Human salivary histatin 1 (Hst1) exhibits a series of cell-activating properties, such as promoting cell spreading, migration, and metabolic activity. We recently have shown that fluorescently labeled Hst1 (F-Hst1) targets and activates mitochondria, presenting an important molecular mechanism. However, its regulating signaling pathways remain to be elucidated. We investigated the influence of specific inhibitors of G protein-coupled receptors (GPCR), endocytosis pathways, extracellular signal-regulated kinases 1/2 (ERK1/2) signaling, p38 signaling, mitochondrial respiration and Na+/K+-ATPase activity on the uptake, mitochondria-targeting and -activating properties of F-Hst1. We performed a siRNA knockdown (KD) to assess the effect of Sigma-2 receptor (S2R) /Transmembrane Protein 97 (TMEM97)-a recently identified target protein of Hst1. We also adopted live cell imaging to monitor the whole intracellular trafficking process of F-Hst1. Our results showed that the inhibition of cellular respiration hindered the internalization of F-Hst1. The inhibitors of GPCR, ERK1/2, phagocytosis, and clathrin-mediated endocytosis (CME) as well as siRNA KD of S2R/TMEM97 significantly reduced the uptake, which was accompanied by the nullification of the promoting effect of F-Hst1 on cell metabolic activity. Only the inhibitor of CME and KD of S2R/TMEM97 significantly compromised the mitochondria-targeting of Hst1. We further showed the intracellular trafficking and targeting process of F-Hst1, in which early endosome plays an important role. Overall, phagocytosis, CME, GPCR, ERK signaling, and S2R/TMEM97 are involved in the internalization of Hst1, while only CME and S2R/TMEM97 are critical for its subcellular targeting. The inhibition of either internalization or mitochondria-targeting of Hst1 could significantly compromise its mitochondria-activating property.
Endocytosis/physiology* ; Histatins/pharmacology* ; Humans ; Membrane Proteins ; Mitochondria/metabolism* ; RNA, Small Interfering/pharmacology* ; Receptors, G-Protein-Coupled/metabolism* ; Receptors, sigma

Objective:To analyze the clinical characteristics and diagnosis and treatment experiences of Langerhans cell histocytosis (LCH) in skull.Methods:Sixteen patients with cranial LCH admitted to our hospital from January 2015 to December 2019 were chosen in our study. Their clinical data, diagnosis and treatment procedures and prognoses were retrospectively analyzed.Results:Among the 16 patients, there were 13 males and 3 females, aged from 1 to 31 years. The clinical manifestations included space-occupying lesions of the skull; and imaging showed bone destruction of the skull, with or without involvement of other bones or organs. All patients were pathologically confirmed to have LCH after surgical total resection of the lesions. Routine whole-body bone scanning was performed after surgery: one was found to have local abnormal metabolic activity and received local radiotherapy; 8 were combined with other bone or organ involvement, and received chemotherapy. All the patients were followed up for 1-5 years, and no recurrence was found, and no one died.Conclusion:Good prognosis can be achieved in cranial LCH patients accepted resection by giving additional treatment according to the results of postoperative reexamination and combination use of standardized radiotherapy and chemotherapy.

Macrophages play an important role in material-related immune responses and bone formation, but the functionality of macrophage-derived extracellular vesicles (EVs) in material-mediated bone regeneration is still unclear. Here, we evaluated intracellular communication through small extracellular vesicles (sEVs) and its effects on endogenous bone regeneration mediated by biomimetic intrafibrillarly mineralized collagen (IMC). After implantation in the bone defect area, IMC generated more neobone and recruited more mesenchymal stem cells (MSCs) than did extrafibrillarly mineralized collagen (EMC). More CD63
Biomimetics ; Bone Regeneration ; Cell Differentiation ; Collagen ; Extracellular Vesicles ; Macrophages ; Osteogenesis

Objective: To evaluate the sensitivity and specificity of exhaled nitric oxide in the diagnosis and therapy of bronchial asthma. Methods: From January 2016 to December 2017, 87 patients diagnosed with bronchial asthma in Luoyang Central Hospital Affiliated to Zhengzhou University were selected as asthma group , and 40 healthy people were selected as healthy control group at the same term.The levels of FeNO, FEV₁/pred, peripheral blood EOS count, ACT score, percentage of sputum EOS were measured. Results: The positive rate of FeNO was 56.3%(49/87) in the asthma group, which in the healthy control group was 5.0%(2/47). The fractional exhaled nitric oxide level of reexamined patients before treatment [(70.9±53.6)ppb] was higher than that after treatment[(12.2±8.7)ppb], and the difference was statistically significant(t=4.323, P=0.001). The fractional exhaled nitric oxide level in the asthma group [(42.4±42.5)ppb] was higher than that in the healthy control group [(9.4±5.8)ppb], and the difference between the two groups was statistically significant(t=2.871, P=0.001). The fractional exhaled nitric oxide level was positively correlated with peripheral blood EOS count (r=0.376, P=0.000) and ACT score (r=0.361, P=0.001), but had no correlation with FEV₁/pred (r=-0.111, P=0.306) and sputum EOS (r=-0.036, P=0.805). Conclusion The measurement of fractional exhaled nitric oxide levels has an important clinical significance in the diagnosis of bronchial asthma.And it has a guiding significance for the evaluation of the therapeutic effect of bronchial asthma.

OBJECTIVE:To provide reference for standardized management of skin test of cephalosporin injection in our hos-pital. METHODS:In retrospective analysis,skin test of 8 kinds of cephalosporin [cefotaxime(1.0 g),cefotaxime(0.5 g),cefoti-am (1.0 g),cefotiam (0.5 g),ceftazidime (0.5 g),cefminox (0.5 g),cefminox (1.0 g),ceftriaxone (1.0 g)] and the cost of skin test, related cost of allergic reaction induced by cephalosporin injection were analyzed statistically during Sept. 1st, 2015-Aug. 31th,2016. The cost of skin test of cephalosporin injection was compared with allergic reaction cost reduced by skin test. RESULTS:The positive rate of 100330 patients who used above 8 kinds of cephalosporin injections was 6.27%;the rate of skin test was 82.49%;the direct cost of skin test was 3434411.72 yuan;the indirect cost was 141985.12 yuan;the cost of pa-tient was 3162901.44 yuan;the cost of medical insurance was 259096.28 yuan;the cost of the whole society was 3576396.84 yuan. From the perspective of the whole society,the cost of skin test of cephalosporin injections was(447049.61±247395.07)yuan, and allergic reaction cost reduced by skin test was (316075.48 ± 260600.49)yuan. The cost of skin test was significantly higher than allergic reaction cost reduced by skin test,with statistical significance(P<0.05). CONCLUSIONS:Skin test of cephalosporin injection is low in positive rate,has high expense. The government standardizes the management of skin test of cephalosporin injec-tion and reduces the economic burden of patients under the premise of ensuring the safety of drug use.

Objective: To investigate the usefulness of loss of CIC expression as the prescreening detection of 1p/19q co-deletion in the diagnosis of oligodendroglial tumors and its prognostic implication. Methods: The retrospective study included 113 oligodendroglial tumors diagnosed in the Department of Pathology, Xuanwu Hospital, Capital Medical University. Expression of CIC protein was detected by immunohistochemistry, and the 1p/19q co-deletion by fluorescence in situ hybridization in all the tumors; and the correlation of the loss of protein and 1p/19q co-deletion with prognosis was assessed. Results: The rate of negative CIC protein expression was 59.3% (67/113) in 113 oligodendroglial tumors. CIC protein expression was differentially lost in various gliomas, 85.7% (42/49) in pure oligodendrogliomas and 39.1% (25/64) in mixed oligodendroglial tumors (P<0.01). The loss of CIC protein expression showed a sensitivity of 76.1% (54/71), specificity 71.1% (27/38), false positive rate of 16.9% (11/65), and a false negative rate of 38.6% (17/44). In 63 cases integrated diagnosis as oligodendroglial tumors with mutant IDH and 1p/19q co-deletion, the loss of CIC protein expression was 81.0% (51/63); the sensitivity and specificity were increased to 81.0% (51/63) and 76.9% (20/26), and the false positive rate and false negative rate decreased to 10.5% (6/57) and 37.5% (12/32), respectively. By using Kaplan-Meier analysis, the CIC negative group showed a trend towards better outcome than the CIC positive group, but there was no statistical difference (overall survival: P=0.218; progression free survival: P=0.249). Conclusions Detection of the lost CIC protein expression can predict the chromosome 1p/19q co-deletion. In oligodendroglial tumors with IDH mutant and 1p/19q co-deletion, there is no relation between prognosis and CIC protein expression.

OBJECTIVETo deepen the understanding of clinical manifestations and treatment of patients with positive lupus anticoagulant (LAC).

METHODSThe clinical data of 2 patients were analyzed and related literature were reviewed.

RESULTSCase 1, a 31-year-old female, diagnosed as lupus anticoagulant positive, secondary to undifferentiated connective tissue disease, was presented with menorrhagia and thrombocytopenia. Anti-nuclear antibody (ANA) was positive 1:1000 (homogeneous type) with anti-double stranded DNA positive, and dRVVT LA1/LA2 was 3.4. Coagulation function was alleviated after treatment with glucocorticoid and total glucosides of paeony. Case 2, a 59-year-old female was presented with gingival bleeding, hematuria with the level of F II:C 13%. dRVVT LA1/LA2 was 2.0. Anti-nuclear antibody (ANA) was positive 1:1000 (type of cytoplasmic granule), anti-double stranded DNA was positive. The patient was diagnosed as hypoprothrombinemia-lupus anticoagulant syndrome (LAHS) and acquired coagulation factor deficiency. The signs of hemorrhage were alleviated after treatment with methylprednisolone 40 mg/day and cyclophosphamide, while the level of F II:C was below normal.

CONCLUSIONSymptoms of patients with positive LAC are variable. The diagnosis relies on history of disease and laboratory test. Currently, there is no standardized treatment. Cases of LAHS should be thoroughly investigated for any known causes and related disorder.

Adult ; Blood Coagulation ; Cyclophosphamide ; therapeutic use ; Female ; Glucocorticoids ; therapeutic use ; Hematologic Tests ; Hemorrhage ; Humans ; Hypoprothrombinemias ; diagnosis ; Lupus Coagulation Inhibitor ; blood ; Methylprednisolone ; therapeutic use ; Middle Aged

Objective To explore the influencing factors on short -term efficacy of intravenous thrombolysis with rt -PA.Methods The clinical data of the 95 acute ischemic stroke(AIS)patients who received thrombolytic therapy were analyze.Multivariate logistic regression analysis was used to determine the possible influencing factors. Results Fifty -six(58.95%)patients had favourable outcomes after thrombolytic therapy for 24 hours.Multivariate logistic regression analysis indicated that diabetes(OR =3.933,95% CI 1.199 ~12.897)and TOAST classification (OR =1.448,95% CI 1.032 ~2.032 )were the independent predictors of short -term outcome.Conclusion Diabetes and TOAST classification are the major influencing factors of short -term efficacy after intravenous thrombolysis with rt -PA.It should pay attention screening patients for intravenous thrombolysis therapy and predicting the efficacy of thrombolysis.

OBJECTIVETo explore the clinical value of immature platelet fraction (IPF), absolute immature platelet fraction (A- IPF) and thrombelastograph (TEG) on assessment of bleeding risk of immune thrombocytopenia (ITP).

METHODStwo hundred and seventy- one patients with ITP were assessed based on ITP-BAT bleeding grading system. IPF, A-IPF were determined in 271 patients ,TEG in 125 patients. The correlations between bleeding grades and IPF, A-IPF, variables of TEG in subgroups were analyzed by statistical method. The predictive value of IPF, A-IPF, and variables of TEG on bleeding risk of ITP patients was evaluated.

RESULTSThere were no significant differences in bleeding degree in all patients with different gender and disease stage (P>0.05). Mild bleeding rate in children was higher than that in adult (P<0.05). PLT inversely correlated with bleeding grade for the entire cohort (P<0.001). In all subjects, PLT< 30 × 10⁹/L and pediatric cohorts with PLT< 30 × 10⁹/L, PLT were negatively correlated with IPF (P<0.05), positive correlated with A-IPF (P<0.001) and the maximum amplitude (MA (P<0.05). Bleeding grades were significantly correlated with IPF, A-IPF, MA in all subjects and patients with PLT< 30 × 10⁹/L (P<0.001). IPF, A-IPF and MA did not correlate with bleeding grades in children with PLT< 30 × 10⁹/L (P>0.05). ROC curve analysis revealed IPF, A-IPF and MA had better predictive value (AUC 0.745, 0.744, 0.813, P<0.001). Multivariate analysis showed that IPF and MA were independence factors for predicting bleeding risk in ITP patients and comprehensive predictive value was higher (AUC 0.846, P<0.001) than single variable.

CONCLUSIONIPF, A-IPF and MA could accurately evaluate bleeding risk in ITP patients. It may be considered as reference index of the treatment and observation index of curative effect.

Adult ; Blood Platelets ; Child ; Hemorrhage ; etiology ; physiopathology ; Humans ; Multivariate Analysis ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; complications ; physiopathology ; ROC Curve

OBJECTIVETo study the expression of autophagy-related proteins (Beclin-1, LC3 and p62) in brain tissue with malformations of cortical development and related molecular pathogenesis.

METHODSThe brain tissue of 18 cases with epileptogenic foci resection, including 6 cases of tuberous sclerosis complex (TSC), 6 cases of focal cortical dysplasia type IIb (FCD IIb) and 6 cases of focal cortical dysplasia type I (FCD I), were retrieved. Immunohistochemical study for Beclin-1, LC3 and p62 proteins was performed. The degree of positivity for Beclin-1 and LC3 proteins was compared. Western blot was used to quantitatively analyze the LC3 protein in focal lesion of each disease groups.

RESULTSImmunohistochemical study showed that the three proteins were mainly expressed in the dysmorphic neurons and balloon cells/giant cells of TSC and FCD IIb. The positivity was more intense in the dysmorphic neurons than the other cell types. Immunostaining for Beclin-1 showed granular or diffuse cytoplasmic positivity, in addition to the strong expression in axons. On the other hand, LC3 showed diffuse or perinuclear cytoplasmic expression. The staining for p62 was mainly cytoplasmic or perinuclear and sometimes nuclear. In FCD type I, only individual cells showed positive expression for the three proteins. The number of Beclin-1 and LC3-positive cells was larger in TSC group, followed by FCD IIb group and FCD I group.And there were significant differences between TSC group and FCD I group, as well as FCD IIb group and FCD I group (P<0.05). Quantitative expression of LC3 protein by Western blot showed smaller amount in TSC group, followed by FCD IIb group and FCD I group.

CONCLUSIONSThe dysmorphic neurons and balloon cells/giant cells of TSC and FCD IIb show abnormality in autophagy, resulting in intracytoplasmic protein accumulation. There are differences in molecular pathogenesis in these cell types.