Diffuse intrinsic pontine glioma( DIPG)is a highly invasive tumor located in the pons (middle)of the brain stem. They are usually diagnosed during childhood and account for 10% -15% of primary brain tumors in children. DIPG has a very poor prognosis. Fewer than 10% of DIPG patients survive more than 2 years after diagnosis. The imaging manifestations of DIPG are typical,and biopsy is only performed in atypi-cal cases. The tissue specimens of newly diagnosed DIPG are very few and limit its molecular biological research. Recent advances in surgical and molecular-analytic techniques have increased the safety of biopsy which has already been used in many clinical trials step by step. The research of DIPG′s molecular pathogenesis and treatment is sure to achieve new breakthroughs.

Early diagnosis and intervention is an important way to delay the progress of Alzheimer′s disease (AD).Compared with cerebrospinal fluid, blood sampling is not invasive and easy to be obtained in clinic practice.In this study, the serum samples of 9 controls, 10 AD and 12 mild cognitive dysfunction (MCI) patients were analyzed and compared through one by one analysis to screen potential markers for AD diagnosis.The experimental results showed that VGFYESDVMGR of α-2-macroglobulin peptide was closely related to the late stage of AD disease, and the large amount degradation of apolipoprotein C-Ⅲ, histone H1.2 and histone H1.4 was significantly related to early stages of AD progression.The characteristics of serum peptidome were different for the early and late AD, and these four proteins may be used as potential biomarkers of AD disease.In addition, the obvious ladder sequence characteristic was observed for apolipoprotein C-Ⅲ and histone H1, which could partly explain why the peptides distribution in different samples was somewhat contingent.On the contrary, the distribution at protein level was more stable.Finally, it was confirmed that the peptides of proteins such as fibrinogen α-chain, thymosin β-4 and patchy proteins were the dominant peptides in all serum samples.Overall, this study showed that the method of using serum peptidomics to diagnose AD was possible.The results may provide evidence and references for the large-scale clinical validation of AD.

OBJECTIVEThis study aimed to evaluate the biological characteristics of a human specifically targeted antimi- crobial peptide C16LL-37 against Streptococcus mutans (S. mutans).

METHODSIn this study, an antimicrobial peptide LL-37, a peptide derived from CSP(C16) (S. mutans competence stimulating peptide), and recombinant peptide C16LL-37 were synthesized by Fmoc-chemistry-based strategy. The selectivity and antibacterial activity of C16LL-37 were identified by the colony counting method on microbial culture plates. After treatment of C16LL-37 at 32 µmol · L⁻¹, the morphological changes in S. mutans were observed by using scanning electron microscopy (SEM). In addition, enzyme-linked immunosorbent assay was used to evaluate the hemolytic activity and antibacterial activity of C16LL-37 under different conditions.

RESULTS1) The minimum inhibitory concentration of C16LL-37 was 16 µmol · L⁻¹, and the minimum bactericidal concentration was 64 μmol ·L⁻¹. 2) The survival rate of S. mutans was 3.46% after C16LL-37 treatment at 64 µmo-L⁻¹ for 30 min, whereas it was 0% at 64 µmol · L⁻¹ for 60 min. The survival rates of four other kinds of bacteria were more than 60% at any time (P < 0.05). 3) The morphological change in S. mutans was observed after C16LL-37 treatment at 32 µmol · L⁻¹ by using SEM. S. mutans presented an irregular shape, rough surface, and evident splitting. 4) The hemolysis rate of C16LL-37 (≤ 64 µmol · L⁻¹) was less than 0.33%. 5) This study showed no significant in- fluence on the antibacterial activity of C16LL-37 under different conditions, such as temperature, pH, salinity, and trypsin at low concentration (P > 0.05).

CONCLUSIONC16LL-37 exhibited obvious specificity for S. mutans, strong antibacterial activity, low toxicity, and high stability. Thus, C16LL-37 has good potential in caries research and clinical application.

Anti-Infective Agents ; pharmacology ; Antimicrobial Cationic Peptides ; pharmacology ; Bacterial Proteins ; Dental Caries ; Enzyme-Linked Immunosorbent Assay ; Humans ; Microbial Sensitivity Tests ; Microscopy, Electron, Scanning ; Peptides ; Streptococcus mutans ; drug effects

OBJECTIVE: To investigate 18β-sodium glycyrrhetinic acid impact on nasal mucosa epithelial cilia in rat models of allergic rhinitis (AR). METHOD: AR models were established by ovalbumin-induction. Wister rats were randomly divided into groups as normal group, model group, budesonide (0.2 mg/kg) group and sodium glycyrrhetinic acid (20 mg/kg and 40 mg/kg) group after the success of AR models. At 2 weeks and 4 weeks after treatment, the behavioral changes of rats were observed and recorded, and nasal septum mucosae were collected after 2 week and 4 week intervention, and the morphological changes of nasal mucosae were observed by electron microscope. RESULT: Model group developed typical AR symptoms, the total score in all animals was > 5. With budesonide and sodium glycyrrhetinic acid treatment, the AR symptoms were relieved, and the total scores were reduced significantly (P < 0.01). Compared with the model group: after 2 weeks' intervention, thick mucous secretions on the top of columnar epithelium cilia in rat nasal mucosa was significantly reduced, and cilia adhesion, lodging, shedding were relieved in budesonide group and sodium glycyrrhetinic acid group, the relieve in budesonide group was slightly better than that in sodium glycyrrhetinic acid group; after 4 week intervention, Cilia adhesion, lodging, shedding were completely vanished, and the cilia were ranged in regular direction in budesonide group and sodium glycyrrhetinic acid group. Cilia in sodium glycyrrhetinic acid (20 mg/kg) group was more orderly, smooth than that in budesonide group and sodium glycyrrhetinic acid group (40 mg/kg), and the condition of cilia in sodium glycyrrhetinic acid group (20 mg/kg) was similar to the normal group. CONCLUSION 18β-sodium glycyrrhetinic acid is effective to restrain the pathological changes of nasal mucosa cilia in rat models of AR.
Animals ; Budesonide ; pharmacology ; Cilia ; drug effects ; Disease Models, Animal ; Glycyrrhetinic Acid ; analogs & derivatives ; pharmacology ; Nasal Mucosa ; drug effects ; Ovalbumin ; Random Allocation ; Rats ; Rhinitis, Allergic ; drug therapy

The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. Antisense EphB4 vectors and shRNA vectors targeting mammalian target of rapamycin (mTOR) were constructed and transfected into A2780 and SKOV3 cells (two ovarian cancer cell lines). The effects of the antisense EphB4 vectors and the shRNA vectors on the proliferation, apoptosis and invasion of ovarian cancer cells were measured, and the expression of EphB4, mTOR and Akt detected. The results showed that transfection with mTOR shRNA could inhibit growth, induce apoptosis, and reduce invasive ability of ovarian cancer cells, which was accompanied by downregulation of EphB4, mTOR and Akt. The inhibitory effects on cell growth caused by mTOR shRNA alone were weaker than those by antisense pEGFP-C1-EphB4. In the antisense pEGFP-C1-EphB4-transfected cells, it was found that EphB4 knockdown could decrease the mTOR expression and slightly reduce the Akt phosphorylation. Significant suppressive effects on cell growth were observed in cells co-transfected with antisense pEGFP-C1-EphB4 and mTOR shRNA. In co-transfection group, the expression levels of EphB4, mTOR and Akt were distinctly lower than those in other groups. It was concluded that suppression of EphB4 may inhibit the growth of ovarian cancer cells by downregulation of the PI3K/Akt/mTOR pathway, and reverse Akt phosphorylation induced by mTOR shRNA. Inhibition of EphB4 and mTOR combined may cooperatively suppress the biological behaviors of ovarian cancer cells.

OBJECTIVE: To observe 18β-glycyrrhetinic acid (GA) impact on ultrastructure of tight junctions (TJs) of nasal mucosa epithelial cells in rats models of allergic rhinitis (AR). METHOD: Ninety-six Wistar rats were randomly divided into control group, model group, loratadine group, and 18β-glycyrrhetinic acid group, and each group had 24 rats. Ovalbumin was used to establish a rat AR model. The behavioral changes and the tight junctions of nasal epithelial were observed and compared in different groups after 2,4,6 and 10 weeks intervention. RESULT: The length of TJs in allergic rhinitis model became shorter, electron-high-density plasma membrane became thicker, number of the integration loci reduced and gap of TJs widened or even ruptured. With the consistent effect of allergens,the changes of TJs in the model group aggravated gradually,and the changes of ultrastructure of TJs in 18β-glycyrrhetinic acid group was relieved apparently compared to model group and even were close to the control model with time. CONCLUSION 18β-glycyrrhetinic acid can recover the ultrastructure of the tight junctions of AR rat nasal epithelial cells.
Animals ; Cell Count ; Epithelial Cells ; ultrastructure ; Glycyrrhetinic Acid ; analogs & derivatives ; pharmacology ; Nasal Mucosa ; cytology ; Ovalbumin ; Rats ; Rats, Wistar ; Rhinitis, Allergic ; drug therapy ; Tight Junctions ; drug effects

Objcetive To investigate the relationship between blood β-hydroxybutyric acid and urine ketone in the di-agnosis of diabetic ketosis (DK). Methods Peripheral blood β-hydroxybutyric acid and urine ketone were detected when the peripheral blood glucose was more than 13.9 mmol/L in patients with diabetes. Results (1) In 81 diabetes pa-tients with blood glucose more than 13.9 mmol/L, the incidence of DK was 13.58% and the incidence of diabetic ke-toacidosis (DKA) was 9.88%. (2) The peripheral blood glucose was positively correlated with β-hydroxybutyric acid (r=0.330, P=0.003), but it was not correlated with urine ketone. (3) The peripheral blood β-hydroxybutyric acid was posi-tively correlated with urine ketone (r=0.516, P=0.000). (4) In patients with DK or DKA, 5.26%(1/19) of those were with urine ketone(-) or (+-), whereas 36.84% (7/19) of those were with blood β-hydroxybutyric acid less than 1 mmol/L. (5) When urine ketone was used as the reference test for diagnosis of DK, the optimal value of blood β-hydroxybutyric acid was 0.35 mmol/L. Conclusion For missed diagnosis of DK may be happend if blood β-hydroxybutyric acid or urine ketone is used alone, the co-monitoring of blood β-hydroxybutyric acid and urine ketone can reduce the inci-dence of missed diagnosis of DK. The urine ketosis may have existed when the blood β-hydroxybutyric acid is slightly elevated (≥0.35 mmol/L). In the situation, the urine ketone should be tested in order to avoid missed diagnosis of DK.

The aim of the present study was to examine the effects of suppression of EphB4 and/or mTOR on the biological behaviors of ovarian cancer cells, and the potential regulatory pathways. Antisense EphB4 vectors and shRNA vectors targeting mammalian target of rapamycin (mTOR) were constructed and transfected into A2780 and SKOV3 cells (two ovarian cancer cell lines). The effects of the antisense EphB4 vectors and the shRNA vectors on the proliferation, apoptosis and invasion of ovarian cancer cells were measured, and the expression of EphB4, mTOR and Akt detected. The results showed that transfection with mTOR shRNA could inhibit growth, induce apoptosis, and reduce invasive ability of ovarian cancer cells, which was accompanied by downregulation of EphB4, mTOR and Akt. The inhibitory effects on cell growth caused by mTOR shRNA alone were weaker than those by antisense pEGFP-C1-EphB4. In the antisense pEGFP-C1-EphB4-transfected cells, it was found that EphB4 knockdown could decrease the mTOR expression and slightly reduce the Akt phosphorylation. Significant suppressive effects on cell growth were observed in cells co-transfected with antisense pEGFP-C1-EphB4 and mTOR shRNA. In co-transfection group, the expression levels of EphB4, mTOR and Akt were distinctly lower than those in other groups. It was concluded that suppression of EphB4 may inhibit the growth of ovarian cancer cells by downregulation of the PI3K/Akt/mTOR pathway, and reverse Akt phosphorylation induced by mTOR shRNA. Inhibition of EphB4 and mTOR combined may cooperatively suppress the biological behaviors of ovarian cancer cells.
Apoptosis ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; genetics ; Female ; Humans ; Ovarian Neoplasms ; pathology ; physiopathology ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; RNA, Small Interfering ; genetics ; Receptor, EphB4 ; genetics ; metabolism ; Suppression, Genetic ; genetics

Objective:To investigate targeted therapy of ovarian cancer with new fusion proteins that were produced by fusing the first 390 amino acids of diphtheria toxin(DT390)to the TMTP1 peptide.Methods:The cisplatin-resistant cell line,C13*,and cisplatin-sensi-tive cell line,OV2008,were selected as models and divided into control,TMTP1,DT390-TMTP1,DT390-biTMTP1,and DT390-triTMTP1 groups.Laser scanning confocal microscopy was used to observe nuclear morphology.3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetra-zolium bromide(MTT)and flow cytometry assays were used to detect cell survival and apoptosis,respectively.The formation of subcu-taneous tumors in nude mice following injection of C13*cells was used to observe the formation and growth of ovarian cancer.Apop-tosis of cells in the subcutaneous tumor tissue was detected by the terminal deoxynucleotidyl transferase dUTP nick-end labeling(TU-NEL)assay.Results:Laser scanning confocal microscopy showed that DT390-biTMTP1 and DT390-triTMTP1 induced nuclear shrinkage and fragmentation.The MTT assay showed that cell survival was obviously reduced with increasing concentrations of DT390-biTMTP1 and DT390-triTMTP1. Flow cytometry revealed that DT390-biTMTP1 and DT390-triTMTP1 significantly increased cell apoptosis (P<0.05).The apoptosis rates of the DT390-biTMTP1 and DT390-triTMTP1 groups were 66.0%±12.0% and 72.9%±4.6%,respectively.These were higher than the 55.5%±8.9% and 65.1%±9.8% obvserved in OV2008 cells.DT390-biTMTP1 and DT390-triTMTP1 significantly in-hibited the tumor formation (P<0.01) and growth (P<0.05), and increased apoptosis (P<0.05) of subcutaneous tumors. However, DT390-TMTP1 had insignificant effects on C13*and OV2008 cells.Conclusions:DT390-biTMTP1 and DT390-triTMTP1 preferentially tar-geted and inhibited ovarian cancer cells.These fusion proteins may be a promising strategy for clinical therapy of ovarian cancer.

Objective:To use inductively coupled plasma mass spectrometry(ICP-MS)to screen biomarkers of element omics of thyroid cancer,and to establish a risk assessment model of element omics of thyroid cancer,so as to provide a basis for the diagnosis and treatment of thyroid cancer.Methods:A total of 200 patients with thyroid cancer who admitted to Beijing Tongren Hospital from February to November 2020 were selected as the thyroid cancer group,and 50 healthy volunteers who underwent physical examinations at hospital during the same period were selected as the healthy control group.The total amount of 28 trace elements,including iodine(I),calcium(Ca),iron(Fe),nickel(Ni),copper(Cu),zinc(Zn),selenium(Se),antimony(Sb),etc.,in their serum were determined by ICP-MS.The content of trace element,thyroid function,free triiodothyronine(FT3),free tetraiodothyronine(FT4),triiodothyronine(T3),tetraiodothyronine(T4),and thyroid volume of ultrasound examination of were analyzed,and then,a risk assessment model of elemental omics of thyroid diseases was established.Results:There were statistically significant differences in the contents of eight trace elements,including I,Ca,Fe,Ni,Cu,Zn,Se and Sb between the thyroid cancer group and the healthy control group(U=2.601,1.972,2.607,2.611,2.603,2.605,2.601,2.605,P<0.05),respectively.The I,Cr and Mn levels of female patients with thyroid cancer appeared increase,while there were significant differences in I,Mn,Fe,Ni,Cu,Zn,Se and Sb contents of male patients between the thyroid cancer group and the health control group(U=2.601,2.608,2.603,2.602,1.973,2.603,2.601,2.602,P<0.05),respectively.In thyroid cancer group,the FT3,FT4,T3,T4 correlated with I content(r=06209,0.5116,0.557,0.5923,P<0.05),respectively.There were correlations in the concentrations between Fe and Zn,between Cr and Mn,between Ca and Zn,between Se and Fe,and between Zn and Se in the thyroid cancer group(r=0.5523,0.5528,0.7158,0.5699,0.6371,0.5420,P<0.05),respectively.High concentrations of I and Mn were risk factors for thyroid cancer.The specificity and sensitivity of the risk assessment model of elemental omics of thyroid cancer were all larger than 95%.Conclusion:In patients with thyroid cancer,both of the serum Ca of female patients and serum Fe of male patients play important role besides cobalt(Co),Ni,Cu,Zn,Se and Sb play role,which can provide basis for the diagnosis and treatment of thyroid cancer.The risk assessment model based on elemental omics of thyroid cancer has favorable diagnostic performance.