. 17.5%, P=0.457). The incidence rates of grade IV bone marrow depression and the termination of the original regi-men owing to severe adverse reactions were both significantly higher in the three-drug group than in the two-drug group (26.3% vs. 7.0%, P=0.036;31.6%vs. 14.7%, P=0.004). Conclusion:The two-drug regimen had almost the same survival rate and lower toxicity compared with the three-drug regimen. When using the TEP/TCE regimen, a close attention should be focused on its adverse reactions. The findings of this work showed that the two-agent regimen should be one of the standard treatments for CSCLC.