大鼠自发性乳腺癌生物学特性的研究

目的:观察大鼠自发性乳腺癌在不同接种途径下、不同性别、不同品系大鼠体内的生长与转移的生物学特性。方法:制备瘤细胞悬液,首先以不同瘤细胞数在大鼠右腋皮下接种,通过成瘤率及存活天数确定最佳细胞数;以2×106个瘤细胞从右腋皮下、腹腔、后肢肌肉和尾静脉分别接种CHN大鼠,以2×106个瘤细胞分别从右腋皮下接种雌性和雄性CHN大鼠,以2×106个瘤细胞从右腋皮下分别接种不同品系大鼠,分别观察大鼠成瘤率及存活天数;分别取肿瘤、浸润与转移肿瘤组织,做组织切片,HE染色后观察。结果:大鼠自发性乳腺癌在动物体内接种不同瘤细胞数与不同途径接种肿瘤成活率均为100%,肿瘤具有浸润性生长与转移特性,但带瘤宿主存活时间有非常显著性差异(P<0.001);不同性别大鼠移植瘤细胞后宿主存活时间、瘤重和瘤系数在雌雄大鼠之间无显著性差异(P>0.05),提示大鼠自发性乳腺癌属于非激素依赖性瘤株;不同品系大鼠接种瘤细胞悬液后肿瘤仅在CHN和CHN×LOU/M F1代大鼠体内生长,表明大鼠自发性乳腺癌是CHN大鼠的一种同基因型瘤株。结论:大鼠自发性乳腺癌无论用哪种方法在CHN大鼠体内接种成活率均为100%,都具有浸润性生长与转移特性,是CHN大鼠的一种同基因型和非激素依赖性新瘤株,对研究乳腺癌具有重要意义。

Study of human B7 homolog 1 expression in patients with hepatitis B virus infection

AIM: To further investigate the role of human B7 homolog 1 (B7-H1) in the mechanism of persistent hepatitis B virus (HBV) infection. METHODS: Peripheral and intra-hepatic B7-H1 expression were compared by flow cytometry and immunochemical staining between two 2 distinct groups, one being chronic HBV tolerance patients (CHB-T) and the other being acute hepatitis B patients (AHB). B7-H1 mRNA expression level was also compared by real time polymerase chain reaction between CHB-T and AHB patients. The location of intra-hepatic B7-H1 and CD40 expression were analyzed by immunofluorescence. The levels of B7-H1 and CD40 expression on cultured myeloid dendritic cells (mDCs) with or without hepatitis B surface antigen (HBsAg) treatment were analyzed dynamically by flow cytometry. Intracellular interferon-γ (IFN-γ) staining and the stimulatory capacity of mDC of cultured mDC with or without HBsAg treatment were also compared by flow cytometry. RESULTS: Peripheral B7-H1 expression on mDCs was increased significantly in AHB compared to CHB-T patients (P < 0.05). In the liver tissues from CHB-T patients, B7-H1 positive cells were almost absent despite a persistently elevated serum HBsAg load. In contrast, there were indeed increased B7-H1-positive cells in situ in the liver tissue from AHB. In vitro analysis showed the parallel upregulation of B7-H1 and CD40 on CD11c+ mDCs after the onset of stimulation. Addition of recombinant hepatitis B surface antigen (rHBsAg) significantly decreased CD40 expression (P < 0.05 at 16 h, 20 h and 24 h time points). B7-H1 expression was also inhibited by rHBsAg, and the inhibition rate of CD40 was greater than that of B7-H1. This preferential inhibition of CD40 expression on mDCs by rHBsAg resulted in the dysfunction of mDCs and T cells in the mixed leucocyte reaction (MLR) system. With rHBsAg pretreatment, in a carboxyfluorescein diacetate succinimidyl ester (CFSE) labeled MLR system at a ratio of 1:5 responder cell-stimulator cell (R/S), the CFSE dim percentage of T cells decreased from 85.1% to 25.4% and decreased from 30.3% to 12.0% at 1:10 R/S. IFN-γ production by CD8+ T cells, in the MLR system, was reduced significantly by HBsAg pretreatment. At ratios of 1:5 R/S, the percentage of IFN-γ and CD8 dual positive T cells decreased from 55.2% ± 5.3% to 15.1% ±3.1% (P < 0.001), and decreased from 35.0% ± 5.1% to 7.3% ± 2.7% at ratios of 1:10 R/S (P < 0.001). CONCLUSION: B7-H1 is not a signature of immune dysfunction, but an inflammation marker. HBsAg regulate immune response by tipping the balance between B7-H1 and CD40.

Carbon isotopic composition and genetic types of natural gas in the Sichuan Basin, China

The origin and genetic types of natural gas in the Sichuan Basin are still disputed.To classify the origin and genetic types in different areas,the paper analyzes the carbon isotopic composition of gases and geologic features in the Sichuan Basin.The results showed that the gas sourced from terrestrial layers is typically characterized by terrestrial origin and was mainly accumulated nearby to form reservoir.The carbon isotopic composition of gas showed a normal combination sequence distribution,suggesting that natural gas in continental strata is not affected by secondary alteration or that this deformation is very weak.The gas source is singular,and only gas from the southern and northern Sichuan Basin shows the characteristic of mixed sources.However,marine gas presents the characteristics of an oil-formed gas.The carbon isotopic composition of natural gas in the western and central part of the basin mostly distributes in a normal combination sequence,and few of them showed an inversion,indicating that the gas perhaps had not experienced secondary alteration.The carbon isotopic composition of marine-origin gas in the southern,northern and eastern Sichuan Basin displays a completely different distribution pattern,which is probably caused by different mixing ratio of gas with multi-source and multi-period.

Genetic Analysis of Grain Yield in a 7×7 Diallel Cross F_2 Population of Wheat(T.Aestivum L.)

In order to obtain genetic information for grain yield, seven genetically diverse wheat cultivars were crossed in an incomplete diallel to study the inheritance of grain yield using F_2 progenies for two years. Significant differences were observed among genotype, year and genotype × year interaction for grain yield, and both general combining ability(GCA) and specific combining ability(SCA) were also highly significant for grain yield, suggesting that the trait was controlled by both additive and non-additive effect. The GCA estimates revealed that the best combiners for grain yield were Yangmai 5 and Ningmai 9. Adequacy tests revealed that data of grain yield was fully adequate for genetic interpretation. Over-dominance genetic effects were important for the expression of grain yield. Grain yield exhibited moderately high value of narrow sense heritability(h_N^2=66.98% and h_N^2=72.37%).

Detection of Homozygous Deletions and Mutations in the CDKN2A Gene in Hydatidiform Moles

OBJECTIVE To investigate homozygous deletions and mutations in the CDKN2A gene(p16 INK4a and p14 ARF gene)in hydatidiform moles. METHODS A total of 38 hydatidiform mole samples and 30 villi samples were examined for homozygous deletions in the CDKN2A gene by PCR and for mutations by DHPLC. RESULTS i)Among 38 hydatidiform mole samples, homozygous deletions in the p16 INK4a exon 1 were identified in 5 cases(13.2%),while no homozygous deletions were found in the p16I NK4aexon 1 of 30 early-pregnancy samples.The rates of those deletions in hydatidiform compared to early-pregnancy villi samples was statistically significant(P=0.036).ii)No homozygous deletions in the p14 ARF exon 1 or p16 INK4a exon 2 were found in any of the hydatidiform moles or early-preganancy samples.iii) In all hydatidiform moles and early-pregnancy villi samples,no mutations were detected by DHPLC. CONCLUSION We suggest there may be a close correlation between homozygous deletions in the CDKN2A gene and occurrence of hydatidiform moles variation in the CDKN2A gene is mainly caused by homozygous deletions,while mutations may be not a major cause.

Phosphatase and Tension Homolog Overexpression in Insulin Resistant Diabetic Adipose Tissue

Objective To investigate the expression of phosphatase and tension homolog(PTEN) in adipose tissue of KKAy diabetic mice, a mouse model of type 2 diabetes. Methods KKAy diabetic mice were fed with high fat diet for 4 weeks. After blood glucose met the criteria of diabetes(over 16.7 mmol/L), mice were randomly divided into 3 groups: a control group(without any treatment), a rosiglitazone group(treated with rosiglitazone 12.5 mg/kg·d once per day), and a metformin group(treated with metformin 3 g/kg·d twice daily). After 4 weeks, we then determined the expression of PTEN and phosphoserine 473-Akt(pS473-Akt) in the epididymal adipose tissue with Western blots. The mice in each group were further divided into the insulin(-) subgroup and insulin(+) subgroup, which were intraperitoneally injected with saline and insulin(5 mU/g body weight), respectively. Results The expression of PTEN was elevated in the epididymal adipose tissue obtained from KKAy diabetic mice compared with that from the C57BL/6J mice(P<0.001). In accordance with the enhanced expression of PTEN, the level of pS473-Akt stimulated by insulin was decreased in the adipose tissue of KKAy mice compared to the C57BL/6J mice(P<0.001). Treatment with the insulin-sensitizing agents, rosiglitazone and metformin did not inhibit the elevated expression of PTEN in adipose tissue of KKAy diabetic mice. Conclusion PTEN may play an important role in the development of insulin resistance in adipose tissue of type 2 diabetes mice model.

Drought Tolerant Criterion of Wheat Genotypes Using Carbon Isotopes Discrimination Technique

Drought tolerant analysis using carbon isotopes discrimination could be useful tool to decide a genotype with high adaptation to drought. A pot culture experiment was conducted in glass house at the IAEA （International Atomic Energy Agency） laboratories, Seibersdorf, Austria to analyze drought tolerant of wheat genotype using carbon isotopes discrimination technique. Four wheat genotypes viz., LU-26s, Bhittai, Roshan, Taifu, were tested. There were two treatments： Control （normal irrigation） and Drought （@ 30% field capacity of the control treatment）. Drought treatment was imposed after two weeks of germination. The experiment was continued up to 10 weeks after germination and was terminated at flowering stage. The data were recorded in terms of plant height, number of tillers, shoot dry weight and carbon isotopic ratio （δ）. Results showed that genotype LU-26s was found to have best performance under drought condition, with minimum decrease in the growth parameters i.e., plant height, number of tillers and shoot dry weight. High CID （carbon isotopes discrimination） values were also observed in genotype LU-26s, showing close positive correlation between SDW （shoot dry weight） and CID. It is therefore concluded that carbon isotopes discrimination can be an important criterion for the selection of wheat genotypes for drought prone areas.

A Microsatellite Genetic Linkage Map of Black Rockfish(Sebastes schlegeli)

Ovoviviparous black rockfish(Sebastes schlegeli) is an important marine fish species for aquaculture and fisheries in China. Genetic information of this species is scarce because of the lack of microsatellite markers. In this study, a large number of microsatellite markers of black rockfish were isolated by constructing microsatellite-enriched libraries. Female- and male-specific genetic linkage maps were constructed using 435 microsatellite markers genotyped in a full-sib family of the fish species. The female linkage map contained 140 microsatellite markers, in which 23 linkage groups had a total genetic length of 1334.1 c M and average inter-marker space of 13.3 c M. The male linkage map contained 156 microsatellite markers, in which 25 linkage groups had a total genetic length of 1359.6 c M and average inter-marker distance of 12.4 c M. The genome coverage of the female and male linkage maps was 68.6% and 69.3%, respectively. The female-to-male ratio of the recombination rate was approximately 1.07:1 in adjacent microsatellite markers. This paper presents the first genetic linkage map of microsatellites in black rockfish. The collection of polymorphic markers and sex-specific linkage maps of black rockfish could be useful for further investigations on parental assignment, population genetics, quantitative trait loci mapping, and marker-assisted selection in related breeding programs.

Effects of environment and genotype-by-environment interaction on phenotype of Rorippa elata(Brassicaceae),an endemic alpine plant in the Hengduan mountains

Global climate change poses a severe threat to mountain biodiversity.Phenotypic plasticity and local adaptation are two common strategies for alpine plant to cope with such change.They may facilitate organismal adaptation to contrasting environments,depending on the influences of the environment or genotype or their interacted effects.In this study,we use an endemic alpine plant(Rorippa elata)in the Hengduan mountains(HDM)to unravel its phenotypic basis of adaptation strategy and evaluate the relative contributions of environment and genotype to its phenotype.We transplanted 37 genotypes of R.elata into two common gardens across low and high elevations(2800 vs.3800 m)during 2021-2022.Nine fitness-related traits were measured,including flowering probability and glucosinolates(GS)content.We estimated the environmental or genotypic contributions to the phenotype and identified the main environmental components.Our results revealed that both environment and genotype-by-environment interactions contributed to the phenotypes of R.elata.Latitudinal heterogeneity was identified as a key factor that explained 24%of the total phenotypic variation.In particular,genotypes of the northern HDM showed significantly higher plasticity in flowering probability than those of the southern HDM.Furthermore,within the southern HDM,GS content indicated local adaptation to herbivory stresses for R.elata genotypes along elevations.In conclusion,our results suggest that R.elata may have adapted to the alpine environment through species-level plasticity or regional-level local adaptation.These processes were shaped by either complex topography or interactions between genotype and mountain environments.Our study provides empirical evidence on the adaptation of alpine plants.