Aim To study the fluorescence spectroscopy of human serum albumin(HSA)and the interaction of aspirin and HSA.Methods The quenching mechanism of the fluorescence of human serum albumin by aspirin was studied with the fluorescence.The interaction dissociation constants KD of human serum albumin and aspirin were determined at different temperatures according to double reciprocal Lineweaver-Burk plot and the main binding force was discussed by thermodynamic equations.The effect of aspirin on human serum albumin was also studied by synchronous fluorescence spectrometry.Results The quenching mechanism of aspirin to human serum albumin was static quenching.The interaction dissociation constants KD at 37℃,25℃ was 1.44?10-3 and 1.96?10-3 mol?L-1 respectively.The thermodynamic parameters of the reaction was-19.73 kJ?mol-1(?H),-16.21 kJ?mol-1(?G),-11.77 kJ?mol-1(?S).Conclusions The main binding force between aspirin and HSA was Van der Waals interaction.Aspirin binding on the human serum albumin could change the serum protein conformation.

Objective:To explore the clinical characteristics and prognostic risk factors of accidental injuries in pediatric intensive care unit(PICU).Methods:The children with accidental injuries admitted to the PICU at Shanghai Children′s Hospital from January 2017 to December 2019 were analyzed retrospectively.The patients were divided into survival group and death group according to outcome.We collected the patients′ clinical data and laboratory indexes, and analyzed the differences between two groups.Multivariate Logistic regression was used to screen the risk factors of death and receiver-operating characteristic(ROC)curve was used to determine the threshold.Results:(1)A total of 253 children were included, accounting for 5.2% of PICU hospitalization during the same period.There were 137 males(54.2%)and 116 females(45.8%), with an average age of 45(19, 96)months.The top three causes of accidental injuries were fall injury in 82 cases(32.4%), traffic accident in 70 cases(27.7%)and foreign body in 39 cases(15.4%). There were 22 cases of deaths, with a fatality rate of 8.7%.(2)The mean arterial pressure at admission in the death group was lower than that in the survival group( P<0.05), and the total length of stay was longer( P<0.001). There were significant differences in Glasgow coma scale, Pediatric Risk of Mortality Scoring Ⅲ(PRISM Ⅲ)between the two groups( P<0.001). In the death group, and the blood lactic acid(LA) significantly increased( P<0.001), hemoglobin and fibrinogen were lower than those in the survival group, the activated partial thromboplastin time and prothrombin time(PT)were significantly prolonged( P<0.001). (3)Multivariate Logistic regression analysis showed that PRISM Ⅲ score, LA and PT were independent risk factors for the death of accidental injuries.The area under curve(AUC)of PRISM Ⅲ score was 0.987(95% CI 0.976-0.999, P<0.001)and the cut-off value was 11 with a sensitivity of 100% and a specificity of 95.7%.The AUC of LA was 0.886(95% CI 0.810-0.961, P<0.001)and the cut-off value was 3.1 mmol/L with a sensitivity of 85.7% and a specificity of 77.3%.The AUC of PT was 0.835(95% CI 0.730-0.941, P<0.001)and the cut-off value was 13.9 s with a sensitivity of 86.6% and a specificity of 72.7%. Conclusion:The main accidental injuries in PICU of our hospital in the past three years are falling injuries, traffic accidents and foreign body injuries.PRISM Ⅲ score, LA and PT at admission are independent risk factors for predicting death.

Aim To compare the interactions of baicalein and baicalin with bovine serum albumin (BSA) and their mechanism. Methods The binding reactions of baicalein and baicalin with BSA and the effects of glucose on them were studied by spectroscopy to compare the binding constants and binding distances of baicalein-BSA and baicalin-BSA,which were calculated according to Lineweaver-Burk equation and F?ster' energy transfer theory. Thermodynamic parameters were used to calculate the types of interaction force between BSA with baicalein or baicalin and the technique of synchronous fluorescence spectra was used to observe the effects of baicalein or baicalin on the conformation of BSA. Results Both the binding constants and binding distances of baicalein-BSA and baicalin-BSA decreased with temperature increasing and were increased by glucose. Relative to baicalein,the binding affinity of baicalin to BSA decreased obviously with an increase in binding distance. Both baicalein and baicalin could form non-covalent compounds with BSA mainly to quench the intrinsic fluorescence of BSA through a static quenching procedure. Baicalein could interact with BSA through hydrogen bonds and Van der Waals force,and baicalin did it mainly through electrostatic force. Though baicalein or baicalin could induce the conformational changes of BSA by binding reaction,only the former reduced the hydrophobicity in microenvironment around the tryptophan moieties of BSA. Conclusions The glycosylation substitution of baicalein molecule can decrease the binding to BSA (baicalin-BSA) and change the types of interaction force. The physiological concentration of glucose increases the binding constants and the number of binding sites of baicalein and baicalin with BSA.

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

Abstract: Due to the continued emergence of multiple variants of SARS-CoV-2, the ongoing pandemic has resulted in severe mortality over the past two years. After the Alpha, Beta, Gamma and Delta variants, the most recent new variant of concern (VOC) strain to emerge is Omicron (B.1.1.529), which evolved as a result of the accumulation of a large number of mutations. The Omicron variant, which has a much higher transmission rate than the Delta variant, soon replaced the Delta variant and others, is now the dominant variant worldwide. The emergence of Omicron poses new challenges for the prevention and control of COVID-19 and has raised a number of concerns worldwide. Recently, cases of Omicron infection have been reported in several parts of China, and therefore this paper provides a comprehensive analysis and summary of the epidemiology and immune escape mechanisms of the Omicron variant. We also suggest some therapeutic strategies against the Omicron variant, including rapid diagnosis, genome analysis of emerging variants, ramping up of vaccination drives and receiving booster doses, updating the available vaccines, designing of multivalent vaccines able to generate hybrid immunity, up-gradation of medical facilities and strict implementation of adequate prevention and control measures need to be given high priority to handle the on-going COVID-19 pandemic successfully.

Objective:To investigate the clinical characteristics and prognosis of children with acute pancreatitis(AP)admitted to the pediatric intensive care unit(PICU).Methods:The etiology, clinical data, imaging features, complications, and outcomes of children with AP admitted to PICU at Shanghai Children′s Hospital from July 2016 to June 2021 were retrospectively analyzed.Results:Totally, 47 patients with AP including 24 males and 23 females were enrolled.The mean age was 84(48, 144)months.Four patients with mild AP, 5 patients with moderately severe AP(MSAP)and 38 cases with severe AP(SAP)were diagnosed.The main etiology was drug-related pancreatitis in 23 cases(48.9%). The organ dysfunction mainly included shock in 28 cases(59.6%), acute respiratory failure in 21 cases(44.7%), gastrointestinal disorders in 30 cases(63.8%), and coagulopathy in 29 cases(61.7%). The complications included capillary leakage syndrome in 18 cases(38.3%)and hyperglycemia(>8 mmol/L)in 24 cases(51.1%). Serum amylase and lipase levels were increased in 47 cases(100%). There were significant differences in blood lactate[3.0(2.1, 4.5)mmol/L vs.1.6(1.1, 3.1)mmol/L, P=0.013], and the use of vasoactive drugs[15.0(0, 75.0)vs.0(0, 8.8), P=0.035] between drug-induced pancreatitis and non-drug-induced pancreatitis.All the patients′conditions were improved after treatment and transferred out of PICU. Conclusion:The etiology of acute pancreatitis in children is mainly drug-related, and the main organ dysfunction includes shock, acute respiratory failure, gastrointestinal dysfunction and coagulation dysfunction.The prognosis of childhood pancreatitis is good.

Objective:To summarize the clinical features, imaging changes, treatment, and prognosis of children with severe autoimmune encephalitis (AE).Methods:A retrospective study was conducted on patients with severe AE admitted to PICU of Shanghai Children’s Hospital from June 2017 to May 2020. Clinical features, treatment protocols and follow-up data were collected.Results:A total of 27 children were included, among which 18 cases (66.7%) were girls. The on-set age was (7.9±3.2) years. Eighteen cases were diagnosed with anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. Fever (77.8%), headache (40.7%) and vomiting (44.4%) were most of prodromal symptoms in children with severe AE. Patients’ neurological symptoms showed seizures (88.9%), mental behavior abnormalities (81.5%), speech disorders (70.4%) and dyskinesia (70.4%). Moreover, epileptic discharge and slow wave activity were critical feature of electroencephalogram (EEG) abnormalities, and the abnormal signal changes on T2-weighted and FLAIR sequence of head MRI were in the posterior horn of the lateral ventricle. In addition, the main comorbidities included refractory status epilepticus (RSE), cardiovascular dysfunction, central hypoventilation syndrome and acute intracranial hypertension syndrome. For patients with central respiratory failure, the median duration of mechanical ventilation was 19.8 (14.8, 29.1) days. According to treatment protocol, the first-line immune treatment included the combination therapies of methylprednisolone, intravenous immunoglobulin (IVIG) and therapeutic plasma exchange (TPE). Eighteen cases were given with methylprednisolone [10-30 mg/(kg. d), 3-5 d] + IVIG (2 g/kg, within 2 d) + TPE, 1 case was treated with methylprednisolone [10-30 mg/(kg·d), 3-5 d] + TPE and 8 cases were given with[10-30 mg/(kg·d), 3-5 d] + IVIG (2 g/kg, within 2 d). Sequential therapy was given with methylprednisolone (1-2 mg/kg), gradually reduced from 3 to 6 months. Finally, 16 children (59.3%) had neurological damages at the first discharge, among which 8 cases (29.6%) were with dyskinesia, 5 cases (18.5%) were with speech disturbance, and 5 cases (18.5%) were with abnormal mental behaviors.Conclusions:The most of first clinical symptom is epileptic seizures in pediatric severe AE, and most of these patients are diagnosed with Anti-NMDA receptor encephalitis. RSE, cardiovascular dysfunction, central respiratory and acute intracranial hypertension syndrome constitute to main organ dysfunctions.

Objective To investigate the association and predictive value of aspartate transaminase to platelet ratio index(APRI) in sepsis-associated liver injury(SALI). Methods We retrospectively ana-lyzed the medical records of patients with sepsis admitted to PICU in Shanghai Children′s Hospital of Shanghai Jiaotong University from April 2015 to March 2017. According to whether liver injury occurred in the sepsis patients during hospitalization,all the patients were divided into SALI group (n=34) and sepsis group(n=222). The clinical characteristics,serological indexes within 24 hours in the PICU,and the ratio of aspartate transaminase to alanine transaminase( AAR) and APRI were collected and analyzed. The receiver operating characteristic( ROC) curve was used to evaluate the power of APRI for the prediction of SALI. Results (1)A total of 256 patients were enrolled in this study. There were 34 cases with SALI,and there were 222 patients with sepsis only,the incidence of SALI was 13. 3％. (2) The values of APRI and AAR were both higher in the SALI group compared with the sepsis group[APRI:7. 12(1. 71,26. 96) vs. 0. 38 (0. 21,0. 83),P<0. 001;AAR:1. 43(0. 94,2. 69) vs. 2. 17(1. 35,2. 96),P<0. 05]. (3)The multivariate Logistic regression analysis showed that total bilirubin, APRI, AAR and platelet were the independent risk factors of SALI(P<0. 05). (4)In addition,the area under the ROC curve(AUC)for the APRI was 0. 891 (95％CI 0. 815-0. 966,P<0. 001),cut-off value was 1. 73,which was superior to total bilirubin(AUC =0. 744,95％CI 0. 634-0. 853,P<0. 001) and platelet(AUC=0. 726,95％CI 0. 611-0. 841,P<0. 001). The clinical sensitivity and specificity of the APRI for identification of SALI from sepsis was 80. 0％ and 92. 2％, respectively. Conclusion APRI is an independently risk factor for the occurrence of SALI and is a precursory marker for SALI.

Objective To investigate the risk factors for spontaneous bacterial peritonitis (SBP) in patients with cirrhotic ascites, and to establish a new model for predicting the development of SBP. Methods A total of 215 patients who were diagnosed with cirrhotic ascites in Hebei General Hospital from September 2016 to September 2020 were enrolled, and according to the presence or absence of SBP, they were divided into SBP group with 55 patients and non-SBP group with 160 patients. Related clinical data were collected and albumin-bilirubin (ALBI) score, Model for End-Stage Liver Disease (MELD) score, MELD combined with serum sodium concentration (MELD-Na) score, and Child-Pugh score were calculated. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups; a multivariate logistic regression analysis was used to screen out independent risk factors, and the receiver operating characteristic (ROC) curve was plotted to evaluate the performance of ALBI score, procalcitonin (PCT), polymorphonuclear neutrophil (PMN) count in ascites, and the ALBI-PMN-PCT combined model in the diagnosis of SBP. Results Compared with the SBP group, the non-SBP group had a significantly higher concentration of Na + ( Z =-3.414, P =0.001) and significantly lower total bilirubin ( Z =-2.720, P =0.007), creatinine ( Z =-1.994, P =0.046), urea nitrogen ( Z =-2.440, P =0.015), C-reactive protein ( Z =-9.137, P < 0.001), PCT ( Z =-8.096, P < 0.001), prothrombin time ( Z =-1.969, P =0.049), international normalized ratio ( Z =-2.073, P =0.038), PMN ( Z =-8.292, P < 0.001), MELD score ( Z =-2.736, P =0.006), MELD-Na score ( Z =-3.188, P =0.001), Child-Pugh score ( Z =-3.419, P =0.001), and ALBI score ( t =-5.010, P < 0.001), and there were also significant differences between the two groups in the presence or absence of gastrointestinal bleeding or hepatic encephalopathy ( χ 2 =16.551 and 8.142, P < 0.001 and P =0.004). The multivariate logistic regression analysis showed that ALBI score (odds ratio [ OR ]=3.460, 95% confidence interval [ CI ]: 1.296-9.240, P =0.013), PMN ( OR =1.012, 95% CI : 1.007-1.017, P < 0.001), and PCT ( OR =6.019, 95% CI : 2.821-12.843, P < 0.001) were independent risk factors for SBP in patients with cirrhotic ascites. The ROC curve showed that ALBI, PCT, PMN, and ALBI-PMN-PCT had areas under the ROC curve of 0.711, 0.866, 0.875, and 0.934, respectively, in the diagnosis of SBP, with sensitivities of 50.91%, 73.36%, 72.73%, and 89.09%, respectively, and specificities of 86.87%, 81.25%, 100.00%, and 91.87%, respectively. The patients with ALBI-PMN-PCT > 0.272 had an increased risk of developing SBP. Conclusion The ALBI-PMN-PCT combined model has a high value in predicting the onset of SBP in patients with cirrhotic ascites.

OBJECTIVE: To investigate the protective effects of curcumin(CUR) and its mechanism on a rat model of neurotoxicity induced by manganese chloride (MnCl₂), which mimics mangnism. METHODS: Sixty male SD rats were randomly divided into 5 groups, with 12 rats in each group. Control group received 0.9% saline solution intraperitoneally (ip) plus double distilled water (dd) H₂O intragastrically (ig), MnCl₂ group received 15 mg/kg MnCl₂(Mn²⁺ 6.48 mg/kg) intraperitoneally plus dd H₂O intragastrically, CUR group received 0.9% saline solution intraperitoneally plus 300 mg/kg CUR intragastrically, MnCl₂+ CUR1 group received 15 mg/kg MnCl₂ intraperitoneally plus 100 mg/kg curcumin intragastrically, MnCl₂+ CUR2 group received 15 mg/kg MnCl₂ intraperitoneally plus 300 mg/kg CUR intragastrically, 5 days/week, 4 weeks. Open-field and rotarod tests were used to detect animals' exploratory behavior, anxiety, depression, movement and balance ability. Morris water maze (MWM) experiment was used to detect animals' learning and memory ability. ICP-MS was used to investigate the Mn contents in striata. The rats per group were perfused in situ, their brains striata were removed by brains model and fixed for transmission electron microscope (TEM), histopathological and immunohistochemistry (ICH) analyses. The other 6 rats per group were sacrificed. Their brains striata were removed and protein expression levels of transcription factor EB (TFEB), mammalian target of rapamycin (mTOR), p-mTOR, Beclin, P62, microtubule-associated protein light chain-3 (LC3) were detected by Western blotting. Terminal deoxynucleotidyl transterase-mediated dUTP nick end labeling (TUNEL) staining was used to determine neurocyte apoptosis of rat striatum. RESULTS: After exposure to MnCl₂ for four weeks, MnCl₂-treated rats showed depressive-like behavior in open-field test, the impairments of movement coordination and balance in rotarod test and the diminishment of spatial learning and memory in MWM (P < 0.05). The striatal TH⁺ neurocyte significantly decreased, eosinophilic cells, aggregative α-Syn level and TUNEL-positive neurocyte significantly increased in the striatum of MnCl₂ group compared with control group (P < 0.05). Chromatin condensation, mitochondria tumefaction and autophagosomes were observed in rat striatal neurocytes of MnCl₂ group by TEM. TFEB nuclear translocation and autophagy occurred in the striatum of MnCl₂ group. Further, the depressive behavior, movement and balance ability, spatial learning and memory ability of MnCl₂+ CUR2 group were significantly improved compared with MnCl₂ group (P < 0.05). TH+ neurocyte significantly increased, the eosinophilic cells, aggregative α-Syn level significantly decreased in the striatum of MnCl₂+ CUR2 group compared with MnCl₂ group. Further, compared with MnCl₂ group, chromatin condensation, mitochondria tumefaction was alleviated and autophagosomes increased, TFEB-nuclear translocation, autophagy was enhanced and TUNEL-positive neurocyte reduced significantly in the striatum of MnCl₂+ CUR2 group (P < 0.05). CONCLUSION Curcumin alleviated the MnCl₂-induced neurotoxicity and α-Syn aggregation probably by promoting TFEB nuclear translocation and enhancing autophagy.
Animals ; Autophagy ; Chromatin ; Curcumin/pharmacology* ; Male ; Mammals ; Manganese/toxicity* ; Rats ; Rats, Sprague-Dawley ; Saline Solution/pharmacology* ; TOR Serine-Threonine Kinases