Clinical value of anal swab positive in COVID-19 patients / 中华微生物学和免疫学杂志

Objective:To investigate the value of anal swabs positive for 2019-nCoV in patients with COVID-19 and the clinical features of the patients.Methods:Throat swabs, sputum and blood samples, and anal swabs were collected from 104 patients with COVID-19 at admission to test for 2019-nCoV nucleic acid. Clinical characteristics and hematological indexes were compared between viral nucleic acid-positive and -negative groups of different sample types. Fifteen patients whose anal swabs were positive for viral nucleic acid were selected to analyze the length of time before the nucleic acid turned negative in different specimens.Results:Compared with the patients having negative anal swab test results, those having positive test results showed decreased lymphocytes, increased lactate dehydrogenase (LDH) and high-sensitivity C-reactive protein (HsCRP), and higher incidence of severe COVID-19. The levels of HsCRP and IL-6 and the incidence of severe COVID-19 were significantly higher in patients with positive throat swab test results than in those with negative results. No significant difference in hematological indexes or the proportion of severe cases was detected between the patients with positive and negative sputum test results. Only 1.92% of the patients had positive blood test results, but all of them were severe patients. The positive rate of sputum test was the highest, which was 46.15%. Patients with positive results of both throat and anal swab test had significantly decreased lymphocytes, increased HsCRP and IL-6 levels, and higher incidence of critical COVID-19. It took longer time for patients to have negative anal swab and sputum test results. Moreover, it should be noticed that the viral nucleic acid in sputum might become positive again after it turned negative.Conclusions:Patients with positive anal swab test results had reduced lymphocytes, enhanced inflammatory response and higher incidence of severe COVID-19, suggesting that a positive anal swab test might be an indicator of severe COVID-19. Moreover, the time of 2019-nCoV nucleic acid turning negative in anal swabs was longer than that in throat swabs. The combined detection of throat swabs and anal swabs would help to predict the occurrence of severe COVID-19.

Psoralen inhibits RAW264.7 differentiation into osteoclasts and bone resorption by regulating CD4+T cell differentiation / 中国中药杂志

This paper aimed to investigate whether psoralen inhibits the differentiation and bone resorption by regulating CD4+T cell differentiation in RANKL-induced osteoclastogenesis in RAW264.7 cells, and elucidate its mechanism for osteoporosis. CD4+T cells were isolated from spleen cells of Balb/c mice by immunomagnetic separation method. The cells were divided into blank control group and psoralen group. The cells were cultured in 24-well plates and cultured for 3 days, and then they were collected for co-culture experiments after 4 days. Co-culture experiments were divided into RAW264.7 cell group, psoralen+RAW264.7 cell group, without psoralen treatment of CD4+T cells+RAW264.7 cell group, psoralen treatment of CD4+T cells+RAW264.7 cell group. After 5 days of co-culture, TRAP staining was used to detect the number of osteoclasts, and after 8 days of co-culture, bone resorption was evaluated by toluidine blue staining. The expressions of RORγt, Foxp3, IL-17, TNF-α, TGF-β and IL-10 in CD4+T cells and osteoclast differentiation-related genes MMP-9, TRAP and Cat-K were detected by Real-time polymerase chain reaction (RT-PCR); ELISA kit was used to detect IL-17, TNF-α, TGF-β and IL-10 and other cytokines levels. Our data confirmed that the psoralen significantly promoted the expression of Foxp3, TGF-β and IL-10 in CD4+T, and inhibited the expression of RORγt, IL-17 and TNF-α in CD4+T, the CD4+T cells without treatment by psoralen can significantly promote RANKL-induced differentiation of RAW264.7 to osteoclasts, and psoralen treatment of CD4+T can significantly inhibit RANKL-induced RAW264.7 osteoclast differentiation and bone resorption. Taken together, psoralen inhibits the differentiation and bone resorption of RAW264.7 into osteoclasts by promoting the development of CD4+ CD25+ Treg/Th17 balance in CD4+T cells to CD4+CD25+T.

Application of osteopontin and Stathmin-1 expression in the evaluation of prostate cancer recurrence and survival prognosis / 中国医师进修杂志

Objective To explore the application of osteopontin (OPN) and Stathmin- 1 expression in the evaluation of prostate cancer recurrence and survival prognosis. Methods One hundred patients with prostate cancer were selected from June 2014 to June 2016. The expressions of OPN and Stathmin-1 in prostate cancer tissue were detected by immunohistochemical staining SP method, and its correlation with clinicopathological features was analyzed. Results The OPN positive expression in prostate cancer tissue was in 76 cases (76.00%, 76/100), the Stathmin-1 positive expression in prostate cancer tissue was in 79 cases (79.00%, 79/100), and both the OPN and Stathmin-1 positive expression in prostate cancer tissue was in 60 cases (60.00% , 60/100). Spearman correlation analysis result showed that the expression of OPN in prostate cancer tissue had positive correlation with Stathmin-1 (r = 0.491, P＜0.01). The positive expressions of OPN and Stathmin-1 in prostate cancer tissue were relative with Gleason score, T staging and bone metastasis (P ＜ 0.05), but were not relative with recurrence (P＞0.05). The survival time in OPN positive expression, Stathmin-1 positive expression and both OPN/Stathmin-1 positive expression was (18.45 ± 2.03), (17.95 ± 2.01) and (15.24 ± 1.72) months;the survival time in OPN negative expression, Stathmin-1 negative expression and OPN/ Stathmin-1 single positive or both negative was (24.67 ± 2.62), (23.79 ± 2.58) and (26.68 ± 2.72) months, and there was statistical difference (P＜0.05). Conclusions The OPN and Stathmin-1 expressions in prostate cancer tissue are high, and show relations with Gleason score, bone metastases and T staging, and no correlation with recurrence. But OPN and Stathmin-1 can provide reference for survival in patients with prostate cancer prognosis assessment.

The 2A protease of enterovirus 71 cleaves nup62 to inhibit nuclear transport / 病毒学报

To study the impact of the enterovirus 71(EV71) on the nuclear transport mechanism,The pGFP-NLS vector with nuclear location signal(NLS) was constructed, RD cells transfected by the pGFP-NLS vector were inoculated with the EV71 or cotransfected by EV71-2A vector. The results showed that GFP protein with NLS was expressed in the cytoplasm due to the inhibition of nuclear transport. In order to further study the mechanism of the EV71 to prevent nuclear transport,Nup62 was detected by Western blotting after RD cells were infected with EV71 or transfected by EV71-2A vector. The results showed that decreased expression of Nup62 could be detected after infection with EV71 and transfection by EV71-2A vector. This study demonstrates that the cleavage of Nup62 by EV71 2A protease may be the mechanism of nuclear transport inhibition.

Degradation of 14-3-3beta appeared in apoptosis cell induced by PrP106-126 polypeptide / 病毒学报

To investigate changes of 14-3-3beta from apoptosis induced by PrP106-126 polypeptide, HeLa cell was incubated with PrP106-126 for 4h or 8h. Nucleus changes and the expression of PARP were detected differently by Hoechst staining and Western blotting. Expressing of protein and mRNA from 14-3-3beta was determined by Western blotting and Real-time PCR. The results show that typical nucleus pyknosis and chip of apoptosis and degradation of PARP were induced by PrP106-126 peptide in HeLa cells. Degradation of 14-3-3beta appeared in apoptosis groups induced by PrP106-126 peptide. However, 14-3-3beta mRNA did not display any changes in apoptosis groups. This study indicated that degradation of antiapoptosis protein 143-3beta induced by PrP106-126 peptide may be one of pathogenesis mechanism of prion disease.

Primary establishment of an alphaLISA assay for detection of HAV IgM / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop an AlphaLISA method for detection of antibody of Hepatitis A virus.</p><p><b>METHODS</b>After hepatitis A virus antigen was concentrated and biotinylated, optimal biotinylated HAV antigen, donor bead, acceptor bead concentration have been explored and determined by both dot-blotting and AlphaLISA methods. 97 samples including 23 serums from patients HAV infected and 70 serums from blood donors have been detected by new AlphaLISA method.</p><p><b>RESULTS</b>The sensitivity and specificity for anti-HAV IgM were 78% and 98.5%, the positive and negative predictive value were 95% and 93%.</p><p><b>CONCLUSION</b>A homogeneous and fast AlphaLISA assay for detection of HAV IgM has been established by preliminary verification on samples.</p>

Human enterovirus 71 (EV71) resisted type I interferon induced antiviral effect / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To investigate human enterovirus 71 (EV71) resistance to type I interferon induced antiviral effect.</p><p><b>METHODS</b>After type I interferons (alpha, beta) were incubated with HeLa cells, recombinant type I herpes simple virus (HSV-1) with green fluorescent protein (GFP) was inoculated onto the HeLa cells. HSV-1 proliferation was observed by GFP expression and PCR. After EV71 was inoculated onto HeLa cells incubated with the same quantity of interferon, proliferation of EV71 were detected by RT-PCR of 2A gene.</p><p><b>RESULTS</b>Recombinant HSV-1 GFP expression and viral DNA replication obviously decreased in HeLa cells incubated with type I interferon (alpha, beta). However, EV71 effectively proliferated in the interferon irritated HeLa cell by RT-PCR.</p><p><b>CONCLUSION</b>HeLa cell irritated by type I interferon (alpha, beta) produced antiviral substance that inhibits HSV-1 proliferation. EV71 resisted the antiviral substance induced by type I interferon and could significantly replicate in the HeLa cells.</p>