Objective: The main ingredients and the inhibitory effects of essential oil of a compound Chinese herbal medicine Weiqi Decoction (WQD) on AGS cell proliferation were to be investigated. Methods: Chemical compounds of WQD essential oil were detected by gas chromatography and mass spectrometry analysis. Cell viability was measured by methyl thiazolyl tetrazolium method. Cell cycle distribution was detected by flow cytometry. Apoptosis and necrosis of AGS cells were determined by Hoechst 33342/propidium iodine staining. Results: Chemical analysis showed that the main ingredients of WQD essential oil were bornylene and 3-n-butylphthalide. Ligustilide, which is the effective compound of Danggui (Radix Angelicae Sinensis), was not detected in WQD essential oil. The essential oil inhibited cell proliferation in a dose- and time-dependent manner, and blocked cell cycle progression at G(2)/M stage. At the concentrations that resulted in significant inhibition of cell proliferation and cell cycle arrest, essential oil induced both apoptosis and necrosis. Conclusion: The results suggest that WQD essential oil contains some effective ingredients for treating chronic atrophic gastritis and functional dyspepsia, and also has an antiproliferative effect on AGS cells through cell cycle arrest and apoptosis promotion in vitro. Therefore, essential oil should be retained as much as possible during stewing this decoction.

Clinical and imageological characteristics of Paget's disease of bone in 7 patients who were treated in our hospital from 1991 to 2007 were analyzed. The data showed (1) 6 patients had the symptoms of bone pain and bone deformity, with more long bones involved; (2) Serum alkaline phosphatase was over normal range; (3) Imageology showed that osteoclasia was usually combined with bone sclerosis; (4) All the patients were treated with bisphosphonates; (5) 2 patients with serious bone deformity were treated with orthomorphia. Paget's disease of bone is a kind of metabolic bone disease with the characteristics of osteoclasia combined with bone sclerosis. The main features are bone pain and bone abnormality. Bisphosphonates are the first choice of drugs. Patients with serious bone deformity should be treated with orthomorphia.

OBJECTIVETo evaluate the erectile and ejaculatory function of sacral tumor patients after sacral nerve root resection and investigate the relationship of erectile and ejaculatory dysfunction (EED) with the level of sacral nerve injury.

METHODSThis retrospective study included 47 male patients aged 16 to 63 (32.6 +/- 6.8) years treated by sacral tumor resection between January 2008 and August 2013. According to the levels of the sacral nerve roots spared in surgery, the patients were divided into four groups: bilateral S1-S3 (n=16), unilateral S1-S3 (n=21), unilateral S1-S2 (n=6), and unilateral S1 (n=4). The patients were followed up for 12 to 41 (27.2 +/- 10.9) months by questionnaire investigation, clinic review, and telephone calls about their erectile and ejaculatory function at 3, 6 and 12 months after surgery and in August 2013.

RESULTSIn the bilateral S1-S3 group, the incidence rates of EED were 31.25% (5/16), 25% (4/16), and 12.5% (2/16) at 3, 6, and 12 months respectively after surgery, with recovery of erectile and ejaculatory function in August 2013. The incidence rates of EED in the unilateral S1-S3 group were 85.71% (18/21), 71.43% (15/21), 52.38% (11/21), and 42.86% (9/21) at 3, 6 and 12 months and in August 2013, respectively; those in the unilateral S1-S2 group were 100% (6/6), 83.33% (5/6), 83.33% (5/6), and 66.67% (4/6) at the four time points; and those in the unilateral S1 group were all 100% (4/4). No statistically significant differences were found in the incidence rate of EED among the patients of different ages or tumor types (P > 0.05).

CONCLUSIONThe incidence of postoperative EED in male patients treated by sacral tumor resection is closely related to the mode of operation. Sparing the S3 nerve root at least unilaterally in sacral tumor resection is essential for protecting the erectile and ejaculatory function of the patient.

Adolescent ; Adult ; Ejaculation ; physiology ; Erectile Dysfunction ; epidemiology ; etiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Organ Sparing Treatments ; Peripheral Nervous System Neoplasms ; surgery ; Postoperative Complications ; epidemiology ; Postoperative Period ; Retrospective Studies ; Sacrum ; Spinal Nerve Roots ; injuries ; surgery ; Surveys and Questionnaires ; Young Adult

OBJECTIVESTry to induce liver-derived dendritic cells proliferation by plasmid-IL-3 and CD40L genes in mice in vivo.

METHODSRapid tail-vein injection of large amounts of plasmid-carrying genes was performed to transfect genes in mice livers. Liver nonparenchymal cells were isolated by Percoll gradient centrifugation. Dendritic cell markers were detected and dendritic cells were sorted out by flow cytometry. Morphology of dendritic cells was studied microscopically (with Giemsa staining) and under scanning electron microscopy.

RESULTSLiver nonparenchymal cells dramatically increased in the liver lobules, portal and periportal areas in the treated group, but not in the control group. B220+/DEC205+ dendritic cells were detected and sorted by flow cytometry. There were more B220+/DEC205+ dendritic cells (16.0%) in the experiment group than those in the control group (1.1%). Morphologically, the sorted B220+/DEC205+ cells showed irregular shaped nuclei, paucity of cytoplasmic granules and extensive cytoplasmic processes.

CONCLUSIONB220+/DEC205+ dendritic cells were expanded in vivo in mice livers by rapid tail-vein injection of plasmid-carrying genes encoding IL-3 and CD40L in a large amount. Inducing liver dendritic cell proliferation in vivo provides a more convenient way for studying the biology of these cells.

Animals ; CD40 Ligand ; genetics ; Dendritic Cells ; cytology ; Flow Cytometry ; Hepatocytes ; cytology ; Interleukin-3 ; genetics ; Male ; Mice ; Mice, Inbred C57BL ; Transfection

OBJECTIVE: To perform the proteome analysis of conditioned medium prepared from mouse embryonic fibroblast feeder layers by 2-dimensional (2D) electrophoresis and mass spectrometry and to find out the possible differentiation-inhibitory factor in conditioned medium. METHODS: Feeder layers were prepared by 60Co gamma-irradiation on mouse embryonic fibroblast. Insulin-transferrin-sodium selenite supplemented medium was used to culture the feeder layers for 24 hours. The condioned medium prepared from mouse embryonic fibroblast feeder layers were made into powder by lyophilization, the redissolved solution was applied to Sephadex G-50 gel filtration chromatography, and then cold acetone was used to precipitate the proteins in the eluted solution. The protein samples were applied to 2D electrophoresis. The 2D images were analyzed by 2D image analysis software. Selected protein spots were digested by trypsin, analyzed by mass spectrometry, and then searched against the NCBInr batabase using Mascot MS/MS Ions Search. RESULTS: The protein samples extracted from mouse embryonic fibroblast feeder layers conditioned medium could be used for 2D electrophoresis. On 2D images, there were (221+/-67) spots. Most of the proteins were located in the region of MW 20 approximately 70 kD, pI 4 approximately 8. Using mass spectrometry, we preliminarily identified 13 spots: 3 keratins, 3 transferrins, 1 trypsin precursor, 2 unknown proteins (3 spots), 1 connexin 46, 1 beta-galactoside binding protein, and 1 secreted protein, acidic and rich in cysteine. CONCLUSION Conditioned medium prepared from mouse embryonic fibroblast feeder layers contain beta-galactoside binding protein and secreted protein, acidic and rich in cysteine.
Animals ; Cells, Cultured ; Culture Media, Conditioned ; chemistry ; Cysteine ; chemistry ; Electrophoresis, Gel, Two-Dimensional ; Embryo, Mammalian ; Fibroblasts ; cytology ; Galactosides ; chemistry ; Mass Spectrometry ; Mice ; Proteome

Objective To investigate the expression of Toll-like receptor 3 (TLR3) on dendritic cells(DCs) derived from peripheral blood mononuclear cells(PBMCs) of chronic hepatitis B(CHB) patients and to explore the mechanism of sustained infection of hepatitis B virus (HBV). Methods Twenty CHB patients were randomly screened in the study,and ten healthy persons were recruited as controls.The monocytes isolated from peripheral blood of candidates were incubated with recombinant human granuloeyte macrophage colony-stimulating factor (rhGM-CSF) and rhIL-4 to induce the DCs generation and proliferation.Then the phenotype of DCs was identified by micro- scope.The expressions of the phenotypes[histocompatibility leukocyte antigen(HLA)-DR,CD80, CD86,CD83]of immature and mature DCs were measured by flow cytometer.Furthermore,the ex- pression of TLR3 on mature DCs(mDC) and immature DCs(imDC) was determined by flow cytometry and Western blot analysis respectively.Results As for healthy volunteers,the expressions of CD80, CD86,HLA-DR and CD83 on DCs at the 7th day,which were(82.35?8.67)%,(79.61?10.08)%, (92.79?8.48)% and (83.76?5.47)% respectively,were significantly higher than those at the 5th day which were(28.31?8.79) %,(31.17?11.23)%,(27.61?10.28)% and (23.46?11.53)% respec- tively(P0.05).The expression of TLR3 on imDC was significantly higher than that on mDC at control group (P0.05).And the expression of TLR3 on imDC in CHB patients group was significantly lower than that of control group(P

OBJECTIVES: To explore the number of necrophagous flies and seasonal distribution of common necrophagous flies at present in Beijing. METHODS: The specimens of necrophagous flies were collected by the methods of animal carcass, trapping and feeding. And the specimens were observed and counted after the classification and preservation. RESULTS: The necrophagous flies in Beijing belonged to 4 families, 9 subfamilies, 21 genera and 46 species, and 12 species of them were the first records in Beijing. The necrophagous flies had the characteristics of regional and seasonal distribution. CONCLUSIONS The data of seasonal distribution of necrophagous flies and common necrophagous flies in Beijing can provide reference for related research.
Animals ; Beijing ; Cadaver ; Diptera/physiology* ; Entomology ; Larva ; Postmortem Changes

Reduced radiosensitivity of lung cancer cells represents a pivotal obstacle in clinical oncology. The hypoxia-inducible factor (HIF)-1α plays a crucial role in radiosensitivity, but the detailed mechanisms remain elusive. A relationship has been suggested to exist between hypoxia and autophagy recently. In the current study, we studied the effect of hypoxia-induced autophagy on radioresistance in lung cancer cell lines. A549 and H1299 cells were cultured under normoxia or hypoxia, followed by irradiation at dosage ranging from 0 to 8 Gy. Clonogenic assay was performed to calculate surviving fraction. EGFP-LC3 plasmid was stably transfected into cells to monitor autophagic processes. Western blotting was used to evaluate the protein expression levels of HIF-1α, c-Jun, phosphorylated c-Jun, Beclin 1, LC3 and p62. The mRNA levels of Beclin 1 were detected by qRT-PCR. We found that under hypoxia, both A549 and H1299 cells were radio-resistant compared with normoxia. Hypoxia-induced elevated HIF-1α protein expression preferentially triggered autophagy, accompanied by LC3 induction, EGFP-LC3 puncta and p62 degradation. In the meantime, HIF-1α increased downstream c-Jun phosphorylation, which in turn upregulated Beclin 1 mRNA and protein expression. The upregulation of Beclin 1 expression, instead of HIF-1α, could be blocked by SP600125 (a specific inhibitor of c-Jun NH2-terminal kinase), followed by suppression of autophagy. Under hypoxia, combined treatment of irradiation and chloroquine (a potent autophagy inhibitor) significantly decreased the survival potential of lung cancer cells in vitro and in vivo. In conclusion, hypoxia-induced autophagy through evaluating Beclin1 expression may be considered as a target to reverse the radioresistance in cancer cells.

Background People recently realized that it is important for artificial vascular biodegradable graft to bionically mimic the functions of the native vessel.In order to overcome the high risk of thrombosis and keep the patency in the clinical small-diameter vascular graft (SDVG) transplantation,a double-layer bionic scaffold,which can offer anticoagulation and mechanical strength simultaneously,was designed and fabricated via electrospinning technique.Methods Heparin-conjugated polycaprolactone (hPCL) and polyurethane (PU)-collagen type Ⅰ composite was used as the inner and outer layers,respectively.The porosity and the burst pressure of SDVG were evaluated.Its biocompatibility was demonstrated by the 3-(4,5-dimethyl-2-thiazol)-2,5-diphenyl-2H tetrazolium bromide (MTT) test in vitro and subcutaneous implants in vivo respectively.The grafts of diameter 2.5 mm and length 4.0 cm were implanted to replace the femoral artery in Beagle dog model.Then,angiography was performed in the Beagle dogs to investigate the patency and aneurysm of grafts at 2,4,and 8 weeks post-transplantation.After angiography,the patent grafts were explanted for histological analysis.Results The double-layer bionic SDVG meet the clinical mechanical demand.Its good biocompatibility was proven by cytotoxicity experiment (the cell's relative growth rates (RGR) of PU-collagen outer layer were 102.8％,109.2％ and 103.5％,while the RGR of hPCL inner layer were 99.0％,100.0％ and 98.0％,on days 1,3,and 5,respectively) and the subdermal implants experiment in the Beagle dog.Arteriography showed that all the implanted SDVGs were patent without any aneurismal dilatation or obvious anastomotic stenosis at the 2nd,4th,and 8th week after the operation,except one SDVG that failed at the 2nd week.Histological analysis and SEM showed that the inner layer was covered by new endothelial-like cells.Conclusion The double-layer bionic SDVG is a promising candidate as a replacement of native small-diameter vascular graft.

AIM To investigate the efficacy and safety of Chinese Jing Liqueur (Curculiginis Rhizoma,Angelicae sinensis Radix,Cistanches Herba,etc.) in relieving main symptoms of patients with Kidney Yang Deficiency Syndrome.METHODS Within eight-week trial,one hundred and twenty patients randomly and equally assigned to control group and experimental group took 50 mL Chinese Jing Liqueur and 50 mL 10％ diluted Chinese Jing Liqueur,respectively.The grading scale for Kidney Yang Deficiency Syndrome and change curve for body surface temperature after drinking were established on day 0,the 4th week day and the 8th week day.The blood samples were collected for blood hemorheologies detection as well.An array of measurements before and after drinking,the Kidney Yang Deficiency Syndrome scores,single symptom scores,body surface temperature and hemorheologies between the two groups were thus compared.RESULTS The experimental group displayed a significantly higher clinical cure rate (31.034％) than the control group (5.172％) (P =0.000),total effective rate (69.966％)than the control group (34.483％) (P =0.000).The experimental group didn't compromise its superiority to the control group if evaluated by improvement in chilly sensation and the cold limbs (35.593％ to 6.667％,P =0.000);and by the fibrinogen level [(2.845 ± 0.724) g/L to (2.500 ± 0.395) g/L,P =0.004)].No significant difference in incidence of adverse reactions between the two groups was observed (P =0.619).Meanwhile,Chinese Jing Liqueur's power in improving the patients' fatigue and weakness of waist and knees,hyposexuality,listlessness,nocturia and lower extremity edema was noticed as well.CONCLUSION For patients with Kidney Yang Deficiency Syndrome,Chinese Jing Liqueur proves its efficacy in improving their main symptoms through enhancing the basic skin temperature and prolonging the duration of skin temperature rise.