Amniotic Fluid ; Gastrointestinal Hemorrhage ; etiology ; Humans ; Infant, Newborn ; Vitamin K Deficiency ; complications

AIM:To investigate the effects of chronic hypoxia on the aggressiveness of MCF-7, a human breast cancer cell line , and the underlying mechanisms .METHODS: MCF-7 cells were cultured under hypoxia ( 1% O2 , 5%CO2 and 94%N2 ) or control (95%O2 and 5%CO2 ) condition.The viability, proliferation, and invasion and migration abilities of the MCF-7 cells were determined by MTT assay , CCK-8 assay, cell counting, and cell invasion and migration assays.Anchorage-independent growth and the alteration of cellular polarization of the MCF-7 cells were tested by soft agar colony formation assay and Matrigel-3D culture assay, respectively.The effects of chronic hypoxia on the growth and metas-tasis of MCF-7 cells in vivo were investigated by xenograft in nude mice .The morphological changes of the MCF-7 cells were observed under an inverted microscope .Hypoxia-induced alterations in the levels of hypoxia inducible factor-1 ( HIF-1 ) and phosphorylated glycogen synthase kinase-3β( p-GSK-3β) as well as epithelial-mesenchymal transition ( EMT) mol-ecules, such as E-cadherin, N-cadherin, vimentin, matrix metalloproteinase ( MMP)-3 and MMP-9, were determined by Western blot .RESULTS:Chronic hypoxia significantly increased the viability , proliferation , and invasion and migration abilities of MCF-7 cells in vitro, enhanced the anchorage-independent growth , facilitated cellular polarization alteration in Matrigel-3D culture, and promoted cancer metastasis in vivo.Hypoxia up-regulated HIF-1, activated GSK-3β, down-regu-lated E-cadherin and increased the protein levels of N-cadherin, vimentin, MMP-3 and MMP-9.CONCLUSION:Chronic hypoxia enhances the aggressiveness of breast cancer cells probably through EMT .

The COVID-19 pandemic has had a major impact in healthcare systems across the world, with many hospitals having to come up with protocols and measures to contain the spread of the virus. This affects various specialties' clinical practices in many ways. Since early 2020 in Singapore, the Department of Otorhinolaryngology at Tan Tock Seng Hospital had to rapidly adapt to this pandemic as we provided services to the main healthcare facility combating the virus in our country. We had to design new workflows and also remain flexible in view of the ever-changing situation. There are 6 important domains for an otolaryngology department or any clinical department in general to consider when making adjustments to their practices in an outbreak: (1) clinical work, (2) education, (3) research, (4) safety of patients and staff, (5) morale of medical staff and (6) pandemic frontline work. We hope that the sharing of our experiences and the lessons learnt will be useful for both our local and international colleagues.
Ambulatory Care ; Biomedical Research ; COVID-19/prevention & control* ; Delivery of Health Care/methods* ; Education, Medical ; Elective Surgical Procedures ; Health Workforce ; Humans ; Morale ; Otolaryngology/methods* ; Otorhinolaryngologic Surgical Procedures ; Personal Protective Equipment ; Personnel Staffing and Scheduling ; SARS-CoV-2 ; Singapore/epidemiology* ; Workflow

COVID-19/transmission* ; Communicable Disease Control/organization & administration* ; Education, Distance/organization & administration* ; Education, Medical/organization & administration* ; Humans ; Otolaryngology/organization & administration* ; Singapore/epidemiology* ; Telemedicine/organization & administration*

OBJECTIVETo observe the efficacy and safety of Danlong Oral Liquid (DOL) combined Western medicine (WM) in treating mild-to-moderate bronchial asthma patients (heat wheezing syndrome) at acute onset.

METHODSTotally 480 mild-to-moderate bronchial asthma patients (heat wheezing syndrome) at acute onset were randomly assigned to two groups in the ratio 3:1, the treatment group (360 cases) and the control group (120 cases). All patients received basic WM treatment. Patients in the treatment group took DOL, 10 mL each time, 3 times per day for 7 days in total, while those in the control group took Kechuanning Oral Liquid (KOL) , 10 mL each time, 3 times per day for 7 days in total. Efficacy for asthma symptoms, lung functions and scores of TCM syndrome and/or main symptoms were evaluated.

RESULTSThe percentage of clinical control and significant effectiveness of asthma symptoms in the treatment group was significantly higher than that of the control group (77.36% vs 56.07%, P < 0.01). The percentage of clinical control and significant effectiveness of lung functions in the treatment group was significantly higher than that of the control group (74.28% vs 50.00%, P < 0.01). The anterior-posterior difference in scores of TCM syndrome was significantly superior in the treatment group than in the control group (-11.26 ± 4.70 vs -9.21 ± 5.09, P < 0.01). The anterior-posterior difference in scores of main symptoms was significantly better in the treatment group than in the control group (-6.58 ± 3.08 vs -5.16 ± 3.45, P < 0.01). The incidence of adverse reactions was significantly lower in the treatment group than in the control group [1.73% (6/346 cases) vs 10.17% (12/118 cases) , P < 0.05].

CONCLUSIONDOL combined WM was superior to KOL in treating mild-to-moderate bronchial asthma patients (heat wheezing syndrome) at acute onset.

Anti-Asthmatic Agents ; administration & dosage ; therapeutic use ; Asthma ; drug therapy ; Biomedical Research ; Drug Therapy, Combination ; methods ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Hot Temperature ; Humans ; Lung ; Medicine, Chinese Traditional ; Phytotherapy ; Respiratory Sounds ; Syndrome

The objective was to identify some biochemical and physical properties for fusion protein IL6D24-PE40KDEL. Edman degradation, SDS-PAGE, peptide mass fingerprinting, Western blot and MTT were used for identification of the protein. The results showed that the sequence of N-terminus is Met-Ile-Asp-Lys-Gln-Ile, Met was added because of prokaryotic expression system; Western blot revealed that the purified protein could react with IL6 and PEA antibody. The purified protein IL6D24-PE40KDEL could kill the multiple myeloma cell lines U266 expressing high affinity IL6R, but it could not kill the cell lines CEM which not expressed IL6R; The molecular weight was 58.7 kD measuring by SDS-PAGE; peptide mass fingerprinting (PMF) confirmed that the construction of IL6D24-PE40KDEL was correct. A novel protein by Peptident database in EXPASY web site was identified. In conclusion, IL6D24-PE40KDEL is a new targeting protein with bioactivity of specific killing effect.
ADP Ribose Transferases ; chemistry ; metabolism ; pharmacology ; Amino Acid Sequence ; Blotting, Western ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Exotoxins ; chemistry ; metabolism ; pharmacology ; Humans ; Interleukin-6 ; chemistry ; metabolism ; pharmacology ; Molecular Sequence Data ; Pseudomonas aeruginosa ; genetics ; metabolism ; Recombinant Fusion Proteins ; chemistry ; metabolism ; pharmacology ; Sequence Analysis, Protein ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

BACKGROUNDThere were only 3 multiple myeloma (MM) cell lines established in China. In this study, we succeeded in establishing a novel MM cell line and analyzed its biological characteristics.

METHODSMononuclear cells isolated from the peripheral blood (PB) and bone marrow (BM) of a patient with advanced MM (lambda light chain type) were cultured in medium. Cell morphology was analyzed by Wright-Giemsa-staining and cytochemical staining, immunophenotyping by flow cytometry and cytogenetic analysis by chromosome RHG-banding technique. Quantitative fluorescent polymerase chain reaction (PCR) was used to detect Epstein - Barr virus (EBV) DNA.

RESULTSThe established cell line could survive and proliferate in the presence of feeder cells or conditioned medium. The cells secreted lambda light chain and were negative for EBV. The Wright-Giemsa-staining showed typical plasmablast or plasma cell morphology. The cytochemical staining of the cells showed the following reactivity patterns: positive for acid phosphatase, negative for myeloperoxidase. The immunoprofile of the cells was concordant with that of MM cells: positive for CD10, CD28, CD38, CD138, CD56, CD49d, CD44, CD54 and CD58, negative for CD19, CD40, CD95, CD95L, CD34, CD2 and CD5. The cytogenetic analysis showed complex chromosome abnormality of i (1q+), 8q+, 13q+, i (17q), i (18q) and +M. There was no difference in morphology, immunophenotype and cytogenetics between cells from PB and BM.

CONCLUSIONSAn MM cell line secreting lambda light chain named CZ-1 was established. The cells from both PB and BM have the same biological characteristics.

Cell Line, Tumor ; Chromosome Aberrations ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; genetics ; pathology ; Polymerase Chain Reaction

OBJECTIVETo find out a possible approach to improve the effectiveness of radiotherapy and chemotherapy for Ewing's sarcoma by constructing a eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia and to evaluate the effects of this HRE regulated HSV-TK system on killing effect of gancyclovir (GCV) on Ewing's sarcoma cell line SK-ES under hypoxic condition.

METHODSThe HRE was synthesized according to the literature and cloned into the enhancer site of pIRES(2)-EGFP vector to obtain the pHRE recombinant plasmid. The HSV-TK was amplified by PCR and cloned into the multiple clone site of pIRES(2)-EGFP and pHRE to obtain pTK and pHRE-TK recombinant plasmid. The human Ewing's sarcoma cell line SK-ES was transfected by pTK or pHRE-TK recombinant plasmid with liposome and then was exposed to normoxic (21% oxygen) or hypoxic (3% oxygen) condition. The expression of enhanced green fluorescent protein (EGFP) was monitored by fluorescent microscopy. The sensitivity of human Ewing's sarcoma cell line SK-ES transfected with pTK or pHRE-TK recombinant plasmid to the anti-tumour drug GCV was determined with the method of tetrazolium (MTT) after treating with GCV for five days.

RESULTS(1) The result of sequencing showed that the recombinant plasmid pHRE contained HRE, and that the recombinant plasmid pTK and pHRE-TK contained HSV-TK gene in the sense direction. (2) Comparison of fluorescent optical density (FOD) showed that (1) the EGFP FOD value of pHRE and pHRE-TK group cells exposed to hypoxia was significantly higher than those exposed to normoxia (P < 0.01); (2) when the cells were exposed to hypoxia, the EGFP FOD value of pHRE and pHRE-TK group cells was significantly higher than that of pTK and empty vector group (P < 0.01); (3) there was no significant difference among the four groups of cells when they were exposed to normoxia (P > 0.05). (3) Comparison of the sensitivity of four groups of cells to GCV showed that (1) the cells in pHRE-TK and pTK groups were much more sensitive to GCV than the cells in pHRE group under hypoxia condition (P < 0.01), the higher the GCV concentration, the greater the difference; (2) the cells of pHRE-TK group were more sensitive to GCV than those in pTK group under hypoxic condition (P < 0.01), but was almost equally sensitive under normoxic condition (P > 0.05); (3) the pHRE-TK group cells had higher sensitivity to GCV under hypoxia than normoxia (P < 0.01) while the pTK group cells had almost the same sensitivity to GCV under hypoxia and normoxia (P > 0.05).

CONCLUSION(1) The eukaryotic expression vector expressing herpes simplex virus-thymidine kinase (HSV-TK) regulated by hypoxia responsive element (HRE) under hypoxia was constructed successfully. (2) HRE could up-regulate expression of EGFP by SK-ES cells under hypoxia condition. (3) HRE could enhance the killing effect of HSV-TK/GCV system on human Ewing's sarcoma cell line SK-ES under hypoxic condition.

Antiviral Agents ; pharmacology ; Cell Hypoxia ; drug effects ; genetics ; Cell Line, Tumor ; Ganciclovir ; pharmacology ; Gene Expression Regulation ; drug effects ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Microscopy, Fluorescence ; Plasmids ; Polymerase Chain Reaction ; Response Elements ; genetics ; Sarcoma, Ewing ; drug therapy ; metabolism ; Simplexvirus ; genetics ; metabolism ; Thymidine Kinase ; genetics ; metabolism ; Transfection

OBJECTIVETo investigate the effects of electromagnetic radiation on the physiological indices and immune function of operators.

METHODSThe general conditions and electromagnetic radiation awareness rate of 205 operators under electromagnetic radiation were evaluated using a self-designed questionnaire. Physical examination, electrocardiography, and routine urine test were performed in these operators. Peripheral blood was collected from the operators under electromagnetic radiation for blood cell counting and biochemical testing, and their peripheral blood lymphocytes were cultured for determination of chromosomal aberrant frequency and micronucleus frequency. The data from these operators (exposure group) were compared with those of 95 ordinary individuals (control group).

RESULTSThe chief complaint of giddiness, tiredness, dizziness, and amnesia showed significant differences between the exposure group and control group (P < 0.01), and the difference in headache became larger with an increase in working years. The awareness rate of electromagnetic radiation damage was significantly higher in the exposure group than in the control group. The difference in bradycardia was significant between the two groups (P <0.01), and the incidence was higher with longer working years. Significant differences between the two groups were also found in the numbers of individuals with elevated alanine aminotransferase, total bilirubin, and direct bilirubin (P < 0.01), populations with increased lymphocyte ratio and decreased neutrophil ratio (P < 0.01), populations with positive occult blood, urobilinogen, and bilirubin tests, and the number of individuals with increased micronucleus frequency of cultured peripheral blood lymphocytes (P < 0.01). In addition, the exposure group had significantly increased complement C3 and C4 (P < 0.01), significantly increased IgG (P < 0.05), and significantly decreased IgM (P < 0.01), as compared with the control group.

CONCLUSIONElectromagnetic radiation may lead to the changes in physiological indices, genetic effects, and immune function and affect the health and immune function in operators. The adverse effects are increased as the working years increase. So it is important to strengthen occupational protection of operators under electromagnetic radiation.

Adult ; Chromosome Aberrations ; radiation effects ; Electromagnetic Radiation ; Female ; Humans ; Lymphocytes ; radiation effects ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Young Adult

This study was purposed to investigate the connexin 43 (Cx43) expression level in acute leukemia bone marrow stromal cells (ABMSCs) and normal bone marrow stromal cells (NBMSCs), and to explore the difference in communicating functions between these cells. The Cx43 expression levels of ABMSCs and NBMSCs were detected by using immunohistochemistry and computer gray scale assay, and the difference of gap junction intercellular communication (GJIC) was examined through dry transfer technique. The results showed that expression level of Cx43 in ABMSCs was lower than that in NBMSCs and its function of GJIC in ABMSCs was also weaker than that in NBMSCs. It is concluded that cell-cell communication function is lowered in ABMSCs.
Acute Disease ; Bone Marrow Cells ; cytology ; metabolism ; pathology ; Cell Communication ; physiology ; Connexin 43 ; metabolism ; Gap Junctions ; metabolism ; Humans ; Leukemia ; metabolism ; Stromal Cells ; cytology ; metabolism ; Tumor Cells, Cultured