Objective To study the effects and the mechanism of astragalus menbranaceus (AM) on neonatal lymphocyte apoptosis in cord blood. Methods Cord blood mononuclear cells (CBMC) of 30 full-term neonates were cultured with pure phytohemagglutinin (PHA), PHA combined with IL-6 (PHA + IL-6) or AM (PHA + AM) respectively for 48 hours. The apoptosis index (AI) of lymphocytes of CBMC after cultivation were determined by acridine orange-ethidium bromide dying method. The positive expression of CD38 antigen and CD25 antigen were detected by indirect immunofluorescence procedure. The levels of IL-6 in the supernatants of CBMC were detected by ELISA . Results (1) The AI of the PHA + AM group (16. 5?3. 5)% and PHA+IL-6 group (16. 9?4. 0)% was lower than that of the pure PHA group (32.4?2.8)% (P0. 05). (2) The positive expression rates of CD38 of the PHA+AM group and the PHA+IL-6 group were lower than those of the pure PHA group (P0. 05). The positive expression rates of CD25 of the PHA + AM group and the PHA + IL-6 group were higher than those of the pure PHA group (P0. 05). The positive expression of CD38 of CBMC had positive correlation, but CD25 had negative correlation with the AI of CBMC(r=0. 68, -0. 65,P

Objective To observe the effect of vitamin A(VitA) on T help 17(Th17) and regulatory T cells (Treg) in the peripheral blood in children with asthma and their dose effect relationship,and to investigate the immunoregulation mechanism of VitA.Methods Twenty children with asthma (asthma group) and 16 healthy children (healthy control group) were selected.Peripheral blood mononuclear cells(PBMC) were isolated from venous blood by density gradient centrifugation in the aseptic condition.Different concentrations of VitA [0.0μmol/L (blank control),0.5μmol/L,1.0μmol/L,2.0μmol/L] were added into the cultures in the asthma group.The healthy control group were not interfered with VitA.The supernatant was collected after 72 h.The levels of interleukin 17 (IL-17),interleukin 10(IL-10) and transforming growth factor-β1 (TGF-β1) in the supernatant were determined by enzyme-linked immunosorbent assay.Results (1) IL-17 levels produced by PBMC in the asthma group were significantly higher than those in the healthy control group [(960.53±75.59) ng/L vs (425.07±70.71) ng/L,P＜0.01],and the levels of IL-10 and TGF-β1 were significantly lower than those in the healthy control group [(53.13±6.94)ng/L vs (84.41±6.02) ng/L,(304.51±51.52) ng/L vs (489.45±73.68) ng/L,all P＜0.01].(2) IL-17 levels produced by PBMC in the 0.5μmol/L,1.0μmol/L and 2.0μmol/L VitA concentration of the asthma group [(588.95±44.18)ng/L,(573.13±27.43) ng/L,(686.71±38.98) ng/L] were significantly lower than those in the blank control group[(960.53±75.59) ng/L,all P＜0.01],and IL-17 levels in the 2.0 μmol/L VitA concentration were significantly higher than those in 0.5μmol/L and 1.0μmol/L concentration groop (P＜0.01).(3) IL-10 levels produced by PBMC in the 0.5μmol/L,1.0μmol/L and 2.0μmol/L VitA concentration of the asthma group [(105.35±10.79) ng/L,(111.21±16.11) ng/L,(81.09±6.05) ng/L] were significantly higher than those in the blank control group[(53.13±6.94) ng/L,all P＜0.01],TGF-β1 levels produced by PBMC in the 0.5μmol/L,1.0μmol/L and 2.0μmol/L VitA concentration of the asthma group[(933.01±73.98) ng/L,(1223.31±105.99)ng/L,(776.98±145.44) ng/L] were significantly higher than that in blank control group[(304.51±51.52) ng/L,all P＜0.01],and the levels of IL-10 and TGF-β1 in the 0.5 μmol/L and 1.0μmol/L concentration group were significantly higher than those in the 2.0μmol/L concentration group(all P＜0.01).The level of TGF-β1 in the 1.0μmol/L concentration group were significantly higher than that in the 0.5μmol/L concentration group (P＜0.01).Conclusions The function of Th17 in children with asthma during asthma attack was enhanced,and the function of Treg cells was reduced.The balance disorder of the functions of Th17 and Treg cells occurred.VitA can reduce the function of Th17 in peripheral blood,and enhance the activity of Treg cells in the children with asthma.The physiological level of VitA has the best effect,if high VitA concentration is high its effect is significantly decreased.

Objective To observe the expression of Toll-like receptor(TLR) on peripheral blood dendritic cells(DC) in children with Henoch-Schtinlein purpura(HSP),and to investigate the pathogenesis of the abnormal expression of TLR in children with HSP.Methods Twenty hospitalized children with HSP in the Affiliated Hospital of Qingdao University Medical College from Dec.2011 to Jul.2012 were enrolled in the study(HSP group).Twenty agemetched healthy children were selected as a healthy control group.Peripheral venous blood was sampled under aseptic condition,peripheral blood mononuclear cells (PBMC) were isolated from density gradient centrifugation,and DC were generated by recombinat human granulocyte-macrophage colony-stimulating factor(GM-CSF),interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α) in vitro.Expressions of CD83,CD86 and TLR2,TLR3,TLR4 in peripheral blood DC were examined by fluorescent activated cell sorter (FACS).Results 1.No significant distinction was found in the expression of the C Ds3 on peripheral blood DC between HSP group and healthy control group(t =0.80,P ＞ 0.05) ;in HSP group had remarkably increased expression of the CD86 on peripheral blood DC than that of the healthy control group (t =9.56,P ＜ 0.01).2.Expression rates of TLR2,TLR3,TLR4 on peripheral blood DC in the HSP group were higher than those in the healthy control group(t =1 1.79,13.29,9.45,all P ＜ 0.01).3.Expression rates of TLR2,TLR3 and TLR4 in HSP group had positive correlation with expression rates of CD86 (r =0.84,P ＜ 0.01 ; r =0.53,P ＜ 0.05 ; r =0.66,P ＜ 0.05).Conclusions Expressions of TLR2,TLR3 and TLR4 on peripheral blood DC significantly increased and were positively correlated with expression of CD86.This implies that TLR and co-stimulatory molecules might participate in the pathogenesis of HSP by mediating signal transduction,leading to abnormity of cytokines,then inducing Th1/Th2 immune imbalance by showing the advantage of Th2 function.

Cephalosporin is still the first-line drug chosen for the treatment of gonorrhea, although it is likely to develop resistance to Neisseria gonorrhoeae. Recently, the resistance or low sensitivity of cephalosporin to Neisseria gonorrhoeae have been reported at home and abroad. The studies of resistance mechanism to Neisseria gonorrhoeae are particularly important. The resistance mechanism is mainly lied in three aspects. The first is the changes in antibiotic target sites, for example, the changes in penicillin-binding proteins would affect the binding of antibiotics to Neisseria gonorrhoeae; the second is the changes in bacterial cell membrane porin which effects drugs influxing; the third is the changes in efflux system which enhances the bacterial efflux capacity. To study and explore the mechanism of Neisseria gonorrhoeae resistance is of great importance for the prevention and treatment of gonorrhea.

Objective:To explore the TCM syndromes and risk factors of patients with anxiety and/or depression after coronary revascularization through real-world data mining based on the national pilot project of Chinese and Western medicine clinical collaboration for major difficult diseases; To provide clinical evidence and guide practice for the diagnosis and treatment of bicardiac diseases after coronary revascularization.Methods:A retrospective multi-center clinical study was conducted. From September 2018 to December 2019, 577 patients who underwent successful percutaneous coronary intervention in the Department of Cardiovascular Medicine of Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Department of Cardiovascular Medicine of the People's Hospital of Liaoning Province were enrolled using the collaborative platform system of TCM and Western Medicine Diagnosis and follow-up for coronary disease. Clinical data database was established. Baseline data, TCM syndrome types and elements, coronary angiography and stent implantation status, relevant disease history, Hamilton Anxiety and Depression Scale, etc. were collected. A combination of postoperative phone calls and outpatient visits was performed, with follow-up every 3 months for a total of 1 year. The TCM treatment patterns and risk factors of patients with anxiety and depression after coronary artery revascularization surgery were analyzed and explored.Results:A total of 577 patients were enrolled and 561 patients were followed up. Age distribution: The age of males and females undergoing coronary revascularization due to ACS was (61.80±11.00) years and (68.37±10.13) years, with no statistical significance between groups ( P>0.05), but the age of onset in males tended to be earlier than in females. The distribution pattern of TCM syndrome elements showed that the most deficiency syndrome elements were qi deficiency (61.75%, 176/285), followed by yin deficiency (28.77%, 82/285). The most common excessive symptom was blood stasis (39.13%, 108/276), and the other syndromes were phlegm turbidity (36.23%, 100/276) and qi depression (20.29%, 56/276), etc. The distribution of TCM syndrome types was as follows: in the population with anxiety and depression state of coronary revascularization, the TCM syndrome types with frequency higher than 10% were successively phlegm and blood stasis with depression syndrome, qi deficiency and blood stasis syndrome, heart, gallbladder and qi deficiency and qi-yin deficiency with blood stasis syndrome. Among the people without anxiety and depression, the TCM syndromes with a frequency higher than 10% were heart blood stasis syndrome and qi deficiency and blood stasis syndrome, with statistical significance ( χ2=12.07, P<0.01). Correlation analysis showed that the number of stents, and LDL-C were positive correlated with anxiety and depression( r values were 0.107, 0.118,respectively, P<0.05), and the uric acid was negative correlation ( r=-0.127, P=0.011). Multivariate Cox proportional hazards regression showed the age [ RR (95% CI)=1.052 (1.012-1.094), P=0.010] and diabetes mellitus [ RR (95% CI)=4.561 (1.028-20.238), P=0.046] at the sixth month of treatment. Conclusions:The age of acute coronary syndrome and coronary revascularization is mainly concentrated in patients aged 60-70 years, and male patients tend to have earlier onset than female patients, and the risk of coronary heart disease is relatively high. Qi-deficiency syndrome accounts for the highest proportion, and the most excessive syndrome is blood stasis syndrome. The TCM syndromes with high frequency of anxiety and depression are phlegm and blood stasis with depression syndrome and qi-yin deficiency with blood stasis syndrome. The number of stents implanted and low density lipoprotein cholesterol are positively correlated with postoperative anxiety and depression. Age and diabetes history are independent risk factors for end-point events at about 6 months after treatment.

Objective: To investigate the pathogenic mechanism of two novel ITGB2 mutations in leukocyte adhesion defect type 1 (LAD1). Methods: The clinical history and blood sample of an 11 years old patient admitted to Affiliated Hospital of Qingdao University in August 2014 were collected. Expression of CD18 (encoded by ITGB2) was analyzed by flow cytometry. Novel ITGB2 mutations were identified by next-generation sequencing technology and confirmed by Sanger sequencing. The functional effect of ITGB2 mutations was detected by PolyPhen2. Expression vectors of both wild type and mutant ITGB2 were constructed and transfected into mammalian cells for analysis of protein stability and subcellular location. Results: The symptoms of the patient (recurrent infections, lowered alveolar ridge and hypodontia) supported the diagnosis of LAD1. Expression of CD18 on the leukocytes was significantly decreased (0.2%) compared with the control samples from the parents (paternal: 99.0%; maternal: 99.1%). The patient was identified to be compound heterozygous for ITBG2 c.954del G (novel mutation) and c.1802C>A (paternal originated). ITGB2 c.954 del G was confirmed to be a harmful frameshift mutation; ITGB2 c.1802C>A was also predicted to be harmful. In terms of protein stability. There was no significant difference between mutant D18 and wild type. However, subcellular location analysis showed the mutant D18 could not locate on cell membrane. Conclusion The compound heterozygous of ITGB2 mutations (c.954del G and c.1802C>A) decreases the expression and impairs the location of CD18 on leukocytes, which leads to LAD1.

Objective:To investigate the predictive value of mucosal vascular pattern (MVP) under narrow-band imaging (NBI) enteroscopy in patients with ulcerative colitis (UC) in clinical remission for histological healing and clinical recurrence.Methods:A total of 142 patients with UC in clinical remission who visited the First Affiliated Hospital of Weifang Medical University from January 2018 to January 2021 were included in the study and underwent colonoscopy. The white light and NBI endoscopic images were collected and biopsies were obtained. The Mayo endoscopic score (MES) was calculated based on white light images, and MVP staging was evaluated based on mucosal vascular patterns under NBI. Nancy index (NI) was used to evaluate histological healing and patients were followed up for 1 year. The Spearman correlation coefficients of MES and MVP with histological healing and recurrence were calculated. The receiver operator characteristic (ROC) curve was plotted and the area under curve (AUC) was applied to evaluate the accuracy of white light and NBI endoscopy for predicting histological healing of UC in clinical remission.Results:According to the MVP criteria, 47 were defined as clear, 63 blurred, and 32 invisible. Spearman correlation analysis showed a significant correlation between MVP under NBI and histological healing ( r=0.549, P<0.001) and a moderate correlation between MES under white light and histological healing ( r=0.462, P<0.001). Spearman correlation analysis showed a moderate correlation between MVP under NBI and clinical recurrence ( r=0.451, P<0.001) and a moderate correlation between MES under white light and clinical recurrence ( r=0.352, P<0.001). AUC of NBI for diagnosing histological healing of UC in clinical remission was 0.809 (95% CI: 0.738-0.879), with a sensitivity of 84.6% (77/91) and specificity of 64.7% (33/51), superior to the white light endoscopy, of which AUC, sensitivity and specificity were 0.763 (95% CI: 0.678-0.848), 81.3% (74/91) and 66.7% (34/51). Conclusion:MVP staging under NBI could predict histological healing of UC patients in clinical remission and is superior to white light endoscopy.

The clinical characteristics, mutation analysis results, and family tree of a patient with autosomal dominant Alport syndrome (ADAS), who had nephrotic syndrome as the first manifestation were examined.The proband was a 11-month-old girl who presented with nephrotic syndromes and gross hematuria.During the treatment course, the patient had steroid resistance and a poor response to Cyclosporine and Cyclophosphamide pulse therapy.Renal biopsy was performed 2 years after disease onset.Under the light microscopy, glomerular segmental mesangio-proliferative lesions were observed.The staining of type Ⅳ collagen showed the loss of the α3 chain in the glomerular basement membrane (GBM) and tubular basement membrane, and α5 chain loss in GBM.Electron microscopy showed uneven thickness of GBM, with obviously delaminated and tearing dense basement membrane (BM) layer, showing a typical lace-like change.The segmental BM was loosened and widened.Her father did not develop microscopic hematuria until 10 years later, while her grandmother had asymptomatic hematuria and proteinuria when the proband was diagnosed.A new mutation in the COL4A4 gene was found in the proband, namely c. 1715delG (p.G572Vfs * 81). Her father and grandmother carried the same mutation, but her mother and sister did not have.The clinical manifestation of ADAS is clinically heterogeneous and its incidence may be higher than what we have expected.

OBJECTIVE: To explore the clinical characteristics and genetic etiology for a child with atypical Hemolytic uremic syndrome (aHUS) in conjunct with nephrotic level proteinuria. METHODS: A child patient who had visited the Affiliated Hospital of Qingdao University on June 25, 2020 was selected as the study subject. Clinical data of the patient was collected. Whole exome sequencing (WES) was carried out for the child, and candidate variant was verified by Sanger sequencing of the child and his parents. RESULTS: The child, an 8-month-old male, had presented mainly with edema, oliguria, hematuria, nephrotic level proteinuria, anemia, thrombocytopenia, increased creatinine and urea, hypercholesterolemia but normal complement levels. Genetic testing revealed that he has harbored compound heterozygous variants of the DGKE gene, namely c.12_18dupGAGGCGG (p.P7fs*37) and c.1042G>T (p.D348Y), which were respectively inherited from his father and mother. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variants were classified as likely pathogenic and variant of uncertain significance, respectively. By combining his clinical manifestations and results of genetic testing, the child was diagnosed with aHUS with nephrotic level proteinuria. CONCLUSION For infants and young children with aHUS in conjunct with nephrotic level proteinuria, variants of the DGKE gene should be screened. Above finding has expanded the mutational spectrum of the DGKE gene.
Infant ; Female ; Humans ; Child ; Male ; Child, Preschool ; Atypical Hemolytic Uremic Syndrome/diagnosis* ; Mutation ; Genetic Testing ; Thrombocytopenia/genetics* ; Proteinuria/genetics*