OBJECTIVE:To study the inhibitory effects and mechanism of lutein on nasopharyngeal carcinoma C666-1 cells. METHODS:C666-1 cells were stimulated by lutein at different concentrations [0(blank control),20,40,80,160 mg/L] for dif-ferent time(0,12,24,48 h). The proliferation rate of cells was determined by CCK-8 assay,and apoptotic rate of cells was deter-mined by TUNEL method;Western blot was adopted to determine the phosphorylation of S6K and S6 proteins of AMPK and mTOR pathway. RESULTS:Compared with blank control group,proliferation rate of C666-1 cells was significantly reduced after treated with lutein(80,160 mg/L)for 48 h and lutein(160 mg/L)for 12,24,48 h(P<0.05). After treated with lutein(80,160 mg/L)for 48 h and lutein(160 mg/L)for 24,48 h,cell apoptosis was significantly increased(P<0.05). Lutein(80,160 mg/L) could promote intracellular AMPK phosphorylation,and inhibits mTOR pathway S6K,S6 protein phosphorylation after 48 h treat-ment (P<0.05). CONCLUSIONS:Lutein can inhibit nasopharyngeal carcinoma C666-1 cell proliferation and induce nasopharyn-geal carcinoma cell apoptosis and inhibit S6K,S6 protein phosphorylation through promoting AMPK phosphorylation.

BackgroundRecent studies showed that diabetic retinal neuropathy is an earlier and more dangerous complication and neurotrophin has a protective effect on retina.ObjectiveThe present study was to observe the changes of brain derived neurotrophic factor (BDNF),its receptor TrkB,signal pathway protein phosphatized extracellular signal-regulated protein kinase1/2 (p-ERK1/2) and c-fos in the retina after injection of BDNF into the vitreous in STZ induced Wistar diabetic rats.MethodsWistar rats aged 9 weeks-old were randomly divided into BDNF injection group,diabetes mellitus (DM) control group and normal control group and 20 rats for each group.STZ was intraperitoneally injected in the rats of BDNF injection group and DM control group to create the experimental DM.BDNF was intravitreously injected in the rats of BDNF group 2 weeks after administration of STZ in three-day interval for 5 times,and BSS containing O.1％ bovine serum albumin (BSA) was used at the same way in the DM control group and normal control group.The retina was isolated for hybridization in situ for BDNF,and TrkB,p-ERK1/2 and c-fos.Levels in retina were detected using sandwich method ELISA.ResultsThe number of BDNF positive cells and the gray scale were lower obviously in the rat retina of DM control group than those of BDNF injection group and normal control group,showing significant differences among the 3 groups ( F =102.36,92.55 ;P＜0.05 ).ELISA assay showed that TrkB,p-ERK1/2 and c-fos values in retina were statistically significantly different among the 3 groups ( F =92.54,95.46,94.84,P＜0.05 ).The TrkB level in retina was statistically reduced,but the p-ERK1/2 and c-fos levels in retina were increased statistically in DM control group compared with BDNF injection group and normal control group( P＜0.05 ).No statistical difference was found in TrkB,p-ERK1/2 and c-fos values between the BDNF injection group and normal control group(P＞0.05).ConclusionsThe injection of BDNF into the vitreous cavity can protect retina from downregulating BDNF and TrkB levels and up-regulating the p-ERK1/2 and c-fos protein levels in the early stage of DM.

Based on previous results of 16S rDNA sequence homologuous and results of physic-biochemical indexes and morphological characteristics in the present work, bacterial strain ge15 isolated from roots of ginseng plants was identified as Stenotrophomonas rhizophila. Confronting incubation results showed that, strain ge15 inhibited the growth of Alternaria panax, Phytophthora cactorum, and Cylindrocapon destructans significantly, and the width of inhibition zone was 13.3, 24.0, 12.0 mm, respectively. Further results showed that the emergence rate and seedling survive rate of ge15 treatment was significantly higher than those of the control, and which was similar to pesticide carbendazol treatment. The ge15 strain has good application potential in ginseng diseases control without contamination.
Alternaria ; growth & development ; physiology ; Antibiosis ; Biological Control Agents ; Hypocreales ; growth & development ; physiology ; Panax ; growth & development ; microbiology ; Phytophthora ; growth & development ; physiology ; Plant Diseases ; microbiology ; Plant Roots ; growth & development ; microbiology ; Seedlings ; growth & development ; microbiology ; Stenotrophomonas ; classification ; cytology ; isolation & purification ; physiology

Panax ginseng is one of the most important traditional Chinese herbal medicine, soil borne diseases influenced the yield and quality severely. In our previous work, endophytic Bacillus subtilis ge25 strain was isolated from ginseng root, and which showed significant antagonistic activity against several most destructive ginseng phytopathogens. In the present work, crude protein and lipopeptid extracts were prepared from LB and Landy supernate by salting out, acid precipitation methods respectively. The antagonistic activity of crude extracts and stability to temperature and protease digestion were examined by ginseng phytopathogen Alternaria panax. Results showed that, the antagonistic activity of crude protein extracts from LB culture was complete and partially lost when treated by high temperature and proteinase K. However, crude lipopeptid from Landy culture showed significant stabile antagonistic activity to them. Acid-hydrolyzation and TLC-bioautography analysis showed, that the crude lipopeptide contained at least one cyclic lipopeptide. In consideration of the stability and perfect antagonistic activity of ge25, further researches will promote the biocontrol of ginseng diseases in the field.
Alternaria ; drug effects ; physiology ; Bacillus subtilis ; metabolism ; physiology ; Bacterial Proteins ; isolation & purification ; metabolism ; pharmacology ; Endopeptidase K ; metabolism ; Endophytes ; metabolism ; physiology ; Fermentation ; Lipopeptides ; isolation & purification ; pharmacology ; Panax ; microbiology ; Plant Roots ; microbiology ; Temperature

Alpha-glycerophosphate oxidase (alpha-GPO) from Enterococcus casseli flavus was successfully isolated and purified by using polyethylene glycol (PEG)/(NH4)2SO4 aqueous two-phase system (ATPS). The results showed that the chosen PEG/(NH4)2SO4 ATPS could be affected by PEG molecular weight, pH, concentration of PEG and (NH4)2SO4, and inorganic salt as well as additional amount of crude enzyme. After evaluating these influencing factors, the final optimum purification strategy was formed by 16.5% (m/m) PEG2000, 13.2% (m/m) (NH4)2SO4, pH 7.5 and 30% (m/m) additive crude enzyme, respectively. The NaCl was a negative influencing factor which would lead to lower purification fold and activity recovery. These conditions eventually resulted in the activity recovery of 89% (m/m), distribution coefficient of 1.2 and purification fold of 7.0.
Ammonium Sulfate ; chemistry ; Glycerolphosphate Dehydrogenase ; chemistry ; isolation & purification ; Molecular Weight ; Polyethylene Glycols ; chemistry ; Water

Objective To explore the significance of 1.5-Tesla MRI in the early diagnosis of prostate cancer,and analyze the imaging features in T2WI which are helpful for the diagnosis on early prostate cancer.Methods The clinical data of patients with PSA ＜ 20 μg/L who received prostate biopsy between Feb.2006 and Jan.2011 were reviewed including pelvic MRI and biopsy results.We retrospectively analyzed T2WI image features and evaluated the signal of peripheral zone,seminal vesicles,prostate capsule and the relationship with their surrounding tissues.The results of the reading were analyzed with reference to histopathologic evaluation of prostate biopies.Results Among 359 patients,137 patients were diagnosed prostate cancer and 332 cases were found with low signal lesions on peripheral zone of T2WI.Univariate logistic regression analysis showed that age and PSA are closely related to the diagnosis of prostate cancer (P ＜ 0.05).Low signal lesions on peripheral zone of T2WI are more helpful for the diagnosis of prostate cancer on the image.Number,shape,margin of lesions,the contour of the prostatic capsule,the boundary of peripheral zone and transitional zone are significantly associated with the diagnosis of prostate cancer.Through multivariate logistic regression analysis,it was found that age (P ＜ 0.05),PSA (P ＜ 0.05),prostate volume (P ＜ 0.05),lesion shape (flaky vs nodular,P ＜ 0.05 ; diffuse vs nodular,P ＜ 0.05) and the contour of the prostatic capsule (disappearing vs completely clear,P ＜ 0.05) are risk factors of prostate cancer.The areas under receiver operating characteristic curve are 0.833 (P ＜ 0.05).Conclusions In T2WI,lesion shape and the contour of the prostatic capsule are risk factors of prostate cancer.Age,PSA,prostate volume are also significantly associated with the diagnosis of prostate cancer.

Objective To evaluate clinical risk factors for Meropenem resistant Pseudomonas aeruginosa (MRPA) infection in surgical intensive care unit. Methods Retrospective case-control study was designed. Resultant MRPA and Meropenem sensitive Pseudomonas aeruginosa ( MSPA) cases were paired with respective control group. Candidate clinical factors including gender, malignancy, duration of ICU stay, length of operation, volume of blood loss, transfusion, kind of incision ( clean/contaminated/ infected) , and previous antibiotics history were examined by bivariates and multivariate regression. Results Risk factors for MRPA infection included duration of ICU stay before infection (odds ratio [ OR] = 2. 234, 95% confidence interval [ CI] = 1. 710 - 2. 918), administration of third generation cephalosporins (OR = 2. 885, 95% CI = 1. 133 - 7. 345) and Imipenem ( OR = 1. 238, 95% CI = 1. 469 - 35. 659). A long ICU stay (OR = 2. 284, 95% CI = 1. 772 - 2. 945), malignancy (OR =4.796, 95%CI = 2.001 -11.491), previous medication of quinolones ( OR = 3.563, 95% CI = 1.499 - 8.468) and third generation cephalosporins (OR = 3. 796, 95% CI = 1. 543 - 9. 343) were associated with MSPA infection. Conclusion MRPA infection was affected by multiple clinical factors. Short ICU stay, restrict use of third generation cephalosporins and Imipenem may help to prevent MRPA infection.

Objective To analyze the prevalence and characteristics of skin diseases in elderly patients with type 2 diabetes mellitus (T2DM).Methods Clinical out patient records for both elderly and non-elderly individuals with T2DM were collected from the dermatology department of Beijing Hospital.Statistical comparisons were performed between the elderly group and the non-elderly group on the prevalence of common skin complications and disease characteristics.Analysis was performed in the elderly group to identify any association of common skin disorders with gender,age,duration of T2DM,or level of glycosylated hemoglobin (HbA1c).Results Among 679 elderly T2DM cases,pruritus was the most common complication (26.1％),followed by eczema (21.t ％) and tinea pedis/onychomycosis (18.3％).Pruritus was more prevalent in the elderly group (177 cases,26.1％) compared to the non-elderly group (34 cases,9.8％) (x2 =6.006,P＜0.01).On the other hand,tinea pedis/onychomycosis was less prevalent in the senior group (124 cases,18.3 ％) than in the non-senior group (112 cases,32.2 ％) (x2 =37.437,P＜0.01).In the senior group,pruritus was more prevalent among male patients (105 out of 358 cases,29.3％) than among female patients(72 out of 321 cases,22.4％) (x2 =4.181,P =0.041).On average,patients with eczema were older[(72.3 ± 2.2)years] and had a longer history of T2DM [(8.3 ± 0.3)years] compared with patients without eczema[age:(65.2±1.9)years,history of T2DM:(5.2±0.1)years] (t=1.250,P=0.023 for age;t=1.680,P=0.033 for disease duration).Patients with tinea pedis/onychomycosis had higher HbA1c levels (6.7 ± 0.4) ％ than patients without tinea pedis/onychomycosis (6.1 ± 0.3) ％ (t =0.790,P＜0.01).Conclusions Pruritus is the most common skin complication in elderly patients with type 2 diabetes and is more common in elderly patients and male patients.Risk factors for developing eczema in elderly patients with type 2 diabetes may include older age and long disease duration.High levels of glycosylated hemoglobin in elderly patients with type 2 diabetes indicates increased risk for foot fungi infections.

Objective To investigate the role of calreticulin (CRT) in the pathogenesis of rheumatoid arthritis (RA) by analyzing its expression in the sera,synovial fluid and synovial membrane in patients with RA.Methods Levels of CRT in the sera from patients with RA,osteoarthritis (OA),systemic lupus erythematosus (SLE),other autoimmune diseases and health control (HC) were detected by enzyme linked immunosorbent assay (ELISA) and Western bloting.CRT levels in synovial fluid from RA and OA patients were measured by ELISA.Pathological methods were employed to analyze the expression and localization of CRT in synovial membrane.ANVOA,SNK-q test were used for statistical analysis.Results CRT was found to be significantly up-regulated in sera from RA [(4.8±2.4) ng/ml] than that from in OA [(3.6±0.9)ng/ml],SLE [(4.0±1.5) ng/ml],other autoimmune diseases [(3.9±0.8) ng/ml] and HC [(3.7±0.6) ng/ml].Only the monomer form of CRT in the serum could be detected.Pathological results showed that in RA synovium,CRT was mainly expressed in the lining and sublining layers,endothelial cells and perivascular areas； while in OA,only few CRT staining was seen in perivascular areas and the synovial lines.Conclusion In RA,increased levels of CRT are detected in the sera.In addition,high expression of CRT is observed in synovium and its distribution are different from OA.Our results suggest that CRT may be involved in RA joint inflammation and pannus formation.CRT may become a potential serological marker in the diagnosis of RA.

Objective To evaluate the effectiveness of treating glioma in combination with drugs multiply by comparing the size of tumor and the survival time of different groups in rat glioma after targeted blood-brain barrier (BBB ) disruption by MRI-guided focused ultrasound.Methods The stereotaxis instruments and the 10 μl gas-tight syringes were used to inject gliosarcoma cells into the targeted area of the brain in 50 male Sprague-Dawley rats.The glioma-bearing rat model was established.Each rat received either:(1 )no treatment (control;n =8);(2)single liposomal doxorubicin (DOX;n = 10);(3)multiple DOX (n =10);(4)single Avastin (AVS)and DOX (n =10);(5)multiple AVS and DOX (n =10).The SonoVue microbubble ultrasonic contrast agent and DOX or AVS were injected into the tail vein respectively on day 12 after implantation.The tumor size was measured by MRI on pre-treatment,immediacy and once a week of post-treatment after targeted BBB disruption by focused ultrasound,and the life span in rat glioma was recorded.Results The mediam survival of different groups in rat glioma(The range of the life span 13-90 d):no treatment (7 d);single DOX (12 d);multiple DOX (1 5 d);single AVS + DOX (22 d), multiple AVS+ DOX (30 d).There was significant difference of the groups on mediam survival comparison (P < 0.01 ).The tumor growth pattern after post-treatment of different groups in rat glioma except control:single DOX was noticeable fast and multiple AVS+DOX was visibly delayed comparable to other groups,and finally the tumor size of multiple AVS + DOX even became small.Conclusions The microbubble blasting enhances the local tissue permeability and promotes the drug delivery of chemotherapy and anti-angiogenesis locally in glioma-bearing rats by MRI-guided focused ultrasound.Especially,the combination with drugs multiply has a synergism efficacy that may enhance the effectiveness of chemotherapy,reduce tumor growth,and even become small of the tumor size,and increase survival time significantly after BBB disruption.