Objective To explore whether Forkhead O transcription factor-3a (FoxO3a) activity affects the capability of sphere-formation of ovarian cancer SKOV3 cell line.Methods Sphere-forming cells (SFCs) were obtained and amplified through suspended culture with conditioned medium of the stem cells in SKOV3 cell line.After SKOV3 cells were transfected with FoxO3a specific siRNA,the protein expressions of FoxO3a and Bmi1 and the ratio of sphere-formation were compared with Western blot and sphere-forming assay,respectively.Results Compared to parental cells,SFCs from SKOV3 cell line had higher ratio of sphere-formation and over-expressed Bmi1 and pFoxO3a.Transfection of FoxO3a specific siRNA down-regulated the protein expression of FoxO3a and upregulated expression of Bmi1 in SKOV3 cells,and enhanced the capability of sphere-formation.Conclusions Silence of FoxO3a leads to enhanced capability of sphere-formation in SKOV3 cell line.

Objective To investigate the relationship between heart rate variability and type 2 diabetes mellitus(T2DM)in Kirgiz and Han,to understand the differences between ethnic groups,at the same time to provide effective method for type 2 diabetic autonomic neuropathy.Methods 24 -h Holter recording was performed in 1 46 individuals.75 cases of T2DMconfirmed by glycolated hemoglobin(HbA1 c)were selected as observation group, Kirgiz 40 people,Han 35 people.While 71 subjects without T2DM were chosen as control group,Kirgiz 34 people, Han 37 people.The HRV indices,including standard deviation (SD)for the time between normal -to -normal complexes in the entire 24 h electrocardiographic recording(SDNN),the average of SD for the time between normal -to -normal intervals for each 5 min period(SDNN5),the percentage of adjacent RR intervals 50 ms apart(PNN50), normal RR intervals divided by RR intervals derived from the triangle index(TRIA),SD of the average normal -to -normal intervals for each 5 min period (SDANN)and root -mean square of differences of successive RR intervals (RMSSD)were compared between these groups.Results There were no significant differences in age,gender, education level and smoking between Kirgiz and Han patients.The differences were statistically significant in SDNN, SDNN5,PNN50,SDANN,TRIA(t =3.392,P =0.001 ;t =3.652,P =0.001 ;t =3.826,P =0.001 ;t =3.235,P =0.002;t =2.378,P =0.02)between patients with or without T2DMin Kirgiz population.In contrast,the mean square root of SDNN,SDNN5,PNN50,TRIA,RMSSD(t =3.498,P =0.001 ;t =3.41 4,P =0.001 ;t =2.1 05,P =0.039;t =2.1 02,P =0.039;t =3.41 3,P =0.001 )were statistically significant in Han population between T2DM patients and nondiabetic patients.There was no deference between Han and Kirgiz patients without T2DM in HRV,whereas the TRIA(t =2.887,P =0.005 )had significant difference in T2DM patients between Han and Kirgiz population. Conclusion The HRV is significantly reduced in the T2DMpatients,especially the Kirgiz.The HRV analysis using 24h Holter is a noninvasive technology to detect the T2DM.

Objective To study the inhibitory effect of panax notoginseng saponins (PNS) on 3-nitrotyrosine (3-NT) formation in brain induced by heme/NO2 -/H2O2 or ONOO - pathways in vitro. Methods According to the two major pathways of 3-NT formation in vivo, the models of protein nitration induced by heme/NaNO2/H2O2 or ONOO-system were established, respectively, in vitro. Bovine serum albumin (BSA)/rat plasma protein or rat brain homogenate protein were utilized as reactive substrates in both systems. Samples were divided into blank-control group, 3-NT group and PNS group (including low-, medium-and high-concentration subgroups). In 3-NT group, samples were exposed to heme/NaNO2/H2O2 or ONOO-system, respectively, at 37℃for 30 min, whereas in PNS group, samples were pre-incubated with PNS (at final concentrations of 50 mg/L, 100 mg/L, and 200 mg/L) at 37℃for 5 min before the nitrating system exposure. The 3-NT level in each group was detected by Western blot assy. Results Compared with the blank-control group, both heme/NaNO2/H2O2 and ONOO-system can induce significant 3-NT generation in BSA/rat plasma protein or rat brain homogenate protein (P<0.05). Compared with model group, PNS pre-treatment markedly inhibited 3-NT expression in BSA/rat plasma protein in a dose-dependent manner (P<0.05), the inhibitory effect of low intervention on the level of 3-NT in rat brain homogenate protein was not significant (P>0.05). Medium- and high-concentrations of PNS pre-treatment markedly inhibited 3-NT accumulation, with maximum effect at the concentration of 200 mg/L (P<0.05). Conclusion Medium- and high-concentrations of PNS can inhibit 3-NT formation in brain tissue mediated by either heme/NO2-/H2O2 or ONOO-pathways, implying that potential neuroprotective action against 3-NT involves pathological conditions, like trauma, stroke, and neurodegenerative diseases.

Objective To investigate the effect of orphan nuclear receptor NR4A1 expression on oxygenglucose deprivation (OGD)-induced apoptosis in cultured rat cerebellar granule neurons and its possible mechanisms.Methods Primary rat cerebellar granule neurons were cultured for 7 to 8 days,and then treated with OGD.The activity of cultured rat cerebellar granule neurons was assessed by methyl thiazolyl tetrazolium (MTT) assay,apoptosis was detected with flow cytometry,the expressions of NR4A1,caspase-3 and cytochrome c were determined by Western blot analysis,NR4A1 mRNA expression was detected with real-time polymerase chain reaction.The rat cerebellar granule neurons were transfected with lentiviral vector-encoding rat NR4A1.The apoptotic rates and expressions of caspase-3 and cytochrome c in rat cerebellar granule neurons transfected with NR4A1 were detected after OGD.Results The activity of rat cerebellar granule neurons decreased significantly,the apoptotic rate increased significantly,the expressions of NR4A1 mRNA and protein as well as caspase-3 and cytochrome c incrased significantly along with the OGD time.NR4A1 was overexpressed,apoptosis rate was significantly reduced in rat cerebellar granule neurons transfected with NR4A1.The expressions of caspase-3 and cytochrome c were significantly reduced in the rat cerebellar granule neurons transfected with NR4A1 after OGD.Conclusions NR4A1 overexpression may reduce OGD-induced apoptosis in rat cerebellar granule neurons by downregulating the expressions of caspase-3 and cytochrome c.

Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.

Transforming growth factor-?_1 (TGF-?_) mRNA or protein expression in renal cortex of diabetic rats was assessed by real-time quantitative RT-PCR with SYBR Green,immunohistochemistry or Western blot.After melatonin treatment,the expressions of TGF-?_1 mRNA and protein were decreased,suggesting that beneficial effect of melatonin may result from its antioxidative property and inhibiting TGF-?_1 expressions.

Objective To compare the effect of Devine surgery with different fixed positions in the treatment of concealed penis.Methods According to the different fixed positions,45 patients with concealed penis in our hospital from January 2010 to February 2015 were divided into two groups:20 patients were treated with the method that fixing the scapus penis to dermis of the penis' root (group A);the other 25 patients were treated with the method that fixing the scapus penis to pectineal fascia (group B).The surgical success rate,the penile length,and the edema time after surgery between two groups were compared.Results There was no significant difference in surgical success rate and edema time between two groups (P ＞ 0.05).But the penile length of group B was longer than that of group A after surgery (P ＜ 0.05).Conclusion In the treatment of concealed penis with Devine surgery,comparing to the method which fixing scapus penis to dermis of penis'root,the method which fixing scapus penis to fascia pectinea is much better in penile length and penile appearance.

Biopsy ; Histological Techniques ; instrumentation ; methods ; Humans ; Indicators and Reagents ; Paraffin Embedding ; Ultrasonics

ObjectiveTo explore the potential value of diffusion weighted imaging(DWI) in the clinical diagnosis of soft-tissue tumors.MethodsDiffusion-weighted MRI was performed in 48 histologically proven soft-tissue tumor cases using diffusion-weighted spin-echo sequences.Of the 48 soft-tissue tumor cases,there were 28 cases of sarcoma and 20 cases of benign tumor.The apparent diffusion coefficient(ADC) values of a large region with no necrosis,scar tissue,hemorrhage or calcification representing the lesion were measured.ADC values of benign tumors,malignant tumors and normal muscles were compared.ResultsThe ADC value of the malignant tumors (0.95 ± 0.31 )× 10-3mm2/s was significantly lower than that of benign lesions( 1.87 ±0.66) × 10-3mm2/s and muscles( 1.62 ±0.50) × 10 -3mm2/s.There was no significant difference in ADC value between benign lesions and muscles.Sensitivity(75％ ) and specificity(89％ ) were obtained when the ADC value was 1.38 × 10-3mm2/s in the differentiation of benign and malignant tumors.ConclusionDiffusion-weighted MRI had potential value in differentiating benign softtissue tumors from malignant ones.

Objective To establish the ischemic precondition ([PC) model of PC12 cell line in vitro, and to explore the effect of nitric oxide (NO) on the IPC cerebral protection. Method PC12 cells were cultured and used for producing the model of ischemie precondition by the way of oxygen-glucose deprivation. Twenty dishes of cells were randomly divided into four groups (5 dishes for each group): control group, ischemic precondition group (IPC),non-ischemic precondition group (NIPC) and L-NAME treatment group (L-NAME). In control group, the cells were in-cubated with low glucose (<1 g/L) and2% FBS medium in normal oxygen; in IPC group, the cells were administrated with oxygen-glucose deprivation (OGD) for 6 hours, and then subjected with reperfuaion before OGD 15 hours; in NIPC group, the cells were treated the same as control group for 6 hours, and then subjected with reperfusion before OGD 15 hours; in L-NAME group, the cells received L-NAME (1 mmol/L) and cocultured for 30 minutes before OGD 6 hours, and then received the same treatment as the IPC group. To test whether the model was established, metabolic rate of MIT, LDH release were measured and the apoptosis rate was detected by flow cytometry following oxygen-glucose deprivation 15 hours. The activity of nitric oxide synthases (NOS) was as-sessed by biochemical assay. One-way ANOVA and LSD multiple comparison test were used to analyze differences among different groups, and P<0.05 was considered different. Results Compared with NIPC group, the metabolic rate of MTT increased (94.9%±35.1%, P<0.05), while LDH release and the cell apoptotic rate decreased significantly in IPC group (279.1%±28.1%, P<0.03). Compared with control group(100.0%± 13.5%),the activities of NOS increased both in NIPC and IPC groups (390.0%±14.6%, P<0.01;126.3% ±10.6%, P<0.01). Moreover, the apoptosis rates in each group (control group, IPC group, NIPC group and L-NAME group) were 5.90, 8.73, 38.62 and 11.73%,respectively. Conclusions IPC reduces the death and apoptosis rate of PC12 cell after oxygen-glucose deprivation injury. NO might be involved, but it is not the only factor.