Objective To prepare the chitosan-potD nanoparticles and to evaluate its protective efficacy against pneumococcal nasopharyngeal colonization. Methods potD gene was amplificated from pneumococcal genome and was inserted into pVAX1 expression vectors to construct pVAX1-potD recombinant plasmid which was then transfected into 293T cell using LipofectAMINE 2000 to analyze transient potD gene expression in vitro by RT-PCR and Western blot. Chitosan-potD nanoparticles were freshly prepared by coacervation methods at each time and the characterizations of the nanoparticles were then evaluated. BALB/c mice were immunized with chitosan-potD, naked potD DNA or pVAX1 for 4 times at two-week intervals. Anti-PotD IgG, IgG1 and IgG2a levels in serum and IgA levels in nasal washes, bronchoalveolar lavage fluids (BALF) and middle ear lavages(MEL) were detected by indirect enzyme-linked immunosorbent assay (ELISA). IL-17A, IL-4 and IFN-γ levels in splenocytes were determined by double sandwich ELISA. Mice were intrannsally challenged with Streptococcus pneumoniae ATCC6303, and Pneumococci were recovered from the nasopharyngeal niche at the fifth day after challenge. Results potD gene was successfully amplificated by PCR and the sequence was confimed to be consistent with that in the Genbank. The pVAX1-potD recombinant plasmid was successfully constructed and was expressed in eukaryocytes in vitro. The mean size and zeta potential of chitosan-potD nanoparticles was 430 nm and + 20.5 mv, respectively. Chitosan-potD nanoparticles were not digested by DNase Ⅰ , while naked potD DNA was completely digested. The levels of antibodies inculding IgG, IgG1, IgG2a, IgA and cytokines including IL-17A, IL-4 and IFN-γ were significantly higher in mice immunized with chitosan-potD nanoparticles than mice with naked potD or pVAX1 ( P <0.05) only. More importantly, much less Pneumococci were recovered from mice immunized with chitosan-potD nanoparticles than the other groups(P <0.05). Conclusion Chitosan-potD nanoparticles significantly enhanced the immunogenicity and protection efficacy of DNA vaccines by intranasal immunization and could be used as a potential mucosal vaccine to prevent pneumococcal infection.

Objective To compare the sedation and anti-inflammatory effects of dexmedetomidine and midazolam on critical ill children with multiple trauma. Methods A prospective randomized controlled trial was conducted. Sixty-five critical ill children with multiple trauma admitted to pediatric intensive care unit (PICU) of Anhui Province Children's Hospital from January 2014 to September 2016 were enrolled, who were randomly divided into dexmedetomidine group (33 cases) and midazolam group (32 cases). Children of both groups received sufentanil for analgesia. Children in dexmedetomidine group firstly received 1.0 μg/kg intravenous bolus of dexmedetomidine for 10 minutes, then continuous infusion of 0.2-0.7 μg·kg-1·h-1, while in midazolam group children received 1-5 μg·kg-1·min-1 of midazolam in continuous infusion. The goal of sedation was to maintain a Richmond agitation-sedation scale (RASS) score of -1 to 0. The level of serum interleukin (IL-6, IL-8, IL-10, IL-1β), tumor necrosis factor-α (TNF-α) and C-reactive protein (CRP) were detected by enzyme linked immunosorbent assay (ELISA) at 24, 48, 72 hours after treatment, and the duration of mechanical ventilation, ratio of continuous renal replacement therapy (CRRT), length of stay in the PICU, ratio of sepsis and multiple organ failure (MOF) and mortality were also recorded. Results Compared with midazolam, dexmedetomidine decreased the level of pro-inflammatory cytokines and increased the level of anti-inflammatory cytokines. At 24 hours after treatment, the levels of serum IL-1β, TNF-α significantly decreased and IL-10 significantly increased [IL-1β (ng/L):6.48±2.89 vs. 8.07±3.14, TNF-α (μg/L): 11.25±5.21 vs. 15.44±5.97, IL-10 (ng/L): 12.10±5.35 vs. 9.58±4.71, all P < 0.05]. At 48 hours after treatment, the levels of serum IL-6, IL-8, IL-1β, TNF-α and CRP significantly decreased and IL-10 significantly increased [IL-6 (ng/L): 209.67±80.49 vs. 336.31±123.94, IL-8 (ng/L):229.09±80.81 vs. 298.28±90.25, IL-1β (ng/L): 7.31±3.02 vs. 8.74±3.17, TNF-α (μg/L): 12.52±4.79 vs. 16.58±5.98, CRP (g/L): 47.82±24.92 vs. 72.35±31.71, IL-10 (ng/L): 12.90±5.42 vs. 10.01±4.79, all P < 0.05]. At 72 hours after treatment, the levels of serum IL-8 and CRP significantly decreased [IL-8 (ng/L): 234.64±96.24 vs. 290.28±103.97, CRP (g/L): 53.24±29.12 vs. 86.58±38.30, both P < 0.05]. Compared with midazolam, dexmedetomidine could significantly reduce the duration of mechanical ventilation (days: 4.7±1.3 vs. 6.6±2.1), length of PICU stay (days: 9.5±2.7 vs. 12.3±3.9, both P < 0.05), and the ratio of sepsis (33.3% vs. 53.1%, P < 0.05). But there were no significant differences in ratio of CRRT (18.2% vs. 18.8%), MOF (9.1% vs. 18.8%) and mortality (6.1% vs. 12.5%) between two groups (all P > 0.05). Conclusion Compared with midazolam, dexmedetomidine had better efficacy in the treatment of severe multiple trauma in children and reduce the level of inflammation.

A HPLC-QTOF MS method was established for analysis of components in Ophiocordyceps sinensis . The HPLC analysis was performed on an Agilent Zorbax SB Aq (150 mmí4.6 mm, 5 μm) with gr adient elution (5 mmol·L-1 ammonium acetate aqueous solution-acetonitrile), flow rate was 0.8 mL·min-1 and detection wave-length was 260 nm. The developed method was successfully applied in analysis of three different samples in-cluding O. sinensis, Hirsutella sinensis ( anamorph of O.sinensis) and Cordyceps militaris. Nine compounds were i-dentified in both O.sinensis and H.sinensis, which eight compounds were identified in C.militaris.

Objective To analyze the etiology and prognosis of children with thrombocytopenia (TP) in pediatric intensive care unit (PICU). Methods The data of children with TP (exclusion of congenital and unknown TP) admitted to PICU of Anhui Provincial Children's Hospital from January 2008 to December 2017 was analyzed retrospectively. According to the age of onset, the children were divided into infantile group (29 days to less than 1 year), early childhood group (1 to less than 3 years), preschool group (3 to less than 6 years), school age group (6 to less than 10 years) and puberty group (more than 10 years). Moreover, according to the lowest platelet count (PLT), the children were divided into PLT≤20×109/L group, PLT (21-50)×109/L group and PLT > (50-100) ×109/L group. The distribution and mortality of TP were analyzed, and the relationship between age, PLT decrease and prognosis were analyzed by Pearson method. Results Among 6 725 children admitted to PICU in our hospital from January 2008 to December 2017, there were 683 children with TP, with the incidence of 10.2%. Among 683 children with TP, there were 387 males and 296 females, with the median age of 2.72 (0.61, 3.08) years, and 92 children died, with a total mortality of 13.5%. Analysis of primary disease showed that TP caused by non-hematological malignant tumor disease accounted for 73.9%, with the mortality of 11.1% (56/505). TP induced by hematological malignant tumor disease accounted for 21.4%, with the mortality of 24.7% (36/146). Pseudothrombocytopenia accounted for 0.6%, with the mortality of 0% (0/4). Other children who gave up treatment accounted for 4.1%. It was shown by further analysis that multiple organ dysfunction syndrome (MODS) caused by TP associated with non-hematological malignant tumor disease accounted for 26.9%, with the mortality of 15.4% (21/136). Sepsis, severe trauma, pneumonia, central nervous system infection and disseminated intravascular coagulation (DIC) accounted for 17.4%, 16.6%, 12.7%, 11.7% and 11.5%; with the mortality of 8.0% (7/88), 2.4% (2/84), 0% (0/64), 20.3% (12/59) and 24.1% (14/58), respectively. The main causes of TP associated with hematological malignant tumor disease were hemophagocytic syndrome [accounting for 27.4%, with the mortality of 32.5% (13/40)] and bone marrow inhibition [accounting for 21.2%, with the mortality of 25.8% (8/31)]. The younger were the children with TP, the higher would be the mortality. The mortality of infantile group was significantly higher than that of early childhood group, preschool group, school age group and puberty group [18.8% (53/282) vs. 14.0% (28/200), 6.7% (7/104), 4.3% (4/92), 0% (0/5), all P < 0.01]. The lower was the PLT, the higher would be the mortality. The mortality of PLT≤20×109/L group was significantly higher than that of PLT (21-50)×109/L group and PLT > (50-100)×109/L group [18.1% (39/215) vs. 13.0% (32/247), 9.5% (21/221), both P < 0.05]. It was shown by correlation analysis that there was no association between age and 28-day death time in children with TP (r = -0.037, P = 0.727), but PLT was positively correlated with 28-day death time in children with TP (r = 0.844, P = 0.010). Conclusions MODS, infection, sepsis, severe trauma and DIC are the common causes of TP in PICU. The younger are the children with TP, the lower is the PLT, and the worse would be the prognosis.

Objective To summarize and analyze the shielding,retention and reentry works of blood donors,and to investigate the feasibility of retention and reentry strategy.Methods The samples of ELISA single reagent reactive/NAT non-reactive and ELISA non-reactive/ NAT reactive were negative by confirmatory tests.Then the blood was weeded out and the donation qualification was reserved.The donors of shielding more than 6 months could propose the reentry application at any blood station in the province,and were allowed to return to the ranks after qualified by routine detection and re-detection by Jiangsu Provincial Blood Center.The unqualified rates were compared between the donors of again blood donation after retention and reentry with the common donors by χ2 test.Results From October 2014 to June 2016,1 615 cases were ELISA single reagent reactive/NAT non-reactive,among which 67 cases were confirmed as positive,42 cases were undetermined and 1 506 cases were negative;831 cases were ELISA non-reactive/ NAT reactive,in which 809 cases were positive by confirmation and 22 cases were negative.A total of 1 528 donors were confirmed as negative and their donation qualifications were reserved,89 donors conducted blood donation again and 79 were qualified in blood detection.The unqualified rate was 11.24%,compared with that of common donors,the difference was statistically significant(P<0.001).Meanwhile,596 donors applied for reentry,among them 218 persons were weeded out by the reentry blood station.In remaining 378 samples sent to Jiangsu Provincial Blood Center,359 samples were qualified and confirmed to the reentry condition.Among them,332 donors conducted blood donation and all were qualified by blood detection.Conclusion The reentry strategy in Jiangsu Province is reasonable and feasible,but the donors retention strategy needs to be further optimized and perfected.

This study was aimed to prepare the spraying agent of prescriptions of Miao nationality herb and investigate the effect of Miao nationality herbs spray for serum SOD, MDA, and expression of Fas and Caspase-3 mRNA in lung tissues of silica-treated rats. The healthy SD rats were divided into 5 groups. Silica dust suspension was used in the model establishment of 4 groups. After the model was successfully established, 3 groups were randomly selected and given glucocorticoids atomization inhalation, Miao nationality herbs spray, Miao nationality herbs spray combined with intragastric administration of herbal medicine, respectively. After 40-day treatment, water-solubletetrazolium salt (WST-1) was used in the detection of serum superoxide dismutase (SOD). Thiobarbituric acid (TBA) was used in the detection of malondialdehyde (MDA). The mRNA expression variance of the Fas and Caspase-3 were detected by RT-PCR. The results showed that compared with the silica dust suspension group, the SOD activity of serum in the Miao nationality herbs spray group was significantly increased (P< 0.05). MDA content and the mRNA of Fas and Caspase-3 were significantly lower in the Miao nationality herbs spray group (P< 0.05). It was concluded that Miao nationality herbs spray group was able to increase the SOD activity of serum, decrease MDA content, and obviously decrease the expression of Fas and Caspase-3 of lung tissues among silica dust suspension rats.

To investigate the risk factors and prognosis of acute kidney injury (AKI) in children with sepsis in pediatric intensive care unit (PICU). Methods A retrospective analysis of clinical data of PICU sepsis children in Anhui Children's Hospital from May 2015 to May 2018 was performed. The children were divided into AKI group and non-AKI group according to whether AKI occurred within 48 hours of PICU [referring to the diagnostic criteria for Kidney Disease: Improving Global Outcomes (KDIGO)]. The general data, physiological data and clinical outcomes of the two groups were compared; Logistic regression analysis was used to analyze the risk factors of AKI in children with sepsis and the prognostic factors. Results AKI occurred in 55 of 127 children with sepsis, the incidence of AKI was 43.3%, and the overall mortality was 28.3% (36/127), with 41.8% (23/55) in AKI group and 18.1% (13/72) in non-AKI group.① Compared with non-AKI group, oxygenation index, albumin, the pediatric critical illness case score (PCIS) and urine volume in AKI group were significantly decreased, while cystatin C, procalcitonin (PCT), prothrombin time (PT), activated partial thromboplastin time (APTT), pediatric multiple organ dysfunction score (P-MODS), the proportions of mechanical ventilation, vasoactive drug use, shock, septic shock and mortality were significantly increased, while there was no difference in age, gender, mean arterial pressure (MAP), white blood cell count (WBC) and C-reactive protein (CRP) between the two groups. Multivariate Logistic regression analysis showed that low serum albumin [odds ratio (OR) = 0.627, 95% confidence interval (95%CI) = 0.495-0.794, P = 0.000] and homocystatin C (OR = 2.641, 95%CI = 1.157-6.032, P = 0.021) were risk factors for AKI in children with sepsis. ② Compared with the survival group of children with sepsis AKI, the proportion of mechanical ventilation, septic shock, vasoactive drug use, positive balance ratio of liquid for 72 hours, 6-hour lactate clearance rate < 10%, and AKI 3-stage patients in the death group of children with sepsis AKI were significantly increased. Multivariate Logistic regression analysis showed that 72-hour positive liquid balance (OR = 8.542, 95%CI = 1.956-37.307, P = 0.004) and 6-hour lactate clearance rate < 10% (OR = 5.980, 95%CI = 1.393-25.676, P = 0.016) were risk factors for the death of children with sepsis AKI. Conclusions Serum albumin and cystatin C should be closely monitored in children with sepsis. Early detection and intervention of positive fluid balance and low lactate clearance rate can reduce the mortality of AKI in children with sepsis.

OBJECTIVE:To prepare the betulinic acid nanoparticles,and characterize them. METHODS:Using ethanol as sol-vent and water as anti-solvent,anti-solvent recrystallization method was used to prepare betulinic acid nanoparticles. Using particle size as indicator,single factor test and orthogonal test were adopted to optimize the mass concentration of betulinic acid solution, anti-solvent-solvent volume ratio,anti-solvent drip rate,reaction temperature and stirring speed in formulation technology of betulin-ic acid nanoparticles,and verification test was conducted. The betulinic acid nanoparticles were characterized by scanning electron microscopy,laser particle size analyzer,Fourier infrared spectrometer and mass spectrum analyzer. RESULTS:The optimal technol-ogy was as follow as betulinic acid solution mass concentration of 3 mg/mL,anti-solvent-solvent volume ratio of 1:1,anti-solvent drip rate of 8 mL/min,reaction temperature of 20 ℃ and stirring speed of 900 r/min. The average size of prepared betulinic acid nanosuspension was(156.0±8.6)nm(n=3)and the particle size was(235.0±12.2)nm(n=3)after freeze-drying,with nearly spherical appearance,uniform size and regular form. Compared with raw material of betulinic acid,the chemical structure of pre-pared betulinic acid nanoparticles did not change,and there were no significant changes in molecular weight and mass ratio. CON-CLUSIONS:Betulinic acid nanoparticles are successfully prepared.

Objective:To investigate the clinical characteristics and genotypes of neurodevelopmental disorders of microcephaly-epilepsy-cortical atrophy caused by VARS1 gene mutation.Methods:The clinical data of a child with VARS1 gene mutation who visited Anhui Children′s Hospital in December 2021 were reviewed and followed up. Using "VARS1" "VARS" "VALYL-tRNA synthetase" "Valyl-tRNA synthetase" as the search terms, the Chinese data bases (China National Knowledge Infrastructure database, Wanfang database) and PubMed database (the database establishment until 2022). Articles related to genetic diseases were searched and clinical phenotypes and genotypes were summarized.Results:This case was a 3-month-old girl. After birth, she suffered from repeated convulsions and feeding difficulties, and gradually developed microcephaly, hypotonia, and overall developmental delay. Brain imaging showed cortical atrophy, corpus callosum dysplasia, and craniosynostosis. The results of whole exome sequencing indicated a new homozygous gene mutation in VARS1: gene mutation in exon 27 of chromosome 6 c.3203C>T(p.Thr1068 Met), which was from the patient 's parents. She took phenobarbital, levetiracetam, and sodium valproate for anti-epileptic treatment regularly, and then convulsed as a seizure every few days. A total of 19 patients were reported in 5 literatures that met the search criteria, all of whom presented with neurodevelopmental disorders. A total of 20 VARS1 gene mutation sites have been found so far. Conclusions:Neurodevelopmental disorders of microcephaly-epilepsy-cortical atrophy should be considered for children with neurodevelopmental disorders such as microcephaly, epilepsy, developmental delay, and cortical atrophy. Gene sequencing plays an important role in the diagnosis of the disease.

Retinoic acid-inducible gene I (RIG-I) is an important pattern recognition receptor that detects viral RNA and triggers the production of type-I interferons through the downstream adaptor MAVS (also called IPS-1, CARDIF, or VISA). A series of structural studies have elaborated some of the mechanisms of dsRNA recognition and activation of RIG-I. Recent studies have proposed that K63-linked ubiquitination of, or unanchored K63-linked polyubiquitin binding to RIG-I positively regulates MAVS-mediated antiviral signaling. Conversely phosphorylation of RIG-I appears to play an inhibitory role in controlling RIG-I antiviral signal transduction. Here we performed a combined structural and biochemical study to further define the regulatory features of RIG-I signaling. ATP and dsRNA binding triggered dimerization of RIG-I with conformational rearrangements of the tandem CARD domains. Full length RIG-I appeared to form a complex with dsRNA in a 2:2 molar ratio. Compared with the previously reported crystal structures of RIG-I in inactive state, our electron microscopic structure of full length RIG-I in complex with blunt-ended dsRNA, for the first time, revealed an exposed active conformation of the CARD domains. Moreover, we found that purified recombinant RIG-I proteins could bind to the CARD domain of MAVS independently of dsRNA, while S8E and T170E phosphorylation-mimicking mutants of RIG-I were defective in binding E3 ligase TRIM25, unanchored K63-linked polyubiquitin, and MAVS regardless of dsRNA. These findings suggested that phosphorylation of RIG inhibited downstream signaling by impairing RIG-I binding with polyubiquitin and its interaction with MAVS.
Adaptor Proteins, Signal Transducing ; metabolism ; Adenosine Triphosphate ; metabolism ; DEAD Box Protein 58 ; DEAD-box RNA Helicases ; chemistry ; genetics ; metabolism ; Dimerization ; Humans ; Mutagenesis, Site-Directed ; Phosphorylation ; Polyubiquitin ; metabolism ; Protein Binding ; Protein Structure, Tertiary ; RNA, Double-Stranded ; metabolism ; Recombinant Proteins ; biosynthesis ; chemistry ; genetics ; Signal Transduction ; Transcription Factors ; metabolism ; Tripartite Motif Proteins ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitination