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OBJECTIVE To study the effects of the curcumin derivative bisdemethoxycurcumin （BC） promoting neuronal differentiation of neuroblastoma cells Neuro-2a （N2a） in mice and its mechanism. METHODS The effects of BC （1， 2， 4， 6， 8， 10 μmol/L） on the viability of N2a cells were detected by MTT assay to determine the concentration range of drug treatment. The control group， retinoic acid （RA） group （10 μmol/L） and BC groups （1， 2 and 4 μmol/L） were set up， and the length of neural protrusions of the differentiated cells was measured and the cell differentiation rate was calculated after 48 h and 72 h of culture. Compared with 0 min group， Western blot was used to detect the phosphorylation levels of protein kinase B （Akt）， extracellular- signal regulated kinase 1/2 （ERK1/2）， and p38 mitogen-activated protein kinase （p38） proteins in cells treated by 4 μmol/L BC for 5， 15， 30， 60， 120 min. After intervention with inhibitors LY294002 （LY） and PD98059 （PD）， the effects of BC on Akt and ERK1/2 protein phosphorylation levels and promoting neural differentiation were further validated. RESULTS According to the MTT experiment， the BC concentrations for subsequent induction of cell differentiation were determined to be 1， 2， and 4 μmol/L. After 48 hours of differentiation， compared with the control group， the cell differentiation rate in RA group and BC 1， 2 and 4 μmol/L groups， the length of cellular neural processes wjxhhxx413@163.com in the BC 4 μmol/L group significantly increased （P＜0.05 or P＜0.01）；after inducing differentiation of BC for 72 hours，compared with the control group， the cell differentiation rate and the length of cellular neural processes in the RA group， the cell differentiation rate in the BC 4 μmol/L group， and the length of cellular neural processes in the BC 2 μmol/L group all significantly increased （P＜0.05 or P＜0.01）.Compared with the 0 min group， the phosphorylation levels of Akt， ERK1/2， and p38 proteins in cells of the 5， 15， 30， 60 and 120 min groups increased to varying degrees after treated by 4 μmol/L BC， and some differences were statistically significant （P＜0.05 or P＜0.01）. After adding the inhibitor LY/PD， compared with the BC group， the phosphorylation level of ERK1/2 protein in the PD+BC group cells were significantly reduced （P＜0.01）， and the cell differentiation rates in the LY group， LY+BC group， PD group， and PD+BC group was significantly reduced （P＜0.01）. CONCLUSIONS BC promotes N2a cell differentiation mainly by increasing cell differentiation rate and neural protrusion length. The mechanism may be related to the activation of mitogen-activated protein kinase/ ERK and PI3K/Akt signaling pathways.
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Objective:To compare the efficacy of modified single-incision and traditional double-incision for flexor hallucis longus tendon transfer in the treatment of Myerson type III chronic Achilles tendon ruptures in the middle-aged and elderly patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 106 patients with Myerson type III chronic Achilles tendon ruptures admitted to Ningbo No.6 Hospital from January 2015 to May 2021, including 54 males and 52 females; aged 48-82 years [(67.2±8.4)years]. Flexor hallucis longus tendon transfer was assigned to 60 patients using modified single-incision (modified single-incision group) and to 46 patients using traditional double-incision (traditional double-incision group). The operation time, intraoperative blood loss as well as American foot and ankle society (AOFAS) ankle-hindfoot score, foot and ankle outcome score (FAOS), visual analogue score (VAS) before operation, at postoperative 12 months and at the last follow-up were compared between the two groups. Time to recover walking and self-care ability, and incidence of complications were compared as well.Results:All patients were followed up for 16-52 months [(37.4±9.5)months]. The operation time and intraoperative blood loss were (51.6±7.4)minutes and (16.6±3.9)ml in modified single-incision group compared to (72.8±7.5)minutes and (32.9±5.2)ml in traditional double-incision group (all P<0.01). There was no significant difference in the AOFAS ankle-hindfoot score, FAOS and VAS between the two groups before operation (all P>0.05). At postoperative 12 months and at the last follow-up, AOFAS ankle-hindfoot score and VAS were not significantly different between the two groups (all P>0.05), but FAOS in modified single-incision group [(112.6±3.0)points, (114.4±3.1)points] was improved significantly compared with traditional double-incision group [(110.8±4.1)points, (112.7±4.3)points] ( P<0.05 or 0.01). At postoperative 12 months and at the last follow-up, the AOFAS ankle-hindfoot score, FAOS and VAS in both groups were improved or decreased significantly compared with those before operation (all P<0.05). All patients recovered their pre-injury daily activities. The time to recover walking and self-care ability were (9.6±2.0)weeks and (12.7±1.7)weeks in modified single-incision group compared to (10.8±1.8)weeks and (13.7±1.9)weeks in traditional double-incision group (all P<0.01). In modified single-incision group, superficial incision infection ( n=1) was found and cured after oral antibiotics. In traditional double-incision group, superficial incision infection ( n=2), deep incision infection ( n=1), deep venous thrombosis ( n=1) and medial plantar nerve injury ( n=1) were found and cured after symptomatic internal medical therapy; claw toe deformity ( n=2) was found and relieved after wearing customized insoles and functional exercise. The incidence of complications was 1.7% (1/60) in modified single-incision group compared to 15.2% (7/46) in traditional double-incision group ( P<0.01). Conclusion:In contrast with traditional double-incision surgery, modified single-incision for flexor longus tendon transfer in the treatment of Myerson type III chronic Achilles tendon ruptures in the middle-aged and elderly patients has advantages such as shorter operation time, less intraoperative blood loss, better functional recovery, faster postoperative recovery and less complications.
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Numerous mutations in the spike protein of SARS-CoV-2 B.1.1.529 Omicron variant pose a crisis for antibody-based immunotherapies. The efficacy of emergency use authorized (EUA) antibodies that developed in early SARS-CoV-2 pandemic seems to be in flounder. We tested the Omicron neutralization efficacy of an early B cell antibody repertoire as well as several EUA antibodies in pseudovirus and authentic virus systems. More than half of the antibodies in the repertoire that showed good activity against WA1/2020 previously had completely lost neutralizing activity against Omicron, while antibody 8G3 displayed non-regressive activity. EUA antibodies Etesevimab, Casirivimab, Imdevimab and Bamlanivimab were entirely desensitized by Omicron. Only Sotrovimab targeting the non-ACE2 overlap epitope showed a dramatic decrease activity. Antibody 8G3 efficiently neutralized Omicron in pseudovirus and authentic virus systems. The in vivo results showed that Omicron virus was less virulent than the WA1/2020 strain, but still caused deterioration of health and even death in mice. Treatment with 8G3 quickly cleared virus load of mice. Antibody 8G3 also showed excellent activity against other variants of concern (VOCs), especially more efficient against authentic Delta plus virus. Collectively, our results suggest that neutralizing antibodies with breadth remains broad neutralizing activity in tackling SARS-CoV-2 infection despite the universal evasion from EUA antibodies by Omicron variant.
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Emerging SARS-CoV-2 variants are threatening the efficacy of antibody therapies. Combination treatments including ACE2-Fc have been developed to overcome the evasion of neutralizing antibodies (NAbs) in individual cases. Here we conducted a comprehensive evaluation of this strategy by combining ACE2-Fc with NAbs of diverse epitopes on the RBD. NAb+ACE2-Fc combinations efficiently neutralized HIV-based pseudovirus carrying the spike protein of the Delta or Omicron variants, achieving a balance between efficacy and breadth. In an antibody escape assay using replication-competent VSV-SARS-CoV-2-S, all the combinations had no escape after fifteen passages. By comparison, all the NAbs without combo with ACE2-Fc had escaped within six passages. Further, the VSV-S variants escaped from NAbs were neutralized by ACE2-Fc, revealing the mechanism of NAb+ACE2-Fc combinations survived after fifteen passages. We finally examined ACE2-Fc neutralization against pseudovirus variants that were resistant to the therapeutic antibodies currently in clinic. Our results suggest ACE2-Fc is a universal combination partner to combat SARS-CoV-2 variants including Delta and Omicron.
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Kashin-Beck disease (KBD) is a disabling osteoarthropathy of unknown cause, which occurs most frequently in children and adolescents and affects many regions in China. In adults KBD patients, the degeneration of weight-bearing joints is the most serious, especially the knee joint. Up to now, there are still a large number of adult patients with KBD arthritis of the knee, and they have not been systematically and effectively treated. The main manifestations are knee joint pain, thickening, deformation and restricted movement. In severe cases, it can lead to knee joint varus, valgus, flexion contracture, and rotational deformity, which seriously affects the quality of life and work ability of the patients. Early diagnosis of KBD arthritis of the knee is difficult and there is no effective treatment, so it needs to be differentiated from knee osteoarthritis (OA). KBD and OA have common pathological characteristics. According to the "Consensus of Four-stepladder Program of Knee 0steoarthritis" (2018), OA treatment is divided into four levels: basic treatment, drug treatment, restorative treatment and reconstruction treatment. In this paper, the treatment of KBD arthritis of adult knee joint is summarized, which is also divided into four steps. It is necessary to make stepwise treatments according to the severity of KBD arthritis of the knee and the differences of patient's own condition, so that the limited medical resources can be efficiently used. At the same time, this paper also systematically summarizes the epidemiological characteristics, pathological features and diagnostic criteria of KBD arthritis of the knee, aiming to provide effective reference and guidance for the treatment of KBD arthritis of the knee.
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Acute respiratory tract infection is the most common infectious disease in children, which seriously threatens children′s health.Rapid and accurate etiological diagnosis is of great significance for the clinical treatment and control of these diseases.Pathogen nucleic acid test was applied and became the main method of respiratory tract infection diagnosis for its high sensitivity and specificity.To regulate the application of pathogen nucleic acid amplification test in respiratory tract infection in children, improve the diagnosis level, expert consensus on nucleic acid amplification test of respiratory pathogens in children was prepared to guide the application and promote pathogens diagnosis ability.
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This study aimed to explore the application value of new biological specimen oral fluid in SARS-CoV-2 nucleic acid and antibody detection. Oral fluid and paired respiratory and blood specimens from 7 confirmed cases of two COVID-19 cluster epidemic were collected in Beijing from October to November 2021. SARS-CoV-2 virus and IgG antibody were detected by real time PCR kits and serum antibody detection reagents, and SARS-CoV-2 IgG antibody in oral fluids was detected by a new established method of magnetic particle chemiluminescence. The results showed that the nucleic acid amplification test of SARS-CoV-2 on nasopharyngeal swabs, throat swabs and oral fluid specimens from 3 confirmed cases of COVID-19 was positive, among which the Ct value for ORF1a/b and N gene of oral fluid samples in 2 cases was close to that of throat swab, and the Ct value of oral fluid sample for 1 case was higher than that of throat swab. The complete genome sequence of one oral fluid specimen was obtained, which belonged to the VOC/Delta variant strain. The SARS-CoV-2 IgG antibodies of the paired oral fluid and serum were all positive, and the S/CO values of oral fluid were all lower than those of serum. The series of oral fluid results showed that SARS-CoV-2 IgG antibody level increased from 11 to 32 days after the onset of the disease.
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Humans , COVID-19/diagnosis , Nucleic Acids , SARS-CoV-2 , Sensitivity and SpecificityABSTRACT
ObjectivesAs the COVID-19 pandemic is still ongoing and SARS-CoV-2 variants are circulating worldwide, an increasing number of breakthrough infections have been detected despite the good efficacy of COVID-19 vaccines. MethodsA prospective, comparative cohort study was conducted in Beijing Ditan Hospital to evaluate the clinical, immunological and genomic characteristics of COVID-19 breakthrough infections. Data on 88 COVID-19 breakthrough cases (vaccinated group) and 41 unvaccinated cases (unvaccinated group) from June 1 to August 20, 2021 were extracted from a cloud database. Among these 129 COVID-19 cases, we successfully sequenced 33 whole genomes, including 16 from the vaccinated group and 17 from the unvaccinated group. ResultsAsymptomatic and mild cases predominated in both groups, but 2 patients developed severe disease in the unvaccinated group. Between the two groups, the median time of viral shedding in the vaccinated group were significantly lower than those in the unvaccinated group (p = 0.003). A comparison of dynamic IgG titres of cases in the two groups indicated that IgG titres in the vaccinated group showed a significantly increasing trend (P =0.028). The CD4+T lymphocyte count was lower in the unvaccinated group, and there was a significant difference between the two groups (p=0.018). In the vaccinated group, the number of moderate cases who received Sinopharm BBIBP (42 cases) was significantly higher than those who received Sinovac Coronavac (p=0.020). Whole-genome sequencing revealed 23 cases of delta variants, including 15 patients from the vaccinated group. However, no significant difference was observed in either the RT-qPCR results or viral shedding time. ConclusionsCOVID-19 vaccine breakthrough infections were mainly asymptomatic and mild, the IgG titres were significantly higher and increased rapidly, and the viral shedding was short. Delta variants may be more likely to cause breakthrough infections, and vaccination may not reduce the viral loads and shedding time.
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Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern (VOCs) continue to wreak havoc across the globe. Higher transmissibility and immunologic resistance of VOCs bring unprecedented challenges to epidemic extinguishment. Here we describe a monoclonal antibody, 2G1, that neutralizes all current VOCs and has surprising tolerance to mutations adjacent to or within its interaction epitope. Cryo-electron microscopy structure showed that 2G1 bound to the tip of receptor binding domain (RBD) of spike protein with small contact interface but strong hydrophobic effect, which resulted in nanomolar to sub-nanomolar affinities to spike proteins. The epitope of 2G1 on RBD partially overlaps with ACE2 interface, which gives 2G1 ability to block interaction between RBD and ACE2. The narrow binding epitope but high affinity bestow outstanding therapeutic efficacy upon 2G1 that neutralized VOCs with sub-nanomolar IC50 in vitro. In SARS-CoV-2 and Beta- and Delta-variant-challenged transgenic mice and rhesus macaque models, 2G1 protected animals from clinical illness and eliminated viral burden, without serious impact to animal safety. Mutagenesis experiments suggest that 2G1 could be potentially capable of dealing with emerging SARS-CoV-2 variants in future. This report characterized the therapeutic antibodies specific to the tip of spike against SARS-CoV-2 variants and highlights the potential clinical applications as well as for developing vaccine and cocktail therapy.
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BackgroundThe SARS-CoV-2 B.1.1.7 variant which was first identified in the United Kingdom (U.K.) has increased sharply in numbers worldwide and was reported to be more contagious. On January 17, 2021, a COVID-19 clustered outbreak caused by B.1.1.7 variant occurred in a community in Daxing District, Beijing, China. Three weeks prior, another non-variant (lineage B.1.470) COVID-19 outbreak occurred in Shunyi District, Beijing. This study aimed to investigate the clinical features of B.1.1.7 variant infection. MethodsA prospective cohort study was conducted on COVID-19 cases admitted to Ditan hospital since January 2020. Data of 74 COVID-19 cases from two independent COVID-19 outbreaks in Beijing were extracted as study subjects from a Cloud Database established in Ditan hospital, which included 41 Shunyi cases (Shunyi B.1.470 group) and 33 Daxing cases (Daxing B.1.1.7 group) that have been hospitalized since December 25, 2020 and January 17, 2021, respectively. We conducted a comparison of the clinical characteristics, RT-qPCR results and genomic features between the two groups. FindingsCases from Daxing B.1.1.7 group (15 [45.5%] male; median age, 39 years [range, 30.5, 62.5]) and cases from Shunyi B.1.470 group (25 [61.0%] male; median age, 31 years [range, 27.5, 41.0]) had a statistically significant difference in median age (P =0.014). Seven clinical indicators of Daxing B.1.1.7 group were significantly higher than Shunyi B.1.470 group including patients having fever over 38{degrees}C (14/33 [46.43%] in Daxing B.1.1.7 group vs. 9/41 (21.95%) in Shunyi B.1.470 group [P = 0 .015]), C-reactive protein ([CRP, mg/L], 4.30 [2.45, 12.1] vs. 1.80, [0.85, 4.95], [P = 0.005]), Serum amyloid A ([SAA, mg/L], 21.50 [12.50, 50.70] vs. 12.00 [5.20, 26.95], [P = 0.003]), Creatine Kinase ([CK, U/L]), 110.50 [53.15,152.40] vs. 70.40 [54.35,103.05], [P = 0.040]), D-dimer ([DD, mg/L], 0.31 [0.20, 0.48] vs. 0.24 [0.17,0.31], [P = 0.038]), CD4+ T lymphocyte ([CD4+ T, mg/L], [P = 0.003]), and Ground-glass opacity (GGO) in lung (15/33 [45.45%] vs. 5/41 [12.20%], [P =0.001]). After adjusting for the age factor, B.1.1.7 variant infection was the risk factor for CRP (P = 0.045, Odds ratio [OR] 2.791, CI [1.025, 0.8610]), SAA (0.011, 5.031, [1.459, 17.354]), CK (0.034, 4.34, [0.05, 0.91]), CD4+ T (0.029, 3.31, [1.13, 9.71]), and GGO (0.005, 5.418, [1.656, 17.729]) of patients. The median Ct value of RT-qPCR tests of the N-gene target in the Daxing B.1.1.7 group was significantly lower than the Shunyi B.1.470 group (P=0.036). The phylogenetic analysis showed that only 2 amino acid mutations in spike protein were detected in B.1.470 strains while B.1.1.7 strains had 3 deletions and 7 mutations. InterpretationClinical features including a more serious inflammatory response, pneumonia and a possible higher viral load were detected in the cases infected with B.1.1.7 SARS-CoV-2 variant. It could therefore be inferred that the B.1.1.7 variant may have increased pathogenicity. FundingThe study was funded by the National Key Research and Development Program (grant nos.2020YFC0846200 and 2020YFC0848300) and National Natural Science Foundation of China (grant no. 82072295).
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Objective:To clarify the infection and epidemic characteristics of the human metapneumovirus (HMPV) in Chinese patients with febrile respiratory syndrome (FRS), and to provide important baseline data for clinical diagnosis, treatment, prevention and control of HMPV-induced respiratory tract diseases in China.Methods:FRS cases from January 2009 to June 2021 in 9 provinces in China, including Beijing, Hebei, Jilin, Shandong, Shaanxi, Xinjiang, Anhui, Guangdong, Hunan were retrospectively analyzed for their respiratory samples, clinical and epidemic data.The respiratory samples were detected for HMPV by quantitative real-time PCR.Results:A total of 11 660 cases were tested for HMPV, involving 296 (2.54%) HMPV-positive cases.Among 296 HMPV-positive cases, 218 were single HMPV infection, and 78/296 (26.35%) were co-infected with one or more respiratory viruses.HMPV mainly affected children under 5 years of age (3.10%), and in this population, the proportion of pneumonia in HMPV co-infection cases was significantly higher than that of single HMPV infection.HMPV could be detected all year round, which was more popular in winter and spring, with the peak of HMPV epidemic in March.Conclusions:HMPV is one of the important pathogens causing acute respiratory infection in children, showing a clear seasonal epidemic.HMPV can be infected alone or in combination with other respiratory viruses, which may increase the risk of pneumonia in children.
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Objective:To understand the etiology and clinical characteristics of hospitalized severe community-acquired pneumonia(SCAP) in Changchun, and provide scientific basis for its etiology diagnosis and targeted treatment.Methods:The study subjects included 618 children with clinical diagnosis of SCAP who were hospitalized from January 2016 to December 2019.We collected pharyngeal swabs and alveolar lavage fluid from children.Virus isolation, bacterial culture, time-of-flight mass spectrometry, PCR/RT-PCR, colloidal gold method and Optochin test were used to detect the antigen, nucleic acid and protein profiles in the specimen.Results:There were more boys than girls in hospitalized children with SCAP.The peak age of onset was 7 to 12 months.Most cases occurred in winter and spring.The highest detection rate of SCAP virus was 56.15%(347/618); 73.49%(255/347) were positive for one virus, among which the top five were respiratory syncytial virus (27.8%), influenza A virus (23.9%), influenza B virus (16.1%), rhinovirus (12.2%) and metapneumovirus (10.2%). Two viruses were positive for 19.88%(69/347); three viruses were positive for 4.32%(15/347); four viruses were positive for 2.31%(8/347). Atypical microbial infections were 29.77%(184/618), of which Mycoplasma pneumoniae accounted for 95.65%(176/184). Bacterial infections were 17.31%(107/618), mainly Streptococcus pneumoniae(39.25%, 42/107) and Staphylococcus aureus(24.30%, 26/107). The mixed infection of multiple pathogens was 7.61%(47/618), among which the mixed infection rates of Mycoplasma pneumonia with Streptococcus pneumoniae, virus were 40.43% and 34.04%, respectively.High fever, faster breathing, and perioral cyanosis were risk factors for SCAP, with OR and 95% CI of 7.71 and 4.56-13.04, 2.43 and 2.02-2.93, 3.53 and 2.56-4.86, respectively.Viral co-infection occurred in 36.96%(34/92) of complications such as heart failure, toxic encephalopathy, and myocardial damage; Mycoplasma pneumoniae and other pathogens co-infected 35.29% of children with pleural effusion. Conclusion:The pathogens of SCAP in Changchun are mainly viruses notably, respiratory syncytial virus is the dominant pathogen, followed by Mycoplasma pneumoniae.The bacterial pathogen is mainly Streptococcus pneumoniae.High fever, faster breathing, and cyanosis around the mouth are risk factors for severe pneumonia.Multi-pathogen mixed infection is prone to serious complications.
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Severe acute respiratory syndrome CoV-2 (SARS-CoV-2) caused the Corona Virus Disease 2019 (COVID-19) cases in China has become a public health emergency of international concern (PHEIC). Based on angiotensin converting enzyme 2 (ACE2) as cell entry receptor of SARS-CoV, we used the hACE2 transgenic mice infected with SARS-CoV-2 to study the pathogenicity of the virus. Weight loss and virus replication in lung were observed in hACE2 mice infected with SARS-CoV-2. The typical histopathology was interstitial pneumonia with infiltration of significant lymphocytes and monocytes in alveolar interstitium, and accumulation of macrophages in alveolar cavities. Viral antigens were observed in the bronchial epithelial cells, alveolar macrophages and alveolar epithelia. The phenomenon was not found in wild type mice with SARS-CoV-2 infection. The pathogenicity of SARS-CoV-2 in hACE2 mice was clarified and the Kochs postulates were fulfilled as well, and the mouse model may facilitate the development of therapeutics and vaccines against SARS-CoV-2.
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This study shows the case of a patient with peroneus brevis tendon rupture in ankle fracture.The patient complained of swelling,pain and activity limitation in her right ankle caused by sprain.She was diagnosed with ankle fracture,supination-adduction (Lauge-Hansen typing) by medical history,physical examination and imaging.The peroneus brevis tendon was noted complete ruptured when suturing the incision after ORIF,which was then repaired by"8" shaped suture.Based on literature review,the injury mechanism of the case may be because of overload inversion force toward to lateral inferior from the fracture of fibula on the peroneus brevis tendon.Additional attention should be paid to patients who suffered from an supination-adduction ankle fracture for whether peroneus brevis tendon tears before and during surgery.
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Objective To understand the clinical features and pathogenic spectrum of encephalitis and menin﹣gitis syndrome in children. Methods A total of 667 cases of children with encephalitis or meningitis diagnosed and documented at Changchun Children′s Hospital from May 2012 to July 2015 were enrolled. A variety of samples in diffe﹣rent types were collected and presented,including 335 cerebrospinal fluid specimens,530 blood samples,and 332 stool samples. All the samples were collected from the patients within 72 hours on admission. Moreover,these samples are analyzed and tested,including PCR for enterovirus(EV),herpesvirus(HSV),mycobacterium tuberculosis( TB)and My﹣coplasma pneumoniae(MP)nucleic acid in cerebrospinal fluid samples;fecal specimens were tested for EV,enterovirus 71(EV71),coxsackievirus A6(CA6),coxsackievirus A16(CVA16),coxsackievirus A10( CVA10)nucleic acids;degenerate primers to amplify Echovirus 30(Echo30). Clinical data of children were collected. Results The peak in﹣cidence of encephalitis and meningitis syndrome was from June to August,age distribution was from 0 to 15 years old, the proportion of children aged from 0－6 accounted for 81. 41﹪;the highest proportion was among 0－1 years old in﹣fants,occupying 32. 38﹪;408 males and 259 females;the main symptoms were fever(586 cases),apathy(337 ca﹣ses),vomiting(307 cases)and headache(203 cases). And clinical signs included drowsiness(103 cases),neck stiff﹣ness(71 cases),meningeal irritation(12 cases),and pathological reflex( 313 cases),etc. The clinical diagnosis included 272 cases of viral encephalitis,332 cases of severe hand,foot and mouth disease complicated by encephalitis, 30 cases of bacterial meningitis,and 33 other cases;the etiological detection included:the positive rates of EV,EBV and Echo30 in cerebrospinal fluid specimens were 59. 72﹪,3. 16﹪ and 70. 00﹪,respectively. And EV71,CVA16,CVA6, EV71+CA16 and EV71+CVA16+CVA6 nucleic acids were detected in fecal samples,in which the highest detection rate was EV71(98. 96﹪). Conclusions In Changchun Children′s Hospital,the children with encephalitis and menin﹣gitis are mainly viral encephalitis. The main symptoms were fever,apathetic,drowsiness,vomiting and headache. Signs included,neck stiffness,meningeal irritation,and pathological reflexes,etc. The main pathogen of the disease is EV71.
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Objective@#To study the application of measles specific IgM and IgG antibody detection in classification of primary vaccination failure (PVF) and secondary vaccination failure (SVF).@*Methods@#Measles surveillance information system was used to collect measles confirmed cases in Tianjin, 2013-2015, and their blood specimens were collected, totally 284 cases were enrolled. Measles IgM and IgG were detected with enzyme-linked immunosorbent assay (ELISA), and the relative avidity index (RAI) was used to express the result of measles avidity. Measles IgM, IgG and IgM/IgG was analyzed with receiver operating characteristic (ROC) curve, the area under the ROC curve (AUC) as evaluation indicators. In addition, compared with a measles outbreak (26 cases) of a middle school in Tianjin in 2016, for making further verification on the diagnostic value of vaccination failure with IgM, IgG and IgM/IgG.@*Results@#The age of cases ranged was 0-58 years old, the interval median (P25, P75) of serum collection after rash onset was 2 (1, 4) days. The positive rate of measles IgM and IgG in acute phase specimens were 76.06% (216 cases) and 88.38% (251 cases). According to the ROC curve analysis, the area under the ROC curve (AUC) of IgM, IgG and IgM/IgG were 0.753, 0.891 and 0.952, indicating that IgM/IgG was the best index to distinguish PVF and SVF. The best cut off value for IgM/IgG was 0.06, the sensibility and specificity were 88.75% and 86.63%. When IgM/IgG >1, 96.30% cases were low-avidity (RAI <40%), only 1 case was equivocal response (RAI: 40%-60%). 97.14% cases were high-avidity (RAI >60%) when IgM/IgG <0.01, only 3 cases were equivocal response (RAI 40%-60%). The threshold of IgM/IgG was used to verify the measles outbreak of a middle school in Tianjin, 2016. In the acute phase specimens, 100% (26 cases) of IgM/IgG were <0.06, 84.62% (22 cases) of IgM/IgG were <0.01.@*Conclusion@#The detection of measles IgM and IgG with ELISA, and IgM/IgG is a valuable diagnostic tool to distinguish PVF and SVF.
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Objective@#To clarify the genotype of varicella-zoster virus (VZV) in Jilin province in 2017, and to discriminate between vaccine strain and wild-type strain.@*Methods@#Vesicle fluid and throat swab samples were collected from 10 individuals with suspected VZV in Jilin province from January to March of 2017. Real-time fluorescent quantitative PCR was used to detect viral nucleic acid. Specific regions of ORF22, ORF38 and ORF62 of VZV were amplified by PCR. Viral genotype was determined by five SNPs of ORF 22 and vaccine strain or wild-type strain was distinguished by four SNPs of ORF 38 and ORF 62. The results were analyzed with MEGA5 and BioEdit software, using the VZV reference strain sequences from GenBank.@*Results@#VZV-positive strains were detected in 10 samples, all belonged to Clade 2. There was a synonymous mutation (C→T) in position 38 048 of JL17-7 strain. The nucleotide homology of ORF22 showed that all 10 samples were on the same branch with the Clade 2 referenced strains. Compared with Clade 2 referenced strains, the homology of nucleotide and amino acid for all 10 samples were 99.5%-100% and 99.3%-100%, respectively. The four specific SNPs of ORF38 and ORF62 in 10 samples were A-T-T-T, which were consistent with wild-type strain.@*Conclusions@#This study reveals that the VZV strains circulating in Jilin province in 2017 were all wild-type strains belonging to Clade 2.
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Objective@#To understand the pathogenic composition of hand, foot and mouth disease (HFMD) in Ningxia Hui Autonomous Region (Ningxia) from 2016 to 2017, and analyze the genetic characteristics of the main pathogens enterovirus (EV)-A71 and coxsackievirus (CV)-A16.@*Methods@#Analysis of the result of nucleic acid testing of HFMD in Ningxia from 2016 to 2017 to determine the pathogenic composition of HFMD. The complete VP1 coding region was amplified by RT-PCR and the gene sequence was determined for the enterovirus strains sent to the National HFMD Network Monitoring Laboratory in Ningxia from 2016 to 2017. BLAST analysis confirmed the serotype of the strain, and the phylogenetic tree was constructed respectively by selecting EV-A71 and CV-A16 isolates.@*Results@#The leading pathogens of HFMD in Ningxia of 2016 and 2017 were other EV (397, 43.72%) and EV-A71 (918, 56.18%) respectively, and the dominant pathogens in different months may differ. The pathogenic composition causing HFMD in the past two years has changed from CV-A16 and other EV to EV-A71 and other EV. The isolated EV-A71 strains were C4a evolutionary branch and the isolated CV-A16 strains were B1b evolutionary branch.@*Conclusions@#Compared to 2016, in 2017 EV-A71, CV-A16 and other EV changed dynamically. Dynamic monitoring of EV-A71 in Ningxia is of great significance to guide the strategy of using EV-A71 vaccine, concentrating medical resources to strengthen the treatment and reduce the mortality rate of severe HFMD cases.
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Objective@#The genetic characteristics of the human adenovirus type 53 (HAdV-53) strains isolated from Taiyuan city of Shanxi Province were studied to obtain the baseline data of their molecular characteristics.@*Methods@#Conjunctival swabs (n=79) were collected from epidemic keratoconjunctivitis (EKC) patients in Shanxi eye Hospital in 2016, and five HAdV-53 strains were obtained after virus isolation and identification based on the three major capsid genes sequences including Penton base, Hexon and Fiber gene. And the corresponding sequences of global epidemic HAdV-53 strains and the strains with the same genetic origin as HAdV-53 were also downloaded from GenBank database, and then the three gene database were established, respectively. With the database, phylogenetic tree was constructed, and the genetic and molecular evolutionary characteristics were analyzed with bioinformatics software.@*Results@#Five HAdV-53 strains in Shanxi Province in 2016 showed high consistency with the HAdV-53 strains prevalent in other countries in 1996-2014 (>99.8%). All HAdV-53 strains were in the same evolutionary branch with their recombinant source genotypes (HAdV-37 and HAdV-8) in Penton base and Fiber gene, respectively, and maintained a high degree of consistency in gene sequences. In Hexon gene, HAdV-53 strains were more closed to its recombinant source genotype HAdV-22, the nucleotide and amino acid sequences between two types were highly homologous, while HAdV-53 and HAdV-22 belonged to different evolutionary branches, and the evolution rate of HAdV-53 based on Hexon gene was 3.51×10-5 substitution/site/year.@*Conclusion@#HAdV-53 has become an important new ocular infectious pathogen of Taiyuan. HAdV-53 strain are relatively conservative and stable based on Penton base, Hexon, and Fiber gene.
ABSTRACT
Objective@#To understand the clinical features and pathogenic spectrum of encephalitis and meningitis syndrome in children.@*Methods@#A total of 667 cases of children with encephalitis or meningitis diagnosed and documented at Changchun Children′s Hospital from May 2012 to July 2015 were enrolled.A variety of samples in diffe-rent types were collected and presented, including 335 cerebrospinal fluid specimens, 530 blood samples, and 332 stool samples.All the samples were collected from the patients within 72 hours on admission.Moreover, these samples are analyzed and tested, including PCR for enterovirus(EV), herpesvirus(HSV), mycobacterium tuberculosis(TB) and Mycoplasma pneumoniae(MP) nucleic acid in cerebrospinal fluid samples; fecal specimens were tested for EV, enterovirus 71 (EV71), coxsackievirus A6 (CA6), coxsackievirus A16 (CVA16), coxsackievirus A10 (CVA10) nucleic acids; degenerate primers to amplify Echovirus 30 (Echo30). Clinical data of children were collected.@*Results@#The peak incidence of encephalitis and meningitis syndrome was from June to August, age distribution was from 0 to 15 years old, the proportion of children aged from 0-6 accounted for 81.41%; the highest proportion was among 0-1 years old infants, occupying 32.38%; 408 males and 259 females; the main symptoms were fever(586 cases), apathy(337 cases), vomiting (307 cases) and headache(203 cases). And clinical signs included drowsiness (103 cases), neck stiffness (71 cases), meningeal irritation (12 cases), and pathological reflex (313 cases), etc.The clinical diagnosis included 272 cases of viral encephalitis, 332 cases of severe hand, foot and mouth disease complicated by encephalitis, 30 cases of bacterial meningitis, and 33 other cases; the etiological detection included: the positive rates of EV, EBV and Echo30 in cerebrospinal fluid specimens were 59.72%, 3.16% and 70.00%, respectively.And EV71, CVA16, CVA6, EV71+ CA16 and EV71+ CVA16+ CVA6 nucleic acids were detected in fecal samples, in which the highest detection rate was EV71(98.96%).@*Conclusions@#In Changchun Children′s Hospital, the children with encephalitis and meningitis are mainly viral encephalitis.The main symptoms were fever, apathetic, drowsiness, vomiting and headache.Signs included, neck stiffness, meningeal irritation, and pathological reflexes, etc.The main pathogen of the disease is EV71.
