Objective To investigate the effects of ischemic preconditioning (IP) on the expression of extracellular signal-regulated protein kinase (ERK) and c-jun N-terminal kinase (JNK) in hippocampus in a gerbil model of ischemia-reperfusion (I/R) injury and the role of ERK and JNK in the mechanism of ischemic cerebral preconditioning. Methods Gerbils of both sexes weighing 50-70kg were randomly divided into 4 groups : ( Ⅰ ) sham operation group; ( Ⅱ ) IP group; (Ⅲ) I/R group and ( Ⅳ ) IP + I/R group. Cerebral ischemia was produced by occlusion of bilateral common carotid arteries and confirmed by isoelectricity on EEG. In sham operation group bilateral common carotid arteries were exposed but not occluded. In IP and I/R groups the animals were subjected to 3 min (IP group) or 5 min (I/R group) cerebral ischemia respectively. In IP + I/R group the animals first underwent 3 min cerebral ischemia followed by 24h reperfusion and then were again subjected to 5 min cerebral ischemia. Open field test was performed to evaluate the behavioral deficit 1,3,5 and 7 days after ischemia. The animals were sacrificed at 15 min, 2, 4, 6h and 1, 3, 5, 7 days after ischemia. The brains were immediately removed for detection of apoptosis (TUNEL) and expression of p-ERK and JNK (immuno-histochemistry) in hippocampal CA1 and CA3 regions and microscopic examination. Results Compared with I/R group the behavioral deficit was significantly decreased and the number of living pyramidal neurons was significantly increased and apoptotic neurons significantly decreased in IP + I/R group. No p-ERK expression was detected in CA1 region in all of the 4 groups but in CA3 region the p-ERK expression was significantly higher in group IP + I/R than in group I/R. The p-JNK expression increased gradually during reperfusion in both CA1 and CA3 regions and was still detectable 7 days after ischemia and was significantly lower in CA1 region in group IP + I/R than in group I/R.Conclusion IP protects hippocampal neurons from I/R injury by inhibiting the expression of p-JNK in CA1 region and enhancing the activity of p-ERK in CA3 region

Objective:To introduce recent progress on diagnosis and treatment of benign symmetric lipomatosis(BSL).Method:Detailed clinical data of 6 patients with BSL were reviewed and analyzed.We present a summary of the clinical symptoms,physical sign,diagnosis and therapeutic methods of BSL. And related literatures were discussed together.Result:All of 6 patients have excessive subcutaneous fat deposit predominantly around neck.One patients had upper extremity localizations. Six patients had the complication of left ventricular diastolic function changes,glucose intolerance or diabetes mellitus, chronic hepatopathy, hyperuricemia and sleep apnea syndrome in one or more. One patients with several symptoms occur simultaneously, another one female patient was accompanied by all symptoms but chronic hepatopathy. Five male patient were alcohol abusers. Tatal neck lipentomy and abstinence from alcohol were performed on 5 patients. One patient refused treatment.During a follow-up of 3 months to 4 years, one patients was relapsed again,and no recurrence was seen in another 4 patients. All pathological results were nonencapsulated fat. Conclusion:BSL is a lipodystrophy caused by diffuse fatty tissue, symmetry deposition in the neck and shoulder subcutaneous fascia space or deep fascial space .The highest incidence favors to middle-aged man who is alcoholist. Chronic alcohol addiction and typical clinical symptoms help to diagnosis BSL. Lipectomy represents a successful procedure in treating BSL.

Objective To evaluate the risk factors of breast cancer in Asian women and to provide evidences for establishing a risk assessment model. Methods Published studies concerning risk factors of breast cancer in Asian women were searched systemically and assessed by NOS (Newcastle-Ottawa Scale) items between 1995 and 2010. RevMan 4.2 software was used for data analysis and for calculating OR and its 95%CI on every risk factor. Results 27 studies including 403170 women were selected for Meta-analysis. According to NOS items, 20 studies were classified as A degree and 7 studies were evaluated as B degree. The risk factors of breast cancer and its pooled odds ratio values with statistical significance were as follows: 3.00 (95%CI: 1.68-5.36) when number of abortions≥3; 2.39 (95%CI: 1.78-3.21 ) when with family history of breast cancer; 1.54(95%CI: 1.30-1.82) when age at first live birth ≥30 (year); smoking was 1.50(95%CI: 1.03-2.20); 1.48(95%CI:1.20-1.83) with no live births; 1.29 (95%CI: 1.12-1.47) with no breast feeding; 1.26 (1.07-1.49)with age at menarche ≤12 (year) and 1.16(95%CI: 1.01-1.32) with alcohol drinking. Conclusion Number of abortions≥3, family history of breast cancer, age at first live birth ≥30 (year) ,smoking, no live births, no breast feeding, age at menarche ≤ 12 (year), and alcohol drinking were among the priorities in the establishment of breast cancer risk assessment model for Asian women.

Objective To observe the clinical efficacy ofChanhou Fujiu decoction plus acupuncture-moxibustion in preventing and treating incomplete drug abortion.Method A total of 204 patients in early-stage pregnancy asking for drug abortion were randomized into group A, group B, and group C, 68 cases in each group. Group A was given oral administration of Mifeprostone and Misoprostol tablets, plus Azithromycin dispersible tablet for prevention of infection. Group B was intervened by orally takingChanhou Fujiu decoction in addition to the treatment given to group A. Group C was intervened by acupuncture-moxibustion therapy based on the treatment given to group B. The vaginal bleeding time and amount, and the recovery of menstruation in the three groups were observed after the intervention, and the clinical efficacies were compared.Result Compared with group A, the vaginal bleeding amount after drug abortion was significantly smaller (P<0.05), the bleeding duration were significantly shorter (P<0.05), and time taken for the recovery of menstruation was markedly shorter (P<0.05) in group B and C, and there were no significant abnormal conditions in the menstrual periods (P>0.05). Compared with group B, group C had significantly smaller amount of vaginal bleeding (P<0.05) and shorter duration of vaginal bleeding (P<0.05), and there were no significant abnormal conditions in the time taken for the recovery of menstruation and menstrual periods (P>0.05). The total effective rate was 75.0％ in group B and 95.6％ in group C, both significantly higher than 58.8％ in group A (P<0.05), and the total effective rate in group C was markedly higher than that in group B (P<0.05). Conclusion Chanhou Fuyuan decoction plus acupuncture-moxibustion can effectively prevent and treat complications of drug abortion, and is an effective method in preventing and treating incomplete drug abortion.

OBJECTIVETo study mechanismt of Fufang Haishe capsule for dementia by observing the effect of it on PC-12 cell apoptosis, which was induced by beta-amyloid protein (Abl-42).

METHODNerve growth factor (NGF) was used to cultivate the PC-12 cells. Fufang Haishe capsule at different concentrations was added into the culture medium so as to identify the nontoxic concentrations with MTT. To analyze the PC-12 cell apoptosis respectively by MTT assay, Flow cytometry (FCM technique) with different concentrations of Fufang Haishe capsule (0.01, 0.1, 1, 5 mg x mL(-1)), adding Ab or not Western blot was used to detect apoptosis which was measured on the implementation of caspase-9 and caspase-3 activity.

RESULTFufang Haishe capsule could significantly inhibit the apoptosis of PC-12 cells induced by Abeta with increased colorimetric MTT asay ( compare among the control group and concentration 0, 0.01, 0.1, 1 and 5 mg x mL(-1) group, which is the same below: 1.75 +/- 0.12, 0.73 +/- 0.35, 0.79 +/- 0.11, 0.83 +/- 0.07, 1.31 +/- 0.07, 1.80 +/- 0.38, P < 0.01) and the decreased apoptosis rate of the cells which was analysed by flow cytometry (1.93 +/- 0.41)%, (46.17 +/- 4.08)%, (35.35 +/- 4.63)%, (28.62 +/- 3.81)%, (15.13 +/- 3.15)%, (7.84 +/- 1.76)%, P < 0.01. In addition, Fufang Haishe capsule inhibited the activity of caspase-9 and caspase-3 of PC-12 cells which was induced by Abeta.

CONCLUSIONFufang Haishe capsule significantly inhibite apoptosis of PC-12 cells induced by Abeta. The mechanism might be that Fufang Haishe capsule decrease the activity of the apoptosis implementing protein,caspase-9 and caspase-3.

Animals ; Apoptosis ; drug effects ; Capsules ; Caspases ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; PC12 Cells ; Rats

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.
Administration, Oral ; Animals ; Arylamine N-Acetyltransferase ; genetics ; metabolism ; Caffeine ; metabolism ; urine ; Cytochrome P-450 CYP1A2 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Female ; Male ; Medicine, Tibetan Traditional ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Theophylline ; urine ; Uracil ; analogs & derivatives ; urine ; Uric Acid ; analogs & derivatives ; urine ; Xanthines ; urine

Endoplasmic reticulum (ER) stress occurs in macrophage-rich areas of advanced atherosclerotic lesions and contributes to macrophage apoptosis and subsequent plaque necrosis. The purpose of the present study was to investigate the effects of caveolin-1 (Cav-1) on ER stress-induced apoptosis in cultured macrophages and the underlying mechanisms. RAW264.7 cells were incubated with thapsigargin (TG) to establish ER stress model. And Cav-1 expression was detected by Western blot. After being pretreated with filipin(III), a caveolae inhibitor, RAW264.7 cells were assayed with flow cytometry and confocal laser scanning microscopy to detect cell apoptosis. Moreover, p38 mitogen-activated protein kinase (MAPK) phosphorylation and C/EBP homologous protein (CHOP) expression were detected with Western blot. The results showed that Cav-1 expression was markedly increased at early stage of TG treatment (P < 0.05) and then decreased with prolonged or high dose TG treatments. The increasing of Cav-1 expression induced by TG in RAW264.7 cells was abolished under inhibition of caveolae by filipin(III) (P < 0.05). The effect of TG on apoptosis of RAW264.7 cells was further augmented after pretreatment with filipin(III) (P < 0.05). Western blotting showed that MAPK phosphorylation induced by TG was inhibited by filipin(III) in RAW264.7 cells (P < 0.05), whereas CHOP remained unchanged (P > 0.05). These results suggest that Cav-1 may play a critical role in suppressing ER stress-induced macrophages apoptosis in vitro, and one of the mechanisms may be correlated with the activation of p38 MAPK prosurvival pathway.
Animals ; Apoptosis ; drug effects ; Caveolin 1 ; genetics ; metabolism ; Cell Line ; Endoplasmic Reticulum Stress ; physiology ; Filipin ; pharmacology ; MAP Kinase Signaling System ; Macrophages ; cytology ; drug effects ; Mice ; Thapsigargin ; pharmacology ; Transcription Factor CHOP ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Dermatomyositis ; diagnosis ; Diagnostic Errors ; Hepatitis B ; diagnosis ; Hepatitis D ; diagnosis ; Humans ; Male ; Middle Aged ; Superinfection ; diagnosis

OBJECTIVETo study the proto-oncogene RON mediated aggression of Raji cells and the inhibitory effects by monoclonal antibody Zt/f2 (2f2).

METHODSThe effects of RON ligand macrophage stimulating protein (MSP) (2.0 nmol/L) and inhibitory Zt/f2 (2F2) (2.0 nmol/L) antibody on proliferation of RON positive Raji cells after treatment for 24 and72 hours were detected by MTT method, colony formation units (CFU) of Raji cells by methylcellulose semi solid culture, Raji cells apoptosis and cell cycle analysis by AnnexinV/PI double staining, expression of RON, apoptosis-related proteins, and cyclins by Western blot.

RESULTS(1)Compared with the cell viability (1.0) and counts of CFU (103.6±7.0) in control group, Raji cells after MSP treatment had better viability (1.35±0.20) and CFU counts (133.7±10.4) (P<0.05), but worse viability (0.68±0.11) and CFU counts (66.3±6.1) after Zt/f2 (2F2) treatment (P<0.05). (2)Percentage of Raji cells apoptosis after Zt/f2 (2F2) antibody treatment (12.16±2.33)% was significantly increased than the control (2.89±1.03)% (P<0.05). The percentage of Raji cells arrested in G0/G1 phase was increased after Zt/f2 (2F2) antibody treatment as compared to the control [ (54.96 ±3.70)% vs (39.10±2.30)%, (P<0.05) ]. (3) High-level of RON phosphorylation and β-catenin expression activated by MSP could be inhibited significantly by Zt/f2 (2F2), which also up-regulated the expression of caspase-3, caspase-8, caspase-9 and PARP and down-regulated anti-apoptotic MCL-1 gene and inhibitor of apoptosis protein XIAP expression, accompanied with G1 phase protein changes accordingly.

CONCLUSIONMSP could aggravate Raji cells proliferation. Inversely, Zt/f2 (2F2) could inhibit proliferation and induce apoptosis by inhibition of RON phosphorylation and up-regulation of apoptosis related proteins.

Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Proto-Oncogenes ; Receptor Protein-Tyrosine Kinases ; metabolism

OBJECTIVETo study the effects of sinomenine (Sin) on cell proliferation, intracellular expression of cyclooxygenase-2 (COX-2), and production of PGE2 in lipopolysaccharide-induced PC-12 cells, To explore the Sin's mechanism on nerve cell.

METHODPC-12 cells were cultured with nerve growing factors (NGF), and pretreated with Sin at various concentrations (0, 3 x 10(-6), 30 x 10(-6), 150 x 10(-6) mol x L(-1)) for 2 hours, then with or without stimulation of lipopolysaccharide (LPS). The proliferation activity of PC-12 cells was determined by 3H-TdR incorporation, and the production of PGE2 in culture supernatants of PC-12 cells was detected with competitive ELISA. Expression of COX-2 mRNA in PC-12 cells was analyzed by semi-quantitative RT-PCR, and expression of COX-2 protein was estimated by Western blot method and cellular enzyme immunoassay. Nuclear factor-kappa B (NF-kappaB) activity in whole-cell extract of PC-12 cells was also measured by an ELISA-based method.

RESULTThe data showed that Sin down-regulated the expression of COX-2 mRNA and protein, and reduced the production of PGE2 in the LPS-stimulated PC-12 cells which correlated with Sin's concentrations positively. In addition, NF-kappaB activity in LPS-stimulated cells was suppressed significantly by Sin. No inhibition of proliferation of PC-12 cells due to Sin treatment was observed.

CONCLUSIONSin mediates the down-regulation of expression of COX-2 and production of induced PGE2 in PC-12 cells by suppressing the activity of NF-kappaB.

Animals ; Cell Proliferation ; drug effects ; Cyclooxygenase 2 ; Dinoprostone ; biosynthesis ; Down-Regulation ; Lipopolysaccharides ; Morphinans ; isolation & purification ; pharmacology ; NF-kappa B ; metabolism ; PC12 Cells ; Plants, Medicinal ; chemistry ; Prostaglandin-Endoperoxide Synthases ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Sinomenium ; chemistry