Serum hsa_circ_0000615 is a prognostic biomarker of sorafenib resistance in hepatocellular carcinoma.

BACKGROUND: Circular RNAs (circRNAs) can shape tumor progression and chemoresistance. How specific circRNAs shape hepatocellular carcinoma (HCC) chemoresistance, however, remains to be fully elucidated. METHODS: In total, serum samples were collected from 202 HCC patients that had completed four sorafenib chemotherapy cycles. Serum hsa_circ_0000615 levels in these patients were quantified via quantitative real-time polymerase chain reaction (qRT-PCR), with demographic details and survival outcomes being recorded for subsequent analyses. RESULTS: We found hsa_circ_0000615 to be significantly upregulated in chemoresistant HCC patients relative to chemosensitive patients, with such upregulation being positively correlated with disease stage. Moreover, the area under the curve (AUC) value for hsa_circ_0000615 was moderately good, and high levels of hsa_circ_0000615 expression were associated with shorter overall survival among chemoresistant HCC patients. CONCLUSION: Our results highlight hsa_circ_0000615 as a promising driver of sorafenib resistance in HCC patients, highlighting it as a promising target for the treatment of this deadly cancer type.

Targeted inhibition of ACLY expression to reverse the resistance of sorafenib in hepatocellular carcinoma.

Resistance to sorafenib has been documented in hepatocellular carcinoma (HCC) patients. We investigated: (i) the correlation between adenosine triphosphate citrate lyase (ACLY) expression and sorafenib resistance in HCC; and (ii) if targeted inhibition could reverse sorafenib resistance. Samples of HCC tissue were obtained from patients and ACLY expression was measured. PET/CT was employed to measure maximum standard unit value (SUVmax) in HCC patients before and after sorafenib treatment. Using HepG2 cells, we created a sorafenib-resistant cell line. Glucose metabolism and lipid synthesis in HCC cells were tested using 14C-glucose. Disulfide-crosslinked polyethylenimine (SS-PEI)-mediated plasmid transfection was carried out, followed by creation of an HCC model in mice. SUVmax of HCC lesions was closely related to ACLY expression. Patients with high ACLY expression were not sensitive to sorafenib therapy. Lipid metabolism was more active in sorafenib-resistant HCC cells. ACLY expression was higher in sorafenib-resistant cells and HCC-cell sensitivity to sorafenib increased after ACLY-knockout. The latter reversed sorafenib resistance in HCC cells more significantly under hypoxic conditions. SS-PEI/proline-modified short hairpin-(psh)RNA-ACLY plus sorafenib inhibited the growth of drug-resistant cells significantly. These data suggest that ACLY downregulation can reverse sorafenib resistance, and that SS-PEI can be used to mediate shRNA-ACLY transfection in HCC treatment.

[Design and application of Checklist for quality control in intensive care unit].

OBJECTIVE: To design a Checklist for quality control in intensive care unit and observe the effect of clinical application. METHODS: By consulting guidelines and literature, such as Critical care medicine professional medical quality control index (2015 edition), the quality control Checklist of intensive care unit was designed. It included four parts: quality control data collection, medical record quality verification, special diagnosis and treatment, and hospital infection prevention and control supervision. Every month, a doctor with a senior professional title served as the quality control director, and was responsible for the quality control of the department's medical care, including collecting data of the past 24 hours during the morning handover, discussing and registering special diagnosis and treatment behaviors that would be performed on the day, and coordinating with the nursing team leader, controlling the quality of the whole department throughout the day, such as supervising each medical staff if they had unreasonable behaviors, checking the running and discharge medical records, and inspecting the status of the staff on duty. The data in 2018, 2019 (Checklist implemented) and 2017 (Checklist not implemented) were retrospectively analyzed, including the status of admitted patients, department management information, length of intensive care unit (ICU) stay, and the incidence of three-tube infection [ventilator-associated pneumonia (VAP), catheter-related bloodstream infection (CRBSI), catheter-associated urinary tract infection (CAUTI)], and standardized mortality, etc. RESULTS: From 2017 to 2019, the number of patients admitted was 373, 446, and 480, with annual growth of 19.57% and 7.62% in 2018 and 2019, respectively, and an increase of 28.69% in 2019 compared with 2017. There was no statistically significant difference in the average age and acute physiology and chronic health evaluation II (APACHE II) of patients in the three years. Compared with 2017, the length of ICU stay of patients in 2018 and 2019 were significantly shortened (days: 8.99±6.12, 9.14±7.02 vs. 10.20±7.21), and the incidence of VAP, CRBSI and CAUTI were significantly reduced [VAP (cases/1 000 ventilation days): 12.97±3.60, 9.62±3.14 vs. 17.48±4.89, CRBSI (cases/1 000 catheter days): 3.75±2.19, 3.87±1.87 vs. 6.19±3.13, CAUTI (cases/1 000 catheter days): 3.29±2.18, 3.28±1.87 vs. 5.61±3.18]. The standardized mortality were also significantly reduced [(77.27±7.24)%, (70.61±7.49)% vs. (84.41±9.05)%], the number of non-compliance with hospital infection prevention per month decreased significantly (person times: 54.00±6.30, 41.08±10.76 vs. 72.08±19.68), and the number of special diagnosis and treatment per month increased significantly (person times: 1 056.67±235.27, 1 361.75±278.48 vs. 722.25±145.96), the rate of etiology submission before antimicrobial treatment [(93.21±3.68)%, (96.59±2.49)% vs. (87.86±5.28)%] and deep vein thrombosis (DVT) prevention rate [(91.13±6.36)%, (96.23±2.99)% vs. (85.58±7.68)%] were significantly improved, and all the differences were statistically significant (all P < 0.05). All medical records in the three years were Grade A, but the average scores in 2018 and 2019 were higher than those in 2017 (96.82±2.84, 96.73±2.94 vs. 93.70±3.33, both P < 0.01). Compared with 2018, the incidence of VAP, the rate of etiology submission before antimicrobial treatment, the DVT prevention rate, and the standardized mortality rate in 2019 were further improved, and the number of non-compliance with hospital infection prevention per month decreased and the number of special diagnosis and treatment per month increased, and the differences were statistically significant (all P < 0.05). CONCLUSIONS: The application of quality control Checklist in intensive care unit can build an effective quality control system, reduce the incidence of three-tube infection, standardized mortality and length of ICU stay, improve the quality control awareness and execution of medical staff, and promote the improvement of medical quality.

Serum Interleukin-6 Concentrations and the Severity of COVID-19 Pneumonia: A Retrospective Study at a Single Center in Bengbu City, Anhui Province, China, in January and February 2020.

BACKGROUND At present, the relationships among COVID-19 disease progression, patient prognosis, and immune status are unclear. This single-center retrospective study evaluated the correlation between serum interleukin-6 (IL-6) levels at admission with the severity of COVID-19 pneumonia, as determined by admission to the intensive Care Unit (ICU). MATERIAL AND METHODS Patients admitted to The First Affiliated Hospital of Bengbu Medical College in Bengbu City, Anhui Province, China, in January and February 2020 for COVID-19 pneumonia were enrolled in this study. COVID-19 infection was confirmed by the detection of SARS-CoV-2 nucleic acid in throat swab samples using real-time fluorescent reverse transcription PCR. Serum IL-6 concentrations at admission were measured by ELISA. Correlations between serum IL-6 concentrations and ICU admission due to the development of severe COVID-19 pneumonia were evaluated. RESULTS This study enrolled 68 patients with novel coronavirus pneumonia. IL-6 concentrations were significantly higher in patients with more severe than less severe COVID-19 pneumonia. Eight of 40 patients with severe COVID-19 pneumonia became critically ill and required ICU admission. IL-6 concentrations were significantly higher in patients with severe COVID-19 pneumonia who were than who were not treated in the ICU. The area under the receiver operating characteristic (ROC) curve (AUC) was 0.816 (P<0.01), indicating that IL-6 was prognostic of disease severity in patients with COVID-19 pneumonia. CONCLUSIONS Serum IL-6 concentration is closely associated with the severity of COVID-19. Continuous monitoring of IL-6 has clinical value in evaluating patient condition.

[Analysis of Relationship between Serum Total Light Chain κ/λ Ratio and Proportion of Bone Marrow Plasma Cells in Patients with IgG type and IgA type Multiple Myeloma].

OBJECTIVE: To explore the relationship between serum total light chain κ/λ ratio (sTLC-κ/λ) and proportion of bone marrow plasma cells (BMPC) in patients with IgG type and IgA type multiple myeloma (MM) and its clinical significance. METHODS: The levels of serum IgG, IgA, κ type and λ type total light chain were detected in 79 newly diagnosed patients with IgG type (n=52) and IgA type (n=27) MM by immuno-nephelometric assay and the sTLC-κ/λ ratio was calculated. The proportion of BMPC was determined by bone marrow smears in the corresponding period, and the changes in sTLC-κ/λ ratio and the proportion of BMPC were observed in 19 patients with IgG type(n=16) and IgA type (n=3) MM undergoing treatment, 26 cases of non-phasmocytic proliferative diseases were enrolled in control group. RESULTS: In MM patients with IgGκ type and IgAκ type, the sTLC-κ/λ ratio was significantly higher than that in the control group (P<0.01), while in MM patients with IgGλ type and IgAλ type, the sTLC-κ/λ ratio was significantly lower than that in the control group (P<0.01). In MM patients with IgGκ, the sTLC-κ/λ ratio was significantly higher than that in MM patients with IgAκ(P<0.01), while the sTLC-κ/λ ratio in MM patients with IgGλ was significantly lower than that in MM patients with IgAλ. The sTLC-κ/λ ratios in MM patients with IgGκ and IgAκ were positively correlated with the concentrations of IgG (r=0.778,P=0.000) and IgA (r=0.601,P=0.039), while the sTLC-κ/λ ratios of patients with IgGλ and IgAλ were negativily correlated with the IgG(r=-0.586,P=0.01) and IgA level(r=-0.718,P=0.003). In addition, a correlation between each type MM was not found except the IgGκ type MM which had a positive correlation between the sTLC-κ/λ ratio and proportion of BMPC (r=0.579,P=0.002). Nonetheless, 18 of 19 patients with IgG type and IgA type MM undergoing treatment showed concordance between the sTLC-κ/λ ratio and proportion of BMPC change. CONCLUSION: There is a lower correlation between the sTLC-κ/λ ratio and the proportion of BMPC in MM patients with IgG type and IgA type, but there is a high concordance between the sTLC-κ/λ ratio and the proportion of BMPC change in the same patient and it suggests that the sTLC-κ/λ ratio plays an important role in the diagnosis and monitoring of IgG type and IgA type MM.

[Circulating levels of Th1- and Th2-chemokines increase in patients with early syphilis].

Objective To study the changes of plasma T helper type I (Th1)-and Th2-chemokine levels and analyze their roles in immune response and pathogenesis of early syphilis. Methods Heparin-anticoagulated peripheral blood was collected from 56 patients with early syphilis (primary syphilis, PS, n=22; secondary syphilis, SS, n=34) and healthy controls (HC, n=20). The levels of plasma Th1 chemokines including monokine induced by interferon-γ (MIG), interferon-γ inducible protein-10 (IP-10), interferon-inducible T-cell α chemoattractant (I-TAC) and Th2 chemokines including thymus-and activation-regulated chemokine (TARC), macrophage-derived chemokine (MDC) were examined using ELISA. Meanwhile, the levels of plasma cytokines (IFN-γ, IL-4 and TNF-α) and C-reactive protein (CRP) were detected. Results The levels of plasma MIG, IP-10 and TARC, MDC in the patients with PS and SS were significantly higher than those in the healthy controls. Moreover, the level of I-TAC in the patients with SS was significantly higher than that in the healthy controls. In particular, the levels of plasma Th1 chemokines (MIG, IP-10 and I-TAC) in the patients with SS significantly increased compared with those with PS. However, no significant difference was observed in the levels of plasma Th2 chemokines (TARC and MDC) between the patients with PS and SS. The correlation analysis showed that there was an obvious positive correlation between IP-10 and MIG, I-TAC, IFN-γ, TNF-α levels in the patients with early syphilis. Furthermore, the levels of MIG and IP-10 were positively associated with plasma CRP in the patients with early syphilis. Conclusion Both Th1 chemokines and Th2 chemokines are involved in immune response of early syphilis.

Morroniside protects SK-N-SH human neuroblastoma cells against H2O2-induced damage.

Oxidative stress-induced cell injury has been linked to the pathogenesis of neurodegenerative disorders such as spinal cord injury, Parkinson's disease, and multiple sclerosis. Morroniside is an antioxidant derived from the Chinese herb Shan-Zhu-Yu. The present study investigated the neuroprotective effect of morroniside against hydrogen peroxide (H2O2)-induced cell death in SK-N-SH human neuroblastoma cells. H2O2 increased cell apoptosis, as determined by flow cytometry and Hoechst 33342 staining. This effect was reversed by pretreatment with morroniside at concentrations of 1-100 µM. The increase in intracellular reactive oxygen species (ROS) generation and lipid peroxidation induced by H2O2 was also abrogated by morroniside. H2O2 induced a reduction in mitochondrial membrane potential, increased caspase-3 activity, and caused downregulation of B cell lymphoma-2 (Bcl-2) and upregulation of Bcl-2-associated X protein (Bax) expression. These effects were blocked by morroniside pretreatment. Thus, morroniside protects human neuroblastoma cells against oxidative damage by inhibiting ROS production while suppressing Bax and stimulating Bcl-2 expression, thereby blocking mitochondrial-mediated apoptosis. These results indicate that morroniside has therapeutic potential for the prevention and treatment of neurodegenerative diseases.

Co-transplantation of MRF-overexpressing oligodendrocyte precursor cells and Schwann cells promotes recovery in rat after spinal cord injury.

Oligodendrocyte (OL) replacement is a promising treatment strategy for spinal cord injury (SCI). However, the poor survival of transplanted OLs or their precursors and inhibition of axonal regeneration are two major challenges with this approach. Our previous study showed that Schwann cells (SCs) promoted survival, proliferation, and migration of transplanted OL progenitor cells (OPCs) and neurological recovery. Remyelination is an important basis for functional recovery following spinal cord injury. It has been reported that myelin gene regulatory factor (MRF), a transcriptional regulator which specifically is expressed in postmitotic OLs within the CNS, is essential for OL maturation and CNS myelination. In the present study, we investigated whether co-transplantation of MRF-overexpressing OPCs (MRF-OPCs) and SCs could improve functional recovery in a rat model of contusional SCI. MRF overexpression had no effect on OPC survival or migration, but stimulated the differentiation of OPCs both in vitro and in vivo. Co-transplantation of MRF-OPCs and SCs increased myelination and tissue repair after SCI, leading to the recovery of neurological function. These results indicate that co-transplantation of MRF-OPCs and SCs may be an effective treatment strategy for SCI.

Knockdown of PKM2 Suppresses Tumor Growth and Invasion in Lung Adenocarcinoma.

Accumulating evidence shows that activity of the pyruvate kinase M2 (PKM2) isoform is closely related to tumorigenesis. In this study, we investigated the relationship between PKM2 expression, tumor invasion, and the prognosis of patients with lung adenocarcinoma. We retrospectively analyzed 65 cases of patients with lung adenocarcinoma who were divided into low and a high expression groups based on PKM2 immunohistochemical staining. High PKM2 expression was significantly associated with reduced patient survival. We used small interfering RNA (siRNA) technology to investigate the effect of targeted PKM2-knockout on tumor growth at the cellular level. In vitro, siRNA-mediated PKM2-knockdown significantly inhibited the proliferation, glucose uptake (25%), ATP generation (20%) and fatty acid synthesis of A549 cells, while the mitochondrial respiratory capacity of the cells increased (13%).Western blotting analysis showed that PKM2-knockout significantly inhibited the expression of the glucose transporter GLUT1 and ATP citrate lyase, which is critical for fatty acid synthesis. Further Western blotting analysis showed that PKM2-knockdown inhibited the expression of matrix metalloproteinase 2 (MMP-2) and vascular endothelial growth factor (VEGF), which are important in degradation of the extracellular matrix and angiogenesis, respectively. These observations show that PKM2 activates both glycolysis and lipid synthesis, thereby regulating cell proliferation and invasion. This information is important in elucidating the mechanisms by which PKM2 influences the growth and metastasis of lung adenocarcinoma at the cellular and molecular level, thereby providing the basic data required for the development of PKM2-targeted gene therapy.

[Th2 differentiation features of Mycobacterium tuberculosis heat resistant antigen-activated human γδT cells and the regulation of transcription factor T-bet/GATA-3 on differentiation].

OBJECTIVE: To investigate Th2 differentiation features of Mycobacterium tuberculosis heat resistant antigens (MTB-HAg)-activated human γδT cells and the regulation of transcription factor T-box expression in T cells (T-bet) and GATA-binding protein 3 (GATA-3) on differentiation. METHODS: Peripheral blood mononuclear cells (PBMCs) were stimulated with MTB-HAg to generate MTB-HAg-activated T cells (MTBAT) and expanded in the neutral condition or Th2 polarizing condition. After restimulation for 6 hours with phorbol myristate acetate (PMA, 10 ng/mL), ionomycin (500 ng/mL) and monensin (2.5 µmol/L), intracellular cytokines (IFN-γ, IL-4) of γδT cells and αßT cells among MTBAT were detected by four-color fluorescence mAb staining combined with flow cytometry. The highly purified γδT cells and CD4⁺ T cells were sorted by flow cytometer from MTBAT that were cultured in neutral and Th2 polarizing conditions for 28 days. The expressions of T-bet and GATA-3 mRNA in purified γδT cells and CD4⁺ T cells were determined by reverse transcription PCR (RT-PCR) technique. RESULTS: γδT cells among MTBAT cultured in the neutral or Th2 polarizing condition predominantly produced IFN-γ, whereas the percentage of IFN-γ⁺ αßT cells significantly decreased in the Th2 polarizing condition as the culture time went by. Compared with the neutral condition, Th0 type (IFN-γ⁺ IL-4⁺) γδT cells significantly increased, and Th2 type (IFN-γ⁻ IL-4⁺) αßT cells also significantly increased in the Th2 polarizing condition. RT-PCR showed that mRNA expression of T-bet was still at a high level in γδT cells that were expanded in the Th2 polarizing condition, but at a low level in Th2 polarized CD4⁺ T cells. Moreover, the mRNA expressions of GATA-3 in both Th2 polarized γδT cells and CD4⁺T cells were up-regulated. CONCLUSION: In the Th2 polarizing condition, the majority of γδT cells in MTBAT still remained Th1 profile, whereas the portion of γδT cells differentiated into Th0 type cells. Both transcription factor T-bet and GATA-3 failed to display a fully cross-regulation function in Th2 polarized γδT cells.