In addition to the structural genes of the coronavirus genome, S, E, M, and N, there are several additional genes called "group-specific or accessory genes". Their gene products are designated as "accessory proteins", as reports to date make it clear that these proteins are not essential for virus replication in vitro. Nevertheless, many of these genes are still maintained in the virus genome under selective pressure, suggesting that they might play a very important role in the survival of the virus in the natural environment of the infected host. This review will summarize the research progress in the functions of coronavirus accessory genes.
Animals ; Coronavirus ; genetics ; metabolism ; Coronavirus Infections ; virology ; Humans ; Open Reading Frames ; Viral Proteins ; metabolism

Objective To identify the subcellular location of S AR S-CoV N protein in mammalian cells. Methods The gene fraction of SARS-CoV was cloned into the PGEFP-C1 plasmid to construct expression vecto r PGEFP-C1/N. The subcellular location of N in A549 and VeroE6 cells was observ ed under fluorescence microscope with the aid of transient transfection techniqu e. The expression of the fusion protein (GFP-N) was detected by Western blot. Results The PGEFP-C1/N was constructed. N protein was localiz ed in the cytoplasm of transfected cells and detected by Western blot. C onclusion N protein was localized in the cytoplasm of mammalian cells.

RhoA, a small GTPase, is involved in a wide array of cellular functions in the central nervous system, such as cell motility, cytoskeleton rearrangement, transcriptional regulation, phagocytosis and cell growth. It is not known how spinal cord injury (SCI) affects the expression of RhoA in different nerve cells. In the present study, we investigated the changes of RhoA expression in remote areas of the injury at the 3rd, 7th and 30th day after SCI, which was established by T10 contusion method. Moreover, we examine its expression profile in neurons, astrocytes and microglia. RhoA was found to be weakly expressed in these nerve cells in normal spinal cord. Western blotting showed that, after SCI, the total RhoA expression was up-regulated, and the RhoA expression was increased and peaked at the 7th day. Double immunostaining revealed specific and temporal expression patterns of RhoA in different nerve cells. The expression of RhoA in neurons started to increase at day 3, peaked at day 7 and then decreased slightly at day 30. Expression of RhoA in astrocytes increased moderately after SCI and peaked at day 7. There was no obvious change in RhoA expression in microglia after SCI in remote areas. This study demonstrated that, after SCI, RhoA expression exhibited different patterns with different nerve cells of spinal cord. RhoA expression patterns also changed with time after SCI, and among different nerve cells in the injured spinal cord. These findings can help us better understand the roles of RhoA in SCI.

Adult ; Anti-Inflammatory Agents ; therapeutic use ; Churg-Strauss Syndrome ; drug therapy ; pathology ; Follow-Up Studies ; Humans ; Male ; Methylprednisolone ; therapeutic use

Objective To silence the expression of tissue factor(TF) gene of human umbilical vein endothelial cell(HUVECs) of the newborns with placental abruption(PA) and normal newborns.Methods There were two groups in the experiment,normal group and PA group.Three different treatments were established in each group:(1) the blank; (2) the false-intervention ; (3) the TF gene silencing.There were three samples in each treatments.After these treatments,the changes of mRNA expression of the HUVECs were observed before and aftcr thc gene silencing and the changes of the immunofluorescence of the TF protein level.Results After amplificated,plasmid DNA were sequenced to show that the pENTRTM/U6-TF-shRNA was the positive clone.After the transfected,the levels of the mRNA of TF decreased from 0.657 ± 0.097 to 0.220 ± 0.030 and 1.323 ± 0.323 to 0.207 ± 0.150 in the normal and PA group respectively.Compared with the normal group,there were significant differences for the levels of TF mRNA in PA group with the blank,(1.323 ± 0.323 vs 0.657 ± 0.097,P =0.023) and the same result for the second management (1.057 ±0.178 vs 0.540 ± 0.079,P =0.01).But there was no significant difference between the normal and PA group after RNA interference gene silencing (0.220 ± 0.030 vs 0.207 ± 0.150,P ＞ 0.05).Meanwhile,there were significant differences among the three managements in the themselves groups of normal and pathological ones(F =19.30,P =0.002 ;F =27.66,P =0.001).Conclusion The vectors are transfected into HUVECs and play the biological function.And they silence the expression of TF mRNA.PENTRTM/U6-TF-shRNA could inhibit the expression of TF mRNA of HUVECs in the PA newborn.

BACKGROUND:It is wel-known that mechanical stimulation could promote fracture healing. However, what kind of mechanical stimulation induced by treadmil exercise can increase the bone conductibility of bone material and promote the healing of bone defect is stil unclear.
OBJECTIVE:To evaluate the influence of indirect mechanical stimulation produced by treadmil exercise on bone defect healing and osteogenesis of bone materials.
METHODS:Sprague-Dawley rats at 12 weeks old were used in this study to establish a bone defect of 3 mm in diameter and height at the left distal femur. Afterwards, calcium sulphate scaffolds were implanted into the defects. The rats were divided into treadmil exercise group and control group. Treadmil exercise was began at 1 week postoperatively, 10 m/min, 45 minutes per day, 5 days per week, for 3 weeks. Control group did not receive any exercise. Micro-computed tomography was used to determine bone formation in the bone defects at 1, 2, 3, and 4 weeks after surgery. The sections of left distal femur were subject to hematoxylin-eosin staining, the new bone formation and degradation of bone materials in the bone defects were observed.
RESULTS AND CONCLUSION:Micro-CT analysis showed that, a smal amount of new bone formed in both treadmil exercise group and control group at 1 week after surgery. In treadmil exercise group, new bone formation was significantly higher than the control group at 2, 3, 4 weeks (P<0.05). At 4 weeks, histological results also confirmed the difference of new bone formation in bone defect between treadmil exercise group and control group. In addition, bone mineral density of treadmil exercise group was higher than that of control group at 2, 3, 4 weeks, but no significant difference was found (P>0.05). The results suggest that moderate treadmil exercise could promote bone defect healing and enhance osteoconductivity of bone substitute.

Puromycin-sensitive aminopeptidase (PSAP) belongs to the M1 family of aminopeptidases, characterized by the N-terminal substrate binding sequence GAMEN, the enzyme activity center HEXXH(X)₁₈E motif, and the C-terminal ERAP-1-like superfamily structural domain. Encoded by the gene NPEPPS located at 17q21.32, PSAP consists of 919 amino acids and is widely distributed throughout the human body, with the highest expression in the brain, followed by the heart and skeletal muscle. It is also found in the liver, renal tubular epithelium, small intestine, large intestine epithelium, and gastric epithelial cells. PSAP primarily relies on its aminopeptidase hydrolytic activity to remove toxic protein aggregates such as Tau, poly Q, and Cu, Zn-superoxide dismutase 1, making it an important factor in the development of diseases such as Alzheimer's disease, Huntington's chorea, and tumors. Existing PSAP inhibitors include bestatin, amastatin, leuhistin, actinonin, and purinomycin, some of which are already available or in clinical trials. This review provides an overview of the structural and biological functions of M1 family aminopeptidases, with a focus on PSAP, to facilitate further research and targeted drug development.

Adult ; Aged ; Biomarkers ; blood ; Female ; Humans ; Laminin ; blood ; Male ; Middle Aged ; Occupational Exposure ; Risk Assessment ; Silicon Dioxide ; adverse effects ; Silicosis ; blood ; diagnosis ; Superoxide Dismutase ; blood ; Transforming Growth Factor beta ; blood

Bridged-Ring Compounds ; poisoning ; Female ; Humans ; Middle Aged ; Occupational Exposure ; Rodenticides ; poisoning

Adolescent ; Anal Canal ; physiopathology ; Child ; Constipation ; physiopathology ; Female ; Gastrointestinal Motility ; physiology ; Humans ; Male ; Rectum ; physiopathology