Objective: To compare the immunomodulatory effects of spore polysaccharides (Gl-SP) and broken spore polysaccharides (Gl-BSP) isolated from Ganoderma lucidum(Leyss et Fr.) Karst. on murine splenic lymphocytes and peritoneal macrophages in vitro. Methods: Mixed lymphocyte culture reaction (MLR), lymphocyte proliferation in the presence or absence of mitogen, and the cytotoxic activity of splenic natural killer (NK) cells were detected with MTT assay in vitro. The percentage of phagocytosis of neutral red (NR) by mouse peritoneal macrophages was detected by colorimetric assay. Splenic T-lymphocyte subpopulations were measured with flow cytometry(FCM). IL-2, IFN-? and TNF-? in the culture supernatants were detected by ELISA and biological assay. Nitric oxide (NO) production was examined by Griess reaction. Results: At the concentration range of 0.2-12.8 mg/L, Gl-SP and Gl-BSP were shown to increase lymphocyte proliferation in the presence or absence of mitogen, enhance NK cytotoxic activity, augment the production of TNF-? and NO in Gl-SP-or Gl-BSP-activated macrophages, as well the percentage of phagocytosis of NR by macrophages in vitro. Both Gl-SP and Gl-BSP could promote MLR, however, at the dose of 12.8mg/L, Gl-BSP showed higher activity than Gl-SP in the proliferation of lymphocytes. These two kinds of polysaccharide could significantly increase the secretion of IL-2 and IFN-? in double-way MLR at the concentrations of 0.2-12.8 mg/L, but Gl-BSP had stronger effects than Gl-SP at the same concentrations. Both Gl-SP and Gl-BSP could increase the ratio of T-lymphocyte subpopulations in double-way MLR. At the concentrations of 0.2-12.8 mg/L or 3.2-12.8 mg/L, Gl-BSP demonstrated more significant activity in increasing the percentage of the CD4+ or CD8+ subset than Gl-SP. At the concentrations of 0.2-0.8 mg/L, the ratio of the CD4+ and CD8+ subset in the Gl-BSP treated group was higher than that of the Gl-SP treated group. Conclusion:Gl-SP and Gl-BSP have sim-ilar immunomodulatory effects in vitro, as though the immunomodulatory effects of Gl-BSP are stronger than that of Gl-SP.

Objective:To provide clinical data for early identification and diagnosis of vascular embolism in children.Methods:We collected and analyzed the clinical data of children with vascular embolism diagnosed at the Children Medical Center of the First Affiliated Hospital of Hunan Normal University from January 2017 to January 2020.Results:A total of 29 children with vascular embolism were included.The male to female ratio was 2.2∶1(20/9); the age range was 1 month to 13 years, and the median age(IQR) was 16 (6-41)months.Among them, 22 cases were diagnosed with venous thrombosis, including 13 children with lower limb venous thrombosis(13/29, 44.8%), and six children with intracranial venous thrombosis(6/29, 20.7%). Arterial embolism was found in six cases, and left atrial appendage thrombosis was found in one case.Severe pneumonia was the most common primary disease(19/29, 65.5%), followed by cardiopulmonary resuscitation(3/29, 10.3%), and Kawasaki disease(3/29, 10.3%). Analysis on the risk factors of vascular embolization diseases, including catheter-related, long-term bed rest, elevated D-dimer, mechanical ventilation, and intravenous hormone administration, showed that 89.2%(25/29)had ≥3 risk factors at the same time, and 82.8%(24/29)had ≥5 risk factors at the same time.Conclusion:In children with vascular thrombotic diseases, deep venous thrombosis, especially lower extremity venous thrombosis, are common.The severe pneumonia is more common in primary disease.Children with multiple risk factors have a higher risk of developing vascular embolism.In clinic, coagulation function should be monitored and local symptoms should be observed for early identification.

Object To study the amount and properties of glycopeptide of Ganoderma lucidum (Leyss ex Fr ) Kaist. cultivated with grass (GLPG) and wood log (GLPW) ; to investigate the feasibility of substitution of grass for wood log to cultivate G lucidum Methods Glycopeptides were extracted and isolated from the fruit body of G lucidum followed by dialysis; the monosaccharid constituent and relative molecular weight of polysaccharides were examined by GC and gel filtration; structure of glycopeptide was determined by IR, 1H NMR and 13 C NMR Results Relative molecular weights of GLPW and GLPG glycopeptides were (Mw) 5 13?10 5 and 5 85?10 5, (M?) 6 21?10 5 and 6 96?10 5, respectively; the glucosidic bonds of both glycopeptides were ? type; both glycopeptides consist 16 amino acids and Rha, Xyl, Fru, Gal and Glc (there was trace amount of Man in GLPW) Conclusion There are less physical and chemical difference between GLPG and GLPW The purification rate of GLPG is 2 8 times to GLPW

Objective To compare the monosaccharide composition and molar ratio of GL-PPT2, GL-PPT3, and GL-PPT4, which are three polysaccharide peptides isolated from the fruit body of Ganoderma lucidum. Methods The polysaccharide peptides were hydrolyzed by triflouroacetic acid, then derivatized by trichloro-aldehyde-1-pheny-3-methyl-5-pyrazolone (PMP), and determined by HPLC method with the 245 nm detection wavelength. Gradient elution with the solution of KH2PO4 (pH 5.0)-acetonitrile (83.5∶16.5) resulted in baseline separation of nine monosaccharides, of which the chromatographic peaks were very clear. Results GL-PPT2, GL-PPT3, and GL-PPT4 comprise the nine monosaccharides of mannose, ribose, rhamnose, glucose, galactose, arabinose, xylose, galactose, and glucuronic acid. Conclusion The ribose in polysaccharide peptides of G. lucidum is first reported.

Objective:To find out the effects of Ganoderma lucidum polysaccharides peptide (Gl-PP) on the invasion of the human lung carcinoma cell (PG cell). Methods: PG cells were pretreated with different concentration Gl-PP in vitro, using cell proliferation assay, cell migration assay, adhesion assay, zymography and RT-PCR, then the effects of Gl-PP on proliferation, motility, adhesion and MMP-9 activity and mRNA expression of PG cells were investigated in vitro. Results: Gl-PP did not directly inhibit PG cell proliferation in vitro. However, pretreated with Gl-PP, PG cells motility was inhibited significantly. PG cells adhesion was also inhibited. The activity of MMP-9 was inhibited in a dose-dependent manner, and the inhibited ratio was 41.53% at a dose of 100 mg/L Gl-PP. The mRNA expression of MMP-9 of pretreated PG cells was inhibited. Conclusion: Gl-PP could suppress invasion of human lung carcinoma cells in vitro.

Objective To investigate the viral prevalence of acute lower respiratory tract infection (ALRTI)in children. Methods Totally 1165 children with clinical diagnosis of ALRTI during the period from August 2007 to September 2008 were involved in our study. The nasopharyngeal aspirate specimen was collected from each patient. RT-PCRs were performed to detect common respiratory tract viruses including respiratory syncytial virus (RSV), rhinovirus (HRV), parainfluenza virus (PIV, type 1 -3 ), influzenza virus type A and B (IFA,IFB), and PCR was used to detect adenovirus (ADV). Results 783 patients were identified to have at least one kind of viral pathogens and the overall positive rate was 67.2%. The most common virus was RSV (27%), followed by HRV ( 17.4% ) and PIV3 ( 13. 9% ). The peak infection seasons were winter and autumn. The etiological spectrum of ALRTI varied in different age groups. Two or more viruses were identified in 284 out of 783 cases ( 36. 3% ). The mixed infection rate was high in infants under 1 year old (63.7%) while it decreased to 8. 5% in children older than 3 years of age. Conclusion RSV, HRV and PIV3 were the most predominant pathogens in children less than 1 year old. The peak infection seasons were winter and autumn. The infection rate and mixed infection rate in infants under 1 year old were highest. The most common style was RSV and HRV mixed infection.

Since heparin-binding protein was first isolated by Shafer in 1984, its bactericidal function and heparin-binding characteristics have aroused the interest of scholars around the world, especially after the recent discovery of the inflammatory chemotactic effect of heparin-binding protein.The use in different fields such as the predictive role of pre-infection is gradually accepted.This review summarized the application of heparin-binding proteins in children with severe infectious diseases.

Objective To analyze the clinical characteristics of fatal cases with confirmed severe ad﹣enovirus pneumonia in children in order to improve the diagnosis and treatment. Methods The fatal cases of severe adenovirus pneumonia admitted to Pediatric Intensive Care Unit of Hunan Provincial People′s Hospital from January 2019 to July 2019 were collected,whose clinical features,diagnosis,treatment,and the causes of death were analyzed retrospectively. Results A total of eight children were enrolled, and the age ranged from 3 months to 3 years old,and five cases were between 6 months to 2 years old. Three cases had underly﹣ing diseases. Adenovirus genotype identification was performed on six patients,and the results showed that all patients were infected with adenovirus type 7. All patients presented persistent high fever,with a peak temper﹣ature between 39. 8℃ to 41. 0℃,which persisted 10 to 37 days. Blood routine test before admission PICU showed that four cases had the decrease in white blood count and hemoglobin concentration,accompanied by﹣increased serum ferritin levels. Seven cases complicated with infection. Four cases had parainfluenza virus type 3 infection. Six cases had bacterial infection,and Gram﹣negative bacilli were predominant. One had fun﹣gal septicemia. Conventional mechanical ventilation were performed in all patients in this group. Four cases in this group died of severe acute respiratory distress syndrome. The other four cases died of disseminated intra﹣vascular coagulation associated with massive gastrointestinal bleeding, pulmonary hemorrhage, heart failure and septic shock combined with multiple organ failure. Conclusion Fatal adenoviruspneumonia mostly app﹣pears in children between 6 months to 2 years old or with underlying diseases. Adenovirus type 7 was the main serotype. The occurrence of reactive hemophagocytic phenomenon should be worsen progression of the disease. Co﹣infection may be an important cause of death.

Objective:To summarize the clinical characteristics of children with Mycoplasma pneumoniae pneumonia(MPP) complicated with pleural effusion, and explore the effect of mixed adenovirus infection on children with MPP complicated with pleural effusion.Methods:The clinical data of children with MPP complicated with pleural effusion diagnosed in Children′s Medical Center at the First Affiliated Hospital of Hunan Normal University (Hunan Provincial People′s Hospital) from January 2013 to December 2019 were collected.MPP cases were divided into single infection group and mixed infection group according to whether mixing adenovirus infection.The clinical characteristics were compared between two groups.Results:A total of 180 children with MPP complicated with pleural effusion were included, the male to female ratio was 1.22∶1 (99/81), the age was 66.13 (44.35, 83.98) months, and the most common cases were children over 5 years old (55.56%). The length of hospitalization was 9.00 (7.00, 12.00) days.Fever (93.33%) and cough (98.33%) were the most common clinical manifestations, and mild increases in C-reactive protein, erythrocyte sedimentation rate and D-dimer were the most common laboratory results.Among included children, right pleural effusion was the most common (54.44%), bilateral pleural effusion accounted for 26.67%, and left pleural effusion accounted for 18.89%.Compared with single infection group, the mixed infection group had a longer hospital stay, a higher proportion of oxygen intake, a higher proportion of gamma globulin use, and a higher value of lactate dehydrogenase and aspartate aminotransferase.The results of multivariate Logistic regression analysis showed that compared with single infection group, although the mixed infection group had a higher proportion of gamma globulin use (36.54% vs.10.93%, P<0.05), the length of hospital stay, clinical manifestations, laboratory examination, chest CT and fiberoptic bronchoscopy showed no statistically significant difference between two groups. Conclusion:MPP complicated with pleural effusion is more common in children over 5 years old, especially in the right side.Mild increases of C-reactive protein, erythrocyte sedimentation rate, and D-dimer are more common.The clinical features of MPP complicated with pleural effusion are similar between mixed adenovirus infection group and single infection group.

Objective:To summarize the clinical characteristics and first aid measures of children with drowning，and to provide scientific basis for the treatment and prevention of drowning in children.Methods:The clinical data of drowning children admitted to the Children's Medical Center of the First Affiliated Hospital of Hunan Normal University（Hunan Provincial People's Hospital）from January 2011 to December 2021 were retrospectively summarized. Baseline characteristics，prehospital emergency care and prognosis of the included children were analyzed.According to the time span, drowning children were divided into two stages, the first stage (2011-2016) and the second stage (2017-2021). According to cerebral performance category assessment scale,drowning children with cardiac arrest were divided into two groups: a group with good neurological prognosis and a group with poor neurological prognosis. The baseline characteristics,pre-hospital emergency care, and prognosis of drowning children during different periods and with different prognosis were analyzed.Results:A total of 53 children were enrolled during the study period，with 24 cases in the first phase（2011-2016）and 29 cases in the second phase（2017-2021）.Boys accounted for 73.6%（39/53）of the cases, the age ranged from 1 month and 24 days to 14 years old，with 1-4 years old being the most common，accounting for 58.5%（31/53）, and the season in which drowning occurred was more common in the summer（39.6%, 21/53）.Six cases (11.3%) had significant neurological sequelae, and the mortality rate was 3.8%（2/53）.There was a decrease in the proportion of drowning with water control in the second stage compared to the first stage（37.5% vs. 13.79%, P=0.046）.Thirty-two（60.4%）children experienced on-site cardiopulmonary resuscitation(CPR) after drowning，with 24 cases in the group with good neurological prognosis and 8 cases in the group with poor neurological prognosis.Children in the poor neurological prognosis group required CPR for a long duration [10.00（1.50, 25.00）min]，had lower body temperature，Glasgow Coma Score，pH and higher blood glucose levels on admission（all P < 0.05）. Conclusion:Drowning is most common in preschool children，common in boys，and drowning prevention measures for young children deserve focused attention. Children found to be in cardiac arrest at the time of drowning have a long duration of drowning and CPR，which predicts a poor neurological prognosis，and timely and effective on-site CPR is the key to reducing the drowning rate.