Objective To observe the changes of melanin granules in superficial corneocytes in vitiligo lesions after irradiation with narrow-band ultraviolet B (NB-UVB) by using an adhesive tape stripping technique.Methods Vitiligo lesions were selected from 6 patients and irradiated with NB-UVB every other day for 31 sessions.Superficial corneocytes were obtained by an adhesive tape stripping technique from the vitiligo lesions and perilesional normal skin before every treatment.The morphology,distribution and color of melanin granules were observed after Masson-Fontana silver staining.The percentage of area occupied by melanin granules in superficial corneocytes were calculated by using the Image-Pro Plus 6.0 software.Statistical analysis was conducted by the SPSS11.5 software.Results There were still a few superficial corneocytes containing melanin granules remaining in the vitiligo lesions before treatment.The percentage of area occupied by melanin granules was (5.31 ± 4.12)％ before treatment,(6.24 ± 2.65)％ after 10 treatment sessions,(10.14 ± 5.73)％ after 20 sessions,and (13.05 ± 6.17)％ after 30 sessions,with significant differences between these time points (F =4.334,P ＜ 0.05).Multiple comparisons revealed a significant increase in the percentage of area occupied by melanin granules after 30 treatment sessions compared with those before treatment and after 10 treatment sessions (both P ＜ 0.01 ).The morphology and color of melanin granules in repigmented lesions after treatment differed from those in perilesional normal skin before treatment.Conclusion The adhesive tape stripping technique may serve as a useful tool for the evaluation of repigmentation in vitiligo lesions after phototherapy.

Objective To evaluate the effects of hydrogen-rich saline on inflammatory responses in the sciatic nerve and spinal cord of rats with neuropathic pain (NP).Methods Ninety male adult Sprague-Dawley rats,weighing 180-220 g,were randomly divided into three groups (n=30 each) using a random number table:sham operation group (group S);NP group;hydrogen-rich saline group (group H).NP was induced by chronic constrictive injury.The animals were anesthetized with intraperitoneal 10％ chloral hydrate 300 mg/kg.The left sciatic nerve was exposed and ligated with 4-0 silk at 1 mm intervals.After ligation of the sciatic nerve,hydrogen-rich saline (0.6 mmol/L) 10 ml/kg was injected intraperitoneally twice a day for 7 consecutive days in group H,and the equal volume of normal saline was given in S and NP groups.At 7,10 and 14 days after ligation,the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured.The rats were then sacrificed,and the left sciatic nerve and L4-6 segments of the spinal cords were removed for detection of the contents of tumor necrosis factor-alpha (TNF-α),interleukin-1 beta (IL-1β) and high mobility group box-1 (HMGB1) at 7,10 and 14 days after ligation.Results Compared with group S,the MWT and TWL were significantly decreased,and the contents of TNF-α,IL-1β and HMGB1 in the left sciatic nerve and spinal cords were increased at each time point in NP and H groups.Compared with group NP,the MWT and TWL were significantly increased,and the contents of TNF-α,IL-1β and HMGB1 in the left sciatic nerve and spinal cords were decreased at each time point in group H.Conclusion Hydrogen-rich saline alleviates NP through inhibiting inflammatory responses in the sciatic nerve and spinal cord of rats.

[ABSTRACT]OBJECTIVETo investigate the efficacy and the related complications of combinedpostauricular methylprednisolone injection and systemic therapy for profound idiopathic sudden hearing loss (ISSNHL). METHODSTotal of 60 patients with ISSNHLwho had received therapy from June 2014 to May 2015 in Beijing Anzhen Hospital affiliated to Capital University of Medical Science,were randomly divided into 2 groups,the systemic application group (30 patients): dexamethasone (DEX) was applied intravenously in dose of 10 mg×5 d, and the postauricular injection group (30 patients): methylprednisolone sodium suecinate was injected subperiosteally near the upper one-thirds of postauricular sulcus every day, 40 mg×5 d.All 60 patients received the same medications for 2 weeks to improve the hearing. Hearing and tympanic membrane were monitered before the injections and two weeks after the termination of injections. SPSS 16.0 software was used to analyze the data.RESULTSThe postauricular injection group: 23 of 30 ears had improvement of hearing. No related complications were reported. The systemic application group: 23 of 31 ears had improvement of hearing (P>0.05) No related complications were reported.CONCLUSIONCombined postauricular methylprednisolone injection and systemic medications therapy can be considered as is an effective therapy for profound idiopathic sudden hearing loss. It can avoid the side-effects of high dose systemic corticostemid treatment. For ISSNHL patients, postauricular methylprednisolone injection may be an appropriate treatment.

Objective To evaluate the anatomical structure of bone in the formation of acetabular teardrop shadow. Methods The acetabular teardrop shadow of the X ray plain film in 100 children and 300 adults, and of CT in 43 cases were analysed 、measured and compared. Results In children, teardrop shadow appeared “U”in 80 00%, 14 00% in teardrop, 6 00% in line. In adult teardrop shadow appeared “U”in 41 33%, 53 33% in teardrop, 5 34% in line. There were significant differences between children and adult in the frequency of “U” and teardrop (? 2=43 34, P

Objective To investigate the effects of staphylococcal enterotoxin A SEA gene on target cells mediated by replicative-deficient recombinant adenovirus vector. Methods Lymphocytes of C57BL/6 mice were infected with various titers of recombinant adenoviruses. Supernatants were collected after 12 h, 24 h, 48 h, 72 h, 96 h, 120 h and 144 h of incubation and analyzed for proliferation of lymphocytes by MTT assay. IL-2 level in the culture supernatants was measured with ELISA. The killing effect of lymphocytes was also observed by MTT assay. Results Proliferation response and elevated levels of IL-2 were observed in experimental group. The killing effect on B16 cells was stronger in experimental group, which seemed to be dose-dependent with the increase of ratio of lymphocytes/target cells. Conclusions SEA gene can be expressed in lymphocytes of C57BL/6 mice mediated by replicative-deficient recombinant adenovirus vector. The expressing products can activate lymphocytes of C57BL/6 mice, which kill B16 cells in vitro.

This study was performed in the Laboratory of Pharmaceutical Department, North China Coal Medical University from November 2005 to May 2006. The effective components in the Ruyi jinhuang san power were extracted with alcohol according to the solubility of the main components in Ruyi jinhuang san. The medical film prepared with chitosan was investigated by comparing with that prepared with Poly (vinyl alcohol). The content of the constituents such as curcumin, rhein, emodin, chrysophanol, and berberine in the film were determined with high pressure liquid chromatography (HPLC). The drug release in vitro of film was determined by Paddle method. Alcohol was an efficient agent for extracting the main components in Ruyijinhuang san, compared to the other solvents. The release rate of rhein in the film was very well, using the release rate of Rbein as an evaluation index. Ruyijinhuang san film prepared with chitosan was flexible and had characteristics of sustained-release in vitro.

Objective To evaluate the effect of hydrogen on the expression of nuclear factor E2?related factor 2 ( Nrf2 ) during endotoxin?induced oxidative injury to macrophages. Methods Normally cultured Raw264.7 cells were divided into 4 groups ( n=36 each) using a random number table: control group (group C), hydrogen?rich saline group ( group H), endotoxin group ( group E) and endotoxin plus hydrogen?rich saline group (group EH). In E and EH groups, endotoxin was added with the final concentration of 1 μg∕ml. Hydrogen?rich saline ( hydrogen concentration 0. 06 mmol∕L) was added in H and EH groups. All the cells were incubated for 48 h. At 6, 24 and 48 h of incubation, cells were collect?ed to detect the activity of reactive oxygen species ( ROS) , and cells were collected and proteins extracted for determination of superoxide dismutase ( SOD) and catalase ( CAT) activities and Nrf2 expression by Western blot. Results Compared with group C, the ROS activity was significantly increased, the levels of SOD and CAT were significantly decreased, and the expression of Nrf2 was significantly up?regulated in E and EH groups (P<0.05), and no significant change was found in the parameters mentioned above in group H (P>0.05). Compared with group E, the ROS activity was significantly decreased, the levels of SOD and CAT were significantly increased, and the expression of Nrf2 was significantly up?regulated in group EH (P<0.05). Conclusion Hydrogen can attenuate endotoxin?induced oxidative injury to macro?phages, and the mechanism may be related to up?regulated expression of Nrf2.

Objective To evaluate the role of nuclear factor erythroid 2?related factor 2 ( Nrf2)∕antioxidant response element( ARE) signaling pathway in inhibition of lipopolysaccharide ( LPS)?induced inflammatory factor release from macrophages by hydrogen. Methods RAW264. 7 macrophages of mice were cultured in 6?well plates (2×106 cells∕well) and were divided into 4 groups (n=24 each) using a random number table: control group ( group C); LPS group; hydrogen?rich saline+LPS group ( group LPS+H2); Nrf2 small interference RNA (siRNA)+LPS+hydrogen?rich saline group (siRNA+LPS+H2 group) . LPS 1 μg∕ml was added in group LPS. In group LPS+H2 , LPS 1μg∕ml was added, and the cul?ture medium was then replaced with the culture medium containing 0. 6 mmol∕L hydrogen?rich saline. In group siRNA+LPS+H2 , after Nrf2?siRN was successfully transfected into the cells, the cells were continu?ously incubated for 24 h, and the culture medium was then replaced with the culture medium containing 0.6 mmol∕L hydrogen?rich saline after LPS 1 μg∕ml was added. At 24 h of incubation, the supernatant was sep?arated for determination of the lactic dehydrogenase (LDH) activity (using colorimetric method) and for detection of the concentrations of tumor necrosis factor?alpha ( TNF?α) , interleukin?1 beta ( IL?1β) , high mobility group box?1 (HMGB1) and IL?6 (by ELISA). The cells were collected for measurement of the proliferation of cells ( by methyl thiazolyl tetrazolium assay) and for determination of the expression of Nrf2 and heme oxygenase?1 ( HO?1) in cells ( by Western blot) . Results Compared with group C, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly in?creased, the proliferation of cells was significantly decreased, and the expression of HO?1 in cells was sig?nificantly up?regulated in LPS and siRNA+LPS+H2 groups, and the expression of Nrf2 in cells was signifi?cantly up?regulated in LPS and LPS+H2 groups (P<0.05). Compared with group LPS, the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly decreased, the proliferation of cells was significantly increased, and the expression of Nrf2 and HO?1 in cells was sig?nificantly up?regulated in group LPS+H2 , and the expression of Nrf2 and HO?1 in cells was significantly down?regulated in group siRNA+LPS+H2 ( P<0.05) . Compared with group LPS+H2 , the LDH activity and concentrations of TNF?α, IL?1β, IL?6 and HMGB1 in the supernatant were significantly increased, the proliferation of cells was significantly decreased, and the expression of Nrf2 and HO?1 in cells was signifi?cantly down?regulated in group LPS+H2+siRNA ( P<0.05) . Conclusion The mechanism by which hydro?gen inhibits LPS?induced inflammatory factor release from macrophages is related to the activation of Nrf2∕ARE signaling pathway in mice.

Aim Bupivacaine is a kind of long-acting amide local anesthetics.This paper aims to explore the effects of bupivacaine on the short-circuit currents in human alveolar epithelial monolayers and study the possible mechanisms.Methods Short-circuit currents were recorded by ussing-chamber setup.Amiloride-sensitive currents were defined as the difference be-tween the total current and the amiloride-resistant cur-rent.ERK1 /2 phosphorylation protein levels were ana-lyzed by Western blot at 0,1 5,30 and 60 min after administration of 1 00 μmol·L -1 bupivacaine.Results Bupivacaine could inhibit the short-circuit currents in H441 monolayers dose-dependently,which could be inhibited by amiloride.Western blot analysis showed that bupivacaine increased the level of ERK1 /2 phos-phorylation.Conclusion These data demonstrate that bupivacaine can reduce the alveolar ion transport by in-hibiting the amiloride-sensitive currents,possibly by the enhancement of ERK1 /2 phosphorylation. The effects of alveolar fluid clearance following application of bupivacaine should be considered clinically when the patient is complicated with lung injury.

Objective To evaluate the role of autophagy in activation of astrocytes in the spinal cord of rats with neuropathic pain (NP).Methods One hundred thirty-six male Sprague-Dawley rats, aged 8-10 weeks, weighing 250-300 g, were randomly divided into 4 groups (n =34 each) using a random number table: sham operation group (group S), group NP, autophagy inhibitor 3-methyladenine (3-MA) group (group MA), and autophagy activator rapamycin group (group R).NP was induced by chronic constriction injury.In 3-MA and R groups, 3-MA 15 mg/kg and rapacymin 10 mg/kg were injected intraperitoneally, respectively, at 1 h before NP.Ten rats were randomly selected, and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured before NP and at 1, 3, 7 and 14 days after NP.Before NP and at 1, 3 and 7 days after NP, 6 rats were randomly sacrificed, the L4-6 segments of the spinal cord was harvested to detect microtubule-associated protein 1 light chain 3 Ⅱ (LC3 Ⅱ), Beclin-1 and glial fibrillary acidic protein (GFAP) expression by Western blot.Results Compared with group S, the MWT was significantly decreased, the TWL was shortened, and the expression of LC3 Ⅱ , Beclin-1 and GFAP was up-regulated at each time point after NP in group NP (P＜ 0.05).Compared with group NP, the MWT was significantly decreased, the TWL was shortened, the expression of LC3 Ⅱ and Beclin-1 was down-regulated, and the expression of GFAP was up-regulated at each time point after NP in group MA, and the MWT was significantly increased, the TWL was prolonged, the expression of LC3 Ⅱ and Beclin-1 was up-regulated, and the expression of GFAP was down-regulated at each time point after NP in group R (P＜0.05).Conclusion Autophagy is involved in the development and maintenance of NP through promoting the activation of astrocytes in the spinal cord of rats.