Host proteins interact with viral elements and affect the life cycle of highly pathogenic avian influenza A virus H7N9.

Host-virus interactions can significantly impact the viral life cycle and pathogenesis; however, our understanding of the specific host factors involved in highly pathogenic avian influenza A virus H7N9 (HPAI H7N9) infection is currently restricted. Herein, we designed and synthesized 65 small interfering RNAs targeting host genes potentially associated with various aspects of RNA virus life cycles. Afterward, HPAI H7N9 viruses were isolated and RNA interference was used to screen for host factors likely to be involved in the life cycle of HPAI H7N9. Moreover, the research entailed assessing the associations between host proteins and HPAI H7N9 proteins. Twelve key host proteins were identified: Annexin A (ANXA)2, ANXA5, adaptor related protein complex 2 subunit sigma 1 (AP2S1), adaptor related protein complex 3 subunit sigma 1 (AP3S1), ATP synthase F1 subunit alpha (ATP5A1), COPI coat complex subunit alpha (COP)A, COPG1, heat shock protein family A (Hsp70) member 1A (HSPA)1A, HSPA8, heat shock protein 90 alpha family class A member 1 (HSP90AA1), RAB11B, and RAB18. Co-immunoprecipitation revealed intricate interactions between viral proteins (hemagglutinin, matrix 1 protein, neuraminidase, nucleoprotein, polymerase basic 1, and polymerase basic 2) and these host proteins, presumably playing a crucial role in modulating the life cycle of HPAI H7N9. Notably, ANXA5, AP2S1, AP3S1, ATP5A1, HSP90A1, and RAB18, were identified as novel interactors with HPAI H7N9 proteins rather than other influenza A viruses (IAVs). These findings underscore the significance of host-viral protein interactions in shaping the dynamics of HPAI H7N9 infection, while highlighting subtle variations compared with other IAVs. Deeper understanding of these interactions holds promise to advance disease treatment and prevention strategies.

Rosmarinic acid ameliorates hypoxia/ischemia induced cognitive deficits and promotes remyelination.

Rosmarinic acid, a common ester extracted from Rosemary, Perilla frutescens, and Salvia miltiorrhiza Bunge, has been shown to have protective effects against various diseases. This is an investigation into whether rosmarinic acid can also affect the changes of white matter fibers and cognitive deficits caused by hypoxic injury. The right common carotid artery of 3-day-old rats was ligated for 2 hours. The rats were then prewarmed in a plastic container with holes in the lid, which was placed in 37°C water bath for 30 minutes. Afterwards, the rats were exposed to an atmosphere with 8% O2 and 92% N2 for 30 minutes to establish the perinatal hypoxia/ischemia injury models. The rat models were intraperitoneally injected with rosmarinic acid 20 mg/kg for 5 consecutive days. At 22 days after birth, rosmarinic acid was found to improve motor, anxiety, learning and spatial memory impairments induced by hypoxia/ischemia injury. Furthermore, rosmarinic acid promoted the proliferation of oligodendrocyte progenitor cells in the subventricular zone. After hypoxia/ischemia injury, rosmarinic acid reversed to some extent the downregulation of myelin basic protein and the loss of myelin sheath in the corpus callosum of white matter structure. Rosmarinic acid partially slowed down the expression of oligodendrocyte marker Olig2 and myelin basic protein and the increase of oligodendrocyte apoptosis marker inhibitors of DNA binding 2. These data indicate that rosmarinic acid ameliorated the cognitive dysfunction after perinatal hypoxia/ischemia injury by improving remyelination in corpus callosum. This study was approved by the Animal Experimental Ethics Committee of Xuzhou Medical University, China (approval No. 20161636721) on September 16, 2017.

Transvaginal three-dimensional color power Doppler ultrasound and cervical MVD measurement in the detection of cervical intraepithelial neoplasia.

OBJECTIVE: To explore the potential correlation between three-dimensional color power Doppler ultrasound (3D-CPA) parameters and high-grade cervical lesions and early cervical cancer microvessel density (MVD) and investigate the role of transvaginal three-dimensional power Doppler ultrasonography in the detection of cervical intraepithelial neoplasia. PATIENTS AND METHODS: Totally 90 subjects were randomly divided into three groups: the control group (n = 30, including patients with chronic cervicitis), the high-grade cervical intraepithelial neoplasia (CIN) group (n = 30, mainly CIN II-III), and the early cervical cancer group (stage Ia-IIa) (n = 30). All patients received preoperative 3D-CPA, and the cervical blood flow was graded. The cervical and intra-mass parameters including vascularization index (VI), flow index (FI), and vascularization-flow index (VFI) were measured. The immunohistochemistry of the anti-CD34 monoclonal antibody was performed for the post-operative specimens obtained from each group. The MVD of the tumors was calculated. The difference of each parameter was compared among these three groups, and the correlations between the ultrasound vascular parameters and MVD were analyzed. The high-grade CIN group was followed up for 6 months after the loop electrosurgical excision procedure (LEEP) conization surgery with 3D-CPA. RESULTS: Compared with the other two groups, the early cervical cancer group had significantly higher VI, FI, and VFI parameters (p < 0.01). Compared with the control group, all of the three parameters of the high-grade CIN group were significantly higher (p < 0.01). The MVD values increased from the control group to the high-grade CIN group, and in turn to the cervical cancer group, with significant differences between each pair (p < 0.05). MVD was positively correlated with the ultrasound parameters VI and VFI (r = 0.723, r = 0.692). There were significant differences among the three groups in terms of vascular morphology and type. However, the ultrasound parameters and vascular types were not significantly different between the postoperative CIN group and the control group. CONCLUSIONS: 3D-CPA can be used to assess blood flow in the cervix. It is particularly useful for the early diagnosis of cervical cancer and CIN and for the postoperative follow-up of CIN.

Feasibility study of using fetal DNA in maternal plasma for non-invasive prenatal diagnosis.

BACKGROUND: The discovery of the presence of fetal genetic material in maternal blood has opened up a new approach to prenatal diagnosis. One approach that has been extensively investigated over the past few decades is the isolation of fetal cells from maternal blood (Herzenberg et al. Proc Natl Acad Sci USA. 1979;76:1453-5; Bianchi et al. Proc Natl Acad Sci USA. 1990;87:3279-83; Cha et al. Prenat Diagn. 2005;25:586-91). As the fetal cells are scarce and the enrichment is of low efficiency, the technique could not be implemented in clinics. In 1997, Lo et al. (Lancet 1997;350(9076):485-7) discovered that cell-free fetal DNA is present in the plasma and serum of pregnant women. This discovery suggests that maternal plasma/serum DNA may be a useful source of material for non-invasive prenatal diagnosis. The objective of our study was to investigate the feasibility of using fetal DNA in maternal plasma for prenatal diagnosis. METHODS: Plasma DNA in 277 blood samples of 40 pregnant women at the gestational period from 5 to 40 weeks and 24 h after delivery were extracted by column separation. FQ-PCR was used to amplify the SRY sequence in 237 plasma samples of 30 pregnant women. Fluorescent PCR was used to amplify 9 short tandem repeat loci simultaneously in 40 plasma samples of 10 pregnant women, and genomic DNA samples from their husbands were amplified by the same method. RESULTS: The fetal SRY sequence could be detected from the 7th week of gestation, with a concentration that increased with progressing gestational age, attaining its highest peak before delivery. Twenty-four hours after delivery, fetal SRY sequence could not be detected in the maternal plasma. The concordance rate of the SRY sequence amplification results of plasma-free DNA, with real fetal gender was 100%. Analysis of maternal plasma samples collected during pregnancy revealed the presence of paternally inherited fetal-specific alleles. Among the 30 collected plasma samples, fetal-specific alleles were detected in 23 plasma DNA samples. The rate of positive results was 76% (23/30), and the frequency of positive results was 6/10 in early pregnancy, 8/10 in middle pregnancy, and 9/10 in late pregnancy. Short tandem repeats could not be detected from the maternal plasma 24 h after delivery. CONCLUSION: Fluorescent PCR can be used for amplification of fetal SRY sequence and STRs in maternal plasma to obtain fetal genetic information, which may have implications for non-invasive prenatal diagnosis of certain hereditary diseases.

[Detection of fetal short tandem repeat genotype in maternal plasma by multiplex fluorescent polymerase chain reaction].

OBJECTIVE: To study the feasibility of using free fetal DNA in maternal plasma for non-invasive prenatal diagnosis. METHODS: Maternal DNA extracted from plasma samples of 10 pregnant women at early pregnancy, medium pregnancy and late pregnancy and their husband's DNA isolated from whole blood samples were used to detect genotype by multiplex fluorescent PCR at nine different polymorphic short tandem repeat(STR) loci. RESULTS: Fetus-specific alleles were found in maternal plasma samples studied. By the application of these polymorphic short repeat sequences, every pregnant women/husband pair was informative in at least three of nine loci. Paternally inherited fetal alleles were detected in 23 of 30 plasma samples. They are 6/10 cases in early pregnancy, 8/10 cases in middle pregnancy and 9/10 cases in late pregnancy respectively. CONCLUSION: Fluorescent multiplex PCR can be used for amplification of male and female fetal STRs in maternal plasma to obtain genetic information, which may have implication for non-invasive prenatal diagnosis of certain hereditary diseases independent of the fetal sex.