Objective:To investigate the effects of down-regulation of FABP5 (fatty acid binding protein 5) on radiation damage of skin cells, and explore underlying mechanism.Methods:A lentiviral vector with down-regulated FABP5 was constructed to infect human immortalized keratinocytes (HaCaT) cells, and the transfection efficiency was examined. The HaCaT cells were divided into blank control group, FABP5 down-regulation group (FABP5), radiation group (IR), and FABP5 down-regulation combined with radiation group (FABP5+ IR). After 6 MV X-ray radiation, cell proliferation viability was measured by CCK-8 assay, cell migration was detected by scratch assay, apoptosis was analyzed by flow cytometry, radiosensitivity was evaluated by cloning formation assay, and the cellular protein expressions of PARP1, γ-H2AX, AKT and p-AKT were detected by Western blot.Results:FABP5 was successfully knocked-down in both RNA level ( t=25.14, P<0.05) and protein level ( t=20.06, P<0.05). The down-regulation of FABP5 decreased the abilities of cells proliferation ( t=3.55, 5.88, 3.18, P<0.05) and migration ( t=15.44, P<0.05), but increased cell resistance to irradiation with a radiosensitization ratio of 0.782. The apoptosis rate of FABP5+ IR group was significantly lower than IR group (22.05±6.71)% vs. (9.82±1.45)%, t=3.08, P<0.05. The protein levels of PARP1 and γ-H2AX in FABP5+ IR group were also lower than those in the IR group 0.04±0.04, 0.11±0.06, 0.26±0.11, 0.22±0.07, 0.21±0.10, 0.52±0.22, 0.57±0.06, 0.43±0.02( t=2.83, 3.07, 4.50, 5.33, P<0.05), while the protein level of p-Akt in FABP5+ IR group was higher than that in IR group ( t=-16.24—3.02, P<0.05). Conclusions:Down-regulation of FABP5 inhibited cell proliferation and migration, increased radioresistance, and reduced radiation-induced apoptosis and DNA damage of skin cells probably through PI3K/AKT signaling pathway.

Objective To investigate the effects of endothelial nitric oxide synthase (eNOS) specific inhibitor L-iminoethyl ornithine hydrochloride (L-NIO) for all the fluorine in vitro cultivation of SH-SY5Y cells apoptosis,eNOS mRNA and protein expression,and effect of nitric oxide (NO) content and nitric oxide synthase (NOS) activity change.Methods The SH-SY5Y cells cultured in vitro were divided into control group,low fluoride group,high fluoride group,L-NIO group,low fluoride with L-NIO group,high fluoride with L-NIO group,n =3.The control group added equal volume culture liquid with the experimental group,the concentration of sodium fluoride (NaF) in the low fluoride group and the high fluoride group were 0.2 and 2.0 mmol/L,respectively.The L-NIO group added 3 μmol/L L-NIO,the low fluoride with L-NIO group and the high fluoride with L-NIO group were added to 0.2 mmol/L NaF and 3 μmol/L L-NIO,2.0 mmol/L NaF and 3 μmol/L L-NIO,respectively.The incubation time was 48 h.The expression level of eNOS protein in cells was detected by Western blotting.The expression level of eNOS mRNA in cells was detected by Real-time fluorescence quantitative PCR method.The apoptosis of cells was detected by flow cytometry,NO content and NOS activity in cell culture liquid were detected by nitrate reductase and colorimetric assay.Results Compared with the control group (1.000 ± 0.026),the expression of eNOS protein in the low and high fluoride groups (1.108 ± 0.071,1.349 ± 0.057) increased (P ＜ 0.05),and the L-NIO group (0.755 ± 0.148) decreased (P ＜ 0.05);compared with the low fluoride group,the high fluoride group increased and the low fluoride with L-NIO group (0.802 ± 0.115) decreased (P ＜ 0.05);compared with the high fluoride group,high fluoride with L-NIO group (0.988 ± 0.135) decreased (P ＜ 0.05).Compared with the control group (1.000 ±0.018),the expression of eNOS mRNA in the low and high fluoride groups (1.809 ± 0.099,2.416 ± 0.295) increased (P ＜ 0.05),the L-NIO group (0.609-± 0.077) decreased (P ＜ 0.05);compared with the low fluoride group,the high fluoride group was elevated(P ＜ 0.05),the low fluoride with L-NIO group (1.040 ± 0.034) decreased (P ＜ 0.05);compared with the high fluoride group,high fluoride with L-NIO group (1.233 ± 0.152) decreased (P ＜ 0.05).Compared with the control group [(1.66 ± 0.07)％],the cell apoptosis rate in the low and high fluoride groups [(8.81 ± 0.27)％,(17.60 ± 0.20)％] increased,L-NIO group [(1.03 ± 0.04)％] decreased (P ＜ 0.05);compared with the low fluoride group,the high fluoride group increased and the low fluoride with L-NIO group [(6.03 ± 0.10)％] decreased (P ＜ 0.05);compared with the high fluoride group,the high fluoride with L-NIO [(12.12 ± 0.08)％] decreased (P ＜ 0.05).Compared with the control group [(2.773 ± 0.145)μmol/L],the content of NO in the cell culture medium in the low and high fluoride groups [(8.251 ± 1.047),(14.287 ± 1.062) μmol/L] increased (P＜ 0.05),and the L-NIO group [(1.648 ± 0.155) μmol/L] decreased (P ＜ 0.05);compared with the low fluoride group,the high fluoride group was elevated (P ＜ 0.05),the low fluoride with L-NIO group [(4.622 ± 0.252) μmol/L] decreased (P ＜ 0.05);compared with the the high fluoride group,high fluoride with L-NIO group [(7.899 ± 0.385) μmol/L] decreased (P ＜ 0.05).Compared with the control group [(0.507 ± 0.041) U/ml],the activity of NOS in the cell culture medium in the low and high fluoride groups [(0.772 ± 0.032),(2.258 ± 0.062) U/ml] increased (P ＜ 0.05),and the L-NIO group [(0.346 ±0.015) U/ml] decreased (P ＜ 0.05);compared with the low fluoride group,the high fluoride group was elevated (P ＜0.05),the low fluoride with L-NIO group [(0.637 ± 0.026) U/ml] decreased (P ＜ 0.05);compared with the the high fluoride group,high fluoride with L-NIO group [(1.161 ± 0.071) U/ml] decreased (P ＜ 0.05).Conclusions Excessive fluoride can lead to overexpression of eNOS protein and mRNA in SH-SY5Y cells,increase of apoptosis rate,increase the content of NO in cell culture and enhance the activity of NOS.After co-culture of L-NIO and fluorine,it can antagonize the damage of fluorine to SH-SY5Y cells and play a certain neuroprotective effect.

Objective To evaluate the clinical value of PET-CT and DWI for the detection of bone marrow infiltration of lymphoma .Methods The bone marrow samples of 93 untreated patients with pathologically diagnosed lymphoma were retrospectively analyzed . 61 patients underwent PET-CT examination, and other 32 underwent DWI examination .With bone marrow biopsy results as “gold standard”, the rates and sites of bone marrow infiltration of various lymphoma subtypes were analyzed , and the detection rates of the two imaging techniques were compared according to different lymphoma subtypes . Results 39 patients were diagnosed as bone marrow infiltration based on pathological examination of bone marrow biopsies from routine sampling sites and bone marrow pathological examination of biopsies guided by PET-CT and DWI.The sensitivity, specificity, accuracy, positive and negative predictive values of PET-CT for lymphoma bone marrow infiltration were 80.8%, 88.6%, 85.3%, 84.0%and 86.1%, respectively; for DWI examination, these rates were 84.6%, 89.5%, 87.5%, 84.6%and 89.5%, respectively.The detection rates of the two imaging techniques for aggressive lymphoma were 37.5%(18/48) and 38.1%(8/21), respectively, which were slightly higher than those for the indolent lymphoma [23.1%(3/13) and 27.3%(31/1)], although the differences were not statistically significant (P=0.521, P=0.660).For both aggressive lymphoma and indolent lymphoma , the detection rates of DWI were numerically slightly higher than those of PET-CT(P=0.963, P=1.000).Conclusions PET-CT and DWI have important and similar diagnostic value for bone marrow infiltration of lymphoma .None of PET-CT and DWI can replace bone marrow biopsy (BMB).However, image-guided bone marrow biopsies can improve the detection rate of bone marrow infiltration of lymphoma .

Objective To evaluate the clinical value of PET-CT and DWI for the detection of bone marrow infiltration of lymphoma .Methods The bone marrow samples of 93 untreated patients with pathologically diagnosed lymphoma were retrospectively analyzed . 61 patients underwent PET-CT examination, and other 32 underwent DWI examination .With bone marrow biopsy results as “gold standard”, the rates and sites of bone marrow infiltration of various lymphoma subtypes were analyzed , and the detection rates of the two imaging techniques were compared according to different lymphoma subtypes . Results 39 patients were diagnosed as bone marrow infiltration based on pathological examination of bone marrow biopsies from routine sampling sites and bone marrow pathological examination of biopsies guided by PET-CT and DWI.The sensitivity, specificity, accuracy, positive and negative predictive values of PET-CT for lymphoma bone marrow infiltration were 80.8%, 88.6%, 85.3%, 84.0%and 86.1%, respectively; for DWI examination, these rates were 84.6%, 89.5%, 87.5%, 84.6%and 89.5%, respectively.The detection rates of the two imaging techniques for aggressive lymphoma were 37.5%(18/48) and 38.1%(8/21), respectively, which were slightly higher than those for the indolent lymphoma [23.1%(3/13) and 27.3%(31/1)], although the differences were not statistically significant (P=0.521, P=0.660).For both aggressive lymphoma and indolent lymphoma , the detection rates of DWI were numerically slightly higher than those of PET-CT(P=0.963, P=1.000).Conclusions PET-CT and DWI have important and similar diagnostic value for bone marrow infiltration of lymphoma .None of PET-CT and DWI can replace bone marrow biopsy (BMB).However, image-guided bone marrow biopsies can improve the detection rate of bone marrow infiltration of lymphoma .

OBJECTIVETo establish a method for determining cyanamide in workplace air by high-performance liquid chromatography (HPLC).

METHODSAir samples were collected from the workplace using the shock absorption tube containing water solution at a rate of 2.8∼3.0 ml/min for 60 min; dansyl chloride was used as a derivatization reagent to conduct pre-column derivatization, and the procedure was as follows: acetone solution (2.5 ml), mixed solution (1.0 ml) containing 0.016 mol/L Na2CO3 and 0.184 mol/L NaHCO3, and 10 mg/ml acetone solution of dansyl chloride (0.5 ml) were added into the samples, and reaction proceeded in a water bath (50 °C) for 1 h. HPLC was performed on an ODS C18 column (250 mm × 4.6 mm, 5 üm) with a mobile phase of acetonitrile-phosphate buffer (35:65) at a flow rate of 1.0 ml/min and a column temperature of 25°C; a fluorescence detector was used at an excitation wavelength of 360 nm and an emission wavelength of 495 nm.

RESULTSThe minimum detectable concentration of cyanamide was 0.05 üg/ml; a good linear relationship was noted when the concentration of cyanamide was 0.2∼100.0 üg/ml; the intraday relative standard deviation (RSD) was 0.28%∼1.18%, and the interday RSD was 0.22∼2.16%; the recovery rate was 95.7%∼103.0%, and the sampling efficiency was 95.8%∼96.9%. Water solution of cyanamide (pH<6.5) could be stable in the dark at room temperature for 7 d.

CONCLUSIONThis method is stable, reliable, easy to operate, and highly sensitive and suitable for determination of cyanamide in workplace air.

To study the in vitro antibacterial activity and biocompatibility of 317L stainless steel containing 4.5% copper alloy (317L-Cu), we produced 317L-Cu stainless steel with epsilon-Cu phase. The cell proliferation of osteoblasts on material surface was detected in vitro. Escherichia coli was cultured with 317L-Cu for evaluating the antibacterial activity. We found that the 317L-Cu could effectively kill the Escherichia coli on material surface. The cell proliferation of osteoblasts on material surface was not different significantly, compared with titanium material. Our study clearly demonstrated that the 317L-Cu alloys had a significant antimicrobial activity and was biocompatible in vitro, so it would be suitable to be used as a new medical material with antibacterial activity.
Anti-Bacterial Agents ; pharmacology ; Biocompatible Materials ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Copper ; chemistry ; pharmacology ; Escherichia coli ; drug effects ; Humans ; Osteoblasts ; cytology ; Prosthesis-Related Infections ; prevention & control ; Stainless Steel ; chemistry ; pharmacology

L-Arabinose isomerase (L-AI) is an intracellular enzyme that catalyzes the reversible isomerization of D-galactose and D-tagatose. Given the widespread use of D-tagatose in the food industry, food-grade microorganisms and the derivation of L-AI for the production of D-tagatose is gaining increased attention. In the current study, food-grade strains from different foods that can convert D-galactose to D-tagatose were screened. According to physiological, biochemical, and 16S rDNA gene analyses, the selected strain was found to share 99% identity with Pediococcus pentosaceus, and was named as Pediococcus pentosaceus PC-5. The araA gene encoding L-AI from Pediococcus pentosaceus PC-5 was cloned and overexpressed in E. coli BL21. The yield of D-tagatose using D-galactose as the substrate catalyzed by the crude enzyme in the presence of Mn2+ was found to be 33% at 40 degrees C.
Aldose-Ketose Isomerases ; biosynthesis ; genetics ; Biotransformation ; Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Galactose ; metabolism ; Genetic Vectors ; genetics ; Hexoses ; metabolism ; Pediococcus ; classification ; genetics ; isolation & purification ; Recombinant Proteins ; biosynthesis ; genetics

OBJECTIVE: To over-express cyclin-dependent kinase 2-associated protein 1 (CDK2-AP1) gene, and investigate its effect on the proliferation and cell cycle regulation in breast cancer cell line MCF-7. METHODS: CDK2-AP1 gene coding region was cloned into lentivirus vector. Lentivirus particles were infected into MCF-7 cells to upregulate the expression of CDK2-AP1 gene. The expression level of CDK2-AP1 was detected at both mRNA and protein levels by real-time PCR and Western blot. MTT assay, colony formatting assay, and flow cytometry were performed to detect the change of proliferation and cell cycle in MCF-7 cells. We examined the expression of cell cycle associated genes (CDK2, CDK4, P16Ink4A, and P21Cip1/Waf1) followed by CDK2-AP1 over-expression by Western blot. RESULTS: CDK2-AP1 gene was up-regulated significantly at both mRNA (6.94 folds) and protein level. MTT based growth curve, colony formatting assay and flow cytometry showed that CDK2-AP1 over-expression lentivirus inhibited the proliferation of MCF-7 cells with statistical difference (P<0.05). In addition, with CDK2-AP1 over-expression, MCF-7 cells were arrested in G1 phase accompanied by apoptosis. Western blot showed that the expression level of P21Cip1/Waf1 and P16 Ink4A was upregulated, while the expression level of CDK2 and CDK4, members of the CDK family, was downregulated. CONCLUSION CDK2-AP1 gene plays a cancer suppressor role in breast cancer. Its function includes inhibiting the proliferation of MCF-7 cells and arresting the cell cycle in G1 phase.
Breast Neoplasms ; Cell Cycle ; Cell Division ; Cell Proliferation ; Cyclin-Dependent Kinases ; Down-Regulation ; Humans ; MCF-7 Cells ; Protein Kinases ; genetics ; metabolism ; Tumor Suppressor Proteins ; genetics ; metabolism

Objective To study the expression of heat shock 27 000 associated protein 1 ( HSPBAP1, GenBank: AK096705) in the brain tissues of patients with drug-refractory epilepsy and discuss its function in the pathogenesis. Methods Fluorescent quantitative polymerase chain reaction ( FQ-PCR) and immunohistochemistry were used to test the expression of HSPBAP1 in the surgically removed brain tissues of patients with drug-refractory epilepsy from the brain bank of our department ( n = 36) , and the results were compared with that of normal controls (n = 8 ). Results The relative expression of HSPBAP1 mRNA in the brains of patients with drug-refractory epilepsy was more than 34. 11 times that of controls, and HSPBAP1 protein expression was significantly increased in temporal lobe cortex (0. 0507?0. 0003, P

Objective To investigate expression of extracelluar signal regulated protein kinase(ERK)and phosphorylation ERK(p-ERK)in the temporal lobe from patients with DR-TLE so as to explore the possible roles of ERK in the pathogenesis of DR-TLE.Methods Expression of ERK was detected with Western blot and immunohistochemistry in 32 patients with DR-TLE(24 temporal lobe,8 hippocampi),as compared with 12 controls(9 temporal lobe,3 hippocampi).Results ERK and p-ERK expression in DR-TLE was significantly higher(0.2266?0.0613,0.2097?0.0183 and 0.1924?0.0054,respectively)than those of controls(0.1840?0.0023,0.1974?0.0056 and 0.1825?0.0063,respectively,all P