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ObjectiveTo introduce the Hr gene of spontaneously mutated SHJHhr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model.Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJHhr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJHhr (abbreviated as C.Cg.SHJHhr) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJHhr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJHhr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. ResultsFrom May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJHhr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). ConclusionC.Cg.SHJHhr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJHhr mice in biomedical research.
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Objective To measure and analyze biological characteristics and aging phenotype of SHJHhr mice and provide basic data for the application of the mouse model in aging mechanisms research and antiaging drug development. MethodsWith ICR mice of the same age as control group, the body mass growth data of SHJHhr mice at the age of 3 to 16 weeks, the reproduction ability of 1 to 4 fetuses and the life cycle of SHJHhr mice were measured. Blood routine (30 items) and serum biochemical indexes (25 items) of 6-week-old SHJHhr mice were measured. The venous blood of 8-week-old SHJHhr mice was collected for flow cytometry analysis to determine the content of immune cells. The aging bone structure of the cancellous bone and bone mineral density of SHJHhr mice aged 4, 8 and 26 weeks were measured by micro-CT. Histopathological changes of bone and joint of 8-week-old mice were observed. ResultsCompared with ICR mice, the female and male body mass of SHJHhr mice were significantly lower at the age of 16 weeks (P < 0.05), and the reproductive performance of female mice was low (P < 0.01) or did not have normal reproductive capacity. The shortest survival time of SHJHhr mice was 57 weeks and the longest was 71 weeks, which was shorter than those of normal ICR mice, showing obvious rapid aging phenomenon. At the same time, some physiological and biochemical indexes of blood and pathological changes of bone and cartilage tissues also showed the accelerated aging and abnormality of animal physiological functions. ConclusionSHJHhr mice have some biological characteristics of rapid aging as well as some physiological and pathological changes caused by aging.
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<p><b>OBJECTIVE</b>To investigate the effects of microRNA(miRNA)-29b on the proliferation and migration of breast cancer cells and its molecular mechanism.</p><p><b>METHODS</b>The recombinant lentiviral expression vector (lenti-miRNA-29b) was constructed and transfected into 293T cells to obtain lentivirus particles that were used to infect breast cancer MCF-7 cells. Transfection efficiency of lenti-miRNA-29b in MCF-7 cells was identified by the expression of green fluorescent protein (GFP). The expression of miRNA-29b was detected by real-time PCR. The cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The bioinformatics softwares were used to predict and screen the downstream target genes regulated by miRNA-29b, which were verified by double luciferase reporter gene assay, RT-PCR and Western blot. The effects of screened target gene RTKN on the growth and migration of MCF-7 cells were verified by RTKN siRNA.</p><p><b>RESULTS</b>Recombinant lentiviral expression vector of miRNA-29b were successfully constructed. About 90% and 60% of the breast cancer cells showed green fluorescence in lenti-miRNA-29b and lenti-miRNA-NC groups, respectively. The expression of miRNA-29b in lenti-miRNA-29b group increased significantly compared with the lenti-miRNA-NC group and blank control group (all<0.05); the proliferation and migration ability of MCF-7 cells significantly reduced compared with the control group (all<0.05). The screening with bioinformatics softwares found that the 3'UTR coding region RTKN had the binding site to miRNA-29b; the dual luciferase reporter gene assay showed that the luciferase activity decreased significantly after the MCF-7 cells were co-transfected with wild type RTKN-WT-3'UTR and miRNA-29b mimics report gene vector (<0.05). The RTKN proteins in MCF-7 cells were significantly decreased after transfection with siRNA-RTKN, and the proliferation and migration ability of MCF-7 cells were significantly reduced (all<0.05).</p><p><b>CONCLUSIONS</b>MiRNA-29b can inhibit the proliferation, invasion and metastasis of breast cancer cells by inhibiting the expression of RTKN.</p>
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Fibromyalgia syndrome after comprehensive treatment of breast cancer is rare and seldom reported. Here we present a case of a 50-year-old female patient,who was admitted to the hospital because of generalized fibromyalgia for 3 months and brain metastasis after the right breast carcinoma surgery for 1 month, and the clinical diagnosis was brain metastasis from breast carcinoma combined with fibromyalgia syndrome. The fibromyalgia were relieved with proper symptomatic treatment but the patient eventually died of tumor progression.
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Female , Humans , Middle Aged , Brain Neoplasms , Mortality , Breast Neoplasms , Mortality , Therapeutics , Carcinoma , Mortality , Therapeutics , Fibromyalgia , Diagnosis , TherapeuticsABSTRACT
Objective To establish a Goto-Kakizaki ( GK) rat model of duodenal-jejunal bypass( DJB) and ob-serve the changes in insulin-resistance after surgery, and to explore the mechanism of DJB surgery in treatment of type 2 di-abetes mellitus.Methods Male Goto-kakizaki diabetic rats(GK,n=36)were used as experiment group and 18 healthy male Wistar rats as blank group.GK rats were randomly divided into two groups: diabetic control group and DJB surgery group ( n=18) .Euglycemic-hyperinsulinemic clamp technique was performed in 6 rats randomly taken from each group at third week, sixth week and ninth week after surgery, respectively.The expression levels of Gck, G6P, PEPCK mRNA in the liver and GLUT4 content on skeletal muscle cell plasma membrance were detected one week after the clamp test. Results In the DJB surgery group at the end of third week and sixth week after surgery, the levels of glucose infusion rates and the expression levels of Gck, G6P, PEPCK mRNA in the liver showed no statistically significant difference as com-pared with the diabetic control group (P>0.05).In the DJB surgery group at the end of 9th week after surgery, the glu-cose infusion rate and expression level of Gck mRNA in the liver were significantly higher, and the expression of G6P and PEPCK mRNA was significantly lower than those in the diabetic control group ( P0.05 for all) .Conclusions Our results indicate that the mechanism of DJB surgery improving blood glucose level may be closely related to the amel-ioration of insulin-resistance in the liver, thereby augmenting glucose uptake and inhibiting gluconeogenesis in liver through the regulation of glucose metabolism-related enzymes.There is no significant improvement in insulin-resistance in the skele-tal muscles during the experiment period.This result implies that the effect of DJB surgery on type 2 diabetes is related to the duration of therapy.
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Objective To investigate the effects of the exercise training on the cognitive function ,choline acetyltransferase (ChAT) activity and acetylcholinesterase (AchE) activity in stroke prone spontaneously hypertensive rat (SHR/SP) vascular de-mentia model .Methods 30 male SHR/SP rats were randomly divided into sham operation group ,model group and exercise group (n=10) .The VD model was established by the fractional ligation of bilateral carotid artery (2-VO) .The sham operation group and the model group were given the normal feeding without intervention after operation ;the exercise group adopted the treadmill exer-cise(DSPT-1) for 8 weeks .After the exercise ,the Morris maze test was conducted for evaluating the cognitive function in each group .The rats were finally killed for detecting the ChAT activity and AchE activity of hippocampus .Results In the positioning navigation training ,the latency period of the sham operation group was significantly short than that of the exercise group and the model group ,but the latency period of the exercise group was obviously short than that of rats in the model group (P<0 .05);in the spatial exploration test ,the rats in the sham operation group had more frequency of crossing platform than the other two groups ,the exercise group had more frequency of crossing platform in platform quadrant than the model group (P<0 .05);the exercise training could increase the ChAT activity and lower the AchE activity of hippocampus .Conclusion The exercise training can improve the function of hippocampal cholinergic system in SHR/SP and then increase the cognitive ability .
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Eestablishing a scientific supervision system is essential to achieve a better environment for teaching and learning and improve teaching quality in medical college.We took undergraduate education evaluation as an important supervising subject and made a scientific evaluation program for teaching,learning,and testing,based on specific features of medical college.The practice of the program proves to be successful.We present the main contents in this article.