OBJECTIVETo explore the effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and the expression of the PCA3 gene in human prostate cancer in vitro.

METHODSLNCaP cells were treated with cyclopamine at the concentrations of 1, 5, 10 and 15 micromol/L for 24, 48 and 72 hours. The inhibitory effects of cyclopamine on the proliferation and apoptosis of the LNCaP cells were detected by MTT and flow cytometry respectively, the morphological changes of the cells observed by Hoechst 33258 staining, and the expression of the PCA3 gene determined by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR).

RESULTSCompared with the blank control group, cyclopamine significantly inhibited the proliferation of the LNCaP cells at 5, 10 and 15 micromol/L (P <0.01), reaching IC50 at 10 micro mol/L at 48 hours. The apoptosis rates of the LNCaP cells at 24, 48 and 72 hours were 37.21%, 57.38% and 57.98% in the 10 micromol/L group and 21. 16% , 71.31% and 72.90% in the 15 micro.mol/L group, significantly different from those in the control (P <0. 01). The cell apoptosis showed a rising trend with the increase of cyclopamine concentration and acting-time, while the expression of the PCA3 gene was decreasing with the increased concentration of cyclopamine, significantly lower than that of the blank control group (P <0.01) , and extremely low in the 10 micromo/L group

CONCLUSIONCyclopamine intervention at 10 and 15 micromol/L for 48 and 72 hours could significantly inhibit the at all time points. Proliferation and induce the apoptosis of LNCaP cells and reduce the expression level of PCA3.

Antigens, Neoplasm ; genetics ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Male ; Prostatic Neoplasms ; genetics ; pathology ; Veratrum Alkaloids ; pharmacology

Arthritis, Gouty ; diagnostic imaging ; pathology ; surgery ; Foot Joints ; diagnostic imaging ; pathology ; surgery ; Hand Joints ; diagnostic imaging ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Radiography

Objective To investigate the value of partial hepatectomy and vascular resection in the treatment of hilar cholangiocarcinoma. Methods Seventy four patients with hilar cholangiocarcinoma who underwent hepatectomy of Chinese People' s Liberation Army from January 2008 through December 2011 were analyzed retrospectively.Results Of the 74 patients,33 underwent radical resection and 19 palliative resection,22 received internal or external drainage.In the radical resection group,the median survival time was 27 months,and the overall survival rate at 1,2 and 3 years were 79％,64％ and 49％.In the palliative resection group,the median survival time was 14 months and the overall survival rate at 1,2 and 3 years were 56％,25％,and 19％.In the drainage group,the median survival time was 9 months and the overall survival rate at 1,2 and 3 years were 23％,15％,0.Conclusions Hepatectomy combined with hilar vascular resection helps increase survival rate of patients in radical excision of hilar cholangiocarcinoma and Surgical resection is the most elective method for treatment of hepatic hilar cholangiocarcinoma,and the radical resection might improve the prognosis of the patients with hilar cholangiocarcinoma.

OBJECTIVETo explore the mechanism of miR-124 inhibiting the proliferative activity of prostate cancer PC3 cells.

METHODSLuciferase reporter gene assay was used to examine the specific binding ability of miR-124 to PKM2 mRNA 3'-UTR. After miR-124 was transfected mimic to PC3 cells, the expression levels of PKM2 mRNA and protein were detected by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, respectively. The effects of miR-124 mimic and PKM2 siRNA on the proliferative activity of the PC3 cells were determined by MTT assay.

RESULTSThe expressions of PKM2 mRNA and protein were upregulated (5.12 +/- 0.35) times and (4.05 +/- 0.20) times respectively in the PC3 cells as compared with those in the RWPE-1 cells (P < 0.05). Luciferase reporter gene assay demonstrated that miR-124 targeted PKM2 3'-UTR. At 24 hours after transfection with miR-124 mimic, the PKM2 protein expression in the PC3 cells was downregulated (0.16 +/- 0.04) times (P < 0.05), while the PKM2 mRNA level was not changed significantly (P > 0.05), as compared with the control group. MTT assay showed that both miRNA-124 mimic and PKM2 siRNA could inhibit the proliferation of the PC3 cells, but the former exhibited a greater inhibitory effect than the latter. After transfection with miR-124 mimic and PKM2 siRNA, the cell growth rates were (66.20 +/- 5.10)% vs (82.10 +/- 6.35)% at 24 hours (P < 0.05) and (49.34 +/- 2.37)% vs (70.10 +/- 5.80)% at 48 hours (P < 0.05).

CONCLUSIONmiR-124 can suppress the proliferation of PC3 cells by regulating the PKM2 gene.

Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; genetics ; Humans ; Male ; Membrane Proteins ; genetics ; metabolism ; MicroRNAs ; genetics ; Prostatic Neoplasms ; genetics ; metabolism ; pathology ; Thyroid Hormones ; genetics ; metabolism ; Transfection

Objective: To explore the effect of moxibustion at Shenque (CV 8) on myocardial structure and function in exercise-induced fatigue rats. Methods: A 12-week treadmill running training was performed to create an exercise-induced fatigue rat model. Sixty eligible male specific-pathogen-free grade Sprague-Dawley rats were randomly divided into a blank group, a control group, a model group, a non-meridian non-acupoint group, a Zusanli (ST 36) group and a Shenque (CV 8) group, with 10 rats in each group. Rats in the blank group did not receive treadmill running training or moxibustion. Rats in the control group did not receive treadmill running training but received mild moxibustion at Shenque (CV 8). Rats in the model group received treadmill running training but no moxibustion. Rats in the non-meridian non-acupoint group, the Zusanli (ST 36) group and the Shenque (CV 8) group received moxibustion at the non-meridian non-acupoint points, Zusanli (ST 36) or Shenque (CV 8) immediately after each treadmill running training, 15 min each time, once a day for 5 consecutive days a week at a 2-day interval, 60 times of moxibustion in total. Left ventricular end-diastolic diameter (LVEDd), left ventricular end-systolic diameter (LVESd), left ventricular diastolic volume (LVDv), left ventricular systolic volume (LVSv), ejection fraction (EF), stroke volume (SV), early diastolic peak flow velocity of mitral valve (E) and late diastolic peak flow velocity of mitral valve (A) of each group before and after the last treadmill running training were measured. Blood was collected 6 h after the last treadmill running training, and serum C-reactive protein (CRP), myoglobin (Mb), creatine kinase-myocardial band (CK-MB), cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels were detected. Finally, the heart was separated, the heart mass (HM) was measured, the cTnT level of the myocardial tissue was detected, the ultrastructural changes of the left ventricular myocardium were observed by transmission electron microscope, the left ventricular fraction shortening (LVFS), E/A and heart mass index (HMI) were calculated. Results: Compared with the same group before treatment, the rat cardiac LVEDd, LVESd, LVDv, LVSv, SV, E and A were significantly increased (all P<0.01), and the rat LVFS, E/A and EF were significantly decreased (all P<0.01) in the model group and the non-meridian non-acupoint group after treatment; the rat cardiac SV, LVDv, LVSv, E and A were all increased (all P<0.01), while E/A and EF were decreased (all P<0.01) in the Zusanli (ST 36) group after treatment; the rat cardiac LVDv, E and A were significantly increased (P<0.01 or P<0.05), and E/A was significantly decreased (P<0.01) in the Shenque (CV 8) group after treatment. After treatment, compared with the blank group, the rat cardiac LVEDd, LVESd, SV, LVDv, LVSv, E, A, the serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and the myocardial cTnT were increased (all P<0.01), and the LVFS, E/A and EF were all reduced (all P<0.01) in the model group; compared with the model group and the non-meridian non-acupoint group, rats in the Zusanli (ST 36) group and the Shenque (CV 8) group showed decreased LVEDd, LVESd, SV, LVDv, LVSv, E, A, serum CRP, Mb, CK-MB, cTnI, cTnT and HMI, and myocardial cTnT (P<0.01 or P<0.05), along with increased LVFS, E/A and EF (all P<0.01); compared with the Zusanli (ST 36) group, Mb and A of the Shenque (CV 8) group were decreased (both P<0.01), while both E/A and EF were increased (P<0.01, P<0.05). Transmission electron microscopy examination showed that myofibrils in the blank group and the control group were neatly arranged with clear light and dark bands; the model group and the non-meridian non-acupoint group showed different degrees of myofibril disintegration and breakage, increased and aggregated mitochondria of different sizes, and increased electron density. The myofibrils in the Shenque (CV 8) group and Zusanli (ST 36) group were arranged neatly with clear light and dark bands, and compensatory hyperplasia of mitochondria. Conclusion: Moxibustion at Shenque (CV 8) and Zusanli (ST 36) both can effectively improve the occurrence of myocardial remodeling in exercise-induced fatigue rats, and the effect of moxibustion at Shenque (CV 8) is better in improving cardiac function.

Objective To explore the diagnostic value of two-dimensional speckle tracking technology in the evaluation of right ventricular systolic function in patients with pulmonary disease .Methods Thirty patients with pulmonary heart disease were divided into two groups:group of compensated pulmonary heart disease(compensated group ) (n =15),group of decompensated pulmonary heart disease (decompensated group)( n =15).30 healthy subjects were enrolled in control group .The displacement of the tricuspid annulus at the midpoint(TADmid),the displacement of the tricuspid annulus at the free wall (TADfre) and the displacement of the tricuspid annulus at the septum (TADsep) were acquired,and simultaneous real-time three-dimensional ultrasound detection of right ventricular ejection fraction (RVEF) were taken.The correlation of TADmid with RVEF and pulmonary artery systolic pressure ( PASP) were analyzed.Results TADmid of the healthy control group,the compensated group and the decompensated group were(17.1 ±3.9)mm, (13.6 ±2.6)mm,and(9.5 ±3.2)mm respectively.TADfre were(21.1 ±3.0)mm,(17.6 ±4.2)mm,and (11.5 ±3.8) mm respectively.TADsep were(12.0 ±2.5) mm,(9.7 ±3.3) mm,and(7.4 ±2.7) mm respectively.RVEF were(56.3 ±8.2)%,(39.6 ±6.4)%,and(28.1 ±5.9)% respectively.PASP were (20.6 ±2.6) mm Hg (1 mm Hg =0.133 kPa), (63.3 ±5.6) mm Hg, and (82.5 ±11.2)mm Hg respectively.There were significant differences of TADmid , RVEF, and PASP among the 3 groups ( F =8.581,7.816,9.300,6.507,10.235, all P <0.05).TADfre, TADmid, TADsep and RVEF were all decreased in the compensated group comparing to the healthy control group ,while PASP was increased.The decrease of TADmid was the most significant ,while that of TADfre was the slightest .There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups(t=2.703,2.536,2.379, 2.817,3.026,all P<0.05).TADfre,TADmid,TADsep and RVEF of decompensated group reduced more significantly than the compensated group , while PASP was increased significantly .There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups(t=2.519,2.493,2.236, 2.621,2.985,all P<0.05);TADfre and TADmid were decreased more apparently in decompensated group than that in compensated group ,and so were TADsep and RVEF,while PASP were increased.There were significant differences of TADfre,TADmid,TADsep,RVEF and PASP between the 2 groups (t =1.947, 2.680,2.016,2.653,2.893,all P<0.05).There was significant positive correlation between TADmid and RVEF measured by real-time three-dimensional ultrasound (r =0.904,P <0.01 ).There was significant negative correlation between TADmid and PASP (r=-0.686,P<0.01).The cut-off point value of TADmid measured by speckle tracking technology for evaluation of RVEF <45% and<30% were 13.65 mm and 9.80 mm,respectively.The sensitivity were 94.4%and 90.0%respectively,and the specificity were 78.6%and 90.0%respectively.Conclusions TADmid is hardly affected by the external factors ,and it can better reflect the changes of right ventricular systolic function for patients with pulmonary heart disease .TADmid is positively correlated with RVEF measured by real-time three-dimensional ultrasound and negatively correlated with PASP.Further more,the correlations is significant.The three parameters can authenticate mutually ,and the combination of them can evaluate the right ventricular systolic function in patients with pulmonary heart disease precisely.

OBJECTIVETo investigate the effects of local injection of insulin on the level of systemic blood glucose and granulation tissue formation of wound in patients with diabetic foot ulcer.

METHODSThirty-two patients with diabetic foot ulcer hospitalized in our wards from June 2009 to June 2010 were divided into insulin (I, n = 16) and control (C, n = 16) groups according to the random number table. For patients in I group, after debridement, one half of calculated dose of insulin diluted with equal amount of normal saline was injected diffusely into the base of the ulcer, and another half dose of insulin was subcutaneously injected into abdominal wall for 7 days, two times a day. For patients in C group, after debridement, primary insulin was subcutaneously injected into abdominal wall, 1 mL saline was subcutaneously injected into basal layer of ulcer for 7 days, two times a day. Before injection and 0.5, 1.0, 2.0, and 4.0 hours after injection (PIH), level of fasting blood glucose was determined. Before injection and on post injection day (PID) 3, 5, and 7, the growth of granulation tissue was assessed, and wound specimens were harvested for observation of CD34 expression and calculation of microvessel density (MVD). Data were processed with t test.

RESULTSThe levels of fasting blood glucose in both groups during observational time points ranged from 6.6 mmol/L to 12.8 mmol/L with a mean of (10.0 ± 2.2) mmol/L, and there was no statistical difference (with t values from 0.000 to 2.209, P values all above 0.05). Growth of granulation tissue in I group was more exuberant from PID 5, especially on PID 7 [(59.06 ± 1.58)%], which was significantly richer than that in C group [(23.61 ± 1.57)%, t = 17.420, P = 0.000]. New vessels were observed in I group from PID 3 as indicated by CD34 expression. There was no obvious difference in the number of MVD between I group and C group on PID 3 (t = 0.247, P > 0.05). The number of MVD per 200 times visual field in I group was respectively 8.34 ± 0.48, 11.22 ± 0.97 on PID 5 and 7, which was respectively higher than that in C group (4.42 ± 0.14, 5.44 ± 1.13, with t value respectively 16.568, 27.664, P values all below 0.01).

CONCLUSIONSLocal injection of insulin has a significant effect on systemic blood glucose in patients with diabetic foot ulcer, and it can promote the growth of granulation tissue and wound healing.

Adult ; Aged ; Aged, 80 and over ; Blood Glucose ; metabolism ; Diabetic Foot ; drug therapy ; metabolism ; Female ; Humans ; Injections ; Insulin ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Wound Healing

OBJECTIVETo isolate viruses from mosquitoes in the south of Xinjiang and identify these viruses primarily.

METHODSA total of 13 491 mosquitoes were collected in the south of Xinjiang from Jul to Aug, 2005. These mosquitoes were divided into 130 groups and grinded respectively. The supernates were inoculated in C6/36 and Vero cells. Viruses isolated were detected, the genomic nucleic types by electrophoresis of viral genomes and the morphologies observed under electronmicroscope.

RESULTSAll 42 viruses were isolated, which caused CPEs on C6/36 but not on Vero cells. 27 viruses showed similar genomic profiles with 12 dsRNA segments. 1 virus displayed genomic profile with 10 dsRNA segments. 5 viruses took on similar genomic profiles with about 4 kbp DNA band. 9 viruses did not get any taxonomy information. Electromicroscopic pictures of these viruses revealed that above four types of viruses had distinguished morphologies indicating different virus species.

CONCLUSIONThere should be several virus species in the mosquitoes in the south of Xinjiang. dsRNA virus with 12 genomic segments should play analysis a predominant role in the south of Xinjiang.

Animals ; Bluetongue virus ; classification ; genetics ; isolation & purification ; Cercopithecus aethiops ; China ; Culicidae ; virology ; Dengue Virus ; classification ; genetics ; isolation & purification ; Genome, Viral ; Insect Viruses ; classification ; genetics ; isolation & purification ; RNA, Double-Stranded ; genetics ; RNA, Viral ; genetics ; Reassortant Viruses ; genetics ; Sequence Analysis, DNA ; Vero Cells

Adult ; Aged ; Female ; Humans ; Joint Instability ; complications ; diagnosis ; physiopathology ; surgery ; Lumbar Vertebrae ; pathology ; physiopathology ; Male ; Middle Aged ; Spinal Canal ; pathology ; physiopathology ; Spinal Stenosis ; complications ; diagnosis ; physiopathology ; surgery

Horseshoe kidney and retrocaval ureter are uncommon congenital anomalies of the genitourinary system that are easily diagnosed by typical imaging features. Both anomalies presenting in one patient is a rare disease characterized by isthmus of horseshoe kidney between the abdominal aorta and inferior vena cava. The clinical diagnosis and treatment of horseshoe kidney with retrocaval ureter remain a challenge. Here, we reported a case of a 44-year-old man with the two anomalies who was preoperatively diagnosed by unenhanced computed tomography scanning immediately after retrograde pyelography. The literatures on such combined anomalies are reviewed and the diagnostic evaluation and surgical management of this rare entity are discussed.
Adult ; Humans ; Kidney ; abnormalities ; surgery ; Male ; Ureter ; abnormalities ; surgery