OBJECTIVESTo investigate the changes of nuclear factor (NF-)κBp65 and inflammatory chemotactic factors including monocyte chemoattractant protein 1 (MCP-1/CCL-2), macrophage inflammatory protein 1α (MIP-1α/CCL-3), glial fibrillary acidic protein (GFAP) in brains of the patients with Alzheimer's disease (AD) and reveal the correlation of these factors.

METHODSTen patients with AD and 8 age-matched control subjects were selected in the study. Immunohistochemistry was performed to determine the protein expression of NF-κBp65, MCP-1, MIP-1α and GFAP. Double-immunohistochemistry was used to detect the expression of GFAP and β-amyloid peptide 1-42 (Aβ(1-42)) in the hippocampus, temporal and frontal cortices.

RESULTSAs compared to age-matched controls (the numbers of the positively stained neuronal cells: 0.31 ± 0.20, 0.25 ± 0.20 and 0.25 ± 0.20, respectively), the immunoreactivities of NF-κBp65 in the hippocampus and the temporal and frontal cortices (numbers of the positively stained cells: 3.6 ± 1.5, 2.2 ± 1.2 and 2.2 ± 1.2, respectively) were significantly increased in AD brains. The levels of MCP-1 and MIP-1α in the hippocampus, and the temporal and frontal cortices (numbers of the positively stained neuronal cells: 8.0 ± 1.3, 8.8 ± 1.0, 9.3 ± 1.4, respectively;and 8.1 ± 1.5, 12.5 ± 1.1, 6.4 ± 1.1, respectively) with AD were significantly higher than those of controls (the numbers of the positive neuronal cells: 4.5 ± 0.9, 4.5 ± 0.6, 4.0 ± 1.8, respectively; and 5.0 ± 1.9, 6.3 ± 2.2, 3.8 ± 1.5, respectively). An increased number of glial cells stained with GFAP were observed to extensively distribute around the senile plaques in AD brains. There were significant correlations between NF-κBp65 and these inflammatory chemotactic factors in AD brains.

CONCLUSIONCorrelative expressions of NF and inflammatory chemotactic factors were found in the brains of AD patients, through a mechanism that may involve the inflammatory response induced by Aβ in the processing of AD.

Aged ; Aged, 80 and over ; Alzheimer Disease ; metabolism ; pathology ; Brain ; metabolism ; pathology ; Chemokine CCL2 ; metabolism ; Chemokine CCL3 ; metabolism ; Female ; Frontal Lobe ; metabolism ; pathology ; Glial Fibrillary Acidic Protein ; metabolism ; Hippocampus ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Male ; Neuroglia ; metabolism ; pathology ; Plaque, Amyloid ; metabolism ; pathology ; Temporal Lobe ; metabolism ; pathology ; Transcription Factor RelA ; metabolism

BackgroundRetinal pigment epithelial(RPE) cells can secrete platelet-derived growth factor (PDGF) and PDGF receptor(PDGFR).Studies have shown that PDGF plays a key role in the formation of proliferative vitreous retinopathy(PVR). ObjectiveThis study was to investigate the proliferation and apoptosis changes of RPE after blockage of the PDGFR-α expression by antisense oligonucleotide ( ASODN ) in vitro. Methods Human RPE cells strain was cultured in low glucose DMEM with 10％ fetal bovine serum.Logarithmic phase cells were collected and incubated in 96-well plate at the density of 5 × 105 cells/hole.PDGFR-α ASODN was transfected into RPE cells at different concentrations for 48 hours.The cells of the blank control group were regularly cultured without any transfection.The changes of PDGFR-α expression were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the proliferation of RPE was detected by MTT as the A490 value.Hoechst 33258 fluorescence staining was used to determine the apoptosis of RPE.Flow cytometry method (FCM) was applied to detect the change of cell cycle and apoptosis rate of RPE cells. ResultsThe A490 values of RPE cells were 1.45±0.12,1.07±0.06,0.65±0.05 in blank control group,1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group with the significant difference(P=0.00 ),and that of 1.0 μmol/L Lipo-ASODN group and 2.0 μ mol/L Lipo-ASODN group were significantly lower than the blank control group ( P =0.00,0.00).Hoechst 33258 staining showed that the apoptosis cells were obviously more in Lipo-ASODN group compared with blank control group.PDGFR-α ASODN transfection induced an increase of percentage of RPE cells in G0/G1 phase( F =206.70,P =0.00),and the apoptosis rates in 1.0 μmol/L Lipo-ASODN group and 2.0 μmol/L Lipo-ASODN group were significantly enhanced in comparison with blank control group ( 37.8 ± 1.3 vs 10.5 ± 0.1,61.2 ± 1.9 vs 10.5 ± 0.1 ) ( F =1808.90,P =0.00 ).Expression intensity of PDGFR-α mRNA in RPE cells in Lipo-ASODN groups was lower. ConclusionsBlocking the PDGFR-α expression with ASODN technology can suppress proliferation and induce apoptosis of RPE cells.Intensity of PDGFR-α mRNA expression in RPE cells is ASODN dose-dependent.ASODN targeted to PDGFR-α offers an experimental basis of the gene therapy for PVR.

The fermentative H2-producing strain Clostridium sp. H-61 was isolated from anaerobic sludge,was used as an original strain which was induced by NTG and UV for increasing and the hydrogen production ability. One of the highest efficient H2-producing mutants was named as HCM-23 with its stable hydrogen production ability. which was measured in the batch culture experiments. With the condition of 10 g/L glucose,its cumulative hydrogen yield and hydrogen production rate was 3024 mL/L and 33.19 mmol H2/g DW?h,69.89% and 68.14% higher than that of the original strain,respectively. The terminal liquid product compositions showed that the mutant HCM-23 fermentation was ethanol type,while the original strain H-61 fermentation was butyric acid type. Varieties of parameters of hydrogen production fermentation studied,including time,carbon source,nitrogen source,glucose concentration,glucose utilization,initial pH and incubation temperature had been studied,indicated the optimum condition of hydrogen production for the mutantHCM-23 as initial pH 6.5,temperature 36 ℃,and the favorite substrate was sucrose. The hydrogen production characters of the mutant and the original strain were different,such as,the growth lag phase and the utilization of inorganic nitrogen source,etc. This work shows a good application potential of NTG-UV combined mutation in the biohydrogen production. And the hydrogen production mechanism and metabolic pathway should be explored furthermore.

Objective To study the effect of β-amyloid peptide (Aβ1-42) on expressions of synaptophysin (Syn), dynamin Ⅰ (Dyn Ⅰ) and adaptor protein 180 (AP180) in human neuroblastoma SH-SY5Y cells. Methods Human SH-SY5Y neuroblastoma cells were treated with 0.01, 0.1, 1, 2 and 5 μmol/L Aβ1-42, and control cells were given no treatment. MTT [3-(4,5-dimethylthiazol- 2- yl) - 2, 5-diphenyltetrazolium bromide]reduction of the cells was measured by spectrophotometry. The protein levels of Syn, Dyn Ⅰ and AP180 in SH-SY5Y cells treated with 0.5 and 1 μmol/L Aβ1-42 were surveyed by Western blotting. The mRNA levels of Syn, Dyn Ⅰ and AP180 were detected by Real-time PCR in SH-SY5Y cells treated with 1 μmol/L Aβ1-42. Results SH-SY5Y cells showed obviously decreased reduction rates of MTT after exposure to Aβ1-42(0.1 μmol/L) as compared with the controls (P＜0.05), and dose-dependent negative correlation was noted in these SH-SY5Y cells. The protein level of Dyn Ⅰ in cells treated with 0.5 μmol/L Aβ1-42 was significantly decreased as compared with that in controls (P＜0.05). The protein and mRNA levels of Syn and Dyn Ⅰ in cells treated with 1 μ mol/L Aβ1-42 Were obviously decreased as compared with those in controls (P＜0.05), but the levels of AP180 were not changed. Conclusion Aβ1-42 reduces the levels of Syn and Dyn Ⅰ in SH-SY5Y cells, which might be a mechanism in eonnection with cognitive deficit of AD.

Abstract: Objective　To explore the spatial epidemiological characteristics of severe cases hand, foot and mouth disease (HFMD) in Guangxi, China, from 2014 to 2018, and to provide a basis for identifying the high-risk regions as well as the prevention and control of severe cases of HFMD in Guangxi. Methods　Spatial-temporal scanning analysis, global and local spatial autocorrelation analysis were used to analyze the spatial clustering of HFMD. The trend surface analysis was used to evaluate the spatial distribution trend of HFMD. Results　From 2014 to 2018, the incidence and severe case fatality rates of HFMD were 3.89/100 000 and 4.23%, respectively. Monte Carlo scanning analysis showed that the first cluster region was Cenxi City, the second cluster was mainly concentrated in northwest of Guangxi, and the aggregation time was mainly concentrated in April to May and August to October. The global spatial autocorrelation analysis showed that the severe HFMD was significant clustering distribution, and the Moran's I coefficients of the sever cases, severe morbidity and severe case fatality rate were 0.088, 0.118, 0.197, respectively (P<0.05). Local spatial autocorrelation analysis showed that hotspots of severe HFMD cases were concentrated in the southern Guangxi, mainly in Lingshan County. Anselin local Moran's I clustering and outlier analysis indicated that 5 high-high (H-H) clustering regions for fatality were Lingshan, Pubei, Zhongshan, Zhaoping and Pinggui County. There were 6 high-high (H-H) clustering regions for severe incidence rate, namely Lingshan, Qinnan, Lingyun, Youjiang, Bama Yao Autonomous and Pinggui County, and 1 high-low (H-L) clustering region, Cenxi County. The trend surface analysis showed that the overall number of severe cases of death decreased from east or west to the middle, and increased from north to middle, and then decreased to south. Conclusions　Severe HFMD cases in Guangxi have obvious spatial-temporal clustering, and the hop spots are mainly concentrated in southern Guangxi. The prevention and control of HFMD in areas with high incidence of severe cases should be strengthened to reduce the burden of HFMD cases.

Objective To evaluate the effect of PTH gene polymorphisms on bone mineral density (BMD) at multiple skeletal sites in normal females.Methods PTH gene phenotype was determined by PCR-RFLP of restriction enzyme Bst BⅠin 596 females aged (46.3?13.7) years (20-80 years),and PCR products with or without enzymolytic site were considered as genotype B or genotype b respectively.BMDs of the anteropesterior spine (AP) and supine lateral spine (Lat) of lumbar vertebrae (L_1-L_4),femoral neck (FN),total hip (T-hip), Ward's triangle (Ward),Trochanter (Troch),forearm [radius+ulna ultradistal (RUUD) and total area of radius + ulna (RUT) ] were measured by DEXA (QDR4500A).Results (1) Hardy-Weinberg equilibrium was evident for PTH polymorphisms.The frequencies of genotype were BB 0.784,Bb 0.208,bb 0.008 and frequencies of alleles B,b were 0.888 and 0.112 respectively in 596 normal females.Frequencies of BB,Bb,bb genotypes were 0.781,0.210,and 0.009 respectively in 347 postmenopausal women and their frequencies of alleles B,b were 0.886,0.114.No significant difference was found between post- and premenopausal women in genotype frequen- cy.(2) Comparing their BMDs of AP,Lat,FN,T-hip,Ward,Troch,RUUD and RUT,there was no significant difference between BB and Bb genotypes of pre- and postmenopansal women groups.(3) Logistic regression analysis failed to show any statistical difference between normal and osteoporosis women with regard to PTH phenotype.Conclusion PTH gene polymorphism has little effect on BMD in normal females.

OBJECTIVETo investigate the effect of tanshinone IIA (TanIIA) on the expression of tissue factor (TF) and matrix metalloproteinase-12 (MMP-12) in RAW264.7 cells and explore the possible mechanism.

METHODSRAW 264.7 cells were incubated with ox-LDL in the presence or absence of different concentrations of tanshinone IIA. At the end of the incubation, the cell proliferation was assessed by MTT assay, and superoxide dismutase (SOD) activity and malondialdehyde (MDA) and TF concentrations in the supernatant were detected by xanthine oxidase method, thiobarbituric acid method and ELISA, respectively. Western blotting was employed to determine MMP-12 expression in the cells.

RESULTSThe cell proliferation was dose-dependently inhibited by TanIIA. SOD activity in the supernatant was increased significantly, while the MDA and TF concentration and MMP-12 expression in cells decreased after treatment of the cells with different concentrations of TanIIA.

CONCLUSIONTanIIA inhibits the cell proliferation and TF and MMP-12 expressions in RAW264.7 cells stimulated by ox-LDL, and these effects may be related with the anti-oxidation property of TanIIA.

Animals ; Cell Line ; Diterpenes, Abietane ; pharmacology ; Lipoproteins, LDL ; adverse effects ; Macrophages ; drug effects ; secretion ; Malondialdehyde ; metabolism ; Matrix Metalloproteinase 12 ; metabolism ; Mice ; Thromboplastin ; metabolism

Absorption ; Adult ; Bone Screws ; Bone Wires ; Calcaneus ; diagnostic imaging ; injuries ; physiopathology ; surgery ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; methods ; Fractures, Bone ; diagnostic imaging ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Recovery of Function ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo identify whether Radix Bupleuri (Bupleurum chinense) was fumed with sulfur.

METHODA static headspace GC-MS method was used to detect sulfur in the fumatory Radix Bupleuri, the authentic samples free of sulfur was detected as reference.

RESULTSulfur was detected in six samples from nine samples collected in different locations.

CONCLUSIONThe method can be used to detect sulfur rapidly in the fumatory Radix Bupleuri with sulfur.

Bupleurum ; chemistry ; Drug Contamination ; Gas Chromatography-Mass Spectrometry ; methods ; Hot Temperature ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Sulfur ; analysis ; Technology, Pharmaceutical

Objective To observe the effects of genistein and 17?-estradiol on microstructure of cancellous bone in ovariectomized (OVX) rats.Methods Ninty 7-month-old SD rats were randomly divided into baseline group,ovariectomized (OVX),sham-operated (SHAM),17?-estradiol treated (10?g?kg~(-1).day~(-1),EST) and genistein treated (5 mg?kg~(-1)?day~(-1),GEN) groups,and were killed at the beginning of the experiment,the 3rd and 15th week after operation.MicroCT scanning was performed on the left tibia in vitro.The regions involving 0.5 mm slice thickness and 1.6 mm distal to the tibial growth plate were selected as the regions of interest.Results At the 3rd week after operation,the tissue bone mineral density (tBMD) and trabecular thickness (sTh.Th) in group GEN were significantly higher than those in OVX and EST groups (all P