Aim To study the effect and molecular mechanism of pretreatment with COPP on hypoxia/reoxygenation injury of H9c2 myocytes.Methods H9c2 myocytes model of hypoxia/reoxygenation injury was established and H9c2 myocytes were given COPP pretreatment before hypoxia/reoxygenation.Treatment with Znpp and all-trans retinoic acid(ATRA)inhibited HO-1 and Nrf2-ARE respectively.The level of LDH and CK in cell supernatants were measured.HO-1mRNA expression was analyzed by RT-PCR.HO-1 and Nrf2 protein expressions were analyzed by Western blot.Results Compared with hypoxia/reoxygenation group,the level of LDH and CK in COPP pretreatment groups decreased significantly and the level of HO-1mRNA,HO-1 protein expression and Nrf2 protein expression in the nucleus significantly increased.Znpp abolished protective effect of COPP pretreatment.ATRA blocked the nuclear accumulation of Nrf2 and decreased HO-1 protein expression that COPP pretreatment induced.Conclusions COPP can induce HO-1 overexpression which has protective effect on hypoxia/reoxygenation injury of H9c2 myocytes.Its mechanism is related to Nrf2-ARE signaling pathway.

Objective Through a survey of medical students in autonomous learning, to understand students' learning motivation and learning strategies and find out the existing problems, and to provide the basis for the school to organize autonomous learning and improve the teaching quality. Methods Convenient sampling method was used to extract 1 500 students' autonomous learning scale who were freshman and sophomore, junior students and 1 467 valid questionnaires were collected. According to the corresponding research situation of Zhejiang Normal University, SPSS 19.0 was used to compare the different types of students in the school, and the t test or q test were also done. Results The situation of college students' autonomous learning was better. The girl's average score was (312.51 ±14.03) which was higher than the boys in the school (286.16 ±13.69)and the girls in the control group (302.67 ±10.37). The difference was statistically significant (P<0.05). The average total score of the junior college students (307.92 ±13.84) was better than that of freshman and sophomore (P<0.05), which suggested that junior students had the strongest independent learning ability. Conclusion Colleges and universities should be based on the actual situation of students and formulate corresponding policies and measures from the autonomous learning motivation, the boys dor-mitory management, to ensure the stability of teaching quality improvement.

Objective To investigate the role of Ca2+/calmodulin-dependent protein kinase Ⅱ on pain behavior in a mouse model of bone cancer pain. Methods 40 male C3H/HeN mice were divided randomly into 5 groups:sham group (S group, n=8) ,control group (C group, n=8) and KN93 treat group (T1, n=8;T2, n=8;T3, n = 8 ). Group C and T were induced mouse models of bone cancer pain by intra-left-femur inoculations of osteolytic NCTC2472 cells while group S were injected only α-MEM. On the 14 d after inoculations,group S and C received intrathecal injection of 20% DMSO 5 μl . While group T1, T2, T3 received intrathecal injection of KN93 15nmol,30nmol,60nmol which dissolved in 5 μl 20% DMSO respectively. Mice received pain behavior tests including quantification of spontaneous flinches, paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) before and at 0.5 h,2 h,4 h,8 h after administration. Results Treatment with KN93(15 nmol) have no effect on bone cancer pain,while treatment with KN93(30 nmol,60 nmol) can dose-dependently reverse quantification of spontaneous flinches, mechanical allodynia and thermal hyperalgesia which were induced by bone cancer pain, At 0. 5h after administration, the quantification of spontaneous flinches of the two groups ( (7.25 + 1.49 ), (4. 12 + 1.36 ) ) were decreased when compared with control group ( 11.62 + 1.92 ),PWMT((1.28 +0.14)g;(1.75 +0.46)g),PWTL((14.64 +2.12) s; (16.85 + 1.61)s)were increased when compared with control group ( (0.47 + 0. 16) g, ( 11.32 + 1.68 ) s) (P ＜ 0.05 ＝. The effect lasts for at least 4 h and disappears at 8 h. Conclusion CaMK Ⅱ may play an important role in the mechanism of bone cancer pain. Intrathecal KN93 injection can effectively attenuated bone cancer pain.

BACKGROUND:Studies have shown that bisphosphonates inhibit osteoclast resorption, but whether cathepsin K, a key cytokine of bone resorption, plays an effect has rarely been reported.
OBJECTIVE:To study the effect of bisphosphonate on capthesin K and bone resorption function during osteoclast differentiation.
METHODS:Osteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groups:control group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture and gene expression of capthesin K was detected by immunofluorescence method at 72 hours of culture. Western blot assay was used to detect capthesin K protein expression at 72 hours of culture.
RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group (P<0.01). Immunofluorescence showed expression of capthesin K was higher in the control group than the alendronate group (P<0.01);furthermore, the protein expression of capthesin K was also lower in the alendronate group than the control group (P<0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting gene expression of capthesin K.

BACKGROUND:Tartrate-resistant acid phosphatase is a specific marker for osteoclast differentiation and bone resorption, which is a sign of osteoclast maturity.
OBJECTIVE:To study the effect of alendronate on tartrate-resistant acid phosphatase related to osteoclast differentiation and bone resorption.
METHODOsteoclasts were cultured by mouse monocyte-macrophage cellline-RAW264.7. The cells were divided into two groupcontrol group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor;alendronate group, treated with 100μg/L receptor activator of nuclear factorκB ligand factor+10-7 mol/L alendronate. Osteoclastogenesis and resorption function of osteoclasts were examined at 7 days of culture. Gene expression of tartrate-resistant acid phosphatase was detected by immunofluorescence method. Western blot assay was used to detect protein expression of tartrate-resistant acid phosphatase.
RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase positive multinuclear cells were observed and resorption lacunae formed in two groups. Control group showed the higher number of tartrate-resistant acid phosphatase positive multinuclear cells and larger size of resorption lacunae than the alendronate group (P<0.01). Immunofluorescence showed expression of tartrate-resistant acid phosphatase was higher in the control group than the alendronate group (P<0.01);furthermore, the protein expression of tartrate-resistant acid phosphatase was also lower in the alendronate group than the control group (P<0.01). These findings indicate that bisphosphonates could strongly inhibit osteoclastogenesis and its resorption function by inhibiting protein expression of tartrate-resistant acid phosphatase.

Objective To evaluated the effect of the regional cooperative rescue model implemented on the length of time from first medical contact (FMC) to balloon dilation (B),economic expense and prognosis in patients with acute coronary syndrome (ACS).Methods Patients with ACS (including ST-segment elevation and non-ST-segment elevation) selected from other hospitals within 24 hours after onset were treated with emergency percutaneous coronary intervention.Patients were divided into two groups, regional cooperative rescue group and control group without the regional cooperative rescue model approved.The lengths of FMC-to-B time and Door-to-B time (from arrival at emergency department or OPD to balloon dilation),time required for patients referred to our hospital,cardiac function,averaged hospital costs,average hospital stay,percentage of medication used and a major adverse cardiac event (MACE) were analyzed.Results Mean FMC-to-B time,Door-to-B time,referral time and time consumed to obtain informed consent were significantly shorter [(106±33) min,(31 ±8) min,(62 ±18,8 ±3) min] vs.[(231 ±35) min,(109 ±26) min,(98 ±31) min,(28 ±11) min,respectively] by implementing the regional cooperative rescue compared with control group,and LVEF was increased,and LVED was deceased inregional cooperative rescue group.The mean costs [(44 123.0 ±3 427.0) yuan vs.(51 587.0 ±5 621.0)] yuan,days of hospital stay [(8.7 ±4.1) vs.(13.2 ±6.4)] and percentage of medication used were significantly decreased in the regional cooperative rescue group.The incidence of MACE inregional cooperative rescue group was 6.2％,whereas the incidence in control group was 16.8％.Conclusions The regional cooperative rescue model can improve the prognosis and decrease the FMC-to-B time,the rate of MACE and financial burden in patients with ACS.

Objective To explore the clinical efficacy of laparoscopy-assisted radical D2 proximal gastrectomy.Methods The clinical data of 22 cases of laparoscopy-assisted D2 radical proximal gastrectomy were analysed retrospectively.Results Laparoscopy-assisted radical proximal gastrectomy was carried out in all of the 22 cases. There was no conversion to open surgery in this series. The mean operation time was 188(162～270) min. The mean operative blood loss was 182(100～260)mL. The number of lymph nodes dissected was 18.6(10～32). The average time for gastrointestinal function recovery was 78(48～140)h. Postoperative anastomotic leakage occurred in one patient who had tumor invasion of distal esophagus, and it healed after 4 weeks of conservative treatment. There were no postoperative deaths in this series. All the 22 cases were followed up for 1～37 months (mean 9.2 months) and revealed no recurrence or metastasis.Conclusions Laparoscopy-assisted radical D2 proximal gastrectomy for early and advanced gastric cancer is feasible, safe, and minimally invasive.

Paeoniflorin and its derivatives are main active compounds inGui-Zhi Fu-Ling Capsule (GZFLC). In this study, molecular imprinted polymer (MIP) was prepared by sol-gel process to obtain paeoniflorin and its derivatives in GZFLC. The static adsorption capacity of MIP was measured by scatchard equation. The results showed that the maximum apparent absorbing capacity of MIP was 52.28 mg·g-1. One-step separation of paeoniflorin from 4 g methanol samples of GZFLC was 197 mg with the purity of 89.3%. It was concluded that paeoniflorin MIP can be used to separate phaoniforin and its analogues from GZFLC.

Objective To study the effects of microemulsion/ethosomes on transdermal absorption properties and efficacy of Huoxue Zhitong Cataplasm. Methods The improved Franz diffusion cells were used for the in-vitro permeation experiment with rat skins as the barriers, which was used to evaluate the transdermal absorption properties. In the erxeriment, the contents of paeonol, eugenol and methyl salicylate were used as markers, and detected by ultra performance liquid chromatography to evaluate the transdermal absorption effects. The anti-inflammatory and analgesia activity were evaluated through the writhing plate experiments. Results The cumulative release rate of paeonol in Huoxue Zhitong Cataplasm, Microemulsion Huoxue Zhitong Cataplasm and Ethosomes Huoxue Zhitong Cataplasm were, in order, 65.30%, 61.30%and 60.20%in 24 h;eugenol were, in order, 51.08%, 54.71% and 55.66% in 24 h; methyl salicylate were, in order, 49.20%, 65.17% and 72.15% in 24 h. Furthermore, Microemulsion Huoxue Zhitong Cataplasm high-dose group and Ethosomes Huoxue Zhitong Cataplasm medium-dose group had good effects on reducing the inflammatory exudate of peritoneal capillary and capillary permeability (P<0.05) in animal models. Conclusion Huoxue Zhitong Cataplasm based on microemulsion/ethosomesnano-technology has good transdermal absorption properties and efficacy.

Objective To explore the correction between serum trace elements,peripheral blood T lymphocyte subsets and children with recurrent respiratory tract infection.Methods 66 children with recurrent respiratory tract infection were selected and designed as observation group, and 66 children with health examination were designed as control group.Serum trace elements and peripheral blood T lymphocyte subsets of two groups were compared,and the detection results of observation group with upper respiratory tract infection and lower respiratory tract infection,at the stage of attack and catabasis were compared as well.Results The serum Zn+,Fe3+,Mg2+ and CD3+,CD4+,CD4 /CD8 of observation group were significantly lower than those of control group, while serum Cu2+ and CD8+ were higher than those of control group.Detection results of observa-tion group with lower respiratory tract infection and at the stage of attack were worse than those of children with upper respiratory tract infection and at the stage of catabasis.Serum trace elements and peripheral blood T lymphocyte subsets was closely correlated with the disease (all P <0.05). Conclusion Serum trace elements and peripheral blood T lymphocyte subsets of children with re-current respiratory tract infection are abnormal,and these indexes are closely correlated with the disease.