Objective The objectives of this study were to investigate the relationship between DKK1 protein expression and lymph node invasion,and prognosis in patients with intrahepatic cholangiocarcinoma.Methods From January 2005 to December 2012,78 patients with intrahepatic cholangiocarcinoma were treated with endoscopic resection.Tissue microarray of intrahepatic cholangiocarcinoma and adjacent tissues were done and biochemical indexes were measured before operation.The patients were recruited every 3 months after operation.The Kaplan-Meier method was used to analyze the recurrence-free survival curve and the overall survival curve.The Cox proportional hazards regression model was used to analyze the multivariate analysis and to determine the factors that affect the disease-free survival(RFS)and overall survival(OS).Results There were significant differences in the expression of GGT,hepatic portal lymph node invasion,Child-Pugh grade,MMP9 and DKK1 protein in the negative and positive expression of intrahepatic cholangiocarcinoma(P ＜ 0.05).The positive expression rate of DKK1 protein in intrahepatic cholangiocarcinoma was 35.90％ (28/78),and the positive expression rate of DKK1 protein in hilar cholangiocarcinoma was 14.10％ (11/78).RFS and OS in 78 cases of intrahepatic cholangiocarcinoma were 51.28％ (40/78) and 50.00％ (38/78),41.03 ％ (32/78) and 38.46％ (30/ 78),25.64％ (20/78) and 23.08％ (18/78) after surgery for 1,3 and 5 years,respectively.The results from univariate and multivariate analysis showed that GGT,CA19-9,CEA,tumor size,DKK1 and hilar lymph node invasion were the prognostic factors of OS in patients with intrahepatic cholangiocarcinoma.CEA,tumor size,DKK1 and hilar lymph node involvement were prognostic factors for RFS in patients with intrahepatic cholangiocarcinoma.DKK1 protein expression positive and negative OS curve or RFS curve showed that 5 years after intrahepatic cholangiocarcinoma patients DKK1 protein expression positive and negative OS were 28.20％ and 20.51％;RFS were 24.36％ and 21.79％.Conclusion The expression of DKK1 protein is closely related to lymph node invasion in patients with intrahepatic cholangiocarcinoma.The expression of DKK1 protein is the prognostic factor of OS and RFS in patients with intrahepatic cholangiocarcinoma.

Confronted with similar problems and difficulties in mainland,Taiwan reformed its medical education. The reform included setting up flexible program,formulating objective and detailed rotary guide,making the best of the resources,implementing medical humanities education based on the reality and forming unique cultivation system and mode. These measures can be taken as references for medical education reform in mainland.

BACKGROUND:Pedicle screw is the major instrumentation of surgery in thoracic spine. However, there have been few reports about pedicle morphology relevant to screw insertion tracts, and few reports comparing the normal adolescents and adolescent idiopathic scoliosis patients. OBJECTIVE:To compare the morphologic characteristics of the thoracic pedicle with regard to safe thoracic pedicle screw placement in normal adolescents and adolescent idiopathic scoliosis patients. METHODS: Thoracic pedicles of thirty-five normal adolescents and thirty-five adolescent idiopathic scoliosis patients were measured with three-dimensional reconstruction CT images. Measured parameters include (1) critical distance: the shortest distance from an entry point to the ventral cortex of the lamina. (2) Safe distance: the distance from the entry point to the tangent of the spinal canal at the medial wal of the pedicle. (3) Pedicle screw length. (4) Pedicle width. (5) Pedicle transverse angle. The dangerous area was defined as the distance between the critical distance and the safe distance. RESULTS AND CONCLUSION: The mean critical distance was (9.2±1.0) mm for the normal adolescents, and (9.4±1.2) mm for the adolescent idiopathic scoliosis patients. Safe distances were significantly less in normal adolescents (14.7±0.8) mm than that of the adolescent idiopathic scoliosis group (15.4±1.4) mm (P < 0.001). The dangerous area was (5.4±0.7) mm for the normal adolescents, which was significantly less than that of the adolescent idiopathic scoliosis patients (6.0±1.0) mm (P < 0.001). Pedicle screw length was (36.6±4.1) mm for the normal adolescents and (37.1±5.3) mm for the adolescent idiopathic scoliosis patients. Pedicle width was (5.8±1.2) mm for the normal adolescents and (5.7±1.7) mm for the adolescent idiopathic scoliosis patients. No significant difference in critical distance, pedicle screw length and pedicle width was found between the two groups (P=0.382, 0.135, 0.293). Pedicle transverse angle decreased gradualy from T1 to T12 in both groups. These results verify that pedicle morphology of many parameters is different between normal adolescents and adolescent idiopathic scoliosis patients, especialy in the apical area of the thoracic curve.

OBJECTIVETo apply needle-knife to treat lumber disc herniation (LDH) and surface electromyography were used to analyze biomechanical characteristic of patient's lumber muscle to make a comprehensive evaluation on its efficacy.

METHODSThirty patients who met the inclusive criteria were selected and treated with needle-knife, once a week for 2 weeks. Visual analogue scale (VAS), ASLR and JOA score before and after treatment were observed. Surface electromyography was applied to test the surface electromyography signals. AEMG, MFs and MPF were calculated before and after the treatment.

RESULTSAfter treatment, VAS was significantly reduced, ASLR, JOA, AEMG and MPF were obviously increased, and the absolute value of MFs was lowed (all P < 0.01).

CONCLUSIONThe needle-knife could significantly relieve lumbar muscle strength, muscle tone and muscle fatigue, improve in the imbalance of lumbar extensor muscle group, leading to the recovery of biomechanical characteristic, and the clinical efficacy is superior.

Acupuncture Therapy ; Adult ; Electromyography ; Female ; Humans ; Intervertebral Disc Displacement ; diagnosis ; physiopathology ; therapy ; Lumbar Vertebrae ; physiopathology ; Male ; Middle Aged

Objective To investigate the correlation between virtual touch tissue quantification (VTQ) and the pathological grading of liver fibrosis in chronic hepatitis B.Methods 64 chronic hepatitis B patients (the chronic hepatitis group) and 40 healthy volunteers (the controlled group) were collected.The patients in the chronic hepatitis group were underwent liver biopsy.According to the hepatic fibrosis degree,the patients in the test group were classified into stage 0,1,2,3 and 4.The liver shear wave velocities (SWV) of all the participant were measured by VTQ.The cut-off values were determined by an analysis of receiver operating characteristic (ROC) curve.Results The mean SWV was (1.04± 0.13)m/s in the controlled group.The SWV in stages 0,1,2,3,and 4 were (1.17 ± 0.08)m/s,(1.33 ± 0.32)m/s,(1.53 ±0.32) m/s,(2.09 ± 0.54) m/s,(2.18 ± 0.70) m/s,respectively.There was a significantly difference in SWV between the controlled group and the chronic hepatitis group (F =34.97,P =0.00).The SWV were significantly different not only between stages 0-2,and 3,but also between 0-2 and 4 (F =8.87,P =0.00).A positive correlation was observed between the liver fibrosis and the SWV in the chronic hepatitis group (r =0.67,P =0.00).When a cut off value was set at 1.43 m/s,area under ROC curve was 0.875.The sensitivity and specificity were 100 ％ and 62.5 ％.Conclusions SWV has a better correlation with liver fibrosis.VTQ can make an accurate assessment for stage 3 and stage 4 of the chronic hepatitis B.Therefore,VTQ can be used as a noninvasive and reliable diagnostic indicator for chronic hepatitis B.

Nerve growth factor is one of the most important factors playing an important role in regulating the growth, development and survival of the neuron. The purified NGF from human placenta has been reported, the tissue from which can be isolated the NGF is very limited. It is important for basic research and clinic application to expression hNGF by genetic engineering. By polymerase chain reaction,gene fragment encoding the mature part of ?-NGF was amplified using the DNA of human placenta as template. The fragment was sequenced and inserted into expression vector pET-15b, and the recombinant expression vector pET15b-NGF was transformed into E.coli BL21(DE3)pLysS. After inducing with IPTG the NGF was higher expressed up to 25% of the total cell proteins. The expression product was purified with metal chelate affinity chromatography on Ni-IDA agarose under denaturing condition. The purity of rNGF was higher than 90% and yield of rNGF was 4.56mg/L expressing bacteria. SDS-PAGE revealed the NGF expression product had a Mr 16kDa. Western-blot displayed the recombinant product had strong immunological activity with rabbit anti-human ?-NGF polyclonic antibodies. The expression products were dealed with solubilizing inclusion bodies and refolding protein. The test of nerve fiber growth of chicken embryo DRG neurons displayed rNGF had biological activity.

To investigate the effects of gossypol acetate on apoptosis in primary cultured cells from patients with acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL) and its synergistic effect with dexamethasone. The apoptosis-inducing effect of gossypol acetate on primary cultured leukemia cells was analyzed by flow cytometry (FCM). The effect of gossypol acetate on survival rates of Raji cells and mononuclear cells (MNC) from normal bone marrow were evaluated by MTT assay. After co-treatment with gossypol acetate and dexamethasone, the apoptosis rate of Raji cells was detected by FCM. The results showed that gossypol acetate was able to induce apoptosis in primary cultured ALL cells at concentrations of ≥ 5 µmol/L. The effect was concentration and time dependent. Apoptosis-inducing concentration in CLL cells was higher than that in ALL cells. After exposing to 50 µmol/L gossypol acetate for 48 h, the apoptosis rate of ALL and CLL cells were (90.4 ± 6.2)% and (51.7 ± 10.3)% separately. No major growth inhibitory effect was observed in MNC from normal bone marrow when they were exposed to gossypol acetate at concentrations lower than 10 µmol/L. After exposing for 48 and 72 h, the IC(50) of gossypol acetate for MNC from normal bone marrow was 7.1 and 9.1 times as much as the IC(50) of Raji cells. Co-treatment with 10 µmol/L gossypol acetate and dexamethasone remarkably increased the apoptosis rate of Raji cells. It is concluded that the gossypol acetate has apoptosis-inducing activity in primary cultured leukemia cells from patients diagnosed as ALL and CLL in vitro. The inhibitory effect of gossypol acetate on MNC from normal bone marrow is less prominent than that on Raji cells. Co-treatment with gossypol acetate and dexamethasone notably amplified the pro-apoptosis activity of the latter in Raji cells.
Apoptosis ; drug effects ; Cell Line ; Dexamethasone ; pharmacology ; Gossypol ; analogs & derivatives ; pharmacology ; Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; pathology ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; pathology ; Tumor Cells, Cultured

OBJECTIVETo investigate the effects of gossypol acetate on proliferation and apoptosis in Raji lymphoblastoid cells and explore the possible mechanism.

METHODSTrypan blue staining and ethyl thiazolyl diphenyl-tetrazolium bromide (MTT) assay were performed to measure the effect of gossypol acetate on the growth of Raji cells. The morphologic changes were observed with Wright's staining assay. Apoptosis was identified by agarose-gel electrophoresis and annexin V-FITC marked flow cytometry (FCM) analysis. The distribution of cell cycle, apoptosis rate, and Bcl-2 protein expression were analyzed by FCM. Caspase-3 activity was detected by colorimetric assay.

RESULTSGossypol acetate inhibited proliferation and induced apoptosis of Raji cells at concentration higher than 5 micromol/L. The effects were both dose- and time- dependent. Cycle analysis indicated the alteration of cell cycle and G0/G1 arrest. The activation of Caspase-3 was observed by colorimetric assay. The results of flow cytometry showed that the down-regulation of Bcl-2 protein expression and the activation of Caspase-3 seemed to occur simultaneously.

CONCLUSIONGossypol acetate can inhibit the growth of Raji cells and induce their apoptosis. The mechanism may be related to the alteration of cell cycle and the down-regulation of Bcl-2 protein expression.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Gossypol ; analogs & derivatives ; pharmacology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism

OBJECTIVE: To determine the therapeutic effect and safety of transcatheter closure of ventricular septal defects (VSD) in 50 patients. METHODS: Fifty patients were diagnosed by transthoracic echocardiography. To perform the operation, transthoracic echocardiography and X ray were used continuously to monitor the procedure. Transthoracic echocardiography and ECG were performed at 1, 3, and 6 months after the operation to evaluate the therapeutic effect. RESULTS: The VSD diameter ranged from 1.8 to 13.4 (5.54 +/- 2. 75) mm. The successful rate of the operation was 96.0%, and the complication rate of the operation was 16.7%. A 3 month follow-up was completed in 20 patients, and the median left ventricle end-diastolic dimension significantly decreased from (40.20 +/- 8.80) mm to (32.90 +/- 8.36) mm (P < 0.001). CONCLUSION Transcatheter closure of ventricular septal defects is a good method with a high success rate of placement, fewer complications, and a good occlusion effect.
Adolescent ; Adult ; Balloon Occlusion ; adverse effects ; instrumentation ; methods ; Cardiac Catheterization ; methods ; Child ; Child, Preschool ; Echocardiography ; Female ; Heart Septal Defects, Ventricular ; diagnostic imaging ; therapy ; Humans ; Male ; Prostheses and Implants ; Treatment Outcome

OBJECTIVETo evaluate the interface characteristics of the new-designed locking plate (LP) and limited contact-dynamic compression plate (LC-DCP) and compare the fracture healing between LP and LC-DCP in a goat tibia fracture model.

METHODSEight-hole LP and LC-DCP were applied to fix fresh goat tibiae in a reproducible manner. The average pressure, force and interface contact area were calculated using Fuji prescale pressure sensitive film interposed among the plate and the bone and image analysis system. Eight-hole LP and LC-DCP were applied to each tibia in a goat tibia fracture model. The fracture healing was evaluated by X-ray photography at postoperative 8 weeks. The goats were sacrificed at postoperative 12 weeks. Three-point bending test was conducted in the tibiae.

RESULTSThe interface contact of LP system was smaller than that of LC-DCP (P < 0.05), while interface contact force of LP system was higher than that of LC-DCP (P < 0.05). Radiographs revealed that the fracture line disappeared in the LP group, while the fracture line was visible in DCP group at postoperative 8 weeks. At postoperative 12 weeks, the bending strength and bending load of fractured tibia were higher in LP group than in DCP group, respectively.

CONCLUSIONThe new-designed locking plate can significantly decrease the contact area on the bone interface, which further provides better fracture healing than conventional plates.

Animals ; Biomechanical Phenomena ; Bone Plates ; Fracture Fixation, Internal ; Fracture Healing ; Goats ; Internal Fixators ; Tibia ; physiopathology ; Tibial Fractures ; physiopathology ; surgery