Nitrites play multiple characteristic functions in invasion and metastasis of hepatic cancer cells, but the exact mechanism is not yet known. Cancer cells can maintain the malignant characteristics via clearance of excess mitochondria by mitophagy. The purpose of this article was to determine the roles of nitrite, reactive oxygen species (ROS) and hypoxia inducing factor 1 alpha (HIF-1 α) in mitophagy of hepatic cancer cells. After exposure of human hepatocellular carcinoma SMMC-7721 cells to a serial concentrations of sodium nitrite for 24 h under normal oxygen, the maximal cell vitality was increased by 16 mg x (-1) sodium nitrite. In addition, the potentials of migration and invasion for SMMC-7721 cells were increased significantly at the same time. Furthermore, sodium nitrite exposure displayed an increase of stress fibers, lamellipodum and perinuclear mitochondrial distribution by cell staining with Actin-Tracker Green and Mito-Tracker Red, which was reversed by N-acetylcysteine (NAC, a reactive oxygen scavenger). DCFH-DA staining with fluorescent microscopy showed that the intracellular level of ROS concentration was increased by the sodium nitrite treatment. LC3 immunostaining and Western blot results showed that sodium nitrite enhanced cell autophagy flux. Under the transmission electron microscopy (TEM), more autolysosomes formed after sodium nitrite treatment and NAC could prevent autophagosome degradation. RT-PCR results indicated that the expression levels of COX I and COXIV mRNA were decreased significantly after sodium nitrite treatment. Meanwhile, laser scanning confocal microscopy showed that sodium nitrite significantly reduced mitochondrial mass detected by Mito-Tracker Green staining. The expression levels of HIF-1α, Beclin-1 and Bnip3 (mitophagy marker molecular) increased remarkably after sodium nitrite treatment, which were reversed by NAC. Our results demonstrated that sodium nitrite (16 mg x L(-1)) increased the potentials of invasion and migration of hepatic cancer SMMC-7721 cells through induction of ROS and HIF-1α mediated mitophagy.
Acetylcysteine ; pharmacology ; Autophagy ; Carcinoma, Hepatocellular ; pathology ; Cell Line, Tumor ; Cell Movement ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Liver Neoplasms ; pathology ; Mitochondrial Degradation ; Neoplasm Invasiveness ; Nitrites ; metabolism ; Reactive Oxygen Species ; metabolism ; Sodium Nitrite ; pharmacology

Honeysuckle flower is a traditional herbal medicine in China Through systemically sorting and studying literature of Chinese medicine, this article pointed out that leech used by the traditional Chinese medicine in ancient time has the features of twist vine, slight purple stem with clothing hair; opposite growing leaves, ovule shape with clothing hair on both side; two flowers growing from one pedicel, labiate corolla with 3.2 cm longth, flower grows from white color to yellow color, each branch axil grows only one pedicel, the involucre is ovoid shape, and the flower season is from mid-March to mid-May. Among all species of caprifoliaceae, only Lonicera japonica Thunb. meets these botanic features. Therefore, L. japonica Thunb. should be used as the orthodox species of herbal honeysuckle flower.
China ; Flowers ; anatomy & histology ; classification ; History, Ancient ; Lonicera ; anatomy & histology ; classification ; Medicine in Literature ; Medicine, Chinese Traditional ; history ; Plants, Medicinal ; anatomy & histology ; classification

Objective To study the clinical significance of antinuclear antibody (ANA),anti-thyroid peroxidase antibody (TPO-Ab) and antithyroglobulin antibody (Tg-Ab),thyroid stimulating hormone (TSH),free triiodothyronine (FT3),free thyroxine (FT4) in patients with recurrent spontaneous abortion (RSA).Methods 46 women with RSA diagnosis in out patient department in Peking University Shenzhen Hospital from March 2015 to December 2016 were recruited as the study group,20 women with normal childbirth history were enrolled as the control group,ANA was detected by indirect immunofluorescence (IIF) assay and ELISA,TSH,FT3 and FT4 were detected by chemiluminescence immunoassay technology.Results The positive rates of ANA(IIF),TPO-Ab and Tg-Ab in patients with RSA were 24％,24％ and 15％ respectively which were significantly higher than those in healthy control (P＜0.05).Serum levels of TSH,TPO-Ab and Tg-Ab in patients with RSA were 2.70 ± 1.38 mIU/L,38.99 ± 10.18 IU/ml and 3.07 ± 1.69 IU/ml respectively,which were significantly higher than those in healthy cases (P＜0.05).Conclusion ANA and antithyroid antibodies were closely related with RSA.It is very important to screen ANA and antithyroid antibodies for etiological diagnosis and appropriate intervention in patients with RSA to reduce the incidence of abortion.

Objective To study the effect of misoprostol used to intenerate cervix and ease pain in hys- teroscopy.Methods 380 cases were divided into two groups randomly;260 cases of them were placed misoprostol 200?g in vagina 2 hours before hysteroscopy and 120 cases treated without drugs served as control.The diameter of cerical,hemorrhage,the rate of PAAS and bellyache were observed.Results The diameter of cerical,hemorrhage. the rare of PAAS and bellyache in the group of misoprostol were lower than those in the group of antitheses(P

Objective To evaluate the distribution and age-related changes of brain iron content in healthy people using MR quantitative susceptibility mapping (QSM).Methods Sixty three healthy right-handed volunteers underwent the routine MRI and SWI scan to get SWI-unfiltered phase images and magnitude images.QSM were reconstructed from the SWI-unfiltered phase images and magnitude images using SMART software.The regions of interest at the bilateral frontal white matter and nuclei (caudate nuclews,globus pallidus,putamen,substantia nigra,dorsal thalamus,red nucleus and dental nucleus) were drawn manually,and the susceptibility was measured.The linear correlation between the susceptibility and iron concentration cited from Hallgren and Sourander's post-mortem brain study was calculated.Wilcoxon test were applied to calculate the difference between the bilateral frontal white matter and bilateral nuclei.The correlation of age with susceptibility of bilateral frontal white matter and bilateral nuclei were analyzed by Spearman correlation analysis.Results The median susceptibility (extent) of frontal white matter,caudate nucleus,globus pallidus,putamen,dorsal thalamus,substantia nigra,red nucleus and dentate nucleus of 63 healthy people were-12.81 (-31.56,8.72),39.27 (22.35,75.13),93.99 (19.19,158.75),29.16 (4.11,81.53),2.91 (-27.80,27.95),83.14 (38.57,185.79),63.49 (13.83,142.09),63.30 (36.78,128.53) ppb (× 10-9),respectively.There was high consistency with Hallgren and Sourander's study (r=0.91,P＜0.05).The susceptibility of globus pallidus was the highest,followed by substantia nigra.The least susceptibility was seen in the frontal white matter.The susceptibility of right caudate nucleus,substantia nigra,red nucleus and dental nucleus was higher than that of left side (Z value was-3.18,-4.44,-3.70 and-2.64,respectively,P＜0.05).The susceptibility of bilateral globus pallidus of the male was significantly different from that of the female (Z value was-2.27 and-2.42,respectively,P＜0.05).There was positive correlation between age and the susceptibility of bilateral caudate nucleus,putamen,red nucleus and dental nucleus (r value was 0.30 to 0.49,P＜0.05).Conclusions QSM based on the SWI-unfiltered phase and magnitude images can clearly display the cerebral nuclei and evaluate the brain iron content accurately,which is consistent with the histopathological results.Iron content of bilateral caudate nucleus,putamen,red nucleus and dental nucleus increase with aging.

Objective To evaluate the plasma D-dimer level in the patients of type 2 diabetes mellitus with hypertension and investigate their correlation.Methods Eighty-five subjects were divided into three groups according to clinical manifestation:control group:20 subjects ; type 2 diabetes mellitus group:21 subjects; type 2 diabetes mellitus combined with hypertension group:44 subjects.The level of plasma D-dimer was measured and the difference was compared between groups.The results were showed as mean ± sd,and the difference was compared using ANOVA Test ( SPSS13.0).Results The plasma D-dimer concentrations in normal control group was ( 102.15 ± 32.48 ) μg/L,in single type 2 diabetes mellitus was ( 148.62 ± 80.99 ) μg/L,while plasma concentrations in type 2 diabetes mellitus combined with hypertension was ( 206.28 ± 92.99 ) μg/L.plasma D-dimer concentration was higher in single type 2 diabetes mellitus than that in normal control cases( P ＜0.05) ;And plasma D-dimer concentration was also found increased in type 2 diabetes mellitus combined with hypertension when compared with control group (P ＜ 0.01 ) ;And there was also significant difference on plasma D-dimer concentration between single type 2 diabetes mellitus and type 2 diabetes mellitus combined with hypertension cases ( P ＜ 0.05 ).Conclusion The plasma levels of D-dimer was increased obviously in single type 2 diabetes mellitus and type 2 diabetes mellitus combined with hypertension,it may be related to the imbalance of coagulation and fibrinolytic system.Monitoring of plasma D-dimer concentration in type 2 diabetes and patients with hypertension may have important clinical implications for the prevention of thrombotic diseases.

Objective To evaluate the application and the good qualities of high-resolution ultrasound and CDFI-guided mammotome minimally invasive biopsy device in the diagnosis and treatment of breast lesions.Methods The common clinical operations and the lesions which were guided mammotome minimally invasive biopsy device by high-resolution ultrasound and CDFI were contrasted.The effects of treatment were evaluated.Results 307 le- sions of 102 patients were removed by this method,and the operational process was successful.Patients' skin lacera- tions were tiny.Only one lesion was clinically diagnosed as mild blood clot under skin,but without other complica- tions.Conclusion Contrasted with the common clinily operations.the high-resolution ultrasound and CDFI-guided mammotome minimally invasive biopsy device in the diagnosis and treatment of breast lesion is effective,and the scar is tiny.It releases patients' pain.

OBJECTIVETo explore the prevalence of IDH gene (IDH1 and IDH2) mutations, types of mutations in patients with acute myeloid leukemia (AML), correlation with the internal tandem duplication(ITD) mutation of FLT3 gene, NPM1 gene mutation and some clinical characteristics.

METHODSThe mutations of IDH1 and IDH2 gene at exon 4, NPM1 gene at exon 12 and FLT3-ITD at exon 14 and 15 in 163 newly diagnosed AML patients were detected by PCR amplification followed by direct sequencing of genomic DNA.

RESULTS(1) IDH mutations were found in 25 patients (25/163), and all were heterozygous, of which IDH1 in 7 patients (4.29%) and IDH2 in 18 (11.04%). A total of 4 types of IDH1 mutations were identified (c.395G→A, p.R132H, n = 4; c.394C→A, p.R132S, n = 1; c.394C→G, p.R132G, n = 1; c.315C→T, n = 1). The IDH1 mutation caused substitutions of residue R132 except for one (c.315C→T). All IDH2 mutations caused changes of R140 (c.419G→A, p.R140Q, n = 18). The incidence of IDH2 mutation was significantly higher than that of IDH1 mutation (11.0% v 4.3%, P = 0.022). Both IDH1 and IDH2 mutation were detected in one patient, while IDH1 was synonymous substitution (c.315C→T). IDH-mutated cases showed a significantly higher frequency of concurrent FLT3-ITD mutation compared with wildtype cases (34.6% vs 11.9%, P = 0.003), so did IDH mutations concurrent NPM1 mutation vs NPM1 wildtype (28.1% vs 12.7%, P = 0.033), of which the frequency of concurrent NPM1 and FLT-ITD mutations cases with the IDH mutation was significantly higher than that of NPM1 and FLT-ITD negative (45.5% vs 11.7%, P = 0.002). IDH mutation incidence was significantly higher in normal karyotype cases than in abnormal ones (20.5% vs 5.8%, P = 0.020). Patients with IDH mutations were significantly older than wildtype patients(P < 0.001), whereas, there were no statistically significant differences in gender, peripheral blood (PB) count at diagnosis between two groups.

CONCLUSIONSThe incidence of IDH mutation is higher in patients with de novo AMLs, of which IDH2 mutation more frequently, and the patients associated with older age, normal karyotype at diagnosis. IDH mutation has a strong association with NPM1 and FLT3-ITD mutations, suggesting that IDH mutation has synergistic effect with the latter gene on leukemogenesis.

Adolescent ; Adult ; Aged ; DNA Mutational Analysis ; Female ; Genotype ; Humans ; Isocitrate Dehydrogenase ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Young Adult

OBJECTIVETo establish a new culture method to induce the differentiation of embryonic pancreatic cells into mature endocrine cells.

METHODSMouse embryos at day 12.5 were used and embryonic pancreata were isolated. The isolated embryonic pancreata were cultured on the filter for 7 days, which floated in the dish containing medium. During culture, the expression of pancreas duodenum homeobox-1 (PDX-1), a pancreatic stem cell marker, was examined at day 1. The expression of neurogenin 3 (Ngn3), a pancreatic progenitor cell marker, was examined at day 3. The expressions of endocrine and exocrine markers, insulin, glucagon, and carboxypeptidase (CPA) were examined at day 7 by immunohistochemistry. The kinetics of pancreatic marker expression during culture was assayed by real-time PCR.

RESULTSMany pancreatic stem cells still existed in embryonic pancreata cultured for 1 day; meanwhile, these pancreatic stem cells proliferated in high rate. A large amount of pancreatic progenitor cells were found in embryonic pancreata cultured for 3 days.Pancreatic stem/progenitor cells differentiated into mature endocrine and exocrine cells in embryonic pancreata after having been cultured for 7 days. Furthermore, the expression pattern of pancreatic marker is consistent with that in vivo.

CONCLUSIONWe successfully established a new culture method, with which embryonic pancreatic cells can efficiently differentiate into mature endocrine cell.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; metabolism ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Embryo, Mammalian ; Endocrine Cells ; cytology ; Female ; Homeodomain Proteins ; metabolism ; Male ; Mice ; Nerve Tissue Proteins ; metabolism ; Pancreas ; cytology ; Trans-Activators ; metabolism

OBJECTIVETo construct a method to culture pancreatic progenitor cells in hanging drop and on floating filter,and to examine if pancreatic progenitor cells can differentiate into mature endocrine cells with this method.

METHODSMurine embryos at day 12.5 were isolated and digested into single cells,which were then cultured in hanging drop for 24h and formed spheres.Spheres were cultured on the filter for 6 days,which floated in the dish containing medium.During culture,the expressions of pancreas duodenum homeobox-1(PDX-1)and neurogenin3(Ngn3)were determined.The expressions of endocrine and exocrine markers,insulin,glucagon,and carboxypeptidase(CPA)were determined on day 7 by immunohistochemistry.Insulin secretion of spheres stimulated by glucose was detected by ELISA.The changes of pancreatic marker expressions during culture were monitored by real-time polymerase chain reaction(PCR).

RESULTSOne day after the culture,there were still a large amount of PDX-1 positive cells in pancreatic spheres,and these cells proliferated.On day 3,high expression of Ngn3 was detected,and the Ngn3-positive cells did not proliferate.On day 7,The expressions of endocrine and exocrine markers in the differentiated pancreatic progenitor cells were detected,which were consistent with that in vivo.Insulin was secreted by spheres upon the stimulation of glucose.

CONCLUSIONIn hanging drop and on floating filter,pancreatic progenitor cells can differentiate into mature endocrine cells.

Animals ; Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Endocrine Cells ; cytology ; Homeodomain Proteins ; metabolism ; Insulin ; metabolism ; Mice ; Pancreas ; cytology ; Stem Cells ; cytology ; Trans-Activators ; metabolism