Objective To analyze the antiviral effect of molnupiravir against influenza virus in vitro and in vivo.Methods The antiviral ability of molnupiravir against influenza virus strain H1N1 PR8 was detected in vitro and in vivo.H uman non-small cell lung cancer cell line(A549 cells)was used as an in vitro model of PR8 infection.The antiviral effects of molnupiravir on virus replication,protein synthesis,and viral particle formation were analyzed using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),Western blot(WB)assay,and plaque assay.PR8 nose-dripping infected C57BL/6 female mice were used as an in vivo infection model.The antivi-ral ability of molnupiravir was evaluated by detecting viral load,pathological changes,and immunohistochemistry in the control group,administration group,inoculation group,and inoculation-administration group.Results In vitro test results showed that molnupiravir had no significant inhibitory effect on influenza virus replication,protein syn-thesis,and virus particle formation(all P＞0.05).In vivo test results showed that compared with mice infected with H1N1 alone,the viral load in the lung tissue of mice treated with molnupiravir declined significantly,and the extent of pathological changes was milder.Immunohistochemical detection showed a significant weakening in nuclear protein(NP)antigen signal,and the expression levels of interferon(IFN)-α,interleukin(IL)-4,and IL-6 in the lungs were lower(all P＜0.01).Conclusion As a precursor with antiviral activity,molnupiravir can not exert anti-viral activity in lung-derived cells cultured in vitro,but can be transformed into an active form in the host,which significantly decreases the ability of H1N1 influenza virus to proliferate in the lungs and thereby alleviates the da-mage of virus to lung tissue.

Objective To analyze the antiviral effect of molnupiravir against influenza virus in vitro and in vivo.Methods The antiviral ability of molnupiravir against influenza virus strain H1N1 PR8 was detected in vitro and in vivo.H uman non-small cell lung cancer cell line(A549 cells)was used as an in vitro model of PR8 infection.The antiviral effects of molnupiravir on virus replication,protein synthesis,and viral particle formation were analyzed using real-time fluorescence quantitative polymerase chain reaction(qRT-PCR),Western blot(WB)assay,and plaque assay.PR8 nose-dripping infected C57BL/6 female mice were used as an in vivo infection model.The antivi-ral ability of molnupiravir was evaluated by detecting viral load,pathological changes,and immunohistochemistry in the control group,administration group,inoculation group,and inoculation-administration group.Results In vitro test results showed that molnupiravir had no significant inhibitory effect on influenza virus replication,protein syn-thesis,and virus particle formation(all P＞0.05).In vivo test results showed that compared with mice infected with H1N1 alone,the viral load in the lung tissue of mice treated with molnupiravir declined significantly,and the extent of pathological changes was milder.Immunohistochemical detection showed a significant weakening in nuclear protein(NP)antigen signal,and the expression levels of interferon(IFN)-α,interleukin(IL)-4,and IL-6 in the lungs were lower(all P＜0.01).Conclusion As a precursor with antiviral activity,molnupiravir can not exert anti-viral activity in lung-derived cells cultured in vitro,but can be transformed into an active form in the host,which significantly decreases the ability of H1N1 influenza virus to proliferate in the lungs and thereby alleviates the da-mage of virus to lung tissue.

Objective·To analyze the diversity and composition of the maternal gut microbiota and vaginal microbiota in late pregnancy,neonatal meconium microbiota and vernix caseosa microbiota,and analyze the similarities,differences and correlations.Methods·This is a prospective study.Maternal stool samples and vaginal swabs in late-pregnancy,and neonatal meconium samples were collected from 11 mother-infant pairs at Xinhua Hospital,Shanghai Jiao Tong University School of Medicine from August to November 2018;the vernix caseosa from three sites(forehead,axilla,and inguinal crease)and meconium samples were collected from 14 healthy newborns at International Peace Maternity and Child Health Hospital,Shanghai Jiao Tong University School of Medicine in December 2018.All births were vaginal deliveries.The 16S rRNA gene V3?V4 region sequencing was used.The diversity,composition and similarities/differences of the maternal gut microbiota,the vaginal microbiota,and the neonatal meconium microbiota from the 11 mother-infant pairs,as well as the neonatal vernix caseosa microbiota and the meconium microbiota from the 14 newborns were analyzed.Results·The number of operational taxonomic units(OTUs),ACE index,Chao1 index,and Shannon index of maternal gut microbiota were all higher than those of vaginal microbiota;the ACE indices and the Chao1 indices of the vernix caseosa microbiota at three sites were all higher than those of meconium microbiota(P<0.01).The β diversity varied among the maternal gut microbiota,vaginal microbiota,and neonatal meconium microbiota(P<0.01).The β diversity of neonatal vernix caseosa microbiota from three sites(forehead,axilla,and inguinal crease)was similar,but different from meconium microbiota(P<0.01).At the phylum level,the dominant bacteria were Firmicutes(52.76%)and Bacteroidetes(41.67%)in the maternal gut microbiota,Firmicutes(74.36%)and Actinobacteria(21.25%)in the maternal vaginal microbiota,and Firmicutes(84.22%)and Proteobacteria(8.80%)in the neonatal vernix caseosa microbiota.The dominant bacterium in the neonatal meconium was Proteobacteria in the two batches of samples(81.11%and 88.72%,respectively).At the genus level,the dominant bacteria were Bacteroides(35.42%)and Faecalibacterium(10.12%)in the maternal gut microbiota,Lactobacillus(69.10%)and Bifidobacterium(11.30%)in the vaginal microbiota,and Lactobacillus(79.81%)and Pseudomonas(3.23%)in the vernix caseosa microbiota.The dominant bacterium in the neonatal meconium was Escherichia in the two batches of samples(55.21%and 31.18%,respectively).Conclusion·The α diversity of maternal gut microbiota is higher than that of vaginal microbiota and neonatal meconium microbiota,and it is higher in neonatal vernix caseosa than that in meconium microbiota.The Firmicutes is the predominant phylum in the maternal late-pregnancy gut microbiota,vaginal microbiota,and neonatal vernix microbiota.Lactobacillus is the predominant genus in both maternal vaginal and neonatal vernix caseosa microbiota.Proteobacteria in phylum and Escherichia in genus are predominant in meconium microbiota.The microbiota composition is similar in vernix caseosa at different body sites,but there are differences between the vernix caseosa microbiota and meconium microbiota.

Objective:To evaluate the effect of electroacupuncture preconditioning on cellular immune function in the patients undergoing thyroid surgery.Methods:Sixty-six patients of either sex, aged 30-55 yr, with body mass index of 18.5-23.9 kg/m 2, of American Society of Anesthesiologists Physical Status classification ⅠorⅡ, were selected and divided into electroacupuncture preconditioning group (group D) and control group (group C) by a random number table method, with 33 cases in each group. Patients in both groups received combined intravenous-inhalational anesthesia. Before anesthesia induction, both Hegu and Neiguan acupoints were stimulated for 30 min using a continuous wave at a frequency of 2/100 Hz, and the stimulation intensity was subjected to patient tolerance in group D. Group C received sham electroacupuncture for 30 min without stimulation. Venous blood samples were collected at 30 min before anesthesia induction (T 0) and 12, 24 and 48 h after anesthesia (T 1-3). The percentages of T lymphocyte subsets (CD3 +, CD4 +, CD8 +, CD4 + /CD8 + ) and natural killer cells and concentrations of interleukin-6 and tumor necrosis factor-alpha in the serum were measured by flow cytometry. Results:Compared with group C, the percentage of CD3 +, CD4 + and natural killer cells and ratio of CD4 + /CD8 + were significantly increased at T 1, 2, and the concentrations of interleukin-6 and tumor necrosis factor-alpha in the serum were decreased at T 1-3 in group D ( P<0.05). Conclusions:Electroacupuncture preconditioning can improve cellular immune function in the patients undergoing thyroid surgery.

Objective To investigate the correlation between the number of dissected lymph nodes ( LNs) and the prognosis of patients with node?negative Siewert type ⅡAEG. Methods 248 patients with Siewert type Ⅱ AEG treated in our hospital between January 1998 and December 2008 were retrospectively assessed. All cases underwent left transthoracic subtotal esophagogastrectomy with conventional two?field lymphadenectomy, and were histopathologically proved to be without lymph node involvement. The prognostic impact of the number of dissected LNs was analyzed. Results The overall median survival time and the 1?, 3?, and 5?year overall survival rates were 64 months, 80. 4%, 60. 8% and 51. 0%, respectively. Cox regression showed that the number of dissected LNs and the depth of tumor invasion were independent prognostic factors. Patients with a high number of negative LNs had better overall survival than patients with a low number of negative LNs ( P<0.05) . The patients had better long?term survival outcomes with more than 10 LN dissected for cases with pT1 tumor ( P<0.001) , and so did those with more than 15 LN dissected for cases with pT2?3 tumor (P=0.003, 0.018, respectively). Conclusion The number of negative lymph nodes and the depth of tumor invasion are independent prognostic factors for node?negative Siewert type ⅡAEG, and adequate lymph node dissection can improve the long?term survival.

Objective To investigate the correlation between the number of dissected lymph nodes ( LNs) and the prognosis of patients with node?negative Siewert type ⅡAEG. Methods 248 patients with Siewert type Ⅱ AEG treated in our hospital between January 1998 and December 2008 were retrospectively assessed. All cases underwent left transthoracic subtotal esophagogastrectomy with conventional two?field lymphadenectomy, and were histopathologically proved to be without lymph node involvement. The prognostic impact of the number of dissected LNs was analyzed. Results The overall median survival time and the 1?, 3?, and 5?year overall survival rates were 64 months, 80. 4%, 60. 8% and 51. 0%, respectively. Cox regression showed that the number of dissected LNs and the depth of tumor invasion were independent prognostic factors. Patients with a high number of negative LNs had better overall survival than patients with a low number of negative LNs ( P<0.05) . The patients had better long?term survival outcomes with more than 10 LN dissected for cases with pT1 tumor ( P<0.001) , and so did those with more than 15 LN dissected for cases with pT2?3 tumor (P=0.003, 0.018, respectively). Conclusion The number of negative lymph nodes and the depth of tumor invasion are independent prognostic factors for node?negative Siewert type ⅡAEG, and adequate lymph node dissection can improve the long?term survival.