Objective To detect serum level of long noncoding RNA ( lncRNA) BC200 in gastric cancer(GC) patients, and investigate its relationship with clinical features , and evaluate its diagnostic value for GC.Methods A case-control study was performed.From November 2014 to July 2015, serum levels of lncRNA BC200 were detected by real-time quantitative polymerase chain reaction in 124 patients with GC , 41 patients with atrophic gastritis and 59 normal controls who were hospitalized in Qilu Hospital of Shandong University.Meanwhile , serum carcinoembryonic antigen ( CEA ) and carbohydrate antigen 72-4 ( CA72-4 ) were detected by electrochemical luminescence immunoassay .Serum levels of lncRNA BC200, before and 3, 7, 10, 30, 100 days after radical operation in another 31 patients with GC were determined.The sensitivity and specificity of serum lncRNA BC200, CEA and CA72-4 were analyzed by using of the receiver operating characteristic ( ROC) curve.The comparison between two groups was performed with Mann-Whitney U test and the comparison among many groups was conducted with Kruskal-Wallis H test.Results Serum levels of lncRNA BC200 in GC patients with stage Ⅰ and Ⅱ[1.041(0.794,1.462)] and stage Ⅲ and Ⅳ[1.290 (0.978,1.794)]were significantly higher than those in patients with precancerous lesion [0.969(0.699, 1.219)]and normal controls[0.801(0.556,1.599)](H =54.68,P<0.000 1).Compared with pre-operation[1.120 (0.859,1.663)], the serum BC200 levels decreased significantly in 10 days [0.903 (0.724,1.182)](U=55.0,P<0.000 1), 30 days[0.759(0.671,1.037)](U=299.0,P=0.026 1), and 100 days[0.478(0.378,0.635)](U=41.0,P<0.000 1) after surgery.The area under the receiver operating characteristics curve ( AUC) of serum lncRNA BC200 was 0.865 for GC diagnosis, which was significantly higher than that of serum CA 72-4 ( AUC =0.699 ) or CEA ( AUC =0.807 ) .The AUC of combined detection of three tests was 0.934.Conclusion Serum lncRNA BC200 levels are significantly increased in GC patients , which may be used as a potential biomarker in GC diagnosis and monitoring .

Objective To investigate the daily total fluoride intake in relating to the prevalence of skeletal fluorosis in two villages in Jiangsu Province,in order to provide the scientific evidences for the control and prevention of endemic fluorosis.Methods Adults sampled from a high-fluoride Village,Wamiao,and a low-fluoride Village,Xinhuai,were surveyed in this study according to the fluoride concentration in their household shallow well.The average daily total fluoride intake from difierent sources and the skeletal fluorosis were investigated in each subject.Then the subjects from two villages were allocated into five subgroups(＜2.00,2.00～,3.00～,4.00～,≥5.00 mg/d),the relation fluoride intake and prevalence of osteofluorosis was analyzed.Results The prevalence of skeletal fluorosis in Wamiao Village was 31.06%(41/132),but no skeletal fluorosis case(0/35)was found in Xinhuai Village.According to the daily total fluoride intake,subjects with higher daily total fluoride intake tended to associated with a higher prevalence of skeletal fluorosis in a significant dose-response relationship(regression equation:y=2.624-6.855x+3.424x2:r=0.997).The benchmark dose lower limitation of daily total fluoride intake with 95% confidencewas 2.50 mg/d calculated according to this dose-response relationship,the reference dose(RfD)was 2.50 mg/d.In Wamiao Village a significant difference was also found between daily total fluoride intake in 41 subjects[(5.09±1.20)mg/d]with X-ray detectable skeletal fluorosis and in 91 subjects[(3.08±1.12)mg/d]without X-ray detectable skeletal fluorosis(t=-9.32,P＜0.01).Conclusions These findings indicate that the daily total fluoride intake has a significant dose-response relationship with the prevalence of skeletal fluorosis in an endemic fluorosis area associated with high-fluoride drinking water;and the RfD in this study was lower than that in the national standard of"Chinese hygienic standard for daily total fluoride intake(WS/T 87-1996)"(3.50 mg/d).

OBJECTIVETo investigate the synergistic effects of carnosic acid (CA) with arsenic trioxide (As₂O₃) on proliferation and apoptosis in HL-60 human myeloid leukemia cells, and the major cellular signaling pathway involved in these effects.

METHODSHL-60 cellular proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Cell cycle distribution and apoptosis were monitored by flow cytometry. The activation of casepase-9, Bcl-2-associated agonist of cell death (BAD), p-BAD, p27, phosphatase and tensin homolog deleted on chromosome ten (PTEN), Akt, p-Akt was assessed by Western blot analysis. The expression of PTEN mRNA was tested by reverse transcription polymerase chain reaction (RT-PCR) analysis.

RESULTSCA reduced HL-60 cell viability in a dose- and time-dependent manner, and induced G1 arrest and apoptosis. Moreover, CA upregulated PTEN expression, blocked the Akt signaling pathway, subsequently inhibited phosphorylation of BAD, reactivated caspase-9, and elevated levels of p27. CA also augmented these effects of As₂O₃.

CONCLUSIONCA might be a novel candidate of the combination therapy for leukemia treatment; these effects were apparently associated with the modulation of PTEN/Akt signaling pathway.

Apoptosis ; drug effects ; Arsenicals ; pharmacology ; Base Sequence ; Blotting, Western ; Cell Cycle ; drug effects ; DNA Primers ; Diterpenes, Abietane ; pharmacology ; Drug Synergism ; HL-60 Cells ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Oxides ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Plant Extracts ; pharmacology ; Proto-Oncogene Proteins c-akt ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects

OBJECTIVETo explore the role of survivin and PI3K/AKT pathway in the pathogenesis of psoriasis vulgaris (PV).

METHODSPlaque-like lesions collected from 22 patients with PV in progressive stage and 18 normal control skin specimens were examined using immunohistochemical staining, Western blotting and real-time quantitative PCR for expressions of survivin, PI3K and AKT in the keratinocytes, and their correlation was analyzed. A small interfering RNA (siRNA) was used to knock down AKT in cultured HaCaT cells, and Western blotting was used to detect the changes in the expression of survivin. RESULTS Compared with normal skin, PV lesions showed obviously up-regulated expressions of survivin, PI3K and AKT in the keratinocytes. Survivin expression was positively correlated with PI3K (r=0.4510, P=0.0351) and AKT (r=0.4423, P=0.0393) in the keratinocytes in PV lesions. In cultured HaCaT cells, siRNA-mediated knockdown of AKT caused down-regulation of survivin expression.

CONCLUSIONSurvivin and PI3K/AKT signaling pathway may participate in the occurrence and progression of PV.

This study purposed to investigate the effects of different oxygen concentrations and reactive oxygen species (ROS) on the biological characteristics of hematopoietic stem cells (HSC) and their possible mechanisms through simulating oxygen environment to which the peripheral blood HSC are subjected in peripheral blood HSCT. The proliferation ability, cell cycle, directed differentiation ability, ROS level and hematopoietic reconstitution ability of Lin(-)c-kit(+)Sca-1(+) BMHSC were detected by using in vitro amplification test, directional differentiation test, cell cycle analysis, ROS assay and transplantation of Lin(-)c-kit(+)Sca-1(+) HSC from sublethally irradiated mice respectively. The results showed that oxygen concentrations lower than normal oxygen concentration, especially in hypoxic oxygen environment, could reduce ROS generation and amplify more primitive CD34(+)AC133(+) HSC and active CD34(+) HSC, and maintain more stem cells in the G(0)/G(1) phase, which is more helpful to the growth of CFU-S and viability of mice. At the same time, BMHSC exposed to normal oxygen level or inconstant and greatly changed oxygen concentrations could produce a high level of ROS, and the above-mentioned features and functional indicators are relatively low. It is concluded that ROS levels of HSC in BMHSCT are closely related with the oxygen concentration surrounding the cells and its stability. Low oxygen concentration and antioxidant intervention are helpful to transplantation of BMHSC.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Cell Differentiation ; Cells, Cultured ; Female ; Hematopoietic Stem Cells ; cytology ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Oxygen ; administration & dosage ; pharmacology ; Reactive Oxygen Species ; metabolism

Objective To investigate the effect of growth differentiation factor 15 ( GDF15 ) downregulation on cell proliferation of human glioblastoma U 87MG cells.Methods Human glioblastoma U87MG cells with stable GDF15 downregulation was used as shGDF 15 group.U87MG cells with scramble knockdown was used as scramble group . Protein expression levels of GDF 15 were determined by Western blot analysis .Growth curve and BrdU incorporation assays were used to observe cell proliferation .Protein expression levels of ERK 1/2 and p-ERK1/2 were determined by western blot analysis .CCK-8 assays were used to observe cell proliferation .Results Compared with scramble cells, GDF15 downregulation significantly promoted cell proliferation ( P<0.05 ) , increased DNA synthesis in S phage ( P<0.01 ) , enhanced activity of ERK pathway and cell tolerance to VM-26 ( P<0.05 ) .Moreover , ERK pathway inhibitor rescued the increased cell proliferation with GDF15 downregulation.Conclusions GDF15decrease DNA synthesis in S phage and cell proliferation of human glioblastoma U 87MG cells through inhibiting ERK pathway .GDF15 is a potential target of chemotherapy sensitivity in glioblastoma clinical treatment .

OBJECTIVETo investigate the effects of carnosic acid (CA) on reversal of the multidrug resistance (MDR) of human leukemia cell line K562/A02 and its mechanism.

METHODSMTT assay was used to determine the sensitivity of K562/A02 cells to adriamycin (ADM) pre-and post-treated with CA. Flow cytometry (FCM) and laser scanning confocal microscopy (LSCM) were used to measure intracellular fluorescence intensity and concentration of ADM respectively. The expression level of mdr1 was detected by semi-quantitative RT-PCR. P-glycoprotein (P-gp) expression was detected by FCM and Western blot.

RESULTSCA decreased IC(50) of ADM in K562/A02 cells from 16.31 µg/mL to 1.35 µg/mL, being a 12.08-fold decrease. The intracellular ADM fluorescence intensity of K562/A02 was increased from 17.05 to 60.53 after treated with CA (P < 0.01). In living K562/A02 cells, after treated with CA, the diffuse distribution of intracellular ADM was recovered in both nuclear and cytoplasm, and the concentration of intracellular ADM increased from 4.93µg/mL to 15.43µg/mL. RT-PCR assay showed that CA inhibited the expressions of mdr1 mRNA in K562/A02 cells (P < 0.01). Mean fluorescence intensity of P-gp detected by FCM in CA-treated K562/A02 was decreased to 22.80 as compared with that in untreated K562/A02 cells (44.40, P < 0.05).

CONCLUSIONCA can reverse the MDR of K562/A02 cells in vitro. The mechanism may be associated with down-regulation of mdr1 and inhibition of P-gp function.

ATP Binding Cassette Transporter, Sub-Family B ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Doxorubicin ; pharmacology ; Drug Resistance, Multiple ; drug effects ; Drug Resistance, Neoplasm ; drug effects ; Humans ; K562 Cells

The therapeutic effect of sustained intravitreal injectable voriconazole microspheres (VCZ-MS) on an experimental endophthalmitis of Aspergillus fumigatus was investigated. VCZ-MS was prepared successfully and its physico-chemical property was also evaluated. Right eyes of albino rabbits were infected with an intravitreal injection of 1 000 CFU x mL(-1) of susceptible Aspergillus fumigatus. All fungal endophthalmitis models were randomly divided into five groups 48 hours later: Group A is control group with no treatment; in group B, vitrectomy was performed combined with intravitreal 3 times injections of 100 microg x 0.1 mL(-1) voriconazole every other day. In group C, D and E, vitrectomy was performed combined with intravitreal injection of 0.5 mg, 1.0 mg and 1.5 mg VCZ-MS respectively. The treatment effect was assessed by slit lamp and indirect ophthalmoscope funduscopy examination, using clinical grading system of inflammation in the anterior chamber and the vitreous opacity. The optical microscopy revealed that microspheres obtained from the experiment design were opaque, discrete and spherical particles with smooth surfaces. The drug content and encapsulation efficiency of microspheres were 29.94% and 73.5%, respectively. Endophthalmitis occurred in all eyes of group A, and rapidly developed to panophthalmitis. The inflammation grade of group B, C, D or E was lower than that of group A (P < 0.05). The grade of vitreous opacity in group C, D, E is lower than group B (P < 0.05). Two eyes in group C developed to panophthalmitis. But in group D and E, all eyes whose inflammation was controlled had no recurrence with vitreous clear. Histopathological examination showed normal structures in the cured eyes, while most uncured eyes were atrophic and with eyeball destroyed. So, it can be safely concluded that the curative effect of intravitreal VCZ-MS is significantly better than that of routine intraocular injection of voriconazole. The optimal dose is the one containing 1.0 mg voriconazole.
Animals ; Antifungal Agents ; administration & dosage ; therapeutic use ; Aspergillosis ; drug therapy ; pathology ; Aspergillus fumigatus ; Delayed-Action Preparations ; Endophthalmitis ; drug therapy ; microbiology ; pathology ; Eye ; microbiology ; pathology ; Female ; Intravitreal Injections ; Lactic Acid ; Male ; Microspheres ; Polyglycolic Acid ; Pyrimidines ; administration & dosage ; therapeutic use ; Rabbits ; Random Allocation ; Triazoles ; administration & dosage ; therapeutic use ; Voriconazole

OBJECTIVE: To develop the visual uroflow scale (VUS), analyze the relationship of VUS score and index of free uroflowmetry, assess urination function preliminarily and improve the work efficiency in the clinic. METHODS: Male lower urinary tract symptoms (LUTS) patients, who attended the Department of Urology in Peking University People's Hospital from March 2016 to March 2017, were assessed for their urination function according to the Visual Uroflow Scale without help from clinicians before undertaking a free uroflowmetry test. And afterwards, a free uroflowmetry was undertaken, and variables including maximal flow rate (Qmax), the average flow rate (Qave) and voiding volume (VV) was obtained. During the study, 124 cases were collected and 53 cases met the inclusion and exclusion criteria and were included in the study cohort. The Spearman correlation analysis was used for analyzing the correlation of VUS scores with free uroflowmetry variables and age. The validity of VUS was evaluated. RESULTS: Most of the patients could choose the very figure matched with self-condition by first instinct without any help from the clinician. The data were analyzed by Spearman correlation analysis. In the present study, voiding time was positively correlated with the VUS score (correlation coefficient, 0.62, P < 0.05). In the present cohort, the patients chose the third and fourth figures to take longer time to urinate, implying worse LUTS situation. Flow time and VUS scores were positively correlated (correlation coefficient, 0.61, P < 0.05). The patients with higher VUS scores would spend more time on urinate, no matter how long urinary hesitation was. Both Qmax and Qave were negatively correlated with the VUS score (correlation coefficient －0.54, －0.62, P < 0.05). The study illustrated that the VUS score suggested that the Qmax basically and further reflected the urination function. And its relationship to age revealed the decreased urination function of aging male, which had reached a consensus. CONCLUSION Development of VUS has helped the clinician assess the urination function preliminarily at the first time. Patients are assessed for a VUS score before getting surgery or receiving the drug for treatment, and can be re-assessed after. The VUS score can provide an objective quantitative basis to evaluate the treatment efficacy. In addition, considering that it is convenient, timesaving and easy to understand, the VUS is available for follow-up.
Cohort Studies ; Humans ; Lower Urinary Tract Symptoms ; Male ; Urination ; Urodynamics

Objective To evaluate the serum levels of GP73 for diagnosing autoimmune liver disease.Methods Three populations were included:80 patients with autoimmune liver disease; 80 patients with chronic hepatitis B,and 80 apparently healthy controls.Results The serum levels of GP73 in patients with autoimmune liver disease (112.3 ± 72.55 ng/ml) were significantly higher than those of patients with chronic hepatitis B (86.44 ± 60.69 ng/ml),and healthy controls (35.84 ± 11.8 ng/ml).However,there were no significant differences was observed in group of subjects with autoimmune hepatitis (107.4 ± 90.6 ng/ml),primary biliary cirrhosis ((89.0 ± 45.38 ng/ml),and primary sclerosing cholangitis (113.3 ± 50.87 ng/ml).Chosen the healthy control as "control group",the patients with chronic hepatitis B as "patients group ",the sensitivity and specificity of GP73 for diagnosing autoimmune liver disease were 75.0％ and 97.53％,respectively.The area of ROC analysis was 0.93(95％ CI;0.89-0.97).Conclusion The serum GP73 levels was significantly increased in patients with autoimmune liver disease,compared with those of healthy controls.For patients with higher levels of serum GP73,diagnosis of autoimmune liver disease should also be considered,especially for those patients with overlap-syndrome,except for chronic virus hepatitis and hepatocellular carcinoma.