The cancer-testis gene,MEIOB,sensitizes triple-negative breast cancer to PARP1 inhibitors by inducing homologous recombination deficiency

Objective:The newly defined cancer-testis(CT)gene,MEIOB,was previously found to play key roles in DNA double-strand break(DSB)repair.In this study,we aimed to investigate the effects and mechanisms of MEIOB in the carcinogenesis of triple-negative breast cancers(TNBCs).Methods:The Cancer Genome Atlas database was used to quantify the expression of MEIOB.Cox regression analysis was used to evaluate the association between MEIOB expression and the prognosis of human TNBC.The effects of MEIOB on cell proliferation and migration in TNBCs were also assessed in vitro.Patient-derived xenograft(PDX)models were used to assess the sensitivity of breast cancers with active MEIOB to PARP1 inhibitors.Results:We confirmed MEIOB as a CT gene whose expression was restricted to the testes and breast tumors,especially TNBCs.Its activation was significantly associated with poor survival in breast cancer patients[overall,hazard ratio(HR)=1.90(1.16–2.06);TNBCs:HR=7.05(1.16–41.80)].In addition,we found that MEIOB was oncogenic and significantly promoted the proliferation of TNBC cells.Further analysis showed that MEIOB participated in DSB repair in TNBCs.However,in contrast to its function in meiosis,it mediated homologous recombination deficiency(HRD)through the activation of poly ADP-ribose polymerase(PARP)1 by interacting with YBX1.Furthermore,activated MEIOB was shown to confer sensitivity to PARP inhibitors,which was confirmed in PDX models.Conclusions:MEIOB played an oncogenic role in TNBC through its involvement in HRD.In addition,dysregulation of MEIOB sensitized TNBC cells to PARP inhibitors,so MEIOB may be a therapeutic target of PARP1 inhibitors in TNBC.     

Drug Resistance to HIV-1 Integrase Inhibitors among Treatment-naive Patients in Jiangsu,China

Integrase strand transfer inhibitors (InSTIs) have been widely used in recent years because of their high genetic barrier to resistance. The World Health Organization (WHO) has recommended dolutegravir (DTG)-containing regimens as the preferred first- and second-line antiretroviral therapy (ART) regimens for people living with human immunodeficiency virus (HIV)[1]. During the long-term treatment process, the appearance of drug resistance mutations to InSTIs is inevitable. A meta-analysis has shown that the resistance rate among InSTI treatment-experienced patients is 3.9％ (Raltegravir, RAL), 1.2％ (Elvitegravir, EVG), and 0.1％ (DTG)[2]. However, resistance to InSTIs has not been reported in treatment-naive populations.     

Establishment of an optimized CTC detection model consisting of EpCAM,MUC1 and WT1 in epithelial ovarian cancer and its correlation with clinical characteristics

Objective: Emerging studies have demonstrated the promising clinical value of circulating tumor cells(CTCs)for diagnosis, disease assessment, treatment monitoring and prognosis in epithelial ovarian cancer. However, the clinical application of CTC remains restricted due to diverse detection techniques with variable sensitivity and specificity and a lack of common standards.Methods: We enrolled 160 patients with epithelial ovarian cancer as the experimental group, and 90 patients including 50 pat...     

PPARγ: the dominant regulator among PPARs in dry eye lacrimal gland and diabetic lacrimal gland

AIM: To determine whether lectin-like ox-LDL receptor (LOX-1) regulates adhesion molecules expression and neutrophil infiltration in A. fumigatus keratitis of C57BL/6 mice.METHODS: C57BL/6 mice were pretreated with a neutralizing antibody to LOX-1 (5 μg/5 μL) or control nonspecific IgG (5 μg/5 μL), LOX-1 inhibitor Poly-I (2 μg/5 μL) or PBS by subconjunctival injection. Fungal keratitis mouse models of C57BL/6 mice were established by scraping corneal central epithelium, smearing A. fumigatus on the corneal surface and covering the eye with contact lenses. The corneal response to infection was assessed via clinical score. The mRNA levels of the adhesion molecules ICAM-1, VCAM-1, P-selectin and E-selectin were tested in control and infected corneas by RT-PCR. The protein levels of ICAM-1 were evaluated by immunofluorescence (IF) and Western blot. Neutrophils were extracted from the abdominal cavity of C57BL/6 mice followed by pretreatment using antibody to LOX-1 (10 μg/mL) or control nonspecific IgG (10 μg/mL), the Poly-I (4 μg/mL) or PBS. The cells were then stimulated with A. fumigatus and tested mRNA and protein levels of LFA-1 using RT-PCR and Western blot. IF and myeloperoxidase (MPO) assays were used to assess neutrophil infiltration in mice corneas.RESULTS: Pretreatment of LOX-1 antibody or the Poly-I reduced the degree of inflammation of cornea and decreased the clinical fungal keratitis score compared with pretreatment of IgG or PBS. And these pretreatment also displayed an obvious decline in the mRNA levels of ICAM-1, VCAM-1, P-selectin, E-selectin and LFA-1 expression compared with control groups . Furthermore, pretreated with LOX-1 antibody or Poly-I, the protein levels of ICAM-1 and LFA-1 also decreased compared with control groups. Neutrophil infiltration in the cornea was significantly reduced after pretreatment of LOX-1 antibody or Poly-I compared with control groups by IF and MPO assays. CONCLUSION: These data provide evidence that inhibition of LOX-1 can decrease the expression of adhesion molecules and thus reduce neutrophil infiltration in A. fumigatus infected corneas of C57BL/6 mice.     

Celastrus orbiculatus Extracts Inhibit Human Hepatocellular Carcinoma Growth by Targeting mTOR Signaling Pathways

Objective: To characterize the molecular mechanism underlying the antineoplastic activity of Celastrus orbiculatus Thunb. extracts(COE). Methods: The human hepatocellular carcinoma HepG2 cells with mammalian target of rapamycin(mT OR) knockdown expressed(HepG 2/mT OR-) were constructed using molecular biological technology. In vitro, the HepG 2/mT OR-cells were treated with COE at various concentrations(10, 20, 40, 80, 160 and 320 μg/mL). Cell viability was determined using 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide(MTT) assays. According to the half-maximal inhibitory concentration(IC50) value(140 mg/L), the concentrations of COE in the subsequent experiment was set to alleviate cytotoxicity. The HepG 2/mT OR-cells were divided into 5 groups: negative control(untreated), COE treatment groups(40, 80, 120 mg/L COE) and positive control group(cisplatin, DDP, 2 mg/L), respectively. Wild-type HepG 2 cells were used as a blank control. The effects of COE on the cell apoptosis were analyzed by flow cytometry and transmission electronic microscopy(TEM), respectively. The protein expression levels of mT OR signal pathways were determined by Western blotting. In vivo, HepG2/mTOR-cells(2×10~6 cell/mice) were subcutaneously injected into the right flank of nude mice. Thirty-six female nude mice were randomly assigned to 6 groups according to body weight(6 mice per group) as follows: solvent vehicle control, Banmao Capsule treated group(BM, 195 mg/kg), Tegafur, Gimeracil and Oteracil Potassium Capsules(10 mg/kg) treated group, and different dosages of COE(10, 20, 40 mg/kg) groups. Tumor growth was monitored and immunohistochemical staining was used to examine the expression of apoptosis-related proteins in tumor tissues. Results: COE inhibited the proliferation significantly in a concentration-dependent manner in HepG 2/mT OR-cells(P0.01). COE significantly induced the apoptosis of HepG 2/mT OR-cells(P0.01), and the apoptotic bodies can be observed under TEM. COE significantly inhibits the proteins expression of mT ORrelated signal pathways. In vivo, COE significantly inhibited tumor growth in nude mice(P0.01). Moreover, the results showed that COE down-regulated the expression of Bcl-2 and Bcl-xL, and up-regulated the levels of Bax and caspase-3 protein(P0.01). Conclusion: COE was a potential chemotherapeutic drug in HCC treatments via targeting mT OR signal pathway.     

Simultaneous Determination of Six Compounds in Rat Plasma by Ultra‑Performance Liquid Chromatography with Tandem Mass Spectrometry: Application in the Pharmacokinetic Study of Qing Gan‑Shu Yu‑Fang

A rapid and high selective ultra?performance liquid chromatography (UPLC) with tandem mass spectrometry method for simultaneous determination of six compounds including albiflorin, paeoniflorin, picroside I, picroside II, saikosaponin A, and saikosaponin D in rat plasma was developed and validated using butyl p-hydroxybenzoate as an internal standard. One-step direct protein precipitation with acetonitrile was used to extract the compounds from the rat plasma samples. Chromatographic separation was achieved using an ACQUITY UPLC BEH C18 column (100 mm × 2.1 mm, 1.7 µm) at a flow rate of 0.4 mL/min, using gradient mode containing 0.1% formic acid in water and acetonitrile were used as the Mobile phase A and B. Electrospray ionization in negative ion mode and multiple reaction monitoring were used to identify and quantify active components. Calibration curves showed good linearity (R2 > 0.9908) over a wide concentration range for all compounds. The intra- and interday precision (relative standard deviation) ranged 2.4%–7.0% and 2.6%–8.0%, respectively. The accuracy (relative error) was from ?13.0% to 13.2% at all quality control levels. The recovery ranged from 81.1% to 92.5%. The validated method was successfully applied to pharmacokinetic study in rats after oral administration of Qing Gan?Shu Yu?Fang. The results show that one can draw a conclusion that these six active ingredients can be quickly absorbed and play a pharmacodynamic role rapidly in vivo.     

CCPG1 involved in corneal Aspergillus fumigatus infection

AIM: To investigate whether non-canonical autophagy transport receptor cell cycle progression 1 (CCPG1) is involved in the corneal antifungal immune response. METHODS: Human corneal epithelial cells (HCECs) and human myeloid leukemia mononuclear cells (THP-1) macrophages stimulated by Aspergillus fumigatus (A. fumigatus) were used as cell models. The expression of CCPG1 mRNA was detected by qRT-PCR. Western blot was used to determine the protein expression of CCPG1 and interleukin-1β (IL-1β). The dectin-1 neutralizing antibody was used to detect the association between dectin-1 and CCPG1. Immunofluorescence was used to observe the colocalization of CCPG1 and C-type lectin-like receptor-1 (CLEC-1) in THP-1 macrophages. RESULTS: The expression of CCPG1 started to increase at 4h after infection and increased in a time-dependent manner in HCECs and THP-1 macrophages. With dectin-1 neutralizing antibody pretreatment, the expression of IL-1β was down-regulated. CCPG1 up-regulation in response to A. fumigatus infection was independent of dectin-1. Immunofluorescence showed the colocalization of CCPG1 and CLEC-1 in THP-1 macrophages. CONCLUSION: As a specific autophagy protein of non-canonical autophagy pathway, CCPG1 is involved in corneal infection with A. fumigatus.     

Heparanase-1 is downregulated in chemoradiotherapy orbital rhabdomyosarcoma and relates with tumor growth as well as angiogenesis

AIM:To determine the role of heparanase-1(HPSE-1)in orbital rhabdomyosarcoma(RMS),and to investigate the feasibility of HPSE-1 targeted therapy for RMS.METHODS:Immunohistochemistry was performed to analyze HPSE-1 expression in 51 cases of orbital RMS patients(including 28 cases of embryonal RMS and 23 cases of alveolar RMS),among whom there were 27 treated and 24 untreated with preoperative chemoradiotherapy.In vitro,studies were conducted to examine the effect of HPSE-1 silencing on RMS cell proliferation and tube formation of human umbilical vein endothelial cells(HUVECs).RD cells(an RMS cell line)and HUVECs were infected with HPSE-1 sh RNA lentivirus at a multiplicity of infection(MOI)of 10 and 30 separately.Real-time PCR and Western blot were applied to detect the m RNA and protein expression levels of HPSE-1.Cell viability of treated or control RD cells was evaluated by cell counting kit-8(CCK-8)assay.Matrigel tube formation assay was used to evaluate the effect of HPSE-1 RNAi on the tube formation of HUVECs.RESULTS:Immunohistochemistry showed that the expression rate of HPSE-1 protein was 92.9%in orbital embr yonal RMS and 91.3%in orbital alveolar RMS.Tissue from alveolar orbital RMS did not show relatively stronger staining than that from the embryonal orbital RMS.However,despite the types of RMS,comparing the cases treated chemoradiotherapy with those untreated,we have observed that chemoradiotherapy resulted in weaker staining in patients’tissues.The expression levels of HPSE-1 declined significantly in both the m RNA and protein levels in HPSE-1 sh RNA transfected RD cells.The CCK-8 assay showed that lentivirus-mediated HPSE-1 silencing resulted in significantly reduced RD cells viability in vitro.Silencing HPSE-1 expression also inhibited VEGF-induced tube formation of HUVECs in Matrigel.CONCLUSION:HPSE-1 silencing may be a promising therapy for the inhibition of orbital RMS progression.     

Estimating the Effects of the COVID-19 Outbreak on the Decreasing Number of Acquired Immune Deficiency Syndrome Cases and Epidemiological Trends in China

<正>Since the first case of acquired immune deficiency syndrome (AIDS) was detected in the United States in 1981,this disease has spread to every corner of the world.Despite the annual incidence of AIDS showing a slight decline since1997,it remains a major cause of infectious disease-related death worldwide.Unlike global epidemiological trends,the number of notified AIDS cases has increased in recent years in China^[1].     

Smart materials in dentistry

Most dental materials are designed to have a relatively 'neutral' existence in the mouth. It is considered that if they are 'passive' and do not react with the oral environment they will be more stable and have a greater durability. At the same time, it is hoped that our materials will be well accepted and will cause neither harm nor injury. This is an entirely negative approach to material tolerance and biocompatibility and hides the possibility that some positive gains can be achieved by using materials which behave in a more dynamic fashion in the environment in which they are placed. An example of materials which have potential for 'dynamic' behaviour exists with structures which are partly water-based or have phases or zones with significant water content and for which the water within the material can react to changes in the ambient conditions. Such materials may even be said to have the potential for 'smart' behaviour, i.e. they can react to changes in the environment to bring about advantageous changes in properties, either within the material itself or in the material-tooth complex. The controlled movement of water or aqueous media through the material may cause changes in dimensions, may be the carrier for various dissolved species, and may influence the potential for the formation of biofilms at the surface. Some of these issues may be closely interrelated. Clearly, materials which do not have the capacity for water transport or storage do not have the potential for this sort of behaviour. Some materials which are normally resistant to the healthy oral environment can undergo controlled degradation at low pH in order to release ions which may prove beneficial or protective. It is doubtful whether such behaviour should be classified as 'smart' because the material cannot readily return to its original condition when the stimulus is removed. Other materials, such as certain alloys, having no means of transporting water through their structure, can display smart behaviour by undergoing predictable changes in structure in response to applied mechanical or thermal stimuli. It has been difficult to harness such behaviour to the benefit of patients but progress in this area is slowly being made.