Objective To investigate the wake-promoting action of median nerve electrical stimulation(MNES) in coma rats induced by traumatic brain injury(TBI) and its influence on the expression of α1-adrenergic receptor(α1 R) in the prefrontal contex (PFC).Methods Seventy-two healthy Sprague Dawley(SD) rats were randomly divided into the control group,sham-stimulated group(TBI),stimulated group (TBI+ MNES) and antagonist group(TBI+ OX1R antagonist +MNES).The control group had no any treatment.The TBI coma rat models were prepared in the other 3 groups.The sham stimulated group had no treatment.The antagonist group was injected with orexin receptor-l(OX1R) antagonist SB334867 into lateral ventricle,and both the antagonist group and stimulated group received MNES treatment.Then the behavior changes of rats in each group were observed and the α1 R expression level in PFC was detected by using the immunohistochemistry technique.Results Thirteen rats in the stimulated group and 8 rats in the antagonist group revived,while only 4 rats in the TBI group.The α1R levels from low to high were the blank control group,sham-stimulated group,antagonist group and stimulated group,showing the increasing trend,and the difference was statistically significant(P＜0.05).Conclusion MNES can improve the rat consciousness level after TBI coma,and its mechanism may be related with up-regulating the α1 R expression level in PFC area,moreover Orexin-A participates in this regulation process.

Objective To investigate the wake-promoting effect of vagus nerve stimulation (VNS) on coma rats after traumatic brain in-jury (TBI), and the related mechanism. Methods A total of 168 healthy Sprague-Dawley rats were randomly divided into blank group, TBI group, antagonist group and VNS group, 42 rats in each group. The latter three groups were established TBI model with impact, and the rats in coma at least 30 minutes were included. VNS group accepted VNS, the antagonist group were injected intralateroventricularly Orexin A receptor 1 (OXR1) antagonist SB334867, and TBI group accepted sham VNS. Their behaviors were observed to determine the level of con-sciousness six, twelve and 24 hours after intervention, while the expression ofγ-aminobutyric acid b1 receptor (GABAb1R) in prefrontal cortex was detected with immunohistochemistry and Western blotting. Results There were 42 rats in the blank group, 11 rats in TBI group, 13 rats in the antagonist group, and 28 rats in VNS group awakened finally. The expression of GABAb1R in prefrontal cortex ranged as TBI group, antagonist group, blank group and VNS group from more to less twelve and 24 hours after intervention under Western blotting (F>60.412, P<0.001), and it ranged as TBI group, antagonist group, VNS group and blank group under immunohistochemistry (H=15.121, P=0.002), with no significant difference among time points (H=3.028, P=0.220). Conclusion VNS can promote waking from coma in rats after TBI, which may relate with the decrease of GABAb1R in prefrontal cortex that induced by Orexin A.

Objective To investigate the drug resistance of side population cells in human gallbladder cancer cell line GBC-SD and explore its mechanism. Methods Drug sensitivity assays of 5chemotherapeutic agents were performed on side population cells (SP) and non-SP cells of GBC-SD.GBC-SD was cultured and then treated with the chemotherapeutic agent gemcitabine. The frequency of SP by FACS was measured. RT-PCR and Western blotting were used to detect the expression of AB-CG2 in both the SP and the corresponding non-SP subsets. Results After 1 d treatment with 4 chemotherapeutic agents (gemcitabine, cisplatin, 5-fluorouracil and mitoxantrone) in IC50 concentration to GBC-SD cell line, the reproductive ability of SP was higher than that of non-SP (P<0.05). However, statistical significance was not achieved when compared with epirubicin (P>0.05). The percentage of SP in GBC-SD treated with chemotherapeutic agent gemcitabine after 3 weeks was sharply elevated by FACS (8.02% ±0.13% vs 0.62% ±0.08%, P<0.05), and the expression of ABCG2mRNA and protein were increased in SP as compared with non-SP. Conclusion SP from human gallbladder cancer cell line GBC-SD, like stem cell, showed a heighten resistance to drugs. Increased expression of ABCG2 was largely responsible for the multi-drug resistance.

ObjectiveTo explore the tumor igenic property of side population cells (SP) from human gallbladder carcinoma cell line GBC-SD. Methods SP and non-SP cells were isolated from GBC-SD staining with Hoechst33342 dye by fluorescence-activated cell sorting (FACS). The soft agar clonal assay and xenograft assay were performed to characterize tumorigenic property of side population cells in vitro and in vivo, respectively. The percentage of SP cells was analyzed by FACS in 5 hu man gallbladder carcinoma specimens. ResultsThe percentage of SP cells accounted for approximately 0.87 ％ of GBC-SD cells. The clone-formed rates of SP was more frequent than that of non-SP cells (14.74％ ± 3.53％ vs 5.17％ ± 1.05％), there was statistically significant difference (t =2.75,P＜0. 05). SP cells could generate tumors with as few as 5 × 103 cells (four of seven animals), whereas at least 1 × 105 non-SP cells were needed to form a tumor (one of seven animals). Re-analysis of SPderived tumors by FACS showed that SP cells under in vivo conditions also have the capacity to regenerate the SP and non-SP fractions. Besides, analysis of Hoechst33342 revealed s small fraction of SP cells, ranging from 0. 27％ to 2.3％ in gallbladder carcinoma specimens. ConclusionSP cells from GBC-SD are highly tumorigenic similar as the cancer stem cells.

OBJECTIVE Although it is generally believed that nicotine accounts for the beneficial effect of smoking on ulcerative colitis, the underlying mechanisms remain not well- understood. Our previous finding that nicotine inhibits inflammatory responses through inducing miRNA-124 prompted us to ask whether the miRNA is involved in the protective action of nicotine on UC. METHODS MiR-124 expression in colon tissues and cells was determined by q-PCR and in situ hybridization. The effect of miR-124 on protective role of nicotine in ulcerative colitis was evaluated in DSS-treated mice and IL-6-treated Caco-2 colon epithelial cells. Expression of p-STAT3/STAT3 was detected by immuno?histochemistry and Western blot analysis. RESULTS miR- 124 expression is upregulated in colon tissues from patients and DSS- induced colitis. Nicotine treatment further elevated miR- 124 level in colon tissues of the mice, in infiltrated lymphocytes and epithelial cells, and augmented miR- 124 expression in lymphocytes isolated from human ulcerative colon tissues. Administration of nicotine also reduced weight loss, improved DAI and decreased HE score in DSS-induced colitis. Moreover, knock?down of miR-124 in vivo significantly diminished the beneficial effect of nicotine, and in vitro on IL-6-treated Caco-2 colon epithelial cells. Further analysis indicated that nicotine inhibited STAT3 activation in vivo and in IL-6-treated Caco-2 colon epithelial cells and Jurkat human T lymphocytes, in which miR-124 knockdown led to increased activation of STAT3. CONCLUSION These data indicated that nicotine exerts its protective action in UC through inducing miR-124 and its effect on STAT3, suggesting that the miR-124/STAT3 system is a potential target for the therapeutic intervention of UC.

In recent years, miR-124 has emerged as a critical modulator of immunity and inflammation. Here, we summarize studies on the function and mechanism of miR-124 in the immune system and immunity-related diseases. They indicated that miR-124 exerts a crucial role in the development of immune system, regulation of immune responses, and inflammatory disorders. It is evident that miR-124 may serve as an informative diagnostic biomarker and therapeutic target in the future.

Objective To investigate the regulation on extracellular signal regulated kinase (ERK )1/2 and p38 signaling pathways and immune expression by moist exposed burn therapy/moist exposed burn ointment(MEBT/MEBO)in diabetic foot and their relationship ,to explore the repair mechanism of MEBT/MEBO on diabetic foot ulcers .Methods Totally 40 diabetic foot pa‐tients were treated by MEBT/MEBO ,to take wound tissue before and after treatment and detect the expression of ERK 1/2 ,p38 , MAPKK6 ,c‐myc ,Akt ,ATF2 ,IgA ,IgM ,IgG ,C3c and C4c by immunohistochemistry ,to investigate their relationship .Results Af‐ter treatment with MEBT/MEBO ,the area of foot wounds in 39 patients was reduced in different degree .Only one patient had no obvious change .14 patients(35 .00% ) were markedly effective ,25 patients(62 .50% ) and 1 patient(2 .50% ) were ineffective .Before andaftertreatment,allpositiveexpression,positiveimmunoreactivity(anyindex)andpositiveexpression(specificindex)ofsignal pathway molecules were statistically significant (P<0 .05) .While the positive rate of molecular expression in wound pathway in‐creased ,the positive expression rate of immune factor increased .Before treatment ,a small amount of immune factors were found in the wound tissue .After treatment ,the immune factors IgA ,IgM ,IgG ,C3c ,C4c were distributed widely and diffusely .Before treat‐ment ,the wound tissue showed a very small number of signal molecules .After treatment ,the signal pathway molecules MEBT/MEBO and p38 ,MAPKK6 ,c‐myc ,Akt ,ATF2 showed broad and diffuse distribution .Conclusion MEBT/MEBO may promote the expression of ERK1/2 and p38 signaling molecules and immune in diabetic foot ,p38 and ERK1/2 signaling pathway may promote the healing of diabetic foot wound by increasing the expression of immune .

OBJECTIVETo compare postoperative blood loss under different negative pressures of drainage after total hip arthroplasty for the treatment of femoral neck fractures.

METHODSFrom January 1st to December 30th 2013, 74 patients with femoral neck fractures treated with total hip arthroplasty were randomly divided into two groups: high negative pressure drainage group and low negative pressure drainage group. In high negative pressure drainage group, there were 34 cases including 10 males and 24 females, with a mean age of (75.94 ± 9.02) years old, and the patients were treated with 60 kPa negative pressure of drainage. In the low negative pressure drainage group, there were 40 cases including 13 males and 27 females, with an average age of (74.93 ± 8.90) years old, and the patients were treated with 30 kPa negative pressure of drainage. The amount of total drainage, total blood loss, and hemoglobin change were compared between these two groups.

RESULTSAll the patients got primary healing without infections. In high negative pressure drainage group,the change of hemoglobin was (41.74 ± 15.69) g/L, total blood loss was (1,217.73 ± 459.50) ml and the drainage volume was (312.94 ± 103.44) ml; while in low negative pressure drainage group,the results were (34.90 ± 12.90) g/L, (904.01 ± 381.58) ml and (129.25 ± 44.25) ml separately. All the results in high negative pressure drainage group were higher than those in the other group. Three days after operation, the change of hemoglobin was (46.00 ± 13.29) g/L and total blood loss was (1,304.72 ± 421.75) ml; while in low negative pressure drainage group, the changes of hemoglobin was (43.87 ± 11.39) g/L and total blood loss was (1,196.78 ± 344.20) ml; there were no statistically significant differences between two groups.

CONCLUSIONWhen placing drainage devices after total hip arthroplasty for the treatment of femoral neck fractures, the level of negative pressure should be chosen according to preoperative level of hemoglobin and HCT in patients. For old patients with femoral neck fracture, low negative pressure is more suitable.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Hip ; methods ; Case-Control Studies ; Female ; Femoral Neck Fractures ; surgery ; Humans ; Male ; Middle Aged ; Negative-Pressure Wound Therapy ; Postoperative Hemorrhage ; prevention & control

Aim To explore the effect of receptor component protein(RCP)in the signal transduction of vascular smooth muscle cell(VSMC)proliferation induced by static pressure.Methods The mouse-derived vascular smooth muscle cell line(A10VSMC)was employed in the experiment.Cells were exposed to static pressure,and MTT assay was used to detect the cell viability.Western blot was used to determine the expressions of PCNA,RCP and p-Akt,RCP mRNA was tested by RT-PCR,and co-immunoprecipitation was used to test the interaction between RCP and G proteins.Results The cell viability,expressions of PCNA and RCP increased with the elevation of static pressure and reached their peaks at 120 mmHg,and after 6 hours they got a plateau.The static pressure significantly increased the level of p-Akt,meanwhile,the binding of RCP and Gαs significantly decreased.However,the binding of RCP and Gβ increased in response to static pressure after stimuli of static pressure,but Gγ was obscure.Conclusion Static pressure can induce VSMC proliferation and expression of RCP,which may involve G protein signal transduction model.

Background Sirius system,a new Scheimpflug camera combined with Placido topography,improved the capability of imaging the anterior eye segment significantly.However,the study of assessing the repeatability and agreement between Sirius and Pentacam is still lack up to now.Objective This study was to evaluate the repeatability and agreement of the anterior ocular segment measuring parameters by Sirius and Pentacam in myopia received laser in situ keratomileusis (LASIK).Methods Thirty-five myopic eyes of 35 patients received LASIK were included in School of Optometry and Ophthalmology Eye Hospital from 2010 May through 2010 July.Corneal power flat keratometry (Kf),step keratometry (Ks),mean keratometry (Km),thinnest corneal thickness(TCT),the location of TCT,anterior chamber depth (ACD) and anterior chamber volume (ACV) were measured by Sirius and Pentacam in all the eyes,respectively.The repeatability of the measuring results were evaluated using intraclass correlation coefficients (ICC) and Cronbach's coefficient alpha (CoA),and the agreement of measuring parameters between Sirius and Pentacam was analyzed using Bland-Altman plot.Results Both Sirius and Pentacam demonstrated high intraobserver repeatability,with all ICC and CoA more than 0.90.No significant differences were found in Kf values and Ks values between the two methods (t =-1.533,-1.750,P＞0.05).Km value was (39.14 ± 1.95) D by Sirius measurement,which was sígnificantly higher than (39.05 ± 1.91) D by Pentacam measurement (t =3.572,P =0.001).The TCT was (457.6 ± 40.9) μm by Sirius method,showing a significant reduce in comparison with (465.4±37.5) μm of Pentacam method (t =-6.689,P＜0.001).A positive correlation was seen in the TCT between the two methods (r=0.988,P＜0.001).The Bland-Alrman plots showed the 95％ CI-21 μm to 6 pm in the TCT value between the two devices.Pairwise comparison of the location of TCT measurements showed significant differences between the two devices (t =-4.132,-5.696,P＜0.001),with a good correlation (r=0.751,0.775) and the 95％ CI (-0.36-0.17 mm,-0.35-0.12 mm).A very good agreement was seen in ACD between the two devices (-0.02-0.12 mm),but the agreement result was not very well in the ACV between the two devices with the 95％ CI (-27.70-6.20 mm3).Conclusions Sirius and Pentacam measurements for anterior ocular segment parameters have a very good repeatability in post-LASIK eyes.In addition,good agreement results are exhibited in corneal power,TCT and ACD between Sirius and Pentacam with an acceptable maximal different value between them.Sirius and Pentacam can be used interchangeably in clinical examination.However,the two devices can not interchangeably for ACV measurement and TCT location.