Objective To explore the role of extracellular signal-regulated kinase (ERK)in central nucleus of amygdala (CeA)in the mechanism of fentanyl-induced hyperalgesia (OIH)in rats. Methods Step 1:12 healthy male Sprague-Dawley rats,weighing 60-100 g,were randomly divided into OIH and Control group.The mechanical paw withdrawal threshold (PWT)and the thermal paw withdrawal latency (PWL)were tested at pre-and post-OIH induction.Then the level of p-ERK in the CeA was analyzed by Western blotting.Step 2:After successful induction of OIH and catheterization in CeA,another 30 SD male rats were randomly divided into 5 groups (n = 6 each):Group OIH;Group OIH+U0124;Group OIH+U0126(0.1 5 nmol);Group OIH+U0126(0.45 nmol)and Group OIH+U0126 (1.5 nmol),then 0.3 μl of DMSO,U0124 (1.5 nmol),U0126 (0.1 5 nmol,0.45 nmol,1.5 nmol)was given through the catheter separately.PWT and PWL were tested before cathe-terization,at pre-OIH induction,post-OIH induction and 0.5 h after CeA drug administration.After the last test of pain threshold,the rats were sacrificed and CeA tissues were sampled for analyzing the expression of p-ERK by western blot.Results In step 1 compared with control group,PWT and PWL of OIH group were sharply decreased post-OIH induction (P <0.05),concomitant increase of the expression of p-ERK in CeA in OIH group was also observed.In step 2,both PWT and PWL were sharply decreased post-OIH induction (P <0.05).Intra-CeA U0126 injection,but not U0124, reversed both behavioral hyperalgesia and molecular activation of ERK in CeA in a dose-dependent manner (P <0.05).Conclusion ERK plays a pivotal role in the maintenance of fentanyl-induced hy-peralgesia.Targeting inhibition of ERK activation in CeA can alleviate fentanyl-induced hyperalgesia.

Objective To investigate the role of extracellular signal-regulated kinase1/2 (ERK1/2) in the central amygdala on fentanyl-induced hyperalgesia in rats.Methods Thirty-two male SpragueDawley rats, weighing 60-100 g, were randomly divided into 4 groups (n =8 each) using a random number table: control group (group C), fentanyl-induced hyperalgesia group (group H), U0124 group (group U1) , and U0126 group (group U2).A catheter was implanted in the central amygdale.In group C, normal saline was injected subcutaneously, and 6.5 h later dimethyl sulfoxide (DMSO) was injected via the catheter.In group H, fentanyl was injected subcutaneously to induce hyperalgesia, and 6.5 h later DMSO was injected via the catheter.In group U1, hyperalgesia was induced, and 6.5 h later ERK1 inhibitor U0124 1.5 nmol was injected via the catheter.In group U2, hyperalgesia was induced, and 6.5 h later ERK1/2 inhibitor U0126 1.5 nmol was injected via the catheter.Mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal threshold (TWT) were measured before fentanyl injection, at 6.5 h after injection, and at 30 min after DMSO or U0124/U0126 administration via the catheter (T0-2).After the last measurement of pain threshold, the rats were sacrificed, and the amygdala tissues were sampled for detection of the expression of phosphorylated ERK1/2 (p-ERK1/2) by Western blot in groups C and H.Results Compared with group C, the MWT and TWT were significantly decreased at T1,2in H and U1 groups, and at T1in group U2 (P＜0.05) , the expression of p-ERK2 was up-regulated (P＜0.05) , and no significant change was found in the expression of p-ERK1 in group H (P＞0.05).Compared with group H,the MWT and TWT were significantly increased at T2 in group U2 (P＜0.05) , and no significant change was found in MWT, TWT in group U1 (P＞0.05).Conclusion ERK2 activation in the central amygdala is involved in the development of fentanyl-induced hyperalgesia in rats.

Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.

AIM: To investigate the effects of human umbilical cord-derived mesenchymal stem cells ( hUC-MSCs) on the proliferation and migration of osteosarcoma cells ( Saos-2 ) and the underlying molecular mechanism. METHODS:hUC-MSCs were isolated and cultured by tissue explants adherent method.The cell surface markers on hUC-MSCs were identified by flow cytometry.The effects of conditioned medium ( CM) from hUC-MSCs ( hUC-MSCs-CM) , re-combinant human interleukin-6 (rhIL-6) and IL-6 neutralizing antibody on the proliferation of Saos-2 cells were detected by CCK-8 assay and cell counting.IL-6 secretion of hUC-MSCs was assayed by ELISA.RT-PCR was used to assess the tran-scription level of proliferation-related genes proliferating cell nuclear antigen ( PCNA) , cyclin D1 and survivin.The migra-tion potential of hUC-MSCs and Saos-2 cells was measured by Transwell assay.RESULTS:hUC-MSCs migrated to Saos-2 cells.hUC-MSCs-CM contained a high concentration of IL-6, up to (1 835.5 ±134.1) ng/L.hUC-MSCs-CM and rhIL-6 promoted the proliferation and migration of Saos-2 cells.Addition of neutralizing antibody against IL-6 in the hUC-MSCs-CM impaired this proliferation and migration of Saos-2 cells.The mRNA expression of PCNA, cyclin D1 and survivin was up-regulated by hUC-MSCs-CM and rhIL-6, while this effect was dramatically attenuated by treatment with IL-6 neutralizing antibody.CONCLUSION:hUC-MSCs migrate to osteosarcoma cells and promote the proliferation and migration of osteo-sarcoma cells through secreting IL-6 in vitro.

A simple preparative strategy was proposed for the preparation of water-soluble, bright blue-emitting carbon nanoparticles ( CNPs ) at different temperatures by hydrothermal treatment using bean curd residue as carbon source for the first time. When the temperature reaches 240 ℃, quantum yield ( QY) can be up to 15. 24%. Most importantly, such fluorescent CNPs can be served as an effective sensing platform for label-free sensitive and selective detection of Fe3+ions with a detection of limit as low as 50 nmol/L, which is nearly 2 orders of magnitude lower than the maximum level of Fe3+ions in the drinking water permitted by the European Community. These outstanding properties give promising potential for reliable detection of Fe3+ in intracellular fluid and water samples, even bioimaging and optical imaging without being hazard in the future.

Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.

Objective To analyze labor duration in nulliparous women and discuss the change about curve of labor duration. Methods Two thousand, one hundred and forty nulliparous, full-term pregnant, singleton, cephalic presentation and vaginal delivery women who delivered at Peking University First Hospital were included. There were 1 300 cases between January 1, and December 31, 2009, while 840 cases between January 1, and May 31, 2013. A retrospective study was conducted. Data on maternal age, gestational age at delivery, body mass index at delivery, newborn weight, time of labor initiation, cervical dilatation and the amount of bleeding within the first 24 h after delivery were recorded. Data were compared by t test and χ2 test. The median time span corresponding to one-centimeter-increase in cervical dilatation was calculated. Results (1)Compared with data from 2009, the maternal age [(29.0±3.0) vs (29.6±2.8) years, t=4.77], incidence of postpartum hemorrhage [1.8%(24/1 300) vs 4.3%(36/840),χ2=11.17], proportion of induced labor [37.3%(485/1 300) vs 64.0%(538/840),χ2=146.23] and proportion of analgesia during labor [54.2%(705/1 300) vs 61.5% (517/840), χ2=11.15] were all higher in 2013 (all P<0.01). (2)The median (minimum–maximum) time span corresponding to a one-centimeter-increase in cervical dilatation was 3-4 cm which corresponded to 2.2 h(0.8-4.3 h), 4-5 cm which corresponded to 1.9 h(0.6-4.0 h), 5-6 cm which corresponded to 1.8 h(0.5-4.0 h), 6-7 cm which corresponded to 1.6 h(0.5-2.0 h), 7-8 cm which corresponded to 1.8 h(0.8-2.0 h), 8-9 cm which corresponded to 1.3 h(0.2-2.5 h), and 9-10 cm which corresponded to 0.6 h(0.1-2.5 h). The curve of labor duration showed a slow uptrend with time on the horizontal axis and cervical dilatation on the vertical axis. Conclusions The curve of labor duration exhibits a slow uptrend with neither an acceleration phase nor a deceleration phase. It is important to redefine the time span of labor duration in China for appropriate clinical treatment.

BACKGROUND: Changes in extracellular of chondrocyte can reflect influence of external force on temporomandibular joint and adaptability of body to external force. OBJECTIVE: To study the effect of cyclic tensile stress on main extracellular matrix of condylar chondrocyte.METHODS: The cyclic tensile stress was exposed to the third passage condylar chondrocyte for 0, 1, 6, 12 and 24 hours, respectively, using a Flexcell Strain Unit-5000T system (10% surface elongation, 6 cycles/min). After mechanical loading, total RNA was extracted from the cells harvested from Six-well BioFlex flexible cell Petri Dish, reverse transcribed, and reverse trabscription-PCR was performed to quantify mRNA levels for type Ⅱ collagen and aggrecan.RESULTS AND CONCLUSION: Compared with the control group (0 hour group), both type Ⅱ collagen and aggrecan mRNA expression was significantly increased after loading for 6 hours (P < 0.05), but began to decrease since 12 hours, and significantly decreased at 24 hours (P < 0.05). Results showed that cyclical tensile stress stimuli can affect the synthesis of main extracellular matrix of condylar cartilage, i.e. the synthesis was gradually enhanced with prolonged stimulation duration, but significantly inhibited in response to further stress stimuli.

Objective To test the central frequency (CF) and transmitter gain(TG) of MRI scanners,and set up their action limits.Methods Three different MRI devices (GE 3.0T HD,GE 1.5T HDi and GE 3.0T 750W) were tested by scanning American College of Radiology (ACR) phantom with the axial T1WI series.In the pre-scanning of T1WI series for the ACR phantom,the CF and TG were recorded.It was tested for eight times when MRI scanners were in good condition.The action limits of CF and TG were calculated based on mean values and standard deviations.Results The mean values of CF for three devices were (127 725 772.38±39.68)Hz,(63 875 740.13± 34.15)Hz,and (127 771 958.38±12.19)Hz,respectively.Their action limits were ≤119.04 Hz,≤68.30 Hz,and ≤36.57 Hz,respectively.The mean values of TG for three devices were (125.25±1.28)dB,(101.75±1.98)dB,and (113.25±0.89)dB,respectively.Their action limits were (125.25±2.56)dB,(101.75±3.96)dB,and (113.25±1.78)dB,respectively.Conclusion The CF and TG for three MRI scanners are all consistent with the action limits in this study.The CF and TG action limits will provide criterions for the clinical quality control.

Aim To investigate the molecular mecha-nism of Gab2 in the invasion and metastasis of breast cancer and provide a theoretical basis for clinical pre-vention of breast cancer invasion and metastasis. Methods The Gab2 , MMP-2 and MMP-9 expressions in 80 cases of breast cancer were detected by immuno-histochemistry . Western blot was used to detect the ex-pression of Gab2 protein in MDA-MB-231 cells and MCF-7 cells. The siRNA plasmid was used to transfect MDA-MB-231 cells. Western blot was used to detect the proteins expression of Gab2 , MMP-2 and MMP-9 . Transwell in vitro experiment was used to detect the in-vasion ability of each group transfected MDA-MB-231 cells, Western blot was used to analyze phosphorylation of Akt and ARK5 induced by epithelial growth factor ( EGF ) in transfected cells ( SiGab2/MDA-MB-231 and Scr/MDA-MB-231 ) . Results The expression of Gab2 protein in invasive ductal carcinoma was signifi-cantly higher than in normal breast tissue ( P<0. 01 ) . The expression of Gab2 was dramatically correlated with lymph node metastasis, ER expression, tumor his-tological grade, MMP-2 and MMP-9 (P<0. 05). The expression of Gab2 protein in MDA-MB-231 cells was higher than in MCF-7 cells. The expression of Gab2, MMP-2 and MMP-9 decreased in SiGab2/MDA-MB-231 cells and the invasion ability of SiGab2/MDA-MB-231 cells was significantly decreased ( P<0. 05 ) , and after 5 minutes’ stimulating by EGF, the phosphoryla-tion of Akt and ARK5 was significantly reduced. Con-clusion Gab2 can promote the invasion and metasta-sis of breast cancer by effecting the expression of MMP-2 and MMP-9 through PI3 K/ Akt /ARK5 signal path-way.