Objective:To detect expression levels of complement regulatory proteins CD55 and CD59 in the peripheral blood of patients with Stevens-Johnson syndrome (SJS) /toxic epidermal necrolysis (TEN), and to preliminarily analyze their potential roles in the occurrence of SJS/TEN.Methods:Hospitalized patients with SJS/TEN (SJS/TEN group) were collected from the Department of Dermatology of the First Affiliated Hospital of Anhui Medical University and the Second Affiliated Hospital of Anhui Medical University from December 2017 to December 2022. Meanwhile, patients with maculopapular exanthema (MPE) and healthy physical examinees were also collected and served as the mild group and healthy control group, respectively. Flow cytometry was performed to determine the proportions of CD4 + T lymphocytes and CD8 + T lymphocytes in peripheral blood mononuclear cells (PBMCs). The expression levels of inflammatory cytokines tumor necrosis factor (TNF) -α, interleukin (IL) -6, IL-17, IL-10, interferon (IFN) -γ, IL-2, and IL-4 were detected using flow cytometric bead array technology. The mRNA expression levels of CD55 and CD59 in PBMCs were detected by real-time fluorescence-based quantitative PCR (qRT-PCR). Flow cytometry was also performed to determine the protein expression of CD55 and CD59 on the surface of CD8 + T lymphocytes. Statistical analyses were carried out using one-way analysis of variance and Tukey's test. Results:Totally, 13 patients with SJS/TEN, 27 patients with MPE, and 40 healthy controls were collected. Among the SJS/TEN patients, there were 8 males and 5 females, with their age being 18 to 84 (47.15 ± 19.99) years, and disease duration being 7.74 ± 2.63 days. No significant differences were observed in the gender distribution or age among the 3 groups (both P > 0.05). The proportions of CD4 + T lymphocytes did not differ among the 3 groups ( F = 3.84, P = 0.051). The proportions of CD8 + T lymphocytes in the peripheral blood were significantly higher in the SJS/TEN group (25.60% ± 4.57%) than in the healthy control group (16.20% ± 6.28%; q = 4.59, P = 0.018). The expression levels of inflammatory cytokines TNF-α, IL-6, IL-10, IL-17, and IFN-γ were significantly higher in the SJS/TEN group than in the healthy control group and mild group (all P < 0.001). In addition, the mRNA expression of CD55 ( F = 9.46, P < 0.001) and CD59 in PBMCs ( F = 15.14, P < 0.001) was significantly lower in the SJS/TEN group than in the mild group and healthy control group. The protein expression levels of CD55 ( F = 51.51, P < 0.001) and CD59 ( F = 31.59, P < 0.001) on the surface of CD8 + T lymphocytes were also significantly lower in the SJS/TEN group than in the other two groups and the healthy control group, respectively. Conclusion:Complement regulatory proteins CD55 and CD59 were downregulated in SJS/TEN patients, which may be associated with the activation of CD8 + T lymphocytes and excessive inflammatory responses.

Abstract: Objective　To compare and analyze the clinical characteristics, diagnosis, and treatment of different subtypes of severe influenza in neonates to provide a reference for the diagnosis and treatment of neonatal severe influenza. Methods A cohort of 40 neonates with severe influenza who were hospitalized in the neonatology ward of Children's Hospital Affiliated to Zhengzhou University between January 2019 to December 2023 were selected and divided into two groups based on the virus subtype: influenza A (n=23) and influenza B (n=17). A retrospective analysis was conducted to compare general information, clinical manifestations, auxiliary examinations, complications, and treatment outcomes of neonates with severe influenza A and B infection. Results The number of days of hospitalization was longer in cases of influenza A than that of influenza B. The proportion of neonates with severe influenza A who exhibited fever was higher than that for influenza B, and a higher percentage of those with fever had peak temperatures ranging from 38.1 ℃ to 39 ℃. Gastrointestinal symptoms, including vomiting and diarrhea leading to dehydration, were more evident in severe influenza B cases. The proportion of influenza A cases with abnormal creatine kinase-MB isoenzyme levels (>25 U/L) was higher than that of influenza B, and the differences were statistically significant (P<0.05). There were no significant differences between the two types of influenza in other clinical manifestations, the incidence of pneumonia/respiratory failure complications, peripheral blood leukocyte count and classifications, the proportion of abnormal aspartate aminotransferase (AST) (>40 U/L), alanine aminotransferase (ALT) (>40 U/L), and creatine kinase (CK) (>200 U/L), and lactate dehydrogenase (LDH) values (all P>0.05). In terms of treatment, neonates treated with Oseltamivir within 48 hours of onset mainly suffered from influenza A. Among those treated with Oseltamivir, the proportion of influenza A cases whose body temperature returned to normal within 24 hours was relatively higher, whereas, for those whose temperature returned to normal within 24-72 hours, the proportion was relatively higher in influenza B cases. These differences were statistically significant (all P<0.05). Conclusions Severe neonatal influenza usually occurs in winter and spring. After severe infection, fever is more obvious in neonates with influenza A, which is more likely to cause myocardial cell damage. Neonates with influenza A can be treated with Oseltamivir earlier and return to normal body temperature faster than those with influenza B after Oseltamivir treatment. Gastrointestinal symptoms are more common in neonates with severe influenza B.

OBJECTIVE: Scutellarin is a primary active composition come from Erigeron breviscapus. It is well known that scutellarin has anti-inflammatory and antioxidant physiological functions. In this study, we detected the effects of scutellarin on hepatocyte cell apoptosis in type 2 diabetes mellitus (T2DM) rats. METHODS: Sprague Dawley (SD) (6-8 weeks, 160-180 g) rats were randomly divided into six groups: control, model, scutellarin low-dose, medium-dose, high-dose treatment, and rosiglitazone positive groups; with 10 SD rats in each group (n = 10). The changes of biochemical factors in serum were detected by automatic biochemical instrument, the pathological changes of liver tissue were detected by hematoxylin and eosin (HE) staining, the apoptosis of liver tissue and cells was detected by tissue staining and flow analyzer, and the expression of apoptosis-related factors were determined by qPCR, Western blot and immunohistochemistry in liver tissues or cells. RESULTS: The results showed that scutellarin decreased the levels of fasting blood glucose, total cholesterol, triglyceride, and low-density lipoprotein and increased the levels of high-density lipoprotein. Meanwhile, scutellarin decreased the levels of alanine transaminase (ALT) and aspartate transaminase (AST) and improved liver function. In addition, scutellarin suppressed the secretion of interleukin-1 (IL-1), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and reduced hepatocyte apoptosis. Furthermore, scutellarin inhibited the expression of cleaved Caspase-3, Bax, and cytochrome C (Cyt-C) and promoted the expression of Bcl-2. CONCLUSION Scutellarin can inhibit the apoptotic pathway, thereby relieving T2DM.

Objective:To analyze the detection strategy and basic detection situation of markers of infectious diseases transmitted by transfusion in blood testing laboratories of blood stations in China.Methods:Based on the data of practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021, the data on the testing strategies and the basic detection information of the markers for the transmission of infectious diseases through transfusion in the member laboratories of the practice comparison working party of Blood Stations in Mainland of China from 2017 to 2021 were collected, and the situation of the selection for testing markers, testing strategy and the testing method and other relevant aspects were sorted out and analyzed by charts.Results:The selection of the testing markers was consistent, but HTLV testing item was added in some member laboratories. The detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously was adopted in 47 member blood stations; 3) NAT method was dominated by mini pool-NAT in member laboratories. The number of members adopting mini-pools of 8 (MP8)-NAT decreased from 17 in 2017 to 14 in 2021, while the number of members adopting mini-pools of 6 (MP6)-NAT increased from 13 in 2017 to 22 in 2021; Roche NAT system accounted for the largest proportion.Conclusions:In order to ensure blood safety and avoid missing detection, the blood stations still adopt the detection strategy of using two ELISA reagents and one nucleic acid testing (NAT) reagent simultaneously; Meanwhile, in order to increase the NAT positive rate, the proportion of mini pool-NAT mainly decreased year by year despite its dominating role, while the proportion of individual donation-NAT increased year by year; NAT method is transiting from mini-pools of 8 (MP8) to mini-pools of 6 (MP6); The proportion of imported NAT system used in NAT laboratory is relatively large.

Objective:To evaluate the role of mitochondrial calcium homeostasis in hyperoxia-induced injury to alveolar epithelial cells typeⅡ(AECⅡ) in rats.Methods:Rat AECⅡcell lines (RLE-6TN) were randomly divided into 3 groups ( n=18 each) using a random number table method: nomoxia group (group C), hyperoxia group (group H), and hyperoxia plus ruthenium red group (group HR). The cell lines were cultured in a conventional incubator for 4 h in group C. The cell lines were incubated in an incubator with 90% O 2 for 4 h in group H. In group HR, ruthenium red 2 μmol/L was added first, and then the cell lines were incubated in an incubator with 90% O 2 for 4 h. After the treatment, the Ca 2+ concentrations in mitochondria were measured using the mitochondrial Ca 2+ specific fluorescent probe Rhod-2-AM, the ROS level was measured using the fluorescent probe DCFH-DA, and the cell morphology and mitochondrial ultrastructure were observed. Results:Compared with group C, the Ca 2+ concentrations in mitochondria and ROS level were significantly increased in H and HR groups ( P<0.05). Compared with group H, the Ca 2+ concentrations in mitochondria and ROS level were significantly decreased in group HR ( P<0.05). Compared group C, the cell volume was reduced, the morphology was shrunken round, the cell arrangement was loose, the mitochondrial double-layer membrane structure was broken, and the mitochondrial cristal fragments were observed in group H. Compared with group H, the cell morphology was significantly improved, and the slight damage to mitochondrial double-layer membrane structure was found, and the structure of mitochondrial cristae was normal in group HR. Conclusion:Mitochondrial calcium homeostasis is involved in the pathophysiological process of hyperoxia-induced injury to AECⅡ in rats.

A case with mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase deficiency(HMGCSD)was related and foreign and domestic reported cases were reviewed.The female proband was 7 months and 16 days old, and admitted to the hospital due to acute onset of " fever for 4 days, wheezing for 3 hours, dyspnea and moaning for 2 hours" . She was mainly manifested as encephalopathy, hepatomegaly, liver function damage, low ketone hypoglycemia, and hyperlipidemia.She died of respiratory and circulatory failure on the third day of hospitalization.Two compound he-terozygous variants in HMGCS2 gene were found by total exome sequencing, namely, c.1061+ 1 G> C and c. 476 G> T. HMGCSD could be diagnosed by gene detection in combination with clinical features of the patient. Thirteen literatures related to HMGCSD were collected, including 26 patients in total, with the age of onset ranging from 3 months to 6 years. The main cause of the disease was insufficient intake, mainly manifested as hypoglycemia accompanied by low ketone, hepatomegaly, liver damage, etc. A high level of urinary 4- hydroxy-6- methyl-2- pyrone might be a strong indicator of HMGCSD. Three died during the acute attack. Up to now, there were 32 mutations in HMGCS2 reported in 26 patients, and the main type was missense mutation. In this article, the second case of HMGCSD in China was identified, and 2 novel variants of HMGCS2 were found, which extended the clinical phenotype and mutation spectrum of HMGCSD.

Objective: To investigate the value of abnormal expression of HLA-DR on peripheral blood monocytes in evaluating the immune function status, clinical prognosis and severity of patients with hand, foot and mouth disease (HFMD). Methods: From June 2017 to October 2018, 100 cases of mild HFMD, 80 cases of severe HFMD, 32 cases of critical HFMD and 40 healthy children (control group) were recruited in this study. The patients were divided into two groups, lower DR group (DR-L, HLA-DR expression<30%) and normal DR group (DR-N, HLA-DR expression>30%) according to the HLA-DR expression on monocytes. Flow cytometry was used to detect the CD14⁺ monocytes expressing HLA-DR and the absolute count of lymphocyte subsets. Immunoturbidimetry was used to detect the levels of IgG, IgM and IgA in plasma samples. Enzyme-linked immunosorbent assay (ELISA) was performed to detect the levels of IFN-γ and IL-10 in plasma samples. Pediatric critical illness score (PCIS) and the pediatric risk of mortality Ⅲ (PRISM Ⅲ) were used to estimate the severity of HFMD. Results: ① There were significant differences in HLA-DR expression on monocytes among children with mild, severe and critical HFMD (F=47.102, P<0.05). Patients with critical HFMD had the lowest HLA-DR expression (P<0.05). ② The numbers of CD14⁺ monocytes, CD3⁺ T cells, CD4⁺ T cells, CD8⁺ T cells, B cells and NK cells in peripheral blood of the DR-L group were significantly lower than those of the DR-N group and the normal group, especially in patients with severe or critical HFMD (P<0.05). ③ There was no significant difference in the level of IgG, IgA or IgM among the DR-L, DR-N and control groups (P>0.05). ④ Compared with the DR-N group, the DR-L group showed decreased IFN-γ level and increased IL-10 level in plasma (P<0.05). The ratio of IFN-γ/IL-10 of the DR-L group was lower than that of the DR-N group and control group (P<0.05). HLA-DR expression was negatively correlated with the concentration of IL-10 in plasma (r=-0.704, P<0.05), and positively correlated with the IFN-γ/IL-10 ratio (r=0.773, P<0.05). ⑤ Compared with the DR-N group, the DR-L group showed lower PCIS and higher PRISM Ⅲ. HLA-DR expression was positively correlated with PCIS (r=0.715, P=0.00) and negatively correlated with PRISM Ⅲ (r=-0.610, P=0.00). ⑥ The incidence of pulmonary edema, pulmonary hemorrhage and cardiopulmonary failure and the mortality of HFMD patients in the DR-L group were significantly higher than those in the DR-N group (P<0.05). Conclusions Patients with severe or critical HFMD had cellular immune dysfunction and abnormal HLA-DR expression on CD14⁺ monocytes. Assessing the expression of HLA-DR on monocytes could be used to evaluate the cellular immunity of patients with severe or critical HFMD. Lower expression of HLA-DR on CD14⁺ monocytes might be associated with severe HFMD and poor prognosis.

Objective To explore the protective effect of magnesium sulfate on the nerve injury in severe hand, foot and mouth disease ( HFMD) caused by enterovirus A71 ( EV-A71) and to investigate its clinical and prognostic effects.Methods A total of 240 cases of severe HFMD with EV-A71 infection and nerve injury were enrolled.According to the random number table method, the patients were randomly divided into conventional treatment group (control group) and magnesium sulfate treatment group ( treatment group), with 120 cases in each group.The control group was given the routine treatment, and the treatment group was given the magnesium sulfate adjuvant treatment on the basis of routine treatment.The neurological symptoms and signs, clinical efficacy and prognosis were observed before and after treatment in the two groups.The blood and cerebrospinal fluid neuron-specific enolase ( NSE), S100-βprotein and neuropeptide Y ( NPY) were analyzed before and after treatment.The amplitude integrated electroencephalogram (aEEG) was used to monitor the abnormal recovery of EEG.The t-test was applied to analyze quantitative data, and the chi-square test was used for qualitative data comparison.Results Among children with severe HFMD, there were 83 cured cases, 29 improved cases and 8 ineffective cases in control group, with the total effective rate of 93.3%; while in the treatment group, 101 cases were cured, 18 cases were improved and 1 case was ineffective, the total effective rate was 99.2%.The therapeutic effects (Z＝2.918, P＝0.004) and the total effective rate ( χ2 ＝4.156, P＝0.041) were statistically significantly different between the two groups.Three days after treatment, the average levels of serum NSE, S100-βprotein and NPY in magnesium sulfate treatment group were significantly lower than those in control group (t＝-7.239,-10.020 and -11.053, respectively, all P＜0.01).Five days after treatment, the average levels of cerebrospinal fluid NSE, S100-β protein and NPY in magnesium sulfate treatment group were significantly lower than those in control group ( t＝-6.546,-13.308 and -10.258, respectively , all P＜0.01).After treatment, the neurological function score in treatment group was significantly lower than that in control group and that before treatment , and the differences were statistically significant ( t ＝-9.473 and 12.162, respectively, both P ＜0.01 ).The recovery time of the main symptoms and signs in treatment group was ( 2.33 ±0.76 ) d, which was significantly shorter than that of control group ([3.21 ±0.82] d), the difference was statistically significant (t＝-12.52, P＜0.05).The average length of hospital stay in treatment group was (5.79 ±1.42) d, which was shorter than that in control group ([ 6.71 ±1.46 ] d ), and the difference was statistically significant ( t＝-4.932, P＜0.05).Of the 240 children with severe HFMD, 194 (80.8%) patients had abnormal aEEG.Before treatment, the aEEG abnormal rates in control group and the magnesium sulfate treatment group were 79.2%(95 cases) and 82.5%(99 cases), respectively, there was no significant difference ( χ2 ＝0.430, P＞0.05); while after treatment for 3 days, 76 cases in treatment group returned to normal, and the recovery rate of aEEG was 76.8%, which was higher than that in control group (52.6%). The difference was statistically significant ( χ2 ＝12.406, P ＜0.05 ).Conclusions Magnesium sulfate adjuvant therapy can reduce the abnormal levels of NSE, S100-βand NPY in blood and cerebrospinal fluid, relieve clinical symptoms, shorten the course of disease and average length of hospital stay, improve the neurological function score, and promote the recovery of abnormal aEEG.Thus, it has neuroprotective effect on severe HFMD with nervous system lesion.

objective To analyze the clinical characteristics of pneumonia in patients with aplastic anemia for improving early prevention and clinical diagnosis. Methods A retrospective analysis was conducted for patients with aplastic anemia treated in our hosipital from June 2013 to April 2018. The clinical data of pneumonia were reviewed and analyzed in terms of radiological findings, pathogen distribution, and related risk factors. Results The clinical manifestations are atypical in patients with aplastic anemia. The imaging findings suggested that the infection was mainly bilateral pulmonary infection. The common pathogens were gram-negative bacteria such as Klebsiella pneumoniae. Disease type, agranulocytosis and low serum albumin level were independent risk factors for pneumonia in patients with aplastic anemia. Regular immunosuppressive therapy is a protective factor for pneumonia in patients with aplastic anemia. Conclusions The clinical manifestations of pneumonia are diverse in patients with aplastic anemia. The pathogens are mainly gram-negative bacteria. Empirical use of β-lactam-β-lactamase inhibitor combinations or quinolones is beneficial for controlling infection. Regular immunosuppressive therapy, recovery of hematopoietic function, nutritional support are effective measures to reduce the incidence of pneumonia in patients with aplastic anemia.

Objective To study the clinical characteristics of outpatients with hand,foot and mouth disease (HFMD) caused by different serotypes of enteroviruses.Methods This was a prospective study.From February 2017 to March 2018,563 outpatients with HFMD were enrolled by systematic sampling in the Department of Infectious Diseases,Henan Children's Hospital.Throat swabs were collected to determine the serotypes via PCR.Demographic,clinical,and laboratory data were collected by standard questionnaire.All cases were followed up twice at 2 and 9 weeks after the initial outpatient visit through telephone interview.A total of 563 cases were enrolled and 555 (98.6％) cases were positive for human enteroviruses,including 338 (60.9％) males.Analyses were stratified by enterovirus serotypes,Chi square test or Fisher's exact test,Rank sum test was used for comparison among different groups.Results The age of 555 cases was 24.2 (16.4,41.3) months.Among them 44.0％ (224 cases) were identified as coxsackievirus (CV)-A6,while 189 cases,35 cases,14 cases and 73 cases were identified as CV-A16,enterovirus (EV)-A71,CV-A10 and other serotypes,respectively.Fever (≥37.5 ℃C) was present in 51.4％(285/555) of laboratory confirmed cases.The proportions of fever in cases of CV-A6 (68.9％(168/244)) and CV-A10 (12/14) were significantly higher than those in cases of CV-A16 (31.7％(60/189),x2=57.344,14.313,both P=0.000),other serotypes (43.8％(32/73),x2=15.101 and 8.242,P=0.000 and 0.004) and EV-A71 (37.1％(13/35),x2=13.506 and 9.441,P=0.000 and 0.002) respectively.There was no significant difference between CV-A6 and CV-A10 in presentation of fever (x2=1.785,P=0.182).There were 359 cases (64.7％) with eruptions in mouth,hands,feet and buttocks.Cases infected with EV-A71 had the highest proportions (74.3％(26/35)) of rash emerging simultaneously in mouth,hands,feet,and buttocks.The proportion in cases of CV-A 16,CV-A6,CVA 10 and other serotype were 73.5％ (139/189),61.9％ (151/244),7/14 and 49.3％ (36/73),respectively.The proportion of rash on other parts of body,such as face,limbs or torso in cases infected with CV-A6 (16.8％ (41/244)) was the higherest and the proportion in cases of CV-A16,EV-A71,CV-A10 or other serotypes were 8.5％(16/189),5.7％(2/35),1/14,6.8％(5/73),respectively.None of these cases developed serious complications.Desquamation occurred in 45.5％ (179/393) cases 7.5 (5.0,9.0) days after disease onset and 13.5％ (53/393) cases showed onychomadesis 31.0 (18.0,33.5) days after disease onset.The proportion of desquamation and onychomadesis associated with CV-A6 (64.2％ (95/148) and 31.8％ (47/148)) was significantly higher than CV-A16 (31.8％ (49/154) and 1.3％ (2/154),x2=33.601 and 52.482,both P=0.000) and other serotypes (38.0％(19/50) and 6.0％(3/50),x2=10.236 and 12.988,P=0.001 and 0.000).Desquamation appeared more in cases of CV-A6 than in cases of CV-A10 (2/11,x2=9.386,P=0.002),with the proportion of onychomadesis higher in CV-A6 than in EV-A71 (3.3％ (1/30),x2=11.088,P=0.001).Conclusion Clinical manifestation such as fever,rash emerging parts,desquamation and onychomadesis are different among outpatient HFMD cases infected with CV-A16,CV-A6,EV-A71,CV-A10 and other enteroviruses.