Objective:To evaluate the role of ZIP7 in sepsis-induced cardiomyopathy in mice.Methods:Ninety wild-type and 90 cardiomyocyte-specific knockout ZIP7 (ZIP7 cKO) male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups using a random number table method: wild-type sham operation group (Sham group) and wild-type sepsis group (Sep group), ZIP7 cKO sham operation group (cKO + Sham group) and ZIP7 cKO sepsis group (cKO + Sep group), with 45 mice in each group. The sepsis-induced cardiomyopathy model was developed using the cecal ligation and puncture in anesthetized mice. Twenty mice were randomly selected to record the survival for 10 days postoperatively. At 18 h after surgery, the left ventricular ejection fraction (LVEF) and fractional shortening (LVFS) were measured by echocardiography, and serum concentrations of cardiac troponin T (cTnT), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. The contents of hydroxyl radical (·OH) and peroxynitrite anion (ONOO -) were determined using a colorimetric assay, the morphology of myocardial mitochondria was observed with a transmission electron microscope, and the expression of dynamin-related protein 1 (Drp1), mitofusin-2 (Mfn2), and optic atrophy 1 (Opa1) in myocardial tissues was detected using Western blot. Results:Compared to Sham group, the survival rate, LVEF and LVFS were significantly decreased, serum concentrations of cTnT, TNF-α and IL-6 were increased, the contents of ·OH and ONOO - in myocardial tissues were increased, the expression of Drp1 was up-regulated, the expression of Mfn2 and Opa1 was down-regulated ( P<0.05), myocardial cells exhibited mitochondrial swelling, and marked destruction of mitochondrial cristae was observed in Sep group, and no significant differences were found in the aforementioned parameters in cKO+ Sham group ( P>0.05). Compared to Sep group, the survival rate, LVEF and LVFS were significantly increased, serum concentrations of cTnT, TNF-α and IL-6 were decreased, the contents of ·OH and ONOO - in myocardial tissues were decreased, the expression of Drp1 was down-regulated, the expression of Mfn2 and Opa1 was up-regulated ( P<0.05), and mitochondrial swelling in myocardial cells was mild, with less dissolution and destruction of mitochondrial cristae in cKO+ Sep group. Conclusions:Myocardial ZIP7 can promote mitochondrial fusion and inhibit mitochondrial fission, potentially contributing to the mechanism of sepsis-induced cardiomyopathy in mice.

Objective:To evaluate the role of ZIP7 in sepsis-induced cardiomyopathy in mice.Methods:Ninety wild-type and 90 cardiomyocyte-specific knockout ZIP7 (ZIP7 cKO) male C57BL/6 mice, aged 6-8 weeks, weighing 20-25 g, were divided into 2 groups using a random number table method: wild-type sham operation group (Sham group) and wild-type sepsis group (Sep group), ZIP7 cKO sham operation group (cKO + Sham group) and ZIP7 cKO sepsis group (cKO + Sep group), with 45 mice in each group. The sepsis-induced cardiomyopathy model was developed using the cecal ligation and puncture in anesthetized mice. Twenty mice were randomly selected to record the survival for 10 days postoperatively. At 18 h after surgery, the left ventricular ejection fraction (LVEF) and fractional shortening (LVFS) were measured by echocardiography, and serum concentrations of cardiac troponin T (cTnT), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) were measured by enzyme-linked immunosorbent assay. The contents of hydroxyl radical (·OH) and peroxynitrite anion (ONOO -) were determined using a colorimetric assay, the morphology of myocardial mitochondria was observed with a transmission electron microscope, and the expression of dynamin-related protein 1 (Drp1), mitofusin-2 (Mfn2), and optic atrophy 1 (Opa1) in myocardial tissues was detected using Western blot. Results:Compared to Sham group, the survival rate, LVEF and LVFS were significantly decreased, serum concentrations of cTnT, TNF-α and IL-6 were increased, the contents of ·OH and ONOO - in myocardial tissues were increased, the expression of Drp1 was up-regulated, the expression of Mfn2 and Opa1 was down-regulated ( P<0.05), myocardial cells exhibited mitochondrial swelling, and marked destruction of mitochondrial cristae was observed in Sep group, and no significant differences were found in the aforementioned parameters in cKO+ Sham group ( P>0.05). Compared to Sep group, the survival rate, LVEF and LVFS were significantly increased, serum concentrations of cTnT, TNF-α and IL-6 were decreased, the contents of ·OH and ONOO - in myocardial tissues were decreased, the expression of Drp1 was down-regulated, the expression of Mfn2 and Opa1 was up-regulated ( P<0.05), and mitochondrial swelling in myocardial cells was mild, with less dissolution and destruction of mitochondrial cristae in cKO+ Sep group. Conclusions:Myocardial ZIP7 can promote mitochondrial fusion and inhibit mitochondrial fission, potentially contributing to the mechanism of sepsis-induced cardiomyopathy in mice.

Objective:To investigate the survival rate change of retinal ganglion cells（RGCs）in a mouse of optic nerve crush（ONC）.Methods:Ninety-seven male C57BL/6J mice（6 to 8 weeks）were selected and divided into normal group（ n=5）, sham-operation group（ n=5）and ONC group（ n=5）according to the random number table. In normal group, both eyes of the mice did not receive any intervention. In sham-operation group, the right eye of the mice received sham operation, while the left eye reveived no intervention. In ONC group, the left eye received ONC, and the right eye received sham operation. In normal group, the density of RGCs in both eyes was quantified and compared. In sham-operatioin group, the density of RGCs in the sham operation eye was calculated and then compared to the average density of RGCs in normal group. In ONC group, the survival rate of RGCs was set as the ratio between the left eye（ONC eye）and the right eye（sham-operation eye）. The survival rate of RGCs in ONC group was compared after crush injury for 5, 10, 20, 30 seconds）（the sacrifice time was set at day 7）, and was compared after sampling on days 3, 4, 5, 7, 14, 30, 60, 90, 180（the duration of crush injury was set as 20 seconds）. Results:In normal group, the density of RGCs in the right eye was（5, 167.3±55.6）cell/mm 2, with no statistical difference from that in the left eye［（5, 199.6±44.8）cell/mm 2］（ P>0.05）. The density of RGCs in normal group and sham-operation group was（5, 183.5±33.4）cell/mm 2 and（5, 151.5±87.6）cell/mm 2, showing no statistical difference（ P>0.05）. The survival rate of RGCs in ONC group after crush injury for 5, 10, 20, 30 seconds was（37.6±1.1）%,（34.0±0.9）%,（33.6±1.6）% and（30.3±0.6）%（ P<0.01）. In comparison, there was statistical difference in the survival rate of RGCs between crush injury for 5 seconds and for 30 seconds（ P<0.01）, but not among other duration of crush injury（ P>0.05）. The survival rate of RGCs in ONC group after sampling on days 3, 4, 5, 7, 14, 30, 60, 90, 180 was（85.4±2.0）%,（67.6±3.1）%,（43.0±1.0）%,（33.6±1.6）%,（22.7±2.0）%,（12.8±0.6）%,（10.4±0.8）%,（8.6±0.5）% and（6.7±0.2）%（ P<0.01）, showing the most obvious drop from day 3 to day 5. Additionally, the curve became flattened after 30 days. Conclusions:In a mouse model of ONC, varying durations of crushing will lead to different damage to RGCs in a progressive mode, indicating that following the primary injury（ONC）, the RGCs suffer secondary injury as well. Therefore, effectively controlling the secondary injury may be the key point of treating optic nerve injuries.

Objective:To investigate the reproduction and prenatal diagnosis of female with Down's syndrome.Methods:Three cases of women with Down's syndrome were analyzed and relative literatures were reviewed.Results:All the three pregnant women had Down's syndrome special features and mental retardation, and their karyotypes of peripheral blood chromosome were all 47,XX,+21. The karyotypes of peripheral blood chromosome of their husbands were all normal. All the three women underwent amniocentesis during the second trimester. The cytogenetic analysis of cultured amniocytes of all the three cases showed normal karyotypes. Chromosomal microarray analysis (CMA) results of cultured amniocytes from all the three cases were successfully analyzed. Case 1 and Case 3 were normal, while 2.1 Mb deletion of 1q21.1-q21.2 was detected in Case 2. Prenatal ultrasound screening of all the three cases showed normal. All the three women delivered at term and their babies at 1-year-old all had normal phenotype.Conclusion:Females with Down's syndrome may be capable of reproduction and give birth to healthy children. It’s necessary to provide sufficient prenatal and postnatal genetic counselling to Down's pregnant women and families, and to offer comprehensive genetic testing to fetuses during pregnancy.

Objective:To investigate the reproduction and prenatal diagnosis of female with Down's syndrome.Methods:Three cases of women with Down's syndrome were analyzed and relative literatures were reviewed.Results:All the three pregnant women had Down's syndrome special features and mental retardation, and their karyotypes of peripheral blood chromosome were all 47,XX,+21. The karyotypes of peripheral blood chromosome of their husbands were all normal. All the three women underwent amniocentesis during the second trimester. The cytogenetic analysis of cultured amniocytes of all the three cases showed normal karyotypes. Chromosomal microarray analysis (CMA) results of cultured amniocytes from all the three cases were successfully analyzed. Case 1 and Case 3 were normal, while 2.1 Mb deletion of 1q21.1-q21.2 was detected in Case 2. Prenatal ultrasound screening of all the three cases showed normal. All the three women delivered at term and their babies at 1-year-old all had normal phenotype.Conclusion:Females with Down's syndrome may be capable of reproduction and give birth to healthy children. It’s necessary to provide sufficient prenatal and postnatal genetic counselling to Down's pregnant women and families, and to offer comprehensive genetic testing to fetuses during pregnancy.

Objective: To investigate the effect of tumor-associated macrophage (TAM) on the anti-tumor function of chidamide and to explore the mechanism. Methods: Mouse macrophage cell linesAna1 and Raw264.7 were cultured in vitro and induced into TAM with tumor supernatant. HDAC enzyme activity was detected after TAM treated with chidamide. The mRNA expressions of cytokines, such as IL-6, IL-12,TNF and IL-1β, in TAM were detected by qPCR. The protein expression of NF-κB and STAT3 in TAM treated with chidamide were detected by Wb. The mixture of TAM and colon cancer CT26 cells was inoculated into nude mice to construct the subcutaneous xenograft model; and the efficacy of chidamide (3.87 mg/kg) on the growth of CT26 xenograft tumors was observed. The protein expressions of PCNA, F4/80, Arg1 and iNos were detected by immunohistochemistry. Results: Chidamide inhibited the proliferation of CT26 cells. In the in vivo experiment, the inhibition rate of chidamide alone on CT26 xenograft was about 18.7%; however, the inhibition rate was up to 57.2% with the presence of TAM. Chidamide could inhibit the activity of HDAC enzyme in TAM, and further increase the Histone acetylation level. Chidamide could affect the expression of nuclear transcription factor NF-κB, inhibit the expressions of Arg1, IL-6 and IL-12, but up-regulate the expressions of iNOS, TNF and IL-1β in TAM. Conclusion: Chidamide can enhance its inhibitory effect on colon cancer CT26 cells via regulating the expression of cytokines and inhibiting the activity of HDAC in TAM.

The abnormality of serine/threonine kinase Aurora-A is seen in many types of cancers. Although in physiological context it has been shown to play a vital role in cellular mitosis, how this oncogene contributes to tumorigenesis remains unclear. Here we demonstrate that Aurora-A overexpression enhances both the expression level and transcriptional activity of c-Myc. The inhibition of c-Myc expression by RNA interference significantly impaired the oncogenic potential of Aurora-A, resulting in attenuated cellular proliferation and transformation rates as well as fewer centrosomal aberrations. Furthermore, downregulation of c-Myc effectively overcame Aurora-A-induced resistance to cisplatin in esophageal cancer cells. Taken together, our results suggest an important role for c-Myc in mediating the oncogenic activity of Aurora-A, which may in turn allow for future targeting of c-Myc as a potential therapeutic strategy for tumors with Aurora-A overexpression.
Cell Line, Transformed ; Cell Proliferation/drug effects ; Cell Transformation, Neoplastic/drug effects/genetics ; Centro ; Chromo ; Cisplatin/pharmacology ; Down-Regulation ; E ; Gene Expression Regulation, Neoplastic/drug effects ; Humans ; Protein-Serine-Threonine Kinases/genetics/*metabolism ; Proto-Oncogene Proteins c-myc/genetics/*metabolism ; RNA, Small Interfering/genetics ; Transcriptional Activation ; Transgenes/genetics

The Coronaviridae family is characterized by a nucleocapsid that is composed of the genome RNA molecule in combination with the nucleoprotein (N protein) within a virion. The most striking physiochemical feature of the N protein of SARS-CoV is that it is a typical basic protein with a high predicted pI and high hydrophilicity, which is consistent with its function of binding to the ribophosphate backbone of the RNA molecule. The predicted high extent of phosphorylation of the N protein on multiple candidate phosphorylation sites demonstrates that it would be related to important functions, such as RNA-binding and localization to the nucleolus of host cells. Subsequent study shows that there is an SR-rich region in the N protein and this region might be involved in the protein-protein interaction. The abundant antigenic sites predicted in the N protein, as well as experimental evidence with synthesized polypeptides, indicate that the N protein is one of the major antigens of the SARS-CoV. Compared with other viral structural proteins, the low variation rate of the N protein with regards to its size suggests its importance to the survival of the virus.
Amino Acid Motifs ; genetics ; Amino Acid Sequence ; Antigens, Viral ; immunology ; Base Composition ; Base Sequence ; Cluster Analysis ; Computational Biology ; DNA Primers ; Enzyme-Linked Immunosorbent Assay ; Genetic Variation ; Molecular Sequence Data ; Nucleocapsid Proteins ; genetics ; immunology ; metabolism ; Phosphorylation ; SARS Virus ; genetics ; Sequence Analysis, DNA