Objective To investigate the effects of apigenin on the migration of vascular smooth muscle cells (VSMC) induced by platelet derived growth factor (PDGF)-BB and the possible molecular mechanism. Methods VSMCs were isolated from thoracic aortas of male Sprague-Dawley rats using enzyme digestion method. Migration of VSMCs was determined by transwell assay. Western blotting was carried out to evaluate phosphorylation of c-jun N-terminal kinase (JNK). Results Treatment with PDGF-BB (20 ng·mL-1 ) significantly promote VSMC migration,the number of migrated cells was 2. 46 times than that of control group. However,after 12. 5 μmol·L-1 apigenin pretreatment,the number of migrated cells was 46. 5% of the PDGF-BB group. Various dose of apigenin can significantly inhibit VSMC migration induced by PDGF-BB,12. 5 μmol · L-1 apigenin treatment significantly inhibited PDGF-BB phosphorylation of JNK. Conclusion Apigenin can suppress the migration of VSMC induced by PDGF-BB. These beneficial effects on VSMC were at least partly mediated by the inhibition of activity of JNK.

Objective:To choose the best vector for the expression of CS3 fimbriae. Methods: The CS3 operon was cloned into different plasmid vectors such as pUC19 and pTrc99A. The expression of CS3 was monitored by whole-cell ELISA and SDS-PAGE analysis. The assembly of CS3 fimbriae was detected with electron microscopy. Results:The expression level of CS3 fimbriae using plasmid pUC19 as carrying vector was the highest, and the insertion orientation of CS3 gene into the plasmid has a little effect on its expression level. The expression of CS3 fimbriae was confirmed by SDS-PAGE analysis and electron microscopy.Conclusions:The promotor of CS3 itself played the key role in the expression of CS3 fimbriae and the copy number of plasmid was the main factor to affect the expression level.

Purpose: The global fight against the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has led to widespread vaccination efforts, yet the optimal dosing schedule for SARS-CoV-2 vaccines remains a subject of ongoing research. This study aims to investigate the effectiveness of administering two booster doses as the third and fourth doses at different intervals to enhance vaccine protection. Materials and Methods: This study was conducted at a military regional hospital operated by the Ministry of National Defense in Taiwan. A cohort of vaccinated individuals was selected, and their vaccine potency was assessed at various time intervals following their initial vaccine administration. The study participants received booster doses as the third and fourth doses, with differing time intervals between them. The study monitored neutralizing antibody titers and other relevant parameters to assess vaccine efficacy. Results: Our findings revealed that the potency of the SARS-CoV-2 vaccine exhibited a significant decline 80 days after the initial vaccine administration. However, a longer interval of 175 days between booster injections resulted in significantly higher neutralizing antibody titers. The individuals who received the extended interval boosters exhibited a more robust immune response, suggesting that a vaccine schedule with a 175-day interval between injections may provide superior protection against SARS-CoV-2. Conclusion This study underscores the importance of optimizing vaccine booster dosing schedules to maximize protection against SARS-CoV-2. The results indicate that a longer interval of 175 days between the third and fourth doses of the vaccine can significantly enhance the neutralizing antibody response, potentially offering improved protection against the virus. These findings have important implications for vaccine distribution and administration strategies in the ongoing battle against the SARS-CoV-2 pandemic. Further research and large-scale trials are needed to confirm and extend these findings for broader public health implications.

This study was undertaken to establish a murine model for unrelated allogeneic umbilical cord blood transplantation (UCBT). The characteristics and percentage of hematopoietic stem/progenitor cells between near-term fetal and neonatal murine peripheral blood (FNPB) and bone marrow (BM) were evaluated by flow cytometry and semisolid methylcellulose culture. BABL/c (H-2(d)) recipient mice conditioned with high dose CTX were transplanted with FNPB form C57BL/6 (H-2(b)) mice and the survival rate, hematopoietic and immunological reconstruction, graft versus host disease (GVHD) and engraftment level were observed. The results showed that the numbers of day 14 CFU-GM and CFU-GEMM in FNPB (176.40 +/- 78.39)% and (141.40 +/- 56.57)%, respectively were much higher than those in BM (75.20 +/- 26.41)% and (68.80 +/- 23.95)%, respectively. Moreover the percentage of Sca-1(+) CD34(+) cell subsets in FNPB (3.63 +/- 1.13)% was also higher than that in BM (1.41 +/- 0.8 7)%. FNPB transplantation improved survival rate and reconstituted hematopoietic and immune function in recipients. There was no evidence of GVHD. Chimeric analysis showed that the proportion of donor cells in BM of recipients was 27.94% at 21 days after transplantation. It was concluded that FNPB contains more hematopoietic stem and progenitor cells with high expansion ability and weak allogeneic immunity, which was similar to human UCB. The murine model for allogeneic UCBT (C57BL/6-->BALB/c) was established successfully.
Animals ; Animals, Newborn ; Fetal Blood ; cytology ; Flow Cytometry ; Graft vs Host Disease ; etiology ; Hematopoietic Stem Cell Transplantation ; Immunity ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Models, Animal ; Transplantation, Homologous

BACKGROUND/OBJECTIVES: The objectives of this study were to investigate the effects of lycopene on the migration, adhesion, tube formation capacity, and p38 mitogen-activated protein kinase (p38 MAPK) activity of endothelial progenitor cells (EPCs) cultivated with high glucose (HG) and as well as explore the mechanism behind the protective effects of lycopene on peripheral blood EPCs. MATERIALS/METHODS: Mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation. EPCs were identified after induction of cellular differentiation. Third generation EPCs were incubated with HG (33 mmol/L) or 10, 30, and 50 microg/mL of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation and apoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performed by replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity was assayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylated and non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and 50 microg/mL of lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantly lower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirm the causal relationship between MAPK inhibition and the protective effects of lycopene against HG-induced cellular injury, we treated cells with SB203580, a phosphorylation inhibitor. The inhibitor significantly inhibited HG-induced EPC injury. CONCLUSIONS: Lycopene promotes proliferation, migration, adhesion, and in vitro vasculogenesis capacity as well as reduces apoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-induced EPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-induced EPC injury by inhibiting p38 MAPKs.
Apoptosis ; Blotting, Western ; Centrifugation, Density Gradient ; Ficoll ; Flow Cytometry ; Glucose* ; Humans ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Protein Kinases ; Signal Transduction ; Stem Cells*

BACKGROUND/OBJECTIVES: The objectives of this study were to investigate the effects of lycopene on the migration, adhesion, tube formation capacity, and p38 mitogen-activated protein kinase (p38 MAPK) activity of endothelial progenitor cells (EPCs) cultivated with high glucose (HG) and as well as explore the mechanism behind the protective effects of lycopene on peripheral blood EPCs. MATERIALS/METHODS: Mononuclear cells were isolated from human peripheral blood by Ficoll density gradient centrifugation. EPCs were identified after induction of cellular differentiation. Third generation EPCs were incubated with HG (33 mmol/L) or 10, 30, and 50 microg/mL of lycopene plus HG. MTT assay and flow cytometry were performed to assess proliferation and apoptosis of EPCs. EPC migration was assessed by MTT assay with a modified boyden chamber. Adhesion assay was performed by replating EPCs on fibronectin-coated dishes, after which adherent cells were counted. In vitro vasculogenesis activity was assayed by Madrigal network formation assay. Western blotting was performed to analyze protein expression of both phosphorylated and non-phosphorylated p38 MAPK. RESULTS: The proliferation, migration, adhesion, and in vitro vasculogenesis capacity of EPCs treated with 10, 30, and 50 microg/mL of lycopene plus HG were all significantly higher comapred to the HG group (P < 0.05). Rates of apoptosis were also significantly lower than that of the HG group. Moreover, lycopene blocked phosphorylation of p38 MAPK in EPCs (P < 0.05). To confirm the causal relationship between MAPK inhibition and the protective effects of lycopene against HG-induced cellular injury, we treated cells with SB203580, a phosphorylation inhibitor. The inhibitor significantly inhibited HG-induced EPC injury. CONCLUSIONS: Lycopene promotes proliferation, migration, adhesion, and in vitro vasculogenesis capacity as well as reduces apoptosis of EPCs. Further, the underlying molecular mechanism of the protective effects of lycopene against HG-induced EPC injury may involve the p38 MAPK signal transduction pathway. Specifically, lycopene was shown to inhibit HG-induced EPC injury by inhibiting p38 MAPKs.
Apoptosis ; Blotting, Western ; Centrifugation, Density Gradient ; Ficoll ; Flow Cytometry ; Glucose* ; Humans ; p38 Mitogen-Activated Protein Kinases ; Phosphorylation ; Protein Kinases ; Signal Transduction ; Stem Cells*

OBJECTIVEThe development status of pulmonary artery is one of the most important criteria for decision-making strategy and predicting postoperative outcome in congenital heart disease with decreased pulmonary blood flow. Currently, Nakata index and McGoon index have been used as morphologic index in evaluating the development status of pulmonary artery. Those indices have some shortcoming. It was recently found that pulmonary veins index is a more precise morphological indicator of pulmonary blood flow and development status of pulmonary vessels. This study aimed to explore an index of evaluating pulmonary blood stream and the development of pulmonary vessels, as a criterion for surgical decision-making strategy.

METHODSThe diameters of left and right pulmonary arteries and pulmonary veins were measured on DSA films in 74 patients with congenital heart disease with decreased pulmonary blood flow, The correlative analysis was done between Nakata index, McGoon index, pulmonary vein index (PVI) and postoperative outcome which were the length of stay in ICU, duration of mechanical ventilation and dose of inotropic drugs.

RESULTSExcellent correlations between the size of pulmonary veins and pulmonary arteries were found, the correlation between left pulmonary veins and distal portion of left pulmonary artery was 0.73, between left pulmonary veins and proximal portion of left pulmonary artery was 0.72, right pulmonary veins and distal portion of right pulmonary artery was 0.67, and right pulmonary veins and proximal portion of right pulmonary artery was 0.71. The length of stay in ICU, duration of mechanical ventilation and dose of inotropic drugs correlated well with PVI (r = -0.51, -0.478, and -0.693). Compared with Nakata index and McGoon index, PVI was a better criterion for evaluating the developmental status of the whole pulmonary vessels. In the right ventricular outlet reconstruction patients, the McGoon index for patients with low cardiac output syndrome (LCOS) was 1.36 +/- 0.51, and 1.97 +/- 0.58 for patients without LCOS (t = 2.347, P < 0.05), the Nakata index for patients with LCOS was 164 +/- 106 mm(2)/m(2) and 269 +/- 124 mm(2)/m(2) for patients without LCOS (t = 2.218, P < 0.05), the PVI for patients with LCOS was 152 +/- 77 mm(2)/m(2) and 273 +/- 125 mm(2)/m(2) for patients without LCOS (t = 2.936, P < 0.01), pulmonary vessel index of patients with LCOS was less than that of those without LCOS. When PVI was < or = 180 mm(2)/m(2), postoperative hemodynamics was unstable, the frequency of low cardiac output syndrome and mortality significantly increased.

CONCLUSIONSThe development of pulmonary arteries and pulmonary veins correlated with each other. PVI is a precise morphological indicator of pulmonary blood flow and development of pulmonary vessels. It is a helpful indicator to decide surgical strategy.

Adolescent ; Child ; Child, Preschool ; Female ; Heart Defects, Congenital ; physiopathology ; Hemodynamics ; Humans ; Infant ; Male ; Pulmonary Artery ; growth & development ; physiopathology ; Pulmonary Veins ; growth & development ; physiopathology

OBJECTIVETo investigate the risk factors of female breast cancer and its potential alteration in Wuhan area.

METHODSA case-control study was conducted on 213 cases with histopathological diagnosis and 430 matched controls, using conditional logistic regression analysis.

RESULTS28 factors such as educational level, history of benign breast disease, age at menarche, age at menopausal, meat and well-done meat intake, soy bean food, fruit, lactation time, body mass index (BMI), juvenile chest X-ray, psychological factor, were associated with breast cancer risk in one-way variance model. Multivariate conditional logistic regression analysis in total significant factors and subgroups showed that the risk factors of breast cancer would include high level education, psychological trauma, history of benign breast disease later age at menopause, more years of menstrual and more years of menstrual before giving first birth, high BMI, well-done meat intake and smoked food. Factors as later menarche, lactate longer, soybean food, fruit, drink tea habit were protective factors for breast cancer. Further breakdown of data showed some difference between premenopausal and postmenopausal women. Risks in premenopausal women were associated with history of benign breast disease, age of menarche, soybean food intake, whereas risks in postmenopausal women were related to age of menopausal, BMI, waist-hip ratio and fruit intake. Both psychological traumatic and duration of lactation were common pre-and postmenopausal risk and protective factors.

CONCLUSIONSDietary habit and endogenous estrogen exposure related factors played important roles on women breast cancer in Wuhan area.

Adult ; Breast Neoplasms ; etiology ; metabolism ; Case-Control Studies ; China ; Estradiol Congeners ; metabolism ; Feeding Behavior ; Female ; Humans ; Logistic Models ; Middle Aged ; Multivariate Analysis ; Risk Factors

OBJECTIVENoninvasive method for estimating the pulmonary vascular resistance (PVR) was used in patients waiting for two staged Fontan procedure to observe the relationship between estimated PVR and surgical results.

METHODSThirty-three candidate patients for two staged Fontan procedures were randomly selected for this trial. Preoperative hemoglobin (HGB), hematocrit (HCT) and pulse oxygen saturation (SpO2) were measured. Estimated PVRs were then calculated by the regression equations. The cases were divided into three groups of low risk group with PVR less than 2.4 wood, high risk group with PVR between 2.4 to 3.2 wood, and extremely high risk group with PVR more than 3.2 wood. The incidences of postoperative low cardiac output and multi-organ failure were compared. Simultaneously, the cases without or with postoperative severe low cardiac output and multi-organ failure after operation were divided into two groups. The preoperative HGB, HCT and estimated PVR among the groups were analyzed.

RESULTSThe rates of postoperative severe low cardiac output and multi-organ failure were 10.0%, 32.4% and 63.6% for the patients of low, high and extremely high risk groups respectively. The difference was significant (P < 0.01). The preoperative HGB, HCT, SpO2 and PVR were all significantly different between the patients without or with postoperative severe low cardiac output and multi-organ failure. The PVR (HGB) were (2.53 +/- 0.56) wood and (3.24 +/- 0.58) wood respectively. The PVR (HCT) were (2.59 +/- 0.58) wood and (3.21 +/- 0.79) wood respectively. The PVR (SpO2) were (2.22 +/- 0.55) wood and (2.93 +/- 0.58) wood, respectively. The differences were all significant (t = 3.25, 2.52 and 3.33 respectively, P < 0.01 or 0.05).

CONCLUSIONSPreoperative estimated PVRs calculated by HGB, HCT and SpO2 were comparable with the postoperative results. Thus, the invasive preoperative estimation of pulmonary vascular resistance could be a method in estimating the surgical indication and predicting the surgical result for two stage Fontan procedure.

Adolescent ; Cardiovascular Abnormalities ; physiopathology ; surgery ; Child ; Child, Preschool ; Contraindications ; Female ; Fontan Procedure ; methods ; Humans ; Male ; Treatment Outcome ; Vascular Resistance

Objective To investigate clinically and in laboratory a genealogical tree with hemoglobin H disease Combining Hemoglobin Q-Thailand and Hemoglobin E Disease.Methods Genealogical laboratory studies were carried out with the following methods:hemoglobin electrophoresis, various biochemical determinations,and DNA analysis.Results Father's genotype of ?-THAL:??/?~Q ?~(4.2); genotype of ?-THAL:?E/N;phenotype:minor ?-THAL carrier combining Hb Q and Hb E multiple heterozygote;mother' s genotype of ct-THAL:--~(SEA)/??;genotype of ?-THAL:?n/?n.According to comprehensive analysis,mother's phenotype:minor ?-THAL,complex minor ?-THAL carrier combining Hb F ? Initial sign of ?-THAL genotype:--~(SEA)/?~Q ?~(4.2);phenotype:deletion type Hb H genotype disease;?- THAL genotype:?E/?E;phenotype:? E homozygote.According to comprehensive analysis:deletion type Hb H combining HbE multiple heterozygote.Youger brother's ?-THAL genotype:--~(SEA)/?~Q ?~(4.2);?-THAL genotype:?n/?n;phenotype:deletion type Hb H disease.Both mother and her youngest son have G6PD deficiency.Conclusions Guangdong Province is an area with high morbidity of ?-THAL and ?-THAL,Hb E and Hb Q as well as G6PD deficiency.There may be some correlation between Hb E and Fib Q in term's of the high morbidity of regional Hb,but the two types of Hb combining Hb H disease are rare in China and the world in point of nonhomologous chromosome.Attention should be paid to the problems of double heterozygote of ?-THAL complex ?-THAL,and THAL complex G6PD deficiency.Data from the study have enriched the scientific information of molecular genetics of erythroeyte thalassemia and of molecular pathology with important significance in genetics guidance and clinical treatment for patients.