Objective To investigate the influence of recombinant neuregulin-1 beta (rhNRG-1β) on neural stem cell proliferation through extracellular regulated protein kinases (ERK) signaling pathway in oxygen and glucose deprivation (OGD) environment.Methods Neural stem cells were obtained from embryonic brain of mice pregnant for 14-17 d,cultured and identified by immunochemical staining through detection of the indicator nestin using the SABC-FITC (POD)double standard kit.Neural stem cells were divided into three experiment groups (OGD group,control group and OGD + rhNRG-1β group).Control group:identified neural stem cells,2 × 107,were cultured for 3 h in the 24-hole culture plate with DMEM/F12 complete culture medium;OGD group:neural stem cells,2 × 107,were cultured in the 24-hole culture plate deprived glucose DMEM/F12 in a wet airtight container (37 ℃ constant temperature),cells were cultured with mixed gas of nitrogen (950 ml/L) and oxygen (50 ml/L) for 1 h,and then the culture medium was replaced with complete culture medium and cultured for 3 h;OGD + rhNRG-1β group:before OGD intervention,100 μg/L rhNRG-1β was given for 3 h.Neurospheres formation:the three groups of stem cells were dispersed into single cells,1 × 106/ml cells were inoculated to culture plates containing cover slips coated with poly lysine,and cultivated for 7 d,and neurospheres formation of the 3 groups of neural stem cells was observed under microscope,which was aimed to record neural stem cells proliferation changes.Colony formation:the three groups of stem cells,vaccinated in 60 mm in a petri dish,2 × 107 in number,were cultivated in complete culture medium for 24 h.The colony formation of the three groups of cells was observed under microscope,and neural stem cells proliferation changes were observed.Western blotting:the change of phosphorylation ERK (pERK) protein of the three groups of stem cells was determined,and the effect of pERK protein expression regulated by rhNRG-1β in mice neural stem cells proliferation through ERK signaling pathway was observed.Results Microscopically the primary cultured stem cells grew in single or in pairs,in a round shape;neural stem cell proliferated in clumps or colony;neural stem cells expressed the specificity of nestin protein markers with fluorescent yellow-green color.The differences in the aspects of the average diameter of neurospheres and neurospheres quantity in the neural stem cells between groups were statistically significant (F =693.66,1 002.09,all P ＜ 0.01),and among them the neuropheres formation of OGD group was significantly suppressed.Formation quantity and average diameter in OGD group [(88.78 ± 7.14) numbers,(62.12 ± 2.52) μm] were significantly lower than those in the control group [(246.34 ± 8.67) numbers,(128.45 ± 2.33) μm] and those in OGD + rhNRG-1β group [(237.87 ± 6.61) numbers,(118.37 ± 2.71) μm,all P ＜ 0.01].The difference of colony formation rate of neural stem cell was statistically significant (F =132.03,P ＜ 0.01),and among them colony formation of OGD group significantly suppressed.Formation rate in OGD group [(11.65 ± 0.94)％] was significantly lower than that in the control group [(33.23 ± 2.93)％] and that in OGD + rhNRG-1β group [(31.42 ± 2.61)％,all P ＜ 0.01].Western blotting showed that the difference of pERK protein expression of neural stem cells between groups was statistically significant (F =63.76,P ＜ 0.01).Relative expression of the pERK protein in OGD group (0.487 ± 0.072) was significantly lower than that in the control group (1.013 ± 0.112) and that in OGD + rh-NRG-1β group (1.752 ± 0.278,all P ＜ 0.01).Conclusion rhNRG-1β preserves neural stem cell proliferation with phosphorylation ERK protein expression up-regulated in oxygen and glucose deprivation environment.

[Objective]To evaluate the effectiveness of recombinant human bone morphogenetic protein-2 in the repair of rabbit rotator cuff injury.[Method]Forty-eight male New Zealand rabbits,aged 8 months,received an rotator cuff acute injury and reconstruction of the insertion of supraspinatus tendon on greater tuberosity of humerus.The rabbits were randomly divided into 3 groups postoperatively:(1) rhBMP-2 group:fibrin sealant(FS) containing rhBMP-2 was applied to the interface between the bone-tendon interface; (2)FS control group:only FS was applied;(3)blank control group:untreated after the surgery.Harvested 36 specimens underwent biomechanical analysis at the 2nd,4th,8th postoperative weeks respectively.Harvested 12 specimens underwent histological analysis at the 8th postoperative week.[Result]Histological examination showed that Sharpey's fibers were found in the interface with the formation of four-layer indirect insertion in the rhBMP-2 group at the 8th postoperative week.In the FS control group and blank group,the tendon-bone interface was filled with granulation tissue and part of Sharpey's fibers and the newly generated bone tissue.Biomechanical analysis displayed that the tensile strength and stiffness of bone-tendon interface increased time-dependently in all groups,and it in the rhBMP-2 group was significantly higher than those in the other two groups at any time-points (both P

Objective To investigate the mechanism of Secretio bufonis injection on the carcinostatic activities in mice bearing intraperitoneal H22 tumor. Methods Secretio bufonis injection (1.5 mL) was injected into the abdomind cavity of mice once 5 d for 3 times. Cyclophosphamide (0.5 mg/0.5 mL) was injected into other groups. NS (0.5 mL) was injected into control groups. Each group were 20 mice. TNF and survival time of mice were observed. Results Mean survival times of mice bearing intraperitoneal H22 tumor were (28.2?5.8)d for Secretio bufonis group,(27.9?9.7)d for Cyclophosphamide group,(20.7?4.2)d for control group. The content of TNF in mice bearing intraperitoneal H22 tumor were 2.53?0.14 for Secretio bufonis group,3.02?0.14 for Cyclophosphamide group,3.32?0.2 for control group. The survival times of mice bearing intraperitoneal H22 tumor were prolonged in the groups of Secretio bufonis (P

Objective　To investigate the epidemic situat ion of the dental fluorosis among the people in Tianjin area.Methods　With the method of random sampling we investigated epidemic situation of the dental fluorosis among the people aged 12 to 15 in Tianjin area.Results　The minimum patient's rate for this disease among those aged 12 to 15 is 32.11 percent while the maximum is 78.09 percent.The l owest dental fluorosis index is 0.851 and the highest is 1.923.The patient's rate and the index are both higher than the average rate and index of the whole cou ntry.Conclusions　Tianjin is one of the areas where the dental fluorosise epidemic mostly occurs.

Objective:To explore effect of relevant training on understanding of theory of naive biology for plant reproduction in 5- and 6-years old,and to attempt to provide demonstration for a debate of different views between domain universality and domain specificity cognitive development.Methods:48 preschoolers who did not comprehend concept of plant reproduction screened from 64 preschoolers ages 5 to 6 were surveyed by clinical method.Half of the 48 children were assigned to intervention group,the other to control group.The effects of intervention were compared by the experimental before-after design.Results:Relevant biological knowledge might widely accelerate 5- and 6-year-olds' understandings for plant reproduction,and 6-year-olds had more enhancement than 5-year-olds.Pass rates of intervention group were more 29.2% than control group in 5 year olds,and more 54.1% in 6 year olds.Conclusion:Acquiring specific knowledge will generally improve preschoolers' cognition on plant reproduction.Children' cognitive development is influenced jointly by both domain-specific and domain-general thinking.

Fraction absorbed (Fa) is an important parameter to describe the absorption level of oral drugs, and an important basis for the development and optimization of the formulation process. Because it is easily confused with the concept of absolute bioavailability, it has not received enough attention from the industry. There are many complex factors affecting Fa. There are three time-related factors that directly affect the extent of Fa: the release time, the absorption time, and the residence time. The relationship between these three time-related factors determines the extent of Fa. Generally, we are more concerned about the apparent factors that affect the extent of Fa, including independent variables and covariates; The independent variables include administered dose, route, dosage form, etc. The covariates are divided into internal and external factors, and external factors include food factors, drug interactions, etc. Internal causes include age, sex, disease, etc. This paper analyzes and systematically combs how independent variables and covariates directly or indirectly affect the three time-related factors by affecting the body's physiology and internal environment, thus changing the complex process of Fa. Understanding this theoretical framework can better optimize the independent variables to reduce the impact of covariates on Fa. In addition, this paper also introduces the latest progress of prediction and evaluation of Fa, including the progress of complex dissolution device and the status of software prediction.

Objective To outline the classification of spinal tuberculosis based on MRI findings (Southern Medical University classification,SMU classification) and explore its use in the diagnosis,surgical protocols.Methods The MRI data from 230 cases with spinal tuberculosis were analyzed retrospectively.Our classification system was based on clinical and radiological criteria (abscess formation,disc degeneration,vertebral collapse,kyphosis,sagittal index,instability and neurological problems).The surgical strategies were made according with this classification.Results Two hundred and thirty cases with spinal tuberculosis were classified into the 5 types.There were signal change type (type Ⅰ) in 28 cases,abscess formation type (type Ⅱ) in 39,vertebral collapse type (type Ⅲ) in 78,canal compression type (type Ⅳ) in 46 and kyphosis type (type Ⅴ) in 39 respectively.In type I lesion,25 patients had been followed up.Twenty patients were treated medically.Recurrence of tuberculosis was found in 2 cases.Surgical meticulous debridements were done in 5 cases without recurrence.In type Ⅱa lesion,6 patients were treated medically.The other 6 patients underwent surgical meticulous debridement with recurrence occurred in one patient.There was no difference between medical and surgical treatment regarding outcomes in patients with type Ⅰ and Ⅱa lesion.In type Ⅱb-Ⅴ,surgical treatments were carried out according to the pathological changes.There was no difference between medical and surgical treatment regarding outcomes in the patients with type Ⅱb-Ⅴ.Conclusion The SMU classification helps in differentiating the various manifestations of spinal tuberculosis and appears to correlate with the surgical treatment of spinal tuberculosis.We believe that this new classification system can be used as a practical guide in the treatment of spinal tuberculosis.

Aged ; Back ; Carcinoma, Medullary ; metabolism ; pathology ; Diagnosis, Differential ; Female ; Humans ; Melanoma ; metabolism ; pathology ; secondary ; surgery ; Melanoma-Specific Antigens ; metabolism ; S100 Proteins ; metabolism ; Skin Neoplasms ; metabolism ; pathology ; surgery ; Thyroid Neoplasms ; metabolism ; pathology ; secondary ; surgery

Objective To detect the expression of MAGE-A9 in human hepatocellular carcinoma (HCC), and to investigate the association between expression of MAGE-A9 and the clinicopathological characteristics of HCC.Methods Reverse transcription-polymerase chain reaction (RT-PCR), one-step quantitative-PCR (qPCR) and immunohistochemistry (IHC) methods were performed to characterize the expression of MAGE-A9 in HCC cell lines and tissues.Kaplan-Meier survival analysis and Cox regression were employed to evaluate the prognosis of 100 HCC patients.Results The expression of MAGE-A9 mRNA in HCC (4.44±0.342) was significantly higher than that in non-cancerous cells and tissues (1.73±0.178) (P ＜ 0.05).The expression level of the MAGE-A9 protein in HCC was related to the pathological grade (P =0.003), portal vein invasion (P =0.001), distant metastasis (P =0.022) and TNM stage (P =0.005).Cox regression analysis revealed that MAGE-A9 expression is an independent prognostic factor for disease-free survival (P =0.006) and overall survival (P =0.022).Conclusion MAGE-A9 is a valuable prognostic biomarker for HCC patients, and its high expression suggests unfavorable survival outcomes in HCC patients.

Objective To explore the methylation status of Rap1 GTPase activating protein (Rap1GAP) promoter in colon cancer, and to provide the oretical basis and research direction for the early diagnosis, targeted therapy, anti-multidrug resistance of colon cancer and so on. Methods The paraffin embedded specimens of 33 patients with colonic adenocarcinoma diagnosed by pathology were analyzed from Department of Pathology of Xinzhou City People′s Hospital from January 2010 to September 2014, including 19 males and 14 females, and aged 41-72 years old. The paraffin embedded specimens of 16 patients with colonic adenoma were enrolled, including 9 males and 7 females, and aged 34-58 years old. 13 normal tissues from the tumor distal margin (from the tumor > 15 cm) were selected. Quantitative methylation specific PCR (q-MSP) was applied to detect methylation level of Rap1GAP gene promoter. The methylation level differences of Rap1GAP gene promoter region among 3 groups or between different clinicopathologic factor subgroups were compared. Results The methylation rates [median (interquartile range)] of Rap1GAP promoter were 65.43 % (50.35 %), 21.37 % (8.39 %) and 17.43 % (15.71 %) in colonic adenocarcinoma group, colonic adenoma group and adjacent normal tissue group, respectively. The methylation rate of colonic adenocarcinoma group was significantly higher than that of colon adenoma group or that of adjacent normal tissue group (P< 0.05). The methylation rates of Rap1GAP promoter in colonic adenocarcinoma were not correlation with age, sex, differentiation and the stage of TNM [ male vs. female: 42.74 % (70.44 %) vs. 21.98%(80.00%);≤60yearsoldvs.>60yearsold:36.26%(62.62%)and26.23%(76.42 %);well-differentiated vs. moderately/poorly-differentiated: 21.98 % (40.32 %) vs. 42.74 % (74.20 %); TNM Ⅰ-Ⅱ vsⅢ-Ⅳ: 25.31 % (48.27 %) vs. 36.26 % (75.55 %); all P> 0.05]. Conclusion The methylation status of RAP1GAP promoter maybe associate with genesis and development of colon cancer, which might be used as a target for early diagnose of colon cancer.