This study was purposed to investigate the effects of 25 Gy gamma-ray irradiation on the CD62p expression, platelet count and the mean platelet volume (MPV) of manually enriched platelet suspension in different time of shelf life at 22 degrees C. Each of 16 bags with plasma-rich platelet was divided into two bags, one of which was exposed to 25 Gy gamma-ray of 137Cs and the other ones was not exposed. 16 bags then were preserved for 72 hours according to AABB standards. The irradiated platelets were regarded as the observation group, and the other ones were regarded as the control group, the expression of p-selectin (CD62p) in the above 2 groups was detected by flow cytometry before irradiation and at 24, 72 hours after irradiation respectively; at the same time, the platelet count and MPV were assayed by using blood cell counter. The results showed that the expression level of CD62p on platelet in irradiated and control groups increased along with the prolonging of preservation time, the expression rate of CD62p on the platelets preserved for 24 hours was higher than that on fresh platelets with significant difference (p<0.05); the expression rate of CD62p on the platelets preserved for 72 hours obviously was enhanced as compared with platelets preserved for 24 hours (p<0.01). There were no significant differences in CD62p expression rate, platelet count and MPV between irradiated and control groups preserved for 24 and 72 hours (p>0.05), however the MPV of irradiated and control groups preserved for 72 hours was higher than that of fresh platelets (p<0.05). It is concluded that the gamma-ray irradiation does not affect the quantity and quality of platelets, but the preservation time for manually enriched platelet suspension should be shortened as far as possible.
Blood Platelets ; metabolism ; radiation effects ; Flow Cytometry ; Gamma Rays ; Humans ; P-Selectin ; metabolism ; radiation effects ; Platelet Count ; Plateletpheresis ; Preservation, Biological ; methods

OBJECTIVETo investigate the effect of G4PAMAM/VEGFASODN compound on expression of VEGF and VEGF mRNA in MDA-MB-231 breast cancer cells and the growth inhibition of vascular endothelial cells.

METHODSThe diameter of G4PAMAM/VEGFASODN granule was measured by transmission electron microscopy, and its stability under different pH was also observed. The efficiency of transfection in vitro was detected by flow cytometer and the positively transfected cells were detected by laser scanning confocal microscope. The survival rate and the inhibitory rate of vascular endothelial cells were determined by MTT assay. The expression of protein VEGF was detected by immunohistochemical method and the expression of VEGF mRNA was detected by RT-PCR.

RESULTThe diameter of G4PAMAM/VEGFASODN granules was about 10 nm and it arranged as homogeneous netlike. Under pH 5-10 G4PAMAM/VEGFASODN presented highly stable and no dissociation was observed with different charge ratios. The 48-hour transfection rate of G4PAMAM/VEGFASODN in charge ratio of 1:40 was significantly higher than that of the lipofectamine group. None of the transfection products in each group showed cell toxicity. The staining of VEGF protein in the cytoplasm of MDA-MB-231 cells after G4PAMAM/ASODN transfection weakened markedly, and the positive expression rate decreased. Meanwhile, the VEGF mRNA expression was also decreased.

CONCLUSIONWith good stability and transfection rate, compound G4PAMAM/VEGFASODN can be a promising new nanometer vector of gene transfer system. The G4PAMAM/VEGFASODN can inhibit the expression of VEGF gene specifically and efficiently, which may be used for in vivo animal experiment.

Angiogenesis Inhibitors ; genetics ; pharmacology ; Breast Neoplasms ; blood supply ; genetics ; pathology ; Cell Line, Tumor ; Dendrimers ; pharmacology ; Humans ; Nanoparticles ; Nylons ; pharmacology ; Oligonucleotides, Antisense ; pharmacology ; RNA, Messenger ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide (G4PAMAM/VEGFASODN) compound on the expressions of vascular endothelial growth factor (VEGF) and its mRNA of breast cancer cells and on the inhibition of vascular endothelial cells.

METHODSWe examined the morphology of G4PAMAM/VEGFASODN compound and its pH stability, in vitro transfection efficiency and toxicity, and the expressions of VEGF and its mRNA. Methyl thiazolyl tetrazolium assay was used to detect the inhibitory function of the compound on vascular endothelial cells.

RESULTSThe compound was about 10 nm in diameter and was homogeneously netlike. From pH 5 to 10, it showed quite a buffered ability. The 48-h transfection rate in the charge ratio of 1:40 was 98.76%, significantly higher than that of the liposome group (P<0.05). None of the transfection products showed obvious toxicity on the cells. The expressions of both VEGF protein and its mRNA after G4PAMAM/VEGFASODN transfection decreased markedly.

CONCLUSIONWith a low toxicity, high safety, and high transfection rate, G4PAMAM/VEGFASODN could be a promising gene vector. Specifically, it inhibits VEGF gene expression efficiently, laying a basis for further in vivo animal studies.

Angiogenesis Inhibitors ; genetics ; Breast Neoplasms ; blood supply ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Dendrimers ; Gene Expression Regulation ; drug effects ; Humans ; Hydrogen-Ion Concentration ; Microscopy, Electron, Transmission ; Nylons ; Oligodeoxyribonucleotides, Antisense ; genetics ; pharmacology ; ultrastructure ; RNA, Messenger ; genetics ; Transgenes ; genetics ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; ultrastructure

BACKGROUND: Human umbilical cord blood mesenchymal stem cells (hUC-MSCs) can repair the injury of nerve cells caused by ischemia and hypoxia, but which state of cells has a better therapeutic efficacy, primary isolation or induced differentiation is not yet known. OBJECTIVE: To compare the therapeutic effects of hUC-MSCs primary cultured and differentiated in a rat model of cerebral infarction.METHODS: After full-term delivery, fetal umbilical cord blood samples were obtained using quadruple bags by means of density gradient centrifugation. hUC-MSCS were induced in the medium containing basic fibroblast growth factor. Sixty rats were equivalently randomized into four groups: sham, model, primary culture and induced differentiation groups. Animal models of cerebral infarction were made in the rats in the latter three groups. Model rats in the primary culture and induced differentiation groups were subjected to tail vein transplantation of hUC-MSCs that were primary cultured or induced to differentiate in vitro at 7 days after modeling. RESULTS AND CONCLUSION: A marked improvement in balance, walking, spatial orientation, and learning and memory abilities was found in the rats after transplantation of hUC-MSCs that were primary cultured or induced to differentiate. Moreover, compared with the primary culture group, a significant improvement was found in the induced differentiation group, including improved pathological injury of the brain, higher expression of CD34 and Ki-67, lower expression of glial fibrillary acidic protein, lower expression of interleukin 1β and tumor necrosis factor β. Compared with the primary culture group, similar infarction size and expression of interleukin-6 were also found in the induced differentiation group. These findings indicate that hUC-MSCs with induced differentiation exhibit better therapeutic outcomes in the recovery of neurological function of cerebral infarction rats.

The aim of this paper was to investigate the potential pharmacological effect of flavonoids in Sophora alopecuroides by network pharmacology. This study predicted the potential targets of 11 flavonoids of S. alopecuroides with help of reversed pharmacophore matching target recognition service platform (PharmMapper). The pathway information was acquired from DAVID and KEGG databases. Cytoscape software was used to construct the "ingredient-target-pathway" network of flavonoids active components of S. alopecuroides. The flavonoids active components of S. alopecuroides play anti-inflammatory, blood sugar regulating and other pharmacological effects by regulating 62 targets (such as INSR，KDR，MET) and intervening 44 pathways, such as B cell receptor signaling pathway, insulin signaling pathway, neurotrophin signaling pathway, and T cell receptor signaling pathway. In this study, the mechanism of "muti components-multitargets-multiple pathway" of flavonoids was studied. It reflects the multi-components, multi-targets and multiple pathway features of traditional Chinese medicine. Meanwhile, it provides a scientific basis for the elucidation the mechanism of S. alopecuroides as a medicine, and the development and utilization resources of S. alopecuroides.

BACKGROUNDThe prognostic values of the coronary computed tomography angiography (CCTA) score for predicting future cardiovascular events have been previously demonstrated in numerous studies. However, few studies have used the rich information available from CCTA to detect functionally significant coronary lesions. We sought to compare the prognostic values of Gai's plaque score and the coronary artery calcium score (CACS) of CCTA for predicting functionally significant coronary lesions, using fractional flow reserve (FFR) as the gold standard.

METHODSWe retrospectively analyzed 107 visually assessed significant coronary lesions in 88 patients (mean age, 59.6 ± 10.2 years; 76.14% of males) who underwent CCTA, invasive coronary angiography, and invasive FFR measurement. An FFR <0.80 indicated hemodynamically significant coronary stenosis. Lesions were divided into two groups using an FFR cutoff value of 0.80. We compared Gai's plaque scores and CACS between the two groups and evaluated the correlations of these scores with FFR. The statistical methods included unpaired t-test, Mann-Whitney U-test, and Spearman's correlation coefficients.

RESULTSCoronary lesions with FFR <0.80 had higher Gai's scores than those with FFR ≥0.80. Gai's score had the strongest correlation with FFR (r = -0.48, P < 0.01) and had a greater area under the curve = 0.72 (95% confidence interval: 0.61-0.82; P < 0.01) than the CACS of whole arteries and a single artery.

CONCLUSIONSBoth CACS in a single artery and Gai's plaque score demonstrated a good capacity to assess functionally significant coronary artery stenosis when compared to the gold standard FFR. However, Gai's plaque score was more predictive of FFR <0.80. Gai's score can be easily calculated in daily clinical practice and could be used when considering revascularization.

Aged ; Computed Tomography Angiography ; Coronary Angiography ; Coronary Stenosis ; pathology ; Coronary Vessels ; pathology ; Female ; Fractional Flow Reserve, Myocardial ; physiology ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Vascular Calcification ; pathology

OBJECTIVE: To study the influence of twin pregnancy by assisted reproductive technology (ART) versus twin pregnancy by spontaneous conception (SC) on neonatal outcomes. METHODS: A retrospective analysis was performed for the clinical data of 3 356 live twins with a gestational age of ≥24 weeks who were born in Nanjing Maternal and Child Health Hospital from 2017 to 2019, with 2 006 twins (1 003 pairs) in the ART group and 1 350 (675 pairs) in the SC group. The two groups were compared in terms of the mother's general information and pregnancy comorbidities and the general information, diseases, and outcomes of neonates. RESULTS: Compared with the SC group, the ART group had a significantly higher maternal age ( CONCLUSIONS Compared with twin pregnancy by SC, twin pregnancy by ART does not increase the neonatal mortality rate and risk of adverse outcomes.
Cesarean Section ; Female ; Humans ; Infant, Newborn ; Pregnancy ; Pregnancy Outcome ; Pregnancy, Twin ; Premature Birth ; Reproductive Techniques, Assisted ; Retrospective Studies

OBJECTIVETo investigate the effect of premature rupture of the membrane (PROM) on neonatal complications in premature infants.

METHODSThe registration information of 7684 preterm infants with gestational age <37 weeks were collected from the cooperative units in the task group between January 1, 2014 to December 31, 2014. Specially trained personnel from each cooperative units filled in the unified form in a standardized format to record the gender, gestational age, birth weight, PROM, placental abruption, antenatal corticosteroid, Apgar score, amniotic fluid pollution, and complications of the infants. The data were analyzed comparatively between the cases with PROM and those without (control).

RESULTSThe preterm mortality rate was significantly lower but the incidences of ICH, NEC, ROP and BPD were significantly higher in PROM group than in the control group (P<0.05). The 95% confidence interval of the OR value was <1 for mortality, and was >1 for ICH, NEC, ROP and BPD. After adjustment for gestational age, birth weight, gender, mode of delivery, placental abruption, placenta previa, prenatal hormones, gestational diabetes mellitus (GDM), gestational period hypertension and 5-min Apgar score <7, the incidences of NEC, ROP and BPD were significantly different between the two groups (P<0.05) with 95% confidence interval of OR value >1, but the mortality rate and incidence of ICH were not significantly different between the two groups (P>0.05).

CONCLUSIONPROM is a risk factor for NEC, ROP and BPD in preterm infants, and adequate intervention of PROM can reduce the incidences of such complications as NEC, ROP and BPD in the infants.

Apgar Score ; Birth Weight ; Female ; Fetal Membranes, Premature Rupture ; pathology ; Gestational Age ; Humans ; Incidence ; Infant, Newborn ; Infant, Newborn, Diseases ; etiology ; Infant, Premature ; Pregnancy ; Risk Factors

Birth Weight ; Female ; Gestational Age ; Humans ; Incidence ; Infant ; Infant, Newborn ; Infant, Very Low Birth Weight ; Pregnancy ; Risk Factors ; Sepsis/etiology*

BACKGROUND: Circulating tumor DNA (ctDNA) is a promising biomarker for non-invasive epidermal growth factor receptor mutations (EGFRm) detection in lung cancer patients, but existing methods have limitations in sensitivity and availability. In this study, we used the ΔCt value (mutant cycle threshold [Ct] value-internal control Ct value) generated during the polymerase chain reaction (PCR) assay to convert super-amplification-refractory mutation system (superARMS) from a qualitative method to a semi-quantitative method named reformed-superARMS (R-superARMS), and evaluated its performance in detecting EGFRm in plasma ctDNA in patients with advanced lung adenocarcinoma. METHODS: A total of 41 pairs of tissues and plasma samples were obtained from lung adenocarcinoma patients who had known EGFRm in tumor tissue and were previously untreated. EGFRm in ctDNA was identified by using superARMS. Through making use of ΔCt value generated during the detection process of superARMS, we indirectly transform this qualitative detection method into a semi-quantitative PCR detection method, named R-superARMS. Both qualitative and quantitative analyses of the data were performed. Kaplan-Meier analysis was performed to estimate the progression-free survival (PFS) and overall survival (OS). Fisher exact test was used for categorical variables. RESULTS: The concordance rate of EGFRm in tumor tissues and matched plasma samples was 68.3% (28/41). At baseline, EGFRm-positive patients were divided into two groups according to the cut-off ΔCt value of EGFRm set at 8.11. A significant difference in the median OS (mOS) between the two groups was observed (EGFRm ΔCt ≤8.11 vs. >8.11: not reached vs. 11.0 months; log-rank P = 0.024). Patients were divided into mutation clearance (MC) group and mutation incomplete clearance (MIC) group according to whether the ΔCt value of EGFRm test turned negative after 1 month of treatment. We found that there was also a significant difference in mOS (not reached vs. 10.4 months; log-rank P = 0.021) between MC group and MIC group. Although there was no significant difference in PFS between the two groups, the two curves were separated and the PFS of MC group tended to be higher than the MIC group (not reached vs. 27.5 months; log-rank P = 0.088). Furthermore, EGFRm-positive patients were divided into two groups according to the cut-off of the changes in ΔCt value of EGFRm after 1 month of treatment, which was set at 4.89. A significant difference in the mOS between the two groups was observed (change value of ΔCt >4.89 vs. ≤4.89: not reached vs. 11.0 months; log-rank P = 0.014). CONCLUSIONS Detecting EGFRm in ctDNA using R-superARMS can identify patients who are more likely sensitive to targeted therapy, reflect the molecular load of patients, and predict the therapeutic efficacy and clinical outcomes of patients.
Adenocarcinoma of Lung/genetics* ; Circulating Tumor DNA/genetics* ; ErbB Receptors/genetics* ; Humans ; Lung Neoplasms/genetics* ; Mutation/genetics* ; Protein Kinase Inhibitors