Objective To investigate the effect of construct the Notch1 (NICD) eukaryotic expression vector on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. Methods Rat BMSCs were experimented as the object. NICD eukaryotic expression vector was constructed. pEGFP-N1-NICD expressing plasmids were used to transfect BMSCs. The study included control group (CON group), empty vector group (VEC group) and the trans-fection group (TRA group). After 48-hour transfection, BMSCs were observed for general morphology. The protein expres-sions of NSE, GFAP and Notch1 were detected by real-time PCR and Western blotting assay respectively. The apoptosis, cy-cle distribution and cell proliferation were evaluated by flow cytometry and MTT assay. Results The DNA sequencing con-firmed that the pEGFP-N1-NICD recombinant plasmid was successfully constructed, and both VEC group and TRA group expressed green fluorescence after 48-hour transfection. The relative expression levels of Notch1 and GFAP mRNA and pro-tein were significantly higher in TRA group than those in VEC group and CON group (P<0.05), and there was no significant difference between VEC group and CON group. After 48-hour transfection, the ratio of living cells was significantly lower in TRA group than that of CON group and VEC group, and the early apoptotic rate and late apoptotic rate were significantly higher in TRA group than those of CON group and VEC group (P<0.05). The late apoptotic rate was significantly higher in VEC group than that of CON group. The proportion of G1/G0 cells was significantly higher in TRA group than that of CON group and VEC group, but S and G2/M cells were significantly lower (P<0.05). The value of growth curve was gradually de-creased in TRA group than that of CON group and VEC group (P<0.05). Conclusion The high expression of NICD gene might induce apoptosis of BMSCs, inhibit the proliferation in part, and induce into glial-like cell differentiation.

0.05). The factor score of "preoccupation"in patients with Ins/Del genotype was higher than that with Ins/Ins and Del/Del genotypes(P=0.052),as well as in male patients of experimental group(P=0.052). CONCLUSION: The results suggest that D?H gene-4784-4803del polymorphism may not be associated with qualitative and quantitative characters of schizophrenia. Male schizophrenia patients with Ins/Del encoding D?H gene-4784-4803del are significantly characterized with the character of "preoccupation".

OBJECTIVE To explore the effects and underlying mechanism of Jieyu Anshen granule (JY) in chronic unpredictable mild stress (CUMS)-treated rats after ischemic stroke. METHODS A rat model of post-stroke depression(PSD)was developed by additional CUMS procedures after middle cere-bral artery occlusion(MCAO).Sprague-Dawley rats were given 1 g·kg-1and 3 g·kg-1of JY by gastrogavage for 4 weeks.Escitalopram(10 mg·kg-1)served as a reference drug.Behavioral tests including sucrose preference test, forced swim test and open-field test were performed to evaluate the antidepressant effects. Levels of norepinephrine (NE), dopamine (DA) and 5-hydroxytryptamine (5-HT) in rat brain were assayed. The anti-inflammatory activity was evaluated by measuring TNF-α and IL-1β in brain. Serum adrenocorticotropic hormone (ACTH) and corticosterone (CORT) were estimated as indices of hypothalamic-pituitary-adrenal (HPA) axis activity. Western blot analysis was used to evaluate hippo-campal expression of the 5-HT1A receptor (5-HT1AR) and brain-derived neurotrophic factor (BDNF). RESULTS PSD rats exhibited decreased sucrose consumption and motor activity, increased immobility time (P<0.01). JY treatment reversed the depressive behaviors in PSD rats (P<0.05, P<0.01). Treat-ment with JY resulted in significantly increased levels of NE, DA and 5-HT in the hippocampus and prefrontal cortex (P<0.05, P<0.01), and increased expression of 5- HT1AR and BDNF in the hippocampus(P<0.01). JY treatment significantly down-regulated the levels of TNF-α and IL-1β in hippocampus andprefrontal cortex (P<0.05). Treatment with JY also resulted in significantly decreased ACTH and CORTin serum which had been increased (P<0.05). CONCLUSION These findings suggest that JY treat-ment could ameliorate PSD, and the effects are likely ascribed to inhibiting HPA axis hyperfunction andinflammatory, up-regulating the levels of neurotransmitters (NE, DA and 5-HT), and the expression ofhippocampal 5-HT1AR and BDNF.

OBJECTIVETo observe and compare the effects of 17beta-estradiol (EST) on the phasic and tonic contractile activities of the uterine smooth muscles of SD rats in vitro.

METHODSDifferent concentrations of 17beta-estradiol were added into the perfusion muscular sockets containing uterine smooth muscles of SD rats, and the activities of muscle contraction were recorded at the same time.

RESULTS17beta-estradiol had obvious depression effects on spontaneous rhythmic contraction of the uterine smooth muscles in a concentration-dependent manner, it could considerably decrease muscular tension, the mean amplitudes and frequencies of contractile waves (P < 0.01); it could also suppress the uterine contraction stimulated by KCl, CaCl2 or prostaglandin F2alpha (PGF2alpha). Based on the contraction of uterine smooth muscle stimulated by KCl, IC50 was 7.278 micromol/L and pD2 was -0.862 when calculated by linear regression method. 17beta-estradiol could also inhibit the maximal CaC12 contraction of uterine smooth muscle in the Ca2+ free Krebs solution, which the ECQ was 1.422 x 10(-3) mol/L, pD2 was 2.847 (control), but the E50 was 3.028 x 10(-3) mol/L, p2 was 2.519 (added with EST) when calculated by linear regression method.

CONCLUSIONThe depression effects of 17beta-estradiol on the spontaneous rhythmic contraction and activated contraction of the uterine smooth muscles of SD rats could be mediated through the blockage of C2+ influx through potential-dependent Ca2+ channels of plasma membrane.

Animals ; Estradiol ; pharmacology ; Female ; Muscle, Smooth ; drug effects ; Myometrium ; drug effects ; Rats ; Rats, Sprague-Dawley ; Uterine Contraction ; drug effects

In this study, about 350 bp cDNA fragment was amplified by PCR. After being sequenced, the AE1-c-end gene fragment was cloned into EcoR I-Pst I site of pGADT7 to form AD ends in the yeast two-hybrid system. The recombinant plasmid was transformed into yeast AH109, and the expression in the yeast was observed. The results demonstrate that AE1-c-end was obtained. pGADT7-AE1-c-end has no toxic effect on the yeast. It can serve as a target gene of yeast two-hybrid system.
Cloning, Molecular ; DNA, Complementary ; Plasmids ; Polymerase Chain Reaction ; Two-Hybrid System Techniques ; Yeasts

OBJECTIVETo investigate the association between high mobility group box1 (HMGB1) and cervical squamous cell carcinoma (CSCC), and explore the role of HMGB1/RAGE pathway in the metastasis of CSCC.

METHODSLevels of HMGB1 mRNA and RAGE mRNA in CSCC and normal cervical tissues were detected by real time quantitative polymerase chain reaction (qRT-PCR), and the level of HMGB1 protein was determined by immunohistochemistry and Western blotting.

RESULTSThe mRNA and protein expression of HMGB1 was significantly higher in CSCC than that in normal cervical tissue (P < 0.05), correlated with stage, invasion and metastasis (P < 0.05), but not with tumor size and differentiation (P > 0.05). The levels of RAGE mRNA and HMGB1 mRNA were both significantly higher (r = 0.663, P < 0.05) in metastatic CSCC in comparison with those in the non-metastatic cases.

CONCLUSIONHMGB1 is involved in the invasion and metastasis of CSCC, and HMGB1/RAGE pathway plays an important role in the metastasis of CSCC.

Blotting, Western ; Carcinoma in Situ ; metabolism ; pathology ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Cervix Uteri ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; HMGB Proteins ; metabolism ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; RNA, Messenger ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; Uterine Cervical Neoplasms ; metabolism ; pathology

Objective To investigate the impact of genetic and environmental factors on mental health status in adolescents twins. Methods A total of 52 pairs of twins aged 11 years were recruited with support from educational committees and school. After the guardians of these twins had signed an informed consent form,the Chinese version growth and the state of health evaluation (Development and Well-Bing Assessment,DAWBA) carries on twins' mental health growth condition investigation to the twins of Age greater than 11-year-old,and focus on a-nalysis of DAWBA youth-assessment version of more than 11-year-old twins. Buccal mucosa samples were collected from all twins for DNA extraction and zygosity identification test. Results The mood, the behavior symptom and the symptom produced the influence of the DAWBA and in the computer diagnosis' s result showed that the intra-pair correlation coefficience of the emotional disorder(rMZ=0, 200, P = 0. 28; r DZ = 0. 198, P = 0. 447) (MZ means was monozygotic,DZ means was dizygotie, r means was correlation), the special phobia symptom(r MZ = 0.440, P = 0.013;rDZ =0.419, P=0.094),the social phobia (rMZ =0.2%, P = 0. 106; r DZ = -0.119, P = 0.648),the depressive symptom (rMZ =0.556, P=0.001; rDZ = -0.254, P=0.325), and the oppositional/con-duct disorder(rMZ =0.503, P = 0.014; rDZ = -0.270, P = 0.295),eating disorder(r MZ =0.764, P=0.046; rDZ = -0.091, P=0.728) in Monozygotic twins was more remarkable than that in dizygotie twin. Conclusion The genetic factors play the vital role in affects in the child twins' mental health growth.

AIM:To investigate the changes of choroidal thickness in patients with severe non-proliferative diabetic retinopathy (NPDR) after vitreous injection of ranibizumab,and to analyze the relationship between the thickness of choroid and the visual acuity of the patients.METHODS:Eighty patients with severe non-proliferative diabetic retinopathy were selected from January 1,2014 to January 1,2017.All patients were divided into observation group and control group according to the random number table,40 cases in each group.The control group was treated in a conventional manner,and the observation group was injected with ranibizumab in the vitreous.The thickness of the choroid in the macular area and the thickness of the retinal neuroepithelium in the macular area were compared between the two groups before and after treatment.The changes of the corrected visual acuity were analyzed at 1mo before and after treatment.The relationship between the thickness of the choroid and the thickness of the retinal neuroepithelium and the best corrected visual acuity were compared.The complications and adverse events were compared between the two groups after 6wk of treatment.RESULTS:The thickness of the choroid and the retinal neuroepithelium in the macular area before were 219.57± 51.24μm and 474.76 ± 95.56μm,respectively,in the observation group and the control group,217.56± 50.36μm and 473.27 ± 96.48μm,respectively.The thickness of the choroid and the thickness of the retinal neuroepithelium in the macular area after treatment were 180.15±42.06μm and 382.18±84.26,202.48±48.28μm and 407.88± 44.25μm,respectively.The difference between the two groups was statistically significant (P＜0.05).The best corrected visual acuity in the observation group and the control group were 0.47±0.19 and 0.53±0.25 respectively (P＜0.05).There was a positive correlation between the choroidal thickness and the best corrected visual acuity in the macular fovea (regression coefficient=1.12,S=0.48,OR=1.376,P＜ 0.05).There was a positive correlation between retinal neuroepithelial thickness and best corrected visual acuity in the macular area (regression coefficient =0.95,S=0.27,OR=1.020,P＜ 0.05).There were 2 eyes (5％) with subconjunctival hemorrhage,1 eyes (2.5％) of glaucoma,1 eyes (2.5％) of vitreous hemorrhage,0 of choroidal detachment,retinal detachment in 0 in observation group at 6wk after treatment.There were 4 eyes (10％) with subconjunctival hemorrhage,2 eyes (5％) of glaucoma,2 eyes of vitreous hemorrhage (5％),2 eyes of choroidal detachment (5％),retinal detachment in 2 eyes (5％) in the control group,and the difference between the two groups on the complications and adverse reactions was statistically significant (P＜0.05).CONCLUSION:The choroidal thickness of patients with macular edema in severe non-proliferative diabetic retinopathy is correlated with the best corrected visual acuity.Intravitreal injection of ranibizumab can effectively reduce the macular fovea choroidal thickness,reduce macular edema and improve vision,and less complications.

Gastric cancer (GC) is the third leading cause of cancer-related deaths worldwide. GC stem-like cells (GCSCs), with unlimited self-renewal, differentiation, and tumor-regenerating capacities, contribute significantly to the refractory features of GC and have gained increasing attention for their role in GC drug resistance, relapse, and metastasis. Therapies targeting GCSCs seem to be one of the most promising methods to improve the outcomes of GC patients. Extensive investigations have attempted to outline the regulatory mechanisms in GCSCs and to develop GCSCs-targeting therapies with which to diminish GC drug resistance, metastasis and relapse. To the best of our knowledge, there is a lack of reviews summarizing these studies. In this review, we systematically recapitulated findings regarding the regulatory mechanisms of GCSCs, as well as therapies that target GCSCs, hoping to support the development of prognostic biomarkers and GCSCs-targeting anticancer therapies in GC.
Biomarkers ; Drug Resistance ; Hope ; Humans ; Neoplasm Metastasis ; Neoplastic Stem Cells ; Recurrence ; Stem Cells* ; Stomach Neoplasms*

OBJECTIVETo explore whether normal platelet contains tissue factor (TF), and the significance of platelet-associated TF (PATF).

METHODSPlatelets were isolated by Sepharose 2B gel column. ELISA was used to detect the TF content in the lysates of washed platelets. Procoagulant activity of PATF was measured by one stage clotting time assay. The mRNA of TF was detected by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSA certain amount of TF antigen (16.37 +/- 6.39) ng/L was detected in the washed-platelet lysates. Upon activation by collagen, platelets released TF and caused a marked increase in TF level in plasma (P <0.05). Resting platelets had no TF procoagulant activity, while procoagulant activity of platelets activated by collagen increased significantly, which could be blocked by TF McAb and poor VII plasma. TF mRNA could not be detected in washed platelets. TF content in platelets from patients with coronary heart disease was significantly higher than that from normal controls (P < 0.05). Resting platelets from the patients showed a higher procoagulant activity, which could be inhibited by TF McAb.

CONCLUSIONPlatelets contain TF and the latter released by activated platelet was functionally active. Platelet itself might not synthesize TF. Protein content and procoagulant activity of PATF in patients with coronary heart disease were higher than that in controls. All these indicate that platelet may be involved in coagulation and thrombosis by releasing TF.

Adolescent ; Adult ; Blood Platelets ; chemistry ; Coronary Artery Disease ; metabolism ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Platelet Activation ; Thromboplastin ; metabolism ; physiology