DNA methylation of DKK-1 may correlate with pathological bone formation in ankylosing spondylitis.

OBJECTIVE: To investigate DNA methylation (DNAm) status of dickkopf-associated protein 1 (DKK-1) in ossified hip capsule synovium and serum among patients with ankylosing spondylitis (AS). METHODS: Western blot was applied to detect the level of DKK-1 protein expression in hip joint capsule tissues from four patients with AS as well as four patients with femoral neck fracture (FNF) caused by trauma as control. DKK-1 gene promoter methylation (GPM) was examined by methylation-specific polymerase chain reaction. Reverse transcription-polymerase chain reaction was performed to examine the messenger RNA (mRNA) levels of DKK-1, ß-catenin, and Wnt3a in both tissue and serum. The DNAm status of serum DKK-1 was measured among 36 patients with AS and syndesmophytes (AS + syndesmophytes group), 40 patients with AS but no syndesmophyte (AS group), and 42 healthy individuals (control group). Also, the serum levels of DKK-1 were measured by enzyme-linked immunosorbent assay. The modified New York criteria (mNYC) together with the modified Stoke Ankylosing Spondylitis Spinal Score (mSASSS) were adopted to examine the radiographic progression of AS. The receiver operating characteristic (ROC) curve was applied to investigate the diagnostic value of the methylation rate of DKK-1 with regard to radiographic progression. RESULTS: The expressions of DKK-1 protein and mRNA in hip joint capsule tissues of AS patients were significantly lower, while DKK-1 GPM rate, ß-catenin mRNA, and Wnt3a mRNA were markedly higher when compared with FNF group. For serum samples, the DKK-1 methylation rate was significantly higher in AS+ syndesmophytes group in contrast to AS group and healthy controls. Serum levels of DKK-1 protein and mRNA in AS with syndesmophytes group were markedly decreased, while ß-catenin mRNA and Wnt3a mRNA expressions were significantly increased than AS with no syndesmophyte group and the healthy control group. AS patients in Grade 4 showed a significantly higher serum DKK-1 GPM rate than those in Grade 3 based on mNYC. Serum DKK-1 GPM level was markedly and positively correlated with mSASSS. Serum levels of DKK-1 in AS+ syndesmophytes group were markedly lower compared with AS but no syndesmophyte group and healthy controls. ROC curve analysis indicated that serum DKK-1 methylation rate serves as a decent indicator for AS radiographic progression. CONCLUSION: DNAm of DKK-1 may correlate with pathological bone formation in AS, which may provide new strategies for the treatment of AS abnormal bone formation.

Driver mutations in ADGRL3 are involved in the evolution of ependymoma.

Although there have been recent advances in the molecular pathology of ependymomas, little is known about the underlying molecular evolution during its development. Here, we assessed the clinical, pathological and molecular evolutionary process of ependymoma recurrence in a 9-year-old patient who had seven recurrences of supratentorial ependymoma and died from intracranial multiregional recurrences at the age of 19 years old. Whole-genome sequencing (WGS) of 7 tumor samples (1 primary and 6 subsequent recurrent tumors) was performed to elucidate the mutation landscape and identify potential driver mutations for tumor evolution. The genetic profiles of the seven tumor specimens showed significant heterogeneity and suggested a highly branched evolutionary pattern. The mutational signatures and chromothripsis changed with treatments. Strikingly, adhesion G protein-coupled receptor L3 (ADGRL3, also known as Latrophilins 3, LPNH3) was found to be consistently mutated during the entire disease process. However, Sanger sequencing of other 78 ependymoma patients who underwent surgery at our institution showed no genetic alteration of ADGRL3, as found in the present case. The mRNA levels of ADGRL3 were significantly lower in ependymomas (n = 36), as compared with normal brain tissue (n = 3). Grade III ependymomas had the lowest ADGRL3 expression. Moreover, ependymomas with lower mRNA level of ADGRL3 had shorter overall survival. Our findings, therefore, demonstrate a rare evolutionary process of ependymoma involving ADGRL3.

Successful treatment of a critically ill patient with acute respiratory distress syndrome from COVID-19 using mechanical ventilation strategy with low levels of positive end-expiratory pressure: A CARE-compliant case report.

INTRODUCTION: Acute respiratory distress syndrome (ARDS) secondary to COVID-19 is different from the ARDS caused by other infections. Conventional mechanical ventilation strategies using high levels of PEEP may not be beneficial and can even be harmful to patient with ARDS from COVID-19. So the ventilation strategies should be adjusted in order to improve the pulmonary ventilation function and oxygenation status, and outcomes of the patient. PATIENT CONCERNS: Herein, we present a 76-year-old male patient with ARDS secondary to COVID-19. We describe our experience with mechanical ventilation strategy and the changes in respiratory mechanics in the patient during treatment. DIAGNOSIS: The patient had tested positive for coronavirus (COVID-19) in nucleic acid test. Chest CT showed multiple ground glass shadows in both lungs. INTERVENTIONS: The patient received mechanical ventilation with low tidal volume and low PEEP. OUTCOMES: After treatment, the patients condition, as well as oxygenation status was improved, and he tested negative for the coronavirus several times. CONCLUSION: This case demonstrated that the low tidal volume with low levels of PEEP ventilation strategy may be more suitable for ARDS from COVID-19.

Combination of levetiracetam and IFN-α increased temozolomide efficacy in MGMT-positive glioma.

PURPOSE: Glioma, especially glioblastoma (GBM), is the most aggressive malignant brain tumor and its standard therapy is often ineffective because of temozolomide (TMZ) resistance. Reversal of the TMZ resistance might improve the prognosis of glioma patients. We previously found that interferon-α (IFN-α) and anti-epileptic drug levetiracetam (LEV) could sensitize glioma to TMZ, respectively. In this study, we further investigated the efficiency of combining of LEV and IFN-α for improving the efficacy of TMZ. METHODS: We evaluated whether LEV and IFN-α could increase TMZ efficacy using colony formation assay and cell viability assay with MGMT-positive and MGMT-negative glioma cell lines in vitro. Subcutaneous xenografts and orthotopic xenografts mice models were used in vivo to observe the tumor growth and mice survival upon treatments with TMZ, TMZ + IFN-α, TMZ + LEV, or TMZ + LEV + IFN-α. The expression levels of MGMT, markers of pro-apoptotic and anti-apoptotic in tumor samples were analyzed by Western blotting. RESULTS: The combinational use of IFN-α, LEV, and TMZ showed the best anti-tumor activity in MGMT-positive cell lines (U138, GSC-1, U118, and T98 G). TMZ + LEV + IFN-α further obviously increased TMZ + LEV or TMZ + IFN-α efficiency in MGMT-positive cell lines, while not in negative cell lines (SKMG-4, U87, U373, and U251) in vitro, which were also observed in subcutaneous mice models (U138, GSC-1 compared to SKMG-4, U87) and orthotopic models (GSC-1) in vivo. Strikingly, the combination of LEV and IFN-α together with TMZ significantly prolonged the survival of mice with orthotopic GSC-1 glioma. Furthermore, we confirmed that the combination of LEV and IFN-α enhanced the inhibition of MGMT and the activation of apoptosis in U138 tumor on the basis of TMZ treatment. CONCLUSIONS: The combination use of LEV and IFN-α could be an optimal method to overcome TMZ resistance through obvious MGMT inhibition in MGMT-positive glioma.

Transferrin receptor 1 targeted optical imaging for identifying glioma margin in mouse models.

OBJECTIVE: Optical molecular imaging technology that indiscriminately detects intracranial glioblastoma (GBM) can help neurosurgeons effectively remove tumor masses. Transferrin receptor 1 (TfR 1) is a diagnostic and therapeutic target in GBM. A TfR 1-targeted peptide, CRTIGPSVC (CRT), was shown to cross the blood brain barrier (BBB) and accumulate at high levels in GBM tissues. In this study, we synthesized a TfR 1-targeted near-infrared fluorescent (NIRF) probe, Cy5-CRT, for identifying the GBM tissue margin in mouse models. METHODS: We initially confirmed the overexpression of TfR 1 in GBM and the tumor-specific homing ability of Cy5-CRT in subcutaneous and orthotopic GBM mouse models. We then examined the feasibility of Cy5-CRT for identifying the tumor margin in orthotopic GBM xenografts. Finally, we compared Cy5-CRT with the clinically used fluorescein sodium in identifying tumor margins. RESULTS: Cy5-CRT specifically accumulated in GBM tissues and detected the tumor burden with exceptional contrast in mice with orthotopic GBM, enabling fluorescence-guided GBM resection under NIRF live imaging conditions. Importantly, Cy5-CRT recognized the GBM tissue margin more clearly than fluorescein sodium. CONCLUSIONS: The TfR 1-targeted optical probe Cy5-CRT specifically differentiates tumor tissues from the surrounding normal brain with high sensitivity, indicating its potential application for the precise surgical removal of GBM.

Skp2 modulates proliferation, senescence and tumorigenesis of glioma.

BACKGROUND: Gliomas represent the largest class of primary central nervous system neoplasms, many subtypes of which exhibit poor prognoses. Surgery followed by radiotherapy and chemotherapy has been used as a standard strategy but yielded unsatisfactory improvements in patient survival outcomes. The S-phase kinase protein 2 (Skp2), a critical component of the E3-ligase SCF complex, has been documented in tumorigenesis in various cancer types but its role in glioma has yet to be fully clarified. In this study, we investigated the function of Skp2 in the proliferation, stem cell maintenance, and drug sensitivity to temozolomide (TMZ) of glioma. METHODS: To investigate the role of Skp2 in the prognosis of patients with glioma, we first analyzed data in databases TCGA and GTEx. To further clarify the effect of Skp2 on glioma cell proliferation, we suppressed its level in glioblastoma (GBM) cell lines through knockdown and small molecule inhibitors (lovastatin and SZL-P1-41). We then detected cell growth, colony formation, sphere formation, drug sensitivity, and in vivo tumor formation in xenograft mice model. RESULTS: Skp2 mRNA level was higher in both low-grade glioma and GBM than normal brain tissues. The knockdown of Skp2 increased cell sensitivity to TMZ, decreased cell proliferation and tumorigenesis. In addition, Skp2 level was found increased upon stem cells enriching, while the knockdown of Skp2 led to reduced sphere numbers. Downregulation of Skp2 also induced senescence. Repurposing of lovastatin and novel compound SZL-P1-41 suppressed Skp2 effectively, and enhanced glioma cell sensitivity to TMZ in vitro and in vivo. CONCLUSION: Our data demonstrated that Skp2 modulated glioma cell proliferation in vitro and in vivo, stem cell maintenance, and cell sensitivity to TMZ, which indicated that Skp2 could be a potential target for long-term treatment.

Tenascin-cmediated vasculogenic mimicry formation via regulation of MMP2/MMP9 in glioma.

Vasculogenic mimicry (VM), the formation of vessel-like structures by highly invasive tumor cells, has been considered one of several mechanisms responsible for the failure of anti-angiogenesis therapy in glioma patients. Therefore, inhibiting VM formation might be an effective therapeutic method to antagonize the angiogenesis resistance. This study aimed to show that an extracellular protein called Tenascin-c (TNC) is involved in VM formation and that TNC knockdown inhibits VM in glioma. TNC was upregulated with an increase in glioma grade. TNC and VM formation are potential independent predictors of survival of glioma patients. TNC upregulation was correlated with VM formation, and exogenous TNC stimulated VM formation. Furthermore, TNC knockdown significantly suppressed VM formation and proliferation in glioma cells in vitro and in vivo, with a reduction in cellular invasiveness and migration. Mechanistically, TNC knockdown decreased Akt phosphorylation at Ser473 and Thr308 and subsequently downregulated matrix metalloproteinase 2 and 9, both of which are important proteins associated with VM formation and migration. Our results indicate that TNC plays an important role in VM formation in glioma, suggesting that TNC is a potential therapeutic target for anti-angiogenesis therapy for glioma.

Ballpoint connector-protected salt-oil-salt liquid phase microextraction for concentration and enrichment of trace anthraquinone compounds in rhubarb.

This study proposed a new ballpoint connector-protected salt-oil-salt liquid phase microextraction for extraction and enrichment of trace rhein and chrysophanol in rhubarb prior to determination of the analytes by high performance liquid chromatography. In this study, a handy ballpoint connector (between ballpoint tip and ink chamber) was used as extraction device, in which its cavity was filled with n-octanol, and the bare n-octanol in its two opening ends was covered with a thin layer of sodium chloride film. The design subtly assembled salt film onto ballpoint connector for extraction and enrichment, which greatly improved the enrichment factors of the target analytes. Moreover, the novel procedure and its extraction mechanism were described and analyzed, and several crucial parameters reflecting the extraction effect were investigated and optimized. Under optimum conditions, high enrichment factors (247 and 127), good linearities with r ≥ 0.9998, limits of detection (0.6-1.1 ng/mL), relative standard deviations of intra- and interday (2.2-8.8% and 4.3-8.9%), and average recoveries (97.6-98.1%), were obtained, respectively. The proposed method can not only eliminate the negative effects from viscosity and ion strength at high salt concentration of sample phase, but also make salting-out effect be focused on small area so as to maximize the extraction effect.

Severe thoracic trauma caused left pneumonectomy complicated by right traumatic wet lung, reversed by extracorporeal membrane oxygenation support-a case report.

BACKGROUND: Double lumen intubation and one-lung ventilation should be applied without delay in cases of traumatic main bronchial rupture. In most cases, when the patients' vital signs have been stabilized, the repair can be performed. However, when one-lung ventilation is complicated by traumatic wet lung, the mortality rate is likely to be much higher. CASE PRESENTATION: In this case, the patient experienced a left main bronchial rupture, bilateral traumatic wet lung, and acute respiratory distress syndrome (ARDS) because of severe thoracic trauma. Though the patient was treated with intubation and mechanical ventilation (MV), his oxygenation was still not stable. Thus, veno-venous extracorporeal membrane oxygenation (V-V ECMO) was initiated; upon improvement of oxygenation, the patient received an exploratory thoracotomy. Unfortunately, the rupture proved to be irreparable, resulting in a total left pneumonectomy. As there was severe ARDS caused by trauma, ECMO and ultra-low tidal volume (VT) MV strategy (3 ml/kg) were utilized for lung protection post-op. ECMO was sustained up to the 10th day, and MV until the 20th day, post-operation. With the support of MV, ECMO and other comprehensive measures, the patient made a recovery. CONCLUSION: V-V ECMO and ultra-low VT MV helped this thoracic trauma patient survive the lung edema period and prevented ventilator associated pneumonia (VAP). In extreme situations, with the support of ECMO, the tidal volume may be lowered to 3 ml/kg.

Non-destructive characterization of egg odor and fertilization status by SPME/GC-MS coupled with electronic nose.

BACKGROUND: Early and non-destructive identification of fertile (F) eggs is a difficult task in the process of breeding laying hens. The odors emitted from unfertilized (UF), infertile (IF), and fertile (F) eggs were characterized by solid-phase microextraction / gas chromatograph-mass spectrometry (SPME/GC-MS) and electronic nose (E-nose) to determine their differences by principal component, partial least squares, and canonical discriminant analyses. RESULTS: A total of 14 volatiles were identified in unhatched shell white Leghorn eggs, such as nonanal, decanal, 6-methyl-5-hepten-2-one, and 6,10-dimethyl-5,9-undecadien-2-one. Cedrene and decanal contributed greatly to the classification of UF and fertilized (Fd)/IF eggs; cedrene, decanal, 1-octanol and hexanal contributed greatly to the distinction between UF and IF eggs; heptanal might be the potential marker to determine F/IF eggs. P40/1, P10/2, P10/1, TA/2, T40/2 and T30/1, P30/1, P40/2, PA/2, T40/2 mostly contributed to the distinction between UF and Fd eggs and between F and IF eggs, respectively. Canonical discriminant analysis presented superior differentiating efficiency for almost all groups, and the odor differences between UF and Fd eggs were significantly larger than the differences between F and IF eggs. CONCLUSION: Solid-phase microextraction / gas chromatograph-mass spectrometer combined with E-nose may have the potential to non-destructively distinguish UF, F, and IF eggs, which will provide a new perspective to understand the differences among them. © 2018 Society of Chemical Industry.

Graphene/dodecanol floating solidification microextraction for the preconcentration of trace levels of cinnamic acid derivatives in traditional Chinese medicines.

A novel graphene/dodecanol floating solidification microextraction followed by HPLC with diode-array detection has been developed to extract trace levels of four cinnamic acid derivatives in traditional Chinese medicines. Several parameters affecting the performance were investigated and optimized. Also, possible microextraction mechanism was analyzed and discussed. Under the optimum conditions (amount of graphene in dodecanol: 0.25 mg/mL; volume of extraction phase: 70 µL; pH of sample phase: 3; extraction time: 30  min; stirring rate: 1000 rpm; salt amount: 26.5% NaCl; volume of sample phase: 10 mL, and without dispersant addition), the enrichment factors of four cinnamic acid derivatives ranged from 26 to 112, the linear ranges were 1.0 × 10-2 -10.0 µg/mL for caffeic acid, 1.3 × 10-3 -1.9 µg/mL for p-hydroxycinnamic acid, 2.8 × 10-3 -4.1 µg/mL for ferulic acid, and 2.7 × 10-3 -4.1 µg/mL for cinnamic acid, with r2 ≥ 0.9993. The detection limits were found to be in the range of 0.1-1.0 ng/mL, and satisfactory recoveries (92.5-111.2%) and precisions (RSDs 1.1-9.5%) were also achieved. The results showed that the approach is simple, effective and sensitive for the preconcentration and determination of trace levels of cinnamic acid derivatives in Chinese medicines. The proposed method was compared with conventional dodecanol floating solidification microextraction and other extraction methods.

[Effect of early intervention of liver-smoothing and blood-activating decoction combined with acupuncture on patients with post-stroke depression].

To study the effect of early intervention of liver-soothing and Blood-activating decoction combined with acupuncture in improving neurological functions, depressive symptom and life quality of patients with post-stroke depression, and compare with fluoxetine hydrochloride. Specifically, 63 patients with post-stroke depression were randomly divided into the traditional Chinese medicine (TCM) acupuncture group (31 cases) and the western medicine group (32 cases). On the basis of the conventional treatment of the primary disease, the TCM acupuncture group was treated with liver-soothing and blood-activating decoction and acupuncture, while the western medicine group was treated with fluoxetine hydrochloride for four weeks. In the follow-up visit six months later, scores of HAMD, NIHSS and SS-QOL were observed. The scores of HAMD and NIHSS of both groups were significantly decreased (P < 0.01), while the scores of SS-QOL increased significantly, with a notable difference compared with that before the treatment (P < 0.01). Specifically, the TCM acupuncture group's was superior to the western medicine group (P < 0.05). The study suggests that the early intervention of liver-soothing and blood-activating decoction combined with acupuncture on patients with post-stroke depression has the effect in relieving depression symptom and improving neurological functions, thereby improving their quality of life and prognosis.

[Study on the natural ligand(s) of triggering receptor expressed on myeloid cell-1 on bacteria cell wall].

OBJECTIVE: To look for the natural ligand(s) of human triggering receptor expressed on myeloid cell-1 (TREM-1), in order to provide the theoretical basis for elucidation of the pathogenesis of sepsis. METHODS: Neutrophils and monocytes isolated from human peripheral blood were treated with heat-inactivated Staphylococcus aureus, Pseudomonas aeruginosa, Mycobacterium tuberculosis, Staphylococcus aureus L-form or Pseudomonas aeruginosa L-form respectively for 24 hours. The cell wall was extracted from Staphylococcus aureus, Pseudomonas aeruginosa and Mycobacterium tuberculosis by ultrasound. Neutrophils and monocytes were isolated and treated with the cell wall respectively for 24 hours. Neutrophils and monocytes were isolated and treated with three main components from bacterial cell wall (polysaccharides, lipids and proteins) respectively for 24 hours. The level of TREM-1 mRNA was measured with fluorescent quantitative polymerase chain reaction (PCR), and the concentrations of tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta) were measured with enzyme-linked immunosorbent assay (ELISA). RESULTS: The TREM-1 mRNA level and the concentrations of TNF-alpha and IL-1 beta in cell supernatant of neutrophils and monocytes were upgraded when treated with cell, cell wall and cell wall polysaccharides of Staphylococcus aureus and Pseudomonas aeruginosa. Compared with the blank control group, the TREM-1 mRNA level of neutrophils and monocytes was upgraded to (3.86+/-0.20)-fold and (5.15+/-0.56)-fold respectively when treated with cell wall polysaccharides of Staphylococcus aureus (both P<0.05); the TREM-1 mRNA level of neutrophils and monocytes was upgraded to (4.03+/-0.15)-fold and (7.22+/-0.73)-fold respectively when treated with cell wall polysaccharides of Pseudomonas aeruginosa (both P<0.05). The effect could be attenuated by the addition of LP17 which could bind TREM-1 ligand. This attenuating effect was not found when the cells were treated with cell, cell wall or cell wall polysaccharides of Mycobacterium tuberculosis. CONCLUSION: The study provides the evidence that TREM-1 natural ligand(s) is present on cell wall of bacteria including Staphylococcus aureus and Pseudomonas aeruginosa, and it might be polysaccharides.

[Simultaneous determination of impurities in K2CrO4 by ICP-AES].

The trace elements, Ca, Mg, Ba, Co, Cd, Cu, Mn, Pb, Sr, Zn, Al, Fe, Ni, Ti, Si and V, in K2CrO4 were simultaneously determined by inductively coupled plasma atomic emission spectrometry (ICP-AES). Optimum spectral lines were selected for each analyzed element. The matrix effect was studied and eliminated through matrix matching. The results show that the linear correlations of standard curves are good (R2 = 0.998 6-1.000) under the determined conditions. The recoveries of standard addition are in the range of 83.7%-113.0%, and the relative standard deviations are lower than 9.48% (n = 9). Detection limits of these impurities are in the range of 0.075-2. 625 mg x kg(-1). The method is rapid, simple, accurate and credible, so it can be used for the determination of impurities in K2 CrO4 product. The results also show that there is relatively high content of Ca, Mg, Al and Si in K2 CrO4 product, so the K2 CrO4 product can't be directly used for the ion membrane electrochemical synthesis of chromic anhydride and the product must be taken for further purification.

Analysis of nonuniformities of sampled fiber Bragg gratings.

We have analyzed the effects of various normally distributed nonuniformities of sampled fiber Bragg gratings on the reflective spectra and group time delay. Through numerical simulations we have drawn the following conclusions: (1) the magnitude of nonuniformity with normal distribution of the fiber's average refractive-index modulation deltaneff greatly influences the characteristics of both reflective spectra and group time delay, whose suggested precision varies from 20% to 10%; (2) the nonuniformities of sampling periods P and sampling lengths L are important factors that influence the characteristics of the group time delay, and the accepted tolerance of dimensional precision of both P and L are +/- 4 microm; and (3) the tolerance of nonuniformity of the sampling period's chirp coefficient is high, and its precision can be as great as 100% with few adverse effects.