Objective:To evaluate the correlation between the prognosis of patients with hilar cholangiocarcinoma and the degree of bile duct dilatation in MRCP .Methods:The clinical data of 89 patients with hilar cholangiocarcinoma undergoing radical operation at Tianjin Nankai Hospital from Jan 2009 to Dec 2013 were analyzed retrospectively.Results:Tumor size ( P=0.024), Bismuth-Corlette classification ( P=0.048) and tumor stage ( P=0.013) were related factors of biliary dilatation. Tumor differentiation ( P=0.002), R 0 resection ( P=0.002) and biliary dilatation ( P<0.001) were independent predictors of disease-free survival (DFS). Conclusion:The imaging evaluation of the degree of biliary dilatation has a certain predictive value for the prognosis of patients with hilar cholangio-carcinoma.

OBJECTIVE: To explore the conversion of resin monomer, the change of inorganic component and the influencing factors on the oxygen inhibition layer formed on the cured surface of resin cement. METHODS: Three kinds of resin cement were divided into three groups: (1) light-cured group: RelyX Veneer, NX3 (light-cured), Variolink N; (2) dual-cured group: RelyX U200 Automix, NX3 (dual-cured), Multilink Speed; (3) chemically-cured group, and the above 3 types of dual-cured resin cement cured without illumination could be used as chemically-cured resin cement. Each sample was provided with and without oxygen exposure of two matching surfaces, cured respectively, and the variables of light intensity and illumination time were set in the light-cured group and the dual-cured group. Scanning electron microscopy was used to observe the samples' surface morphology. Energy dispersive spectrometer was used to analyze the samples' composition of surface elements. Confocal Raman spectroscopy was used to measure the monomer conversion of resin cement and to obtain the thickness of the oxygen inhibition layer. RESULTS: (1) On the surface of cured resin cement, the weight percentage of oxygen element in the aerobic side was higher than that in the anaerobic side (P < 0.05), and the weight percentage of inorganic element was lower than that in the anaerobic side (P < 0.05). (2) The surface monomer conversion of the cured resin cement on the aerobic surface was significantly lower than that on the anaerobic surface (P < 0.05), and the surface monomer conversion on the aerobic surface and the anaerobic surface was the lowest in the chemically-cured group (P < 0.05), the dual-cured group was the highest (P < 0.05), and the light-cured group was between them. With the increase of light intensity or illumination time, the surface monomer conversion increased (P < 0.05). (3) The thickness of the oxygen inhibition layer was the thickest in the chemically-cured group [(40.27±2.81) μm](P < 0.05), the thinnest in the dual-cured group [(21.87±5.42) μm](P < 0.05) and light-cured group [(23.73±3.84) μm] was between them. With the increase of light intensity or illumination time, the thickness of the oxygen inhibition layer of resin cement decreased (P < 0.05). CONCLUSION When resin cement is exposed to oxygen, it will form an oxygen inhibition layer, its surface's inorganic filler is less, the surface monomer conversion is lower. The surface monomer conversion and the thickness of oxygen inhibition layer are affected by curing mode and illumination factors.
Materials Testing ; Oxygen ; Resin Cements

Objective To study the effect of Numb overexpression on invasion and migration in human bladder cancer cell line T24,and its related mechanism.Methods In June 2013,the plasmids including Numb-ORF plasmid and its blank vector pCMV6-Entry were transfected into T24 cells,and set the blank control as usual.The expressions of Numb and MMP-9 (Matrix metalloproteinase-9) were detected by Real-Time PCR and Western Blot.Then T24 cell invasion and migration were measured respectively by Transwell assay and Scratch Wound Healing Assay.Results Compared to negative control group,which transfected with blank-vector plasmid and untransfected group,Numb expression level in Numb-ORF group was significantly higher (P＜0.01).Meanwhile,MMP-9 level in Numb-ORF group was decreased,△ CT-value as follows:Numb-ORF group was 8.423±0.202,negative control group was 7.294±0.138,untransfected group was 7.220±0.118 (P＜0.01).In Scratch Wound Healing Assay,the repair ratio of Numb-ORF group was less than negative control group and untransfected group.The repair ratio in 12 hours as follow:Numb-ORF group was 0.525±0.037,negative control group was 0.693±0.034,untransfected group was 0.701 ±0.038(P=0.004).In Transwell assay,the number of invasion cells in Numb-ORF group was 12.6±3.2,less than in control group (130.8±9.2) and untransfected group (132.2± 10.6) (P＜0.05).Conclusions Overexpression of Numb by transfection significantly decreases the capability of T24 cell invasion and migration.This suppressed effect may be due to the down-regulation of MMP-9 expression in human bladder cancer.

OBJECTIVETo investigate the factors influencing the pregnancy outcomes of artificial insemination with donor sperm (AID), improve the pregnancy rate, and evaluate the safety of the offspring.

METHODSWe retrospectively analyzed 7,761 cycles of AID for 5,109 infertile couples performed between July 1, 2005 and June 30, 2013 in the Center of Reproductive Medicine of Shenyang No 204 Hospital, the outcomes of pregnancy, and the incidence of birth defects.

RESULTSTotally, 2 252 clinical pregnancies were achieved by AID, in which the pregnancy rate per cycle was 29. 02% and the cumulative pregnancy rate was 44. 08%. The clinical pregnancy rate was remarkably higher in the females of ≤ 35 years than in those of > 35 years old (30.31% vs 20.18%, P < 0.01), in the women with < 5-year infertility than in those with > 5-year infertility (30.83% vs 28.16%, P < 0.01), and in the patients of the ovarian stimulation group than in those of the natural cycle group (33.22% vs 28.68%, P < 0.01) The clinical pregnancy rate was the highest in the first treatment cycle (29.87%), with statistically significant difference from the fourth cycle (23.61%) (P < 0.05), but not between the other cycles (P > 0.05). There were 28 cases of birth defects in the offspring (1.40%), including 6 cases (21.43%) involving the cardiovascular system, 4 (14.29%) involving the musculoskeletal system, 3 (10.71%) involving the urogenital system, 3 (10.71%) involving the central nervous system, 2 cases (7.14%) of cleft lip and palate, 2 (7.14%) involving the respiratory system, 2 (7.14%) involving the gastrointestinal digestive system, and other anomalies.

CONCLUSIONFemale age, infertility duration, and ovarian stimulation treatment are important factors influencing the clinical pregnancy rate of AID. Artificial insemination with cryopreserved donor sperm does not increase the incidence of birth defects, which is considered as a relatively safe technique of assisted reproduction.

Adult ; Cryopreservation ; Female ; Humans ; Infertility ; Insemination, Artificial, Heterologous ; methods ; Male ; Maternal Age ; Ovulation Induction ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate ; Retrospective Studies ; Semen Preservation ; methods ; Spermatozoa ; Time Factors

Objective To investigate the effect of intermittent hypoxia (IH) with pulmonary emphysema on the ex-pression of hypoxia inducible factor-1α(HIF-1α),Bax and Bcl-2, and the mechanism underlying the role of HIF-1αin he-patocyte apoptosis thereof. Methods Sixty rats were randomly divided into four groups: normal control group, rats were treated normally;IH group, rats were treated by 30 s nitrogen and then 90 s air, and rats were treated by from 9:00-17:00 daily;pulmonary emphysema group, rats were treated by smudging for half an hour, twice a day (8:00 and 18:00);IH with pul-monary emphysema group, rats were treated by 30 s nitrogen and then 90 s air from 9:00-17:00 daily. After exposure four-teen weeks, rats were killed. qRT-PCR assay was conducted to detect the expression of HIF-1α mRNA, Bax mRNA and Bcl-2 mRNA in live tissues. Results The expressions of HIF-1αmRNA, Bax mRNA and Bax/Bcl-2 were significantly higher in IH with pulmonary emphysema group than those in control group,IH group and pulmonary emphysema group (P<0.05). The expression level of Bcl-2 mRNA was significantly lower in IH with pulmonary emphysema group than that of con-trol group and pulmonary emphysema group (P < 0.05), but no significant difference compared with that of IH group (P >0.05). The levels of HIF-1αand Bax were positively correlated with the level of Bax/Bcl-2 (r=0.732 and 0.699),but the lev-els of HIF-1αand Bax were negatively correlated with the level of Bcl-2 (r=-0.705). Conclusion The expression levels of HIF-1αmRNA, Bax mRNA and Bcl-2 mRNA were over-regulated in hepatocytes induced by intermittent hypoxia with pul-monary emphysema. The HIF-1αexpression was correlated with Bax and Bcl-2, suggesting that HIF-1αmay promote the hepatocyte apoptosis through transcriptional co-activators, Bax and Bcl-2.

Objective To investigate the effect of Notch1 gene dowuregulated on migration and proliferation of human prostate cancer cell line PC-3.Methods The small interfering RNA (siRNA) targeted Notch1 gene or negative control sequences was transfected into PC-3 cells.The expression of Notch1 or Hes1 gene was detected by Real-Time PCR and Western Blot.Then ability of migration or proliferation was measured by Transwell assay or MTS Assay.Results Compared with negative control group (36.097±1.941) and untransfected group (38.762±1.897),Notch1 expression level in siRNA group (3.960±0.510) was significantly reduced (P ＜ 0.01).Meanwhile,Hes1 level in siRNA group was decreased,expression in three groups as follows:siRNA group was 1.690±0.994,negative control group was 8.776±0.916,untransfected group was 9.803±1.001 (P ＜ 0.01).In Transwell assay,the number of migration cells in siRNA group was 657.867±27.610,more than that in the negative control group (158.533±18.263) and untransfected group (146.933±15.733) (P ＜ 0.01).In MTS assay,there was no significant difference among three groups at 0 h point,however,siRNA group was significantly raised at the time points of 24,48 and 72 h (P ＜ 0.01).Conclusions Downregulation of Notch1 gene by transfection of the siRNA-Notch1 sequences significantly promoted ability of migration or proliferation in PC-3 cells,and the effect may be due to the down-regulation of Hes1 expression.

Objective To observe the effect of exposure of emphysema with intermittent hypoxia on oxidative stress injury of myocardial cells in rats. Methods Sixty male Wistar rats were divided randomly into four experimental groups(each n=15). The normal control group was bred normally. The emphysema group was exposed to cigarette smoke twice a day(once 30 minutes). The intermittent hypoxia(IH)group was exposed to intermittent hypoxia circumstance 8 hours/day,and the emphysema with IH group was exposed to cigarette smoke twice a day (once 30 minutes)and intermittent hypoxia circumstance 8 hours/day. Each group was exposed for 8 weeks. At the beginning of 9 weeks,the blood gas analysis was performed in 5 rats selected randomly from each group,and the rest rats were sacrificed and their hearts and lungs were taken. Under light microscope,the lung tissues stained with hematoxylin-eosin(HE)were examined. The lung pathology and the results of blood gas analysis showed that the emphysema with IH rat model was established successfully. The levels of malonaldehyde(MDA)and superoxide dismutase(SOD)in rat myocardium were measured by enzyme-linked immunosorbent assay(ELISA),and the subunit p22phox mRNA expressions of nicotinamide adenine dinucleotide phosphate(NADPH)-oxidase were detected by real-time reverse transcription-polymerase chain reaction(RT-PCR). Results Compared with the normal group, the MDA levels and p22phox mRNA expressions were increased obviously in emphysema group, IH group and emphysema with IH group〔MDA(μmol/g):2.93±0.54, 3.58±0.63, 4.51±0.72 vs. 1.75±0.56, p22phox mRNA:0.043±0.004,0.067±0.015,0.123±0.016 vs. 0.018±0.002,all P<0.05〕,but the activities of SOD were decreased significantly(U/mg:36.07±4.79,33.51±7.12,24.29±5.36 vs. 46.08±5.12,all P<0.05). In emphysema with IH group,the increase of MDA levels and p22phox mRNA expressions and decrease of SOD levels were more remarkable compared with those in emphysema group and IH group(all P<0.05). The expression of p22phox mRNA was positively correlated with MDA level(r=0.734,P<0.001). Conclusion The myocardial tissue oxidative stress injury in rats induced by emphysema with intermittent hypoxia exposure is more serious than that induced by exposure of either emphysema or intermittent hypoxia alone,NADPH oxidase possibly being the important medium of myocardial cell response to oxidative stress.

Objective To assess the efficacy of one-stage posterior median incision via costotransverse joint in treating thoracic spinal tuberculosis.Methods Thirty three patients with tuberculosis of thoracic spine undergoing one-stage posterior thoracic spine debridement,bone grafting fusion and posterior instrumentation from July 2011 to October 2013 were included in the study.There were 18 males and 15 females.The age was from 17 to 39 years old with an average of 29.5.The course was from 5 to 11 months with an average of 7.3.The Cobb angle was from 19° to 42° with an average of 29.8°.There were 5 in upper thoracic spine,17 in middle thoracic spine and 11 in lower thoracic spine.6 were in Frankel scale grade C,11 were in grade D and 16 were in grade E before surgery.Postoperative kyphosis correction,recovery of neurological function and bone fusion were observed.Results All the surgeries were completed successfully.The operation time was from 120 to 230 min with an average of 183.The blood loss during operation was from 480 to 700 ml with an average of 530.All 33 patients were followed-up for 12-36 months with an average of 22.7 months.The Cobb angle was from 8° to 15° with an average of 10.2°,the correction rate was 77％.The Frankel scale of 11 patients recovered from D to E,2 recovered from C to D and 4 recovered from C to E.The postoperative kyphosis correction and Frankel scale were significantly improved,all patients had a 100％ bone fusion rate and there were no internal fixation loosened or shift of graft bone at the last follow-up.ESR and CRP were returned to the normal range and the tuberculosis symptoms disappeared.Conclusions One-stage posterior median incision via costotransverse joint can complete surgery by the same position and the same incision in treating thoracic spinal tuberculosis with safety and good clinical efficacy.

Objective To study the effect of Numb gene on cell cycle and proliferation in human renal carcinoma cells and its related mechanism. Methods Renal carcinoma cells Caki-1 were taken as research objects, and the Numb-ORF plasmid transfected cells, negative control group and blank control group were set respectively. The expression levels of Numb and cyclin D1 were detected by real-time PCR and Western blot. The cell cycle was analyzed respectively by flow cytometry. Cell proliferation was assessed by comparing with absorbance at 490 nm using a micro-plate reader. Results Compared to the negative control group (5.05±0.45) and blank control group (5.13±0.31), the △Ct value of Numb in the Numb-ORF group (1.92±0.39) was significantly lower. The protein levels in the Numb-ORF group, the negative control group and the blank control group were 6.67±0.83, 3.08±0.47, 2.85±0.36, respectively (P = 0.00). Meanwhile, the △Ct value of cyclin D1 in the Numb-ORF group (6.20±0.87) was higher than that in the negative control group (4.35±0.51) and the blank control group (4.56±0.31) (P = 0.02), and the protein level in the Numb-ORF group (5.85±0.72) was lower than that in the negative control group (10.04±0.83) and the blank control group (11.88±1.26) (P = 0.00). The ratio of G0/G1 cells in the Numb-ORF group was (54.29±4.15) %, the negative control group was (38.69±2.60) % and the blank control group was (41.28±1.29) % (P = 0.00). In proliferation assay, compared with the negative control group (0.93±0.10) and the blank control group (1.02±0.06), the A value at 24 h in the Numb-ORF group (0.67±0.07) was significantly reduced (P = 0.00). Conclusion Numb gene could increase the cell percentage in G0/G1 phase and inhibit proliferation of renal carcinoma cells via down-regulating the expression of cyclin D1.

Aim The effects of midazolam on the whole-cell sodium currents in rat sympathetic neurons were studied to explore the mechanisms where by midazolam mediates hypotension. Methods Whole-cell patch-clamp recordings were performed on enzymatically isolated rat superior cervical sympathetic neurons. Results Midazolam dose-dependently blocked the whole-cell sodium currents evoked by a voltage step to 0 mV from a holding potential of -80 mV with a mean drug concentration required to produce 50% current inhibition (IC50) values of 18.35 ?mol?L-1; Clinically relevant concentration of midazolam(0.3 ?mol?L-1) reduced sodium peak currents by 19.98%(P