Hepatitis B surface antigen (HBsAg) carrying preS sequences could be an ideal candidate for a new hepatitis B virus (HBV) vaccine with higher efficacy. Here we report the success in achieving efficient and stable expression of hepatitis B virus S antigen and preS1 epitope fusion protein (S/preS1) in CHO cells. The HMRCHEF53u/Neo-S/preS1 expression vector carrying S/preS1 gene was constructed and transfected into CHO-S cells. A stable and high-expression CHO cell line, named 10G6, was selected by ELISA and limiting dilution analysis. Western blotting analysis showed S/preS1 expressed from 10G6 cells possessed both S and preS1 antigenicity. 10G6 cells displayed characters of favorable growth and stable S/preS1 expression in repeated batch cultures as evaluated by viable cell density, viability and S/preS1 concentration. And cultivation of 10G6 cells in fed-batch mode resulted in S/preS1 production at 17-20 mg/L with viable cell density at 7 x 10(6)-10 x 10(6) cells/mL.
Animals ; CHO Cells ; Cricetulus ; Epitopes ; biosynthesis ; genetics ; Hepatitis B Surface Antigens ; biosynthesis ; genetics ; immunology ; Hepatitis B Vaccines ; biosynthesis ; genetics ; Hepatitis B virus ; Protein Precursors ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Objective To study the toxic effects of 5-amionlevulinic acid-based photodynamic therapy (ALA-PDT) on human peripheral blood mononuclear cells (PBMCs), cord blood mononuclear cells (CBMCs) and peripheral blood stem cells (PBSCs), and furthermore, to understand the possible causes of this response. Methods We used MTT assay to detect the survival rate of PBMCs, CBMCs and PBSCs after treated by ALA-PDT under the optimum experiment conditions with U937 as control;Annexin V-FITC/PI was used to detect the pattern of cell death induced by ALA-PDT. By using flow cytometry, we detected intracellular PpIX fluorescence intensity. Results After ALA-PDT treatment the survival rate of PBMCs had no significant change;however in PBSCs and CBMCs, the survival rate reduced to 70%, and the survival rate of leukemia cell U937 was the lowest, about 30%. After incubation with ALA,except for PBMCs, intraceUuiar PplX fluorescence intensity of the other two kinds of normal haemocytes and U937 increased obviously. These results combined with the flow cytometry suggested that the main pattern of cell death here was apoptosts. Conclusion Under the optimum experiment conditions, ALA-PDT has a slight effect on normal haemocytes but excellent depletions of leukemia cells. Therefore, it can effectively purify autologons bone marrow or stem cell grafts.

A flow controlling system for pulsed inhaled nitric oxide has been developed and tested, and here its features and initial animal experiments and clinical applications are described. The physical characteristic test indicates that the practical released dose of NO gas is very close to the theoretical flow of NO gas at variant pressures. Animal experiments demonstrate that inhaled NO gas concentration is lower than the concentration of theoretical inhalation, but the variance is not remarkable (p>0.05). When sixteen cases with CHD and PH were chosen to inhale NO gas (15 ppm, 15 min) PAP and PVR of all cases were reduced after inhalation of NO gas from 617 +/-51.3 dyn x s x cm(-5), 54.4+/-13.1 mmHg to 417+/-36.9 dym x s x cm(-5), 33.8+/-12.3 mmHg (PVR, p<0.01; PAP, p<0.01) respectively. When gas inhalation was stopped, these values returned to their base lines after a short period of time. All these show that the pulsed inhaled NO flow controlling instrument in accordance with the requirements of the designing, can be widely used in clinical diagnoses and treatments and will be a new tool offered for the treatments of the patients with PH.
Administration, Inhalation ; Animals ; Blood Pressure ; Cardiac Output ; Child ; Equipment Design ; instrumentation ; Female ; Heart Defects, Congenital ; physiopathology ; therapy ; Humans ; Hypertension, Pulmonary ; physiopathology ; therapy ; Male ; Nebulizers and Vaporizers ; Nitric Oxide ; administration & dosage ; analysis ; Rabbits ; Vascular Resistance

Objective To analyze the prognostic factors in patients with initially diagnosed bone-only metastatic nasopharyngeal carcinoma (NPC). Methods We collected the data of 68 patients with initially diagnosed bone-only metastatic NPC admitted to The Affiliated Tumor Hospital of Shantou University Medical College from 1997 to 2015. Forty-nine patients received chemoradiotherapy. The Kaplan-Meier method was used to calculate the overall survival rate;the log-rank test was used for univariate prognostic analysis;the Cox model was used for multivariate prognostic analysis. Results The median follow-up was 953 months. The 1-, 2-, 3-, and 5-year overall survival ( OS) rates were 53%, 38%, 21%, and 15%, respectively. The median OS time was 134 months. The univariate prognostic analysis showed that spinal metastases, the number of bone metastases, lactic dehydrogenase level before treatment, the radiotherapy technology and dose for primary tumor, and the short-term outcome of primary tumor were associated with OS ( P=002, 001, 000, 002, 002, 001 ) . The multivariate prognostic analysis showed that ≤3 bone metastases, dose to primary tumor>65 Gy, and intensity-modulated radiotherapy ( IMRT) were favorable prognostic factors for OS ( P=003,002,004) . Conclusions For patients with initially diagnosed bone-only metastatic NPC, active treatment ( IMRT, dose to primary tumor>65 Gy) should be considered for those with ≤3 bone metastases to achieve a complete response of primary tumor.

Optical mapping of the membrane potential with voltage-sensitive dyes is an advanced approach that involves in many theories such as molecular photonica, physiology and computer science. Compared with the conventional techniques of membrane patch-clamp and microelectrode, the optical mapping system can measure not only the changes of a single membrane potential from multiple sites simultaneously but also the conduction properties of the cell populations. So this technique provides an important method for studying the electrophysiology of the small cell, the tiny neurite and the cardiac arrhythmia, etc. Because this technique can also avoid the electromagnetic interferences, it uniquely provides an ideal means for studying the mechanisms such as cardiac defibrillation. In this paper the principles and the system structure of the optical mapping are introduced, its applications and future developments are also presented at the same time.
Animals ; Brain ; physiology ; Electrodes ; Electrophysiology ; methods ; Fluorescent Dyes ; Humans ; Membrane Potentials ; physiology ; Myocytes, Cardiac ; physiology ; Optics and Photonics

Objective To explore the influencing factors of the onset of autism spectrum disorder in male children.Methods Totally 151 male children with autism spectrum disorder were selected as case group and 119 healthy male children matched with the age of the case group in the same administrative region were taken as the control group.All children were assessed with the questionnaire for children's autism etiology and risk factors.Results (1) The differences in children having anorexia and partial eclipse (x2 =50.763,P＜0.01),father's age during pregnancy (x2 =11.441,P=0.043),place of pregnancy (x2 =50.763,P＜0.01),hypertension of pregnancy (x2 =5.693,P=0.026),intrauterine hypoxia (x2 =9.332,P=0.002),umbilical cord around the neck(x2 =18.483,P＜0.01),parents smoking and drinking history during pregnancy (x2 =13.660,P=0.008),parental smoking (x2 =12.901,P=0.005) and alcohol consumption (x2 =8.386,P=0.039) during pregnancy,birth height of child (x2 =8.870,P=0.031),amniotic fluid pollution (x2 =4.561,P=0.043),participation time of artificial feeding,major caregivers,delayed development indicators in infants and young children and whether or not the harmonious parent-child relationship were statistically significant(P＜0.05).(2) Children with anorexia and partial diet (OR =12.284,95％ CI =2.768-54.507),living in rural areas during pregnancy (OR =17.251,95％ CI =1.899-1 56.745),parents' history of smoking and drinking (OR =6.191,95％ CI =1.678-22.838),and intrauterine hypoxia during pregnancy (OR=38.859,95％CI=2.944-512.930) may be risk factors for male autism spectrum disorder.Conclusion To correct children's anorexia bias,improve the living environment in pregnancy,reduce pregnancy complications and avoid exposure to tobacco and alcohol pollution during maternal pregnancy can be an effective entry point for the prevention and control of autism spectrum disorders in male children.

Objective: Through the study of angiotensinⅡ- type 2 receptor agonist (AT2R) after pretreatment of mechanical ventilation lung injury (VILI) in rats model, to clarify the role of angiotensin Ⅱ - type 2 receptor agonist (C21) in alleviating VILI inflammation and the damage of immune function and its possible mechanism. Methods: In this experiment, the acute lung injury model was established by mechanical spring-volume ventilation in SD rats, and C21 pretreatment was performed to observe the pathological condition of lung tissue in rats with different ventilation duration, and to detect the inflammatory changes of BALF lavage fluid. Flow cytometry was used to detect the CD68+/iNOS+ labeled M1 type AMφ and the CD68+/Arg-1+ labeled M2 type AMφ in alveolar lavage fluid. Results: The mechanical VILI rat model was successfully established. The pathological injury score of the mechanical ventilation 4 h model, the wet/dry weight of lung tissue, the number of cells and protein in BALF lavage fluid were increased significantly, the levels of TNF-α and IL - 1 were increased significantly, the levels of IL-4 and IL-10 were decreased significantly, and the level of inflammatory reaction decreased with the increase of ventilation time. The M1/M2 ratio in the 4 h ventilation model group was the highest, which was significantly different from the control group (P<0.05). Compared to the model group, the C21 pretreatment significantly reduced the levels of proinflammatory factors TNF-αand IL-1, and increased the levels of anti-inflammatory factors IL-4 and IL-10 in BALF (P<0.01). After the intervention of AT2R agonist at 4 h, 6 h and 8 h, the M1/M2 ratio of the model was lower than that of the model without AT2R agonist at 4 h, 6 h and 8 h. Conclusion Mechanical ventilation for 4 h in SD rats can establish a mechanical ventilation lung injury model. AT2R agonist C21 can promote the polarization of macrophages to m2-type, and C21 pretreatment may alleviate VILI inflammation and immune damage by altering the polarization of macrophages.

In contrast to the well-established genomic 5-methylcytosine (5mC), the existence of N⁶-methyladenine (6 mA) in eukaryotic genomes was discovered only recently. Initial studies found that it was actively regulated in cancer cells, suggesting its involvement in the process of carcinogenesis. However, the contribution of 6 mA in tongue squamous cell carcinoma (TSCC) still remains uncharacterized. In this study, a pan-cancer type analysis was first performed, which revealed enhanced 6 mA metabolism in diverse cancer types. The study was then focused on the regulation of 6 mA metabolism, as well as its effects on TSCC cells. To these aspects, genome 6 mA level was found greatly increased in TSCC tissues and cultured cells. By knocking down 6 mA methylases N6AMT1 and METTL4, the level of genomic 6 mA was decreased in TSCC cells. This led to suppressed colony formation and cell migration. By contrast, knockdown of 6 mA demethylase ALKBH1 resulted in an increased 6 mA level, enhanced colony formation, and cell migration. Further study suggested that regulation of the NF-κB pathway might contribute to the enhanced migration of TSCC cells. Therefore, in the case of TSCC, we have shown that genomic 6 mA modification is involved in the proliferation and migration of cancer cells.
AlkB Homolog 1, Histone H2a Dioxygenase/metabolism* ; Carcinoma, Squamous Cell/pathology* ; Cell Line, Tumor ; Cell Movement/genetics* ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Site-Specific DNA-Methyltransferase (Adenine-Specific)/metabolism* ; Tongue Neoplasms/metabolism*