AIM: To investigate whether melatonin improve the learning and memory dysfunction in the amnesic rats induced by amyloid β-peptide 25-35 (Aβ25-35) via cholinergic nervous system or not. METHODS: The amnesic model in adult rats was induced by injection of Aβ25-35 into hippocampus; Morris water maze was used to determine the effects of Aβ25-35 and melatonin on the learning and memory. The activity of the choline acetyltransferase and acetylcholinesterase were determined by immunohistochemistry and spectrophotometry respectively. RESULTS: Injection of Aβ25-35 20 μginto the adult rats hippocampus induced learning and memory dysfunction, and a decrease in the number of ChAT-immunoreactive neurons in hippocampus. Melatonin (0.1, 1, and 10 mg·kg-1, ig X 10 d) improved the Aβ25-35-treated rats cognitive function and increased the number of ChAT-immunoreactive neurons in hippocampus. CONCLUSION: Improvement of the cholinergic dysfunction by melatonin in adult rats induced by amyloid β-peptide 25-35 may be via cholinergic nervous system.

Carrier Proteins ; metabolism ; physiology ; HIV-1 ; physiology ; Models, Biological ; Proteins ; metabolism ; physiology ; Retroviridae Infections ; virology

NAC transcription factors involved in plant growth and development, as well as responses to biotic and abiotic stress. RNAi Vectors for SmNAC transcription factors of Salvia miltiorrhiza was constructed by using Gateway cloning technology, in order to further study the function of SmNAC1 transcription factor. According to Gateway cloning technology, the specific fragments of SmNAC1 containing attB adapter was amplified by PCR using ultra-fideling phusion polymerase of NEB. By the BP recombination reaction, the PCR product containing attB was transferred to an donor vector (pENTR/SD/D-TOPO). Finally, SmNACi specific gene was cloned into pK7GWIWG2D plant expression vectors by LR recombination reaction. Experimental results showed that Gateway cloning technology provide a rapid and highly efficient way to clone the interested gene.
Cloning, Molecular ; methods ; Genetic Vectors ; genetics ; Plant Proteins ; genetics ; Polymerase Chain Reaction ; RNA Interference ; Reproducibility of Results ; Salvia miltiorrhiza ; genetics ; Transcription Factors ; genetics

Objective To investigate the feasibility of ultrasmall superparamagnetic iron oxide- enhanced(USPIO)-enhanced MR imaging for monitoring synovitis of antigen-induced arthritis in rabbit model and explore the optimal MR imaging sequences.Methods Nine female white rabbits with antigen(0.5 ml mBSA,2 mg/ml)induced arthritis of the right knees were used in the study.The left knees of these rabbits and both knees of another 3 rabbits served as the control.Nine to 28 days(mean 21.3 d)after successful model induction,all knees were imaged before and 24 h after intravenously injection of USPIO (0.3 ml/kg),among which 2 rabbits were also imaged at 48 and 72 h after administration of USPIO respectively.The MR protocol included spin-echo(SE) T_1WI,fast spin-echo(FSE)T_2WI,gradient echo (GRE)T_2~* WI and short tau inversion recovery(STIR).Images were analyzed quantitatively and qualitatively based on signal characteristics and patterns of the synovium.Paired t-test was used for the analysis of the signal intensity of inflammatory synovial membrane before and 24 h after injection of USPIO. MR findings were correlated with histopathology.Results Arthritis was successfully induced in all 9 right knees with intraarticular injection of mBSA.Pathological examination revealed hyperplasia of synovium with infiltration of USPIO-loaded-macrophages.MR depicted synovial thickening(thickness 2.07?0.97 mm) and joint effusion.Synovium and joint fluid appeared as slightly hypo- or iso-intense on T_1 WI and hyper- intense on T_2 WI or T_2~* WI.Twenty four hours after USPIO injection,significant T_1 enhancement(ASNR 41.91%?27.94%),negative T_2 and T_2~* enhancement(△SNR -34.92%?11.77% and -57.24%? 16.05%)were demonstrated in the region of synovial inflammation respectively.The signal at 48 h and 72 h changed less than that at hour 24.No signs of arthritis occurred in all left knees and in all knees of the artificial model group.Conclusion Iron oxide phagocytized into macrophages can be a root cause resulted in signal change on USPIO-enhanced MR images.The gradient echo sequence should be the optimal sequence to be used in USPIO-enhanced MR imaging in antigen-induced arthritis.

Aim To observe the expression of mesen-cephalic astrocyte-derived neurotrophic factor(MANF) in synovial membrane and serum of rats with adjuvant arthritis (AA) and to analyse the relationship between MANF expression and arthritis. Methods AA models were prepared by injecting Freund complete adjuvant (FCA) into SD rats. The swelling of the secondary joint was measured by foot volume measurement. The severity of AA was recorded by arthritis index (AI). Synovial pathological changes were observed by HE staining. The protein and mRNA levels of MANF,BiP and CHOP extracted from synovial tissues in different periods of AA rats were detected by Western blot and reverse transcription-polymerase chain reaction (RT-PCR), respectively. The levels of MANF, C-reactive protein (CRP), interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) in serum were detected by enzyme-linked immunosorbent assay (ELISA) and then the relationship between MANF level and inflam-matory factors were explored. Results AA rat model was established successfully. The expression of BiP significantly increased in synovial tissue on d 2 after CFA injection,and decreased until d 28. The expres-sion of MANF slightly increased on d 2,then remained stable,and significantly increased on d 14, and then decreased gradually. The expression of CHOP kept to rise slowly at a low level. The level of MANF in serum markedly increased on d 14,then gradually decreased, but it was still higher than the normal level on d 28. The level of CRP exhibited similar trend with MANF. Correlation analysis showed that MANF had a negative correlation with arthritis symptoms, IL-1β and TNF-α in the secondary inflammatory period of AA rats. Con-clusions Arthritis induces the expression and secre-tion of MANF,and the level of MANF is closely relat-ed to the progression and severity of arthritis.

OBJECTIVETo inquire into the relationship between lipoprotein lipase (LPL) gene D9N, N291S and S447X polymorphisms and the development of cardiovascular diseases in children with obesity.

METHODSThe polymerase chain reaction (PCR) and restriction fragment length polymorphism (RLFP) techniques were used to detect three common mutations of LPL gene exon D9N, N291S and S447X in 157 obese children and 175 normal controls. Plasma lipid and lipoprotein levels between children with different genotypes were compared.

RESULTSThe D9N and N291S gene mutations were not detected in either the obese or the control groups. There were no significant differences in the frequency of S447X gene mutation between the two groups. There were no significant differences in the levels of plasma lipid and lipoprotein between children with S447 and X447 genotypes.

CONCLUSIONSD9N and N291S gene mutations may not be risk factors associated with cardiovascular diseases in children with obesity. S447X gene mutation might not play an important role in the development of cardiovascular diseases in childhood.

Adolescent ; Cardiovascular Diseases ; etiology ; genetics ; Child ; Female ; Humans ; Lipoprotein Lipase ; genetics ; Male ; Mutation ; Obesity ; genetics ; Risk Factors

OBJECTIVETo assess the potential impact of warming climate in winter to the scale and severity of schistosomiasis epidemics in China.

METHODSBased on the data of mean temperature and monthly minimum temperature in January from 126 (out of 733) weather observation stations in China, the trend and oscillation amplitude was analyzed. The impact of warming climate in winter to the scale of schistosomiasis spreading was assessed, using the indices of 0 degrees C mean temperature and -4 degrees C mean monthly minimum temperature in January. Correlation between these two indices was analyzed.

RESULTSEvidences showed the warming climate in winter was seen all over China with the mean monthly minimum temperature and the mean temperature in January had increased 1.3 degrees C and 0.9 degrees C since 1986. The contours of mean monthly minimum temperature -4 degrees C and mean temperature 0 degrees C in January moved 1 - 2 latitudes northward.

CONCLUSIONThe warming climate phenomenon seen in winter was considered favorable to the living of snails in winter, resulting in the possible increase of northward spreading of snails.Thus, monitoring of snails should be paid special attention.

Animals ; China ; epidemiology ; Climate ; Disease Reservoirs ; Female ; Health Education ; Humans ; Male ; Prevalence ; Schistosoma japonicum ; physiology ; Schistosomiasis japonica ; epidemiology ; prevention & control ; Seasons ; Snails ; parasitology ; physiology ; Temperature

WRKY transcription factor is one of the Zinc finger proteins which contains a highly conserved WRKY domain and is a family of the plant-specific transcription factor. The plasmid pET28a-SmWRKY1 harboring Salvia miltiorrhiza WRKY1 (SmWRKY1) gene was successfully transformed and expressed in Escherichia coli BL21 (DE3). The conditions on protein expression of SmWRKY1 in E. coli, including induction duration, temperature, IPTG concentration and the E. coli concentration were optimized. The results showed that the highest protein expression of SmWRKY1 was obtained at 24 hours after the E. coli was cultured in the presence of 0.2 mol x L(-1) IPTG at 20 degrees C with A600 values of 1.0-1.5. This recombinant histidine-tagged protein was expressed at 2.454 g x L(-1) as inclusion body, which was first extracted using urea, and then purified by Ni2+ affinity chromatography and identified by SDS-PAGE. The expression of SmWRKY1 in E. coli was further confirmed by western blotting analysis.
Blotting, Western ; Cloning, Molecular ; DNA-Binding Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; chemistry ; genetics ; metabolism ; Molecular Weight ; Plant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; chemistry ; genetics ; isolation & purification ; metabolism ; Salvia miltiorrhiza ; genetics

Adult ; Aged ; Amifostine ; therapeutic use ; Benzene ; poisoning ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; chemically induced ; drug therapy

Objective To investigate the relationship between hemoglobin and blood pressure of pregnant women in Zhoushan islands, so as to provide scientific evidence for the etiological study of gestational hypertension. Methods A retrospective study was conducted among 1 383 pregnant women who received perinatal care in Zhoushan Maternal and Child Health Hospital of Zhejiang Province from January 2017 to June 2018. Pregnant women were monitored for hemoglobin content and blood pressure in the early, middle and late pregnancy. The multivariate linear regression was used to analyze the relationship between hemoglobin content and blood pressure in different pregnancy. Results The incidence of anemia in early, middle and late pregnancy was 7.74%, 25.45% and 15.76% respectively. The multivariate linear regression showed that hemoglobin levels during pregnancy had effects on systolic blood pressure in early, middle and late pregnancy, and the earlier hemoglobin levels were monitored, the more obvious the effect on systolic blood pressure was.With the increase of hemoglobin level, systolic blood pressure increased, such as the effect of hemoglobin on systolic blood pressure in early pregnancy, mid-pregnancy and late pregnancy. Hemoglobin of first trimster had the greatest effect (β=0.10, P<0.001), Hemoglobin of second trimester had no obvious effect, and that of third trimester had the second effect (β=0.04, P=0.027).Hemoglobin levels and diastolic blood pressure levels were similar to their relationship with systolic blood pressure. Conclusions Hemoglobin levels during pregnancy have significant effects on systolic and diastolic blood pressure in first, second and third trimsters of pregnancy. Regular measurement of hemoglobin levels during pregnancy can improve the health of pregnant women.