ObjectiveTo investigate the therapeutic effect of Dayuanyin on acute lung injury induced by H1N1 infection and decipher the potential mechanism. MethodThe constituents in Dayuanyin were analyzed by ultra-high performance liquid chromatography-quadrupole-exactive orbitrap mass spectrometry （UHPLC-Q-Exactive Orbitrap MS）. Forty-eight female BALB/c mice were randomized into normal， model， oseltamivir （19.5 mg·kg^-1）， and low-， medium-， and high-dose （2.73， 5.46， 10.92 g·kg^-1） Dayuanyin groups. The normal and model groups were administrated with deionized water by gavage， and the other groups were administrated with the corresponding drugs by gavage. On day 3 of drug administration， the normal group received nasal inhalation of normal saline， and the other groups were inoculated intranasally with A/RP/8/34 （H1N1） for the modeling of influenza virus infection. Mice were administrated with drugs continuously for 7 days and weighed daily. Sampling was performed 12 h after the last administration， and the lung tissue was weighed to calculate the lung index. Hematoxylin-eosin staining was performed to observe the pathological and morphological changes of the lung tissue and bronchi. The cytometric bead array （CBA） was used to measure the serum levels of interferon-gamma （IFN-γ）， C-X-C motif ligand 1 （CXCL1）， tumor necrosis factor-alpha （TNF-α）， chemokine ligand 2 （CCL2）， interleukin-12p70 （IL-12p70）， chemokine ligand 5 （CCL5）， interleukin-1β （IL-1β）， chemokine （C-X-C motif） ligand 10 （CXCL10）， granulocyte-macrophage colony-stimulating factor （GM-CSF）， interleukin-10 （IL-10）， interferon-beta （IFN-β）， interferon-alpha （IFN-α）， and interleukin-6 （IL-6）. According to the results of mass spectrometry and network pharmacology， we analyzed the mechanism of Dayuanyin in treating acute lung injury caused by H1N1. The protein levels of extracellular signal-regulated kinase 1/2 （ERK1/2）， p38 mitogen-activated protein kinase （p38 MAPK）， nuclear factor-kappa B （NF-κB）， and their phosphorylated forms were determined by Western blot. The mRNA levels of myeloid differentiation factor 88 （MyD88）， Toll-like receptor 3 （TLR3）， Toll-like receptor 7 （TLR7）， and Toll-like receptor 8 （TLR8） in the lung tissue were measured by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. ResultA total of 57 compounds， including paeoniflorin and baicalein， were detected in Dayuanyin. Compared with the normal group， the model group showed decreased body weight （P<0.01）， lung edema and hemorrhage， increased lung index （P<0.01）， and elevated levels of IFN-γ， IL-12p70， CCL5， IL-1β， CXCL10， GM-CSF， IFN-β， and IL-6 （P<0.01）. Compared with the model group， Dayuanyin attenuated alveolar wall thickening， capillary congestion， and immune cell infiltration， reduced the alterations in body weight and lung index （P<0.01）， and down-regulated the protein levels of IFN-γ， IL-12p70， CCL5， IL-1β， CXCL10， GM-CSF， IFN-β， and IL-6 （P<0.01）. A total of 57 key genes were predicted by network pharmacological analysis， of which the MAPK signaling pathway was the main target signaling pathway. Compared with the normal group， the model group showed up-regulation in the protein levels of phosphorylation （p）-ERK1/2， p-p38 MAPK， and p-NF-κB （P<0.01） and the mRNA levels of TLR7， TLR8， MyD88， and TLR3 （P<0.05， P<0.01）. Compared with the model group， Dayuanyin lowered the phosphorylation levels of ERK1/2， p38 MAPK， and NF-κB p65 in a dose-dependent manner （P<0.01） and down-regulated the mRNA levels of TLR3， TLR7， TLR8， and MyD88 （P<0.01）. ConclusionDayuanyin can prevent and control H1N1 infection-induced acute lung injury by inhibiting the TLR/MAPK/NF-κB signaling pathway.

Objective:To investigate the epidemiological and clinical characteristics of human parainfluenza viruses (HPIVs) infection among hospitalized children with community-acquired pneumonia (CAP) in China, and provide basic data for diagnosis, treatment and prevention of HPIVs infection.Methods:From November 2014 to February 2020, 5 448 hospitalized children with CAP were enrolled in 14 hospitals in 11 provinces and municipalities directly under the Central Government in southern China and northern China. Nasopharyngeal aspirates or throat swabs were collected, and the nucleic acids of 18 types respiratory viruses including HPIV1-4 were screened by suspension array technology. Demographic data and clinical information were collected for statistical analysis.Results:The total detection rate of HPIVs in 5 448 children with CAP was 8.83% (481/5 448), and the detection rate in males was higher than that in females (62.79% vs. 37.21%; χ2=0.000, P=0.992). The detection rate of HPIVs in 1~< 3 years age group was higher than that in other age groups, and the difference was statistically significant ( χ2=61.893, P<0.001). The detection rate of HPIVs in the northern region was higher than that in the southern region (9.02% vs 8.65%), but the difference was not statistically significant ( χ2=0.239, P=0.625). The prevalence of HPIV1-4 in northern and southern China was not completely same. HPIV1 was mainly prevalent in autumn in both northern and southern regions. HPIV2 was prevalent in summer in northern China, and the detection rate was low in southern China. HPIV3 reached its peak in both spring and summer in both northern and southern China, but its duration was longer in southern China than in northern China. HPIV4 is mainly popular in autumn in both southern China and northern China. Among 481 children infected with HPIVs, 58.42% (281/481) were infected with HPIV alone, and the main clinical manifestations were cough (90.75%) and fever (68.68%). Out of the HPIV-positive cases, 42.62% (205/481) were co-infected with another type of HPIV or a different virus, while 11.43% (55/481) had co-infections with two or more different viruses. HPIV3 was the most common type of co-infection with other viruses. HPIV3 infection accounted for the largest proportion (76.80%) in 47 HPIVs-positive children with severe pneumonia. Conclusions:HPIVs is one of the most important pathogens causing CAP in children in China, and children under 3 years of age are the main populations of HPIVs infection. The prevalence characteristics of all types of HPIVs in children in the north and south are not completely same. HPIV3 is the dominant type of HPIV infections and causes more severe diseases.

ObjectiveTo explore the correlation of sleep quality with negative life events and coping styles in adolescents based on structural equation modeling， and to provide references for improving the adolescents' sleep quality. MethodsFrom December 2021 to May 2022， a total of 767 junior middle school students from three schools in Chongqing were enrolled， and assessed using Pittsburgh Sleep Quality Index （PSQI）， Adolescent Self-rating Life Events Checklist （ASLEC） and Simplified Coping Style Questionnaire （SCSQ）. Then the structural equation model was applied to discuss the correlation of sleep quality with negative life events and coping styles in adolescents. ResultsA total of 222 adolescents （28.94%） were found to have sleep disorders. PSQI score was positively correlated with ASLEC score and negative coping dimension score of SCSQ （r=0.612， 0.590， P<0.01）， and negatively correlated with positive coping dimension score of SCSQ （r=-0.435， P<0.01）. The structural equation model of the relationship between negative life events， coping styles and sleep quality denoted that negative life events exhibited both direct and indirect positive effects on sleep quality （β=0.448， 0.322， P<0.05）， positive coping style had direct negative effects on sleep quality （β=-0.368， P<0.05）， and negative coping style had direct positive effects on sleep quality （β=0.442， P<0.05）. ConclusionNegative life events and negative coping style cause adverse effects on adolescents' sleep quality， while positive coping style exerts positive effects on adolescents' sleep quality.

Objective:To analyze the influence of factors before initiation of extracorporeal cardiopulmonary resuscitation (ECPR) on the prognosis of patients, so as to explore the intervention timing and improvement strategy of ECPR.Methods:A retrospective analysis was performed on 29 patients who underwent ECPR in the First Affiliated Hospital of Hunan Normal University (Hunan people's Hospital)from July 2018 to April 2021. Patients were divided into the survival group ( n = 13) and death group ( n = 16) according to whether they survived at discharge. The duration of conventional cardiopulmonary resuscitation (CCPR), initial heart rate before ECPR, the ratio of out-of-hospital cardiac arrest (OHCA) and in-hospital cardiac arrest (IHCA), and the ratio of transported cases outside the hospital were compared between the two groups. According to different CCPR time, the patients were divided into the ≤45 min group, 45-60 min group and >60 min group to compare the hospital survival and sustained return of spontaneous circulation (ROSC) rate . According to the location of cardiac arrest, the patients from emergency department and other department were divided to compare the survival of IHCA. Results:The total survival rate was 44.83%, the average duration of extracorporeal membrane oxygenation (ECMO) was 114 (33.5, 142.5) h, and the average duration of CCPR time was 60 (44.5, 80) min. The duration of ECMO was longer in the survival group than in the death group ( P = 0.001). The duration of CCPR (the time from CPR to ECMO) in the survival group was significantly shorter than that in the death group ( P = 0.010). Patients with defibrillatory rhythm had higher hospital survival rate ( P = 0.010). OHCA patients had higher mortality than IHCA patients ( P = 0.020). Mortality of patients transferred from other hospitals was higher ( P = 0.025). Hospital survival and ROSC decreased in turn by CCPR duration ≤ 45 min, 45-60 min, and > 60 min ( P = 0.001). The location of CA occurrence had no impact on the hospital survival rate of IHCA patients ( P=0.54). Conclusions:Hospital survival of ECPR is higher than that of CCPR. ECPR is effective for refractory cardiac arrest. The prognosis of ECPR is significantly related to the duration of CCPR, initial heart rate, and location of CA. Education and team training should be strengthened to improve the survival rate of ECPR.

Objective To analyze the clinical features of patients with coronavirus disease 2019 (COVID-19) in Shanghai and to investigate the risk factors for disease progression to severe cases. Methods The clinical data of 292 adult patients with COVID-19 hospitalized in Shanghai Public Health Clinical Center from January 20, 2020 to February 10, 2020 were retrospectively analyzed, including 21 severe patients and 271 mild patients. The demographic characteristics, epidemiological history, history of underlying diseases and laboratory examinations were compared between the two groups. Measurement data were compared using t test or Mann-Whitney U test. The count data were compared using hi-square test. The binary logistic regression equation was used to analyze the risk factors for the progression of patients to severe cases. Results Among the 292 patients, 21 were severe cases with the rate of 7.2% (21/292). One patient died, and the mortality rate was 4.8% in severe patients. The severe patients aged (65.0±15.7) years old, 19 (90.5%) were male, 11 (52.4%) had underlying diseases, 7 (33.3%) had close relatives diagnosed with COVID-19. The mild patients aged (48.7±15.7) years old, 135 (49.8%) were male, 74 (27.3%) had underlying diseases, 36 (13.3%) had close relatives diagnosed with COVID-19. The differences between two groups were all significant statistically ( t =-4.730, χ 2 =12.930, 5.938 and 4.744, respectively, all P <0.05). Compared with the mild patients, the levels of absolute numbers of neutrophils, alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, creatinine, serum cystatin C, C reactive protein (CRP), procalcitonin , D -dimer, pro-B-type natriuretic peptide (proBNP), serum myoglobin, creatine kinase (CK), creatine kinase isoenzyme (CK-MB), serum troponin I (cTnI) in severe patients were all significantly higher ( U =2 091.5, 1 928.0, 1 215.5, 729.0, 1 580.5, 1 375.5, 917.5, 789.5, 1 209.0, 1 434.0, 638.0, 964.5, 1 258.0 and 1 747.5, respectively, all P <0.05), while the levels of lymphocyte count, albumin, transferrin, CD3 + T lymphocyte count, CD8 + T lymphocyte count and CD4 + T lymphocyte count in severe patients were all significantly lower ( U =1 263.5, t =4.716, U =1 214.0, 962.0, 1 167.5 and 988.0, respectively, all P <0.05). Further logistic regression analysis showed that the albumin (odds ratio ( OR )=0.806, 95% CI 0.675-0.961), CRP ( OR =1.016, 95% CI 1.000-1.032), serum myoglobin ( OR =1.010, 95% CI 1.004-1.016), CD3 + T lymphocyte count ( OR =0.996, 95% CI 0.991-1.000) and CD8 + T lymphocyte count ( OR =1.006, 95% CI 1.001-1.010) at admission were independent risk factors for the progression of COVID-19 patients to severe illness (all P <0.05). Conclusions Severe cases of patients with COVID-19 in Shanghai are predominantly elderly men with underlying diseases. Albumin, CRP, serum myoglobin, CD3 + T lymphocyte count and CD8 + T lymphocyte count could be used as early warning indicators for severe cases, which deserve more clinical attention.

The ongoing pandemic of Coronavirus disease 19 (COVID-19) is caused by a newly discovered β Coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). How long the adaptive immunity triggered by SARS-CoV-2 can last is of critical clinical relevance in assessing the probability of second infection and efficacy of vaccination. Here we examined, using ELISA, the IgG antibodies in serum specimens collected from 17 COVID-19 patients at 6-7 months after diagnosis and the results were compared to those from cases investigated 2 weeks to 2 months post-infection. All samples were positive for IgGs against the S- and N-proteins of SARS-CoV-2. Notably, 14 samples available at 6-7 months post-infection all showed significant neutralizing activities in a pseudovirus assay, with no difference in blocking the cell-entry of the 614D and 614G variants of SARS-CoV-2. Furthermore, in 10 blood samples from cases at 6-7 months post-infection used for memory T-cell tests, we found that interferon γ-producing CD4
Adaptive Immunity/physiology* ; Adult ; Aged ; Antibodies, Neutralizing/blood* ; COVID-19/immunology* ; Cohort Studies ; Female ; Humans ; Immunoglobulin G/blood* ; Male ; Middle Aged ; SARS-CoV-2/immunology* ; T-Lymphocytes/physiology* ; Time Factors ; Viral Proteins/immunology*

Objective To investigate the predominant genotypes and epidemiological characteristics of human adenovirus (HAdV) in pediatric community-acquired pneumonia (CAP) in China.Methods This was a repeated cross sectional study.Between November 2014 and November 2016,nasopharyngeal aspirates (NPAs) or throat swabs from each hospitalized pediatric patients diagnosed as CAP in 12 hospitals in Northern and Southern China were collected.Respiratory specimens were screened for 18 respiratory viruses including HAdV by using Luminex xTAG RVP Fast V2 multiplex Assay.Typing of HAdV and analysis for the epidemiological characteristic of HAdV were performed.Results (1) A total of 2 723 hospitalized pediatric patients with CAP were enrolled in this study and 156 (5.7％,156/2 723) respiratory specimens were positive for HAdV,and 74 (6.6％,74/1 128) and 82 (5.1％,82/1 595) were in Northern and Southern China,respectively.There was no significant difference in the positive detection rate between the Northern and Southern China.(2) In Northern China,the HAdV positive rate of children at the age of ＜6 months,6 months-＜1 years,1-＜3 years,3-＜5 years and ≥5 years was 5.9％(6/101),6.7％(7/104),10.3％(34/331),4.1％(11/266) and 4.9％(16/326),respectively,and the incidence of HAdV infection peaked in children aged 1-3 years (x2=11.511,P=0.021).While in Southern China the HAdV positive rate of children at the age of ＜6 months,6 months-＜1 years,1-＜3 years,3-＜5 years and ≥5 years was 2.2％ (7/312),4.6％ (12/259),6.3％ (31/494),7.3％ (18/245) and 4.9％(14/285),respectively.There was no significant difference in the positive detection rate among age groups.(3) In 2015,the highest detection rate of HAdV in northern China was 12.5％ (25/200) in winter,and in Southern China was 6.7％ (35/525) in spring and 5.3％ (19/357) in summer.(4) In 108 cases of HAdV positive specimens typing was done and 80 in cases classification was successfully performed.Totally 7 genotypes of HAdV,including HAdV-3 (n=32),HAdV-7 (n=9),HAdV-1 (n=12),HAdV-2 (n=15),HAdV-5 (n=10),HAdV-6 (n=1) and HAdV-4 (n=1),were detected.The predominant HAdV genotypes were HAdV-3 (30.8％,8/26) and HAdV-7 (26.9％,7/26) in Northern China,while HAdV-3 (44.4％,24/54) and HAdV-2 (22.2％,12/54) were the most prevalent genotypes in Southern China.Conclusions HAdV is an important viral pathogen in pediatric CAP.The predominant HAdV genotypes and peak seasons of HAdV infections were different between Northern and Southern China.The predominant HAdV genotypes were HAdV-3 and HAdV-7 in Northern China,while HAdV-3 and HAdV-2 in Southern China.The peak season of HAdV infections was winter in Northern China.However,HAdV infections are more common in spring and summer in Southern China.

Objective To study any changes in the surface electromyography of the paraspinal muscles before and after surgery in patients with degenerative lumbar kyphosis.Methods Thirty patients with lumbar degenerative kyphosis (LDK) were enrolled and underwent pedicle subtraction osteotomy.Surface electromyograms were recorded before the operation and 3 months afterward to observe the myoelectric activity of the paraspinal muscles.The electromyograms were also compared with those of healthy counterparts.Results The preoperative average EMG (AEMG) values in the LDK group in static standing were significantly higher than those of the control group.After the operation the values had decreased significantly,but were still significantly higher than in the control group.The preoperative AEMG values of the LDK group from the L1-2 segment during torso stretching were significantly lower than those of the control group.They increased significantly after the operation and were no longer significantly different from the control groups' readings.The pre-and post-operative AEMG values from the L4-5 segment were not significantly different and remained significantly lower than in the control group.During lifting and holding the mean power frequency slopes of the LDK group were significantly lower than those of the controls both before and after the operation,though the postoperative slopes were significantly higher than the preoperative ones.Conclusion Patients with lumbar degenerative kyphosis have obvious paraspinal muscle weakness and severe fatigue.Early pedicle subtraction osteotomy can significantly improve the functional status of the paravertebral muscles in the short term.

Objective To explore the relationship of adolescents internalizing problem behavior with genetic and environmental factors. Methods The internalizing problem behavior of 74 twin pairs from Chongqing(aged from 12 to18 years) were investigated using The Youth Self-Report (YSR). The family adaptability and cohesion evaluationscale 2nd edition Chinese version (FACES Ⅱ-CV), Family Stresses Questionnaire (FSQ), the Parenting Styles and Dimensions Questionnaire (PSDQ), Stressful Life Event (SLE) and Simplified Coping Style Questionnaire (SCSQ ) were applied to assess their environment factors and coping style. The blood sample was collected to verify zygosity of twins. Structural equation modeling (SEM)was performed to evaluate the relationship of adolescents internalizing problem behavior with the additive genetic factors(A),shared environment factors(C)and individual specific environmental factors(E). Results The scores of PSDQ and FSQ FACES Ⅱ-CV SCSQ were no significantly different between homozygotic and dizygotic twins(P>0.05).There were significantly negative correlation between internalizing problem behavior and family cohesion (r=0.223, P=0.011).The scores of adolescent internalizing problem behavior were positively correlated with the scores of family stresses(r=0.232, P=0.008), the scores of Authoritarian-parenting-style(r=0.206, P=0.018), the scores of negative coping style (r=0.408, P=0.001);the scores of adolescents internalizing behavioral problems were not significantly correlated with the scores of the stress life events, and the scores of the parenting education level and occupation.The effects of A and E on adolescent internalizing problem behavior were 0.51(95%CI:0.27~0.69)and 0.49 (95% CI: 0.31~0.73) respectively. Conclusion Environmental factors and genetic factors contribute equally to adolescents internalizing problem behavior, which is affected by the authoritarian-parenting-style, family stresses, family cohesion and negative coping style.

Objective To reveal the pathophysiological changes of sepsis, the plasma metabonomics of septic rats was determined and differential metabolites were determined by gas chromatography-mass spectrometry (GC-MS). Methods Male Spraguee-Dawley (SD) rats about 8 weeks were randomly divided into sham group (n = 18) and sepsis group (n = 24). Cecal ligation and puncture (CLP) was used to build sepsis model, while cecum was kept intact only in the sham group. 6 hours after the operation, rats were anesthetized, and blood was harvested through heart thoracotomy. Then the plasma metabonomics was detected by GC-MS and metabolic profile analysis was performed to find the relative differential metabolites.Results Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) showed that the metabolic profiling of the sepsis group was significantly different from the sham group. 259 kinds of metabolites were got by GC-MS, and 69 kinds of differential metabolites were found between sham group and sepsis group, in which 23 differential metabolites were related to amino acid metabolism. Compared with sham group, the contents of putrescine, N-glutamic acid, hydroxynorvaline, 3-cyanuric acid, D-alanyl-D-alanine and urea in the plasma of septic rats increased significantly, which ratios of sepsis/sham group were 10.876, 6.394, 2.800, 2.226, 1.323, 1.203, respectively (allP < 0.05). On the other hand, the contents of oxygen generation of proline, citrulline, glutamine, su-beta-hydroxy aspartic acid, citric acid, N-methyl-DL-alanine, serine, lysine, threonine, N-formyl-L-methionine, methionine, alanine, nicotinuric acid, N-methyl-L-glutamic acid, trans-4-hydroxy-L-proline, proline, L-glutamic acid in the plasma of septic rats decreased significantly, which ratios of sepsis/sham group were 0.858, 0.853, 0.834, 0.816, 0.816, 0.814, 0.813, 0.801, 0.793, 0.792, 0.774, 0.766, 0.748, 0.727, 0.716, 0.674, 0.603, respectively (allP < 0.05).Conclusions Through the GC-MS analysis of plasma metabonomics of septic rats, we found abnormal energy metabolism changes. The content of amino acid in plasma might be a method to evaluate the energy metabolism status of sepsis.