[Abstract ] Objective High-mobility group box-1 (HMGB1) is abundantly released in the epileptogenic brain tissue , but few reports are seen about the effect of HMGB 1 on the expression of P-glycoprotein ( P-gp) in the vascular endothelial cells of the epi-leptogenic tissue .This study is to explore whether HMGB 1 can regulate P-gp expression in the brain microvascular endothelial cells of the mouse in vitro . Methods Immortalized brain microvascular endothelial bEnd .3 cells of the mouse were cultured in vitro and al-located to different concentration groups ( treated with culture medium containing 10 , 100 , 500 , and 1000 ng/mL HMGB1 for 8 hours), treatment duration groups (treated with culture medium containing 100 ng/mL HMGB1 for 4, 8, 16, 24, and 32 hours), and a control group ( treated with culture medium without HMGB 1 ) .The mRNA expression of P-gp-encoding gene-multidrug resistance gene 1a (mdr1a) was detected by real-time qPCR, and its protein expression determined by Western blot and immunocytochemistry . Results The results of qPCR manifested that the expressions of mdr 1a mRNA were 1.646 ±0.176, 1.777 ±0.135, 1.617 ±0.043, and 1.398 ±0.182 in the 10, 100, 500, and 1000 ng/mL HMGB1 groups, respectively, significantly higher than 1.030 ±0.284 in the control group (P<0.05), and so were those in the 4, 8, 16, 24 h, and 32 h groups (2.655 ±0.112, 2.168 ±0.212, 1.823 ± 0.232, 1.418 ±0.376, and 1.445 ±0.123) than in the control (1.010 ±0.164) (P <0.05).Western blot showed a significant increase in the P-gp protein expression in all the concentration groups (P<0.05) as well as in the 8 h and 16 h treatment duration groups as compared with the control group (P<0.05).Immunocytochemis-try also revealed a higher P-gp expression in the HMGB1-treated than in the control cells (P<0.01). Conclusion HMGB1 can upregu-late the expressions of mdr1a mRNA and P-gp protein in the brain microvascular endothelial cells of the mouse , which may associated with drug resistance of central nervous system diseases , especially that of epilepsy .

Objective To observe the roles of nuclear factor-κB (NF-κB) and hypoxia-inducible factor-1 o (HIF-1 α) in hippocampal neurodegeneration of status epilepticus (SE) rats, and explore whether HIF-1α activation is regulated by NF-κB.Methods A total of 110 male Sprague-Dawley rats were randomly divided into seven groups : (1) Control group treated with saline (control, n =15), (2) sham group implanted cannula into lateral ventricle and treated with saline (sham, n =15), (3) SE group treated with pilocarpine (SE, n =20), (4) NF-κB activity inhibitor pyrrolidine dithiocarbamate (PDTC) group treated only with PDTC (PDTC, n =15), (5) SE + PDTC group treated with pilocarpine plus PDTC (SE + PDTC, n =15), (6) SE + HIF-1o siRNA group implanted cannula into lateral ventricle and treated with pilocarpine plus HIF-1 α siRNA (SE + HIF-1α siRNA, n =15), (7) SE + control siRNA group implanted cannula into lateral ventricle and treated pilocarpine plus control siRNA (n =15).SE was induced by injecting lithium chloride and pilocarpine.The seizure of rats was observed.The protein expressions of NF-κB and HIF-1 α in hippocampus of rats were examined by Western blotting.The degenerating neurons in hippocampus were detected by Fluoro-Jade C (FJC) staining.Results Twenty-four hours after termination of SE, the nuclear protein expressions of NF-κB and HIF-1α in hippocampus of rats were increased in SE group (0.57 × 0.06, 0.47 ± 0.07) compared with those in control group (0.23 ± 0.03, 0.20 ± 0.03;P ＜0.05);and compared with SE group PDTC significantly decreased the nuclear protein expressions of NF-κB and HIF-1 α in SE + PDTC group (0.23 ± 0.03, 0.14 ± 0.03;P ＜ 0.05);in SE + PDTC group the numbers of FJC positive cells in CA1 area (28.33 ±5.03) were decreased compared with that in SE group (76.67 ± 13.32);HIF-1 o siRNA injected into lateral ventricle of rats significantly decreased the expression of HIF-1α in hippocampus (0.22 ±0.03) and the number of FJC positive cell in CA1 area (27.34 ±7.02) in SE + HIF-1α siRNA group compared with those in SE group (0.39 ±0.06, 76.67 ± 13.32;P ＜0.05).Conclusions These data suggest that SE can result in activation of NF-κB/HIF-1o pathway in brain.Inhibition of the pathway can attenuate hippocampal neurodegeneration caused by SE, which has the brain protective effect.

Air ; analysis ; Cyclohexanones ; analysis ; Electrophoresis, Capillary ; Reproducibility of Results ; Workplace

Objective To investigate the expression profile of human genes in response to acute sodium arsenite treatment by cDNA microarray. Methods The RNA was purified from the L-02 cells without and with arsenite sodium induction for 2 hours, 15 hours and 24 hours, respectively. Results The hybridization patterns were different between every interval of arsenite induction. Expression of hCYR61 increased after 2 hours' induction, but decreased after 15 hours and 24 hours. Expression of metallothionein Ⅳ and Ⅲ elevated at the whole induction phase. HSP86 was up-regulated after 15 hours and 24 hours' induction, but it did not alter at two hours' induction. Conclusion When exposed to arsenite, the cells are under a meet-an-emergency situation to synthesize the most necessary protein and inhibit synthesis of unessential proteins.

AIM: To study the activation effects of maize pollen polysaccharides(PPM) on human thoracic cavity macrophage (hTM). METHODS: Activities of lactate dehydrogenase(LDH) and acid phosphatase (ACP) in the hTM were detected by automatic biochemical analyzer, and the level of tumor necrosis factor alpha(TNF-?) and interleukin-6(IL-6) in the hTM was analyzed by radioimmunoassay, after hTM were cultured with 0.312,0.625, 1.250, 2.500, 5.000 mg/mL PPM-RPMI 1640 for 24 and 48 hours in vitro. RESULTS: The activities of LDH and ACP increased in the hTM induced by PPM, and the levels of TNF-? and IL-6 in the hTM induced by PPM increased markedly too. And the induced expression effect of TNF-? and IL-6 is associated with the concentration of PPM, and time for PPM inducing. CONCLUSION: PPM can induce cytokines secretion in hTM,and activate hTM in vitro.

To introduce professor Gao Yuchun's clinical experience and treating characteristics of facial paralysis treated with acupuncture and moxibustion. Professor Gao pays attention to yangming when he selects acupoints for clinical syndrome, and directs acupoints selection based on syndrome differentiation in different levels of jingjin, meridians and zangfu; he praises opposing needling technique and reinforcing the deficiency and reducing the excess highly; the acupuncture manipulation is gentle,shallow and slow for reducing the healthy side and reinforcing the affected side, and through losing its excess to complement its deficiency; besides, he stresses needle retaining time and distinguishes reinforcing and reducing. Facial paralysis is treated with key factors such as acupoints selecting based on yangming, acupuncture manipulation, needle retaining time, etc. And the spleen and stomach is fine and good at transportation and transformation; the meridians is harmonious; the qi and blood is smooth. The clinical efficacy is enhanced finally.
Acupuncture Points ; Acupuncture Therapy ; history ; methods ; Aged, 80 and over ; China ; Facial Paralysis ; history ; therapy ; History, 20th Century ; History, 21st Century ; Humans

OBJECTIVE Bergapten (BG), is a furanocoumarin derived from herbal and citrus extracts can act as antioxidant and selective anticancer agents.The current study aimed to investigate whether bergapten would attenuate immunosenescence and to exploreits immunomodulatory effects on immune responses in D-galactose-induced aging BALB/c mice.METHODS Firstly,mice were given D-galactose(180 mg·kg-1)subcutaneous injections for 30 d.To evaluate the establishment of the aging-related effect in mice, serum samples of BALB/c mice were collected from tail vein. Aging BALB/c mice were freely divided into three groups: negative control group received 1% Tween 80 solution only, named D-gal group. Positive groups were received BG administration at the dose of 20 and 100 mg·kg-1, named D-gal+BG(20)group and D-gal+BG(100)group,respectively.Effects of bergapten on T lympho-cyte proliferation and flow cytometry were assessed by using the splenic cell suspension. Enzyme linked immunospot kits were used to quantitatively determine interferon-γ(IFN-γ)and interleukin-4(IL-4) levels of the isolated serum. Immunophenotype was determined by using mixture of antibodies includ-ing anti-CD3,anti-CD4,and anti-CD8.RESULTS Bergapten(20 mg·kg-1)therapy can modulate immu-nity against viral epidemics and attenuate aging-induced immune deficiency(P<0.01),which was correlat-ed with the decline in the activation of the Th and Tc responses in D-galactose induced aging BALB/c mice.According to the in vivo results,bergapten exposure up-regulated the secretion of IFN-γ and IL-4 in T-helper 1(Th1)and T helper 2(Th2)cells(P<0.05,P<0.01).Additionally,BG(20 mg·kg-1)restored antigen-specific CD4+and CD8+T cells in aging models (P<0.05, P<0.01), which may help to curing chronic infections. CONCLUSION The beneficial effect of bergapten in D-galactose induced aging BALB/c mice may be due to the Th and Tc responses activation.

OBJECTIVETo determine the classification of masticatory myospasm by analyzing characteristics of clinical appearances.

METHODSThirty-six cases of masticatory myospasm from 2000 to 2010 were included. The clinical data of these patients were analyzed, including patient information, patient history, clinical characteristics, severity and the frequency of myospasmodic movement, electromyogram (EMG), and the efficacy of botulinum toxin injection treatment.

RESULTSThere were 11 males and 25 females, aged from 15 to 71. According to the clinical manifestation and EMG findings, patients could be divided into two groups: 18 cases were classified as jaw closing type which involved masseter and/or temporalis muscles presenting as trismus and acute pain, the other 18 cases were jaw opening type which involved lateral pterygoid muscles complaining difficulty in jaw closing and teeth clenching. The jaw closing type was often seen in patients of 20 to 50 years old, the jaw opening was frequently seen in patients over 50 years old. Jaw closing type was attacked intermittently and unilaterally, but jaw opening was often attacked continually and bilaterally. The rating scale of the severity of spasmodic movement was not different between the two types, but the frequency of spasmodic attack was much higher for jaw opening type (P < 0.05). The EMG of jaw closing type was classified into persistent, rhythmic and irregular type. The EMG of jaw opening type was classified into spontaneous and exercise-induced type. Twelve cases were treated by botulinum toxin injection that could significantly relieve symptoms.

CONCLUSIONSMasticatory myospasm can be classified into jaw closing and jaw opening types. Jaw closing type involves masseter and/or temporalis muscles and jaw opening type involves lateral pterygoid muscles. Botulinum toxin injection was the most effective therapy for the masticatory myospasm.

Adolescent ; Adult ; Aged ; Botulinum Toxins, Type A ; therapeutic use ; Electromyography ; Female ; Humans ; Jaw ; physiopathology ; Male ; Masseter Muscle ; physiopathology ; Masticatory Muscles ; physiopathology ; Middle Aged ; Pterygoid Muscles ; physiopathology ; Spasm ; classification ; drug therapy ; physiopathology ; Temporal Muscle ; physiopathology ; Young Adult

Objective To discuss the feasibility of single hole thoracoscopy of pleural fibreboard endarterectomy surgical treatment on chronic tuberculous empyema. Methods Retrospective analysis of minimally invasive treatment of 52 cases of chronic tuberculous empyema form January 2013 to May 2016, 50 cases applied single hole thoracoscopy surgery, video-assisted mini-thoracoscopy for another 2 cases. Results There was no death, operation time 60 ~ 240 min, average 160 min, bleeding 150 ~ 2000 ml, average 350 ml, postoperative chest tube drainage time 3 ~ 21 d, average 7 d, postoperative persistent leakage in 3 patients, 3 cases of atelectasis, incisional infection in 1 case, pleural effusion in 1 case, 3 cases of arrhythmia. All the cured patients are received the corresponding treatment, the follow-up of 3 ~ 36 m, the chest CT scan show no atelectasis. Conclusion Under the condition of strict selection of indication, single hole thoracoscopy of pleural fibreboard endarterectomy in treatment of chronic tuberculous empyema is safe and feasible, so it is worthy of making further clinical promotion and application.

OBJECTIVETo research the effects of glycogen synthase kinase (GSK3β) overexpression and GSK3β inhibitor SB-216763 on the proliferation of hepatic oval cells in rats and its regulatory mechanisms by Wnt signaling pathway.

METHODSThe hepatic oval cells WBF-344 were divided into the blank control group, GSK3β over-expression group, DMSO control group and GSK3β inhibitor groups, while the inhibitor groups set up three concentration gradients, that was 1, 5, 10 µmol/L. Using the GSK3β over-expression lentivirus, which had been identified correctly, and SB-216763 dealt with the cells WBF-344. The cells morphology of each group was observed under the phase contrast inverted microscope, and the expression of fluorescence in the lentivirus-transfected group was observed under the fluorescent microscope. The proliferation of each group cells was tested by CCK8 kits. The cells' apoptosis was detected by AnnexinV-FITC/PI kits. The expression of GSK3β, β-catenin and cyclin D1 were detected by Western blot.

RESULTSThe cells of GSK3β over-expression group were fewer and obvious aging. However, in each inhibitor added group, the cells' division and proliferation was vigorous, and the condition was good. Moreover, the cells' proliferation was getting stronger with the concentration of SB-216763 increasing. A large number of green fluorescence was expressed in the lentivirus-transfected cells. The cells' proliferation in GSK3β over-expression group restrained (t = 7.178, P < 0.01, as compared with control), while the cells' proliferation was vigorous in inhibitor groups (F = 45.030, P < 0.01, as compared with control). Flow Cytometry showed that the cells apoptosis was significant in GSK3β over-expression group. Western blot showed that the expression of GSK3β was increased, while the expression of β-catenin and cyclin D1 was decreased in the over-expression group. The expression of GSK3β had no significant difference among the control group and inhibitor groups. However, the expression of β-catenin and cyclin D1 was significantly increased with the concentration of SB-216763 increasing.

CONCLUSIONSThe overexpression of GSK3β can inhibit the Wnt signaling pathway, thus restrain the cells' proliferation and promotes apoptosis. SB-216763 can activate the Wnt pathway, thus promotes cells' proliferation.

Animals ; Cell Line ; Cell Proliferation ; drug effects ; Cyclin D1 ; metabolism ; Glycogen Synthase Kinase 3 ; metabolism ; Glycogen Synthase Kinase 3 beta ; Glycogen Synthase Kinases ; metabolism ; Hepatocytes ; drug effects ; Indoles ; pharmacology ; Male ; Maleimides ; pharmacology ; Rats ; Transfection ; Wnt Signaling Pathway ; beta Catenin ; metabolism