OBJECTIVE: Over-expression of thrombin in ovarian cancer cells is associated with poor prognosis. In this study, we investigated the role of thrombin in inducing epithelial-mesenchymal transition (EMT) in SKOV3 epithelial ovarian cancer cells. METHODS: After thrombin treatment SKOV3 cells were subjected to western blots, reverse-transcription PCR, and enzyme-linked immunosorbent assay to quantify EMT-related proteins, mRNA expression of SMAD2, DKK1, and sFRP1, and the secretion of matrix metalloproteinases (MMPs) and cytokines. Meanwhile, invasion ability was evaluated using transwell assays. RESULTS: The results indicated a dose- and time-dependent down-regulation of E-cadherin and upregulation of N-cadherin and vimentin in thrombin-treated SKOV3 cells, compared with the thrombin-free control group (p<0.05). There was a dose- and time-dependent increase in the levels of SMAD2 and DKK1 mRNAs and a decrease in the levels of sFRP1 mRNA in thrombin-treated SKOV3 cells compared to control cells (p<0.05). Thrombin-treated SKOV3 cells exhibited increased secretion of MMP-9, MMP-2, interleukin (IL)-8, and IL-6 and increased invasion compared to untreated cells (p<0.05). Thrombin altered the morphology of SKOV3 cells to a spindle-like phenotype. Addition of hirudin to thrombin-treated cells reversed the effects of thrombin. CONCLUSION: Thrombin induced EMT and promoted the invasion of SKOV3 cells, possibly via distinct signaling pathways. Hirudin inhibited the effects of thrombin, suggesting that anticoagulant therapy could be a novel therapeutic strategy for ovarian carcinoma.
Blotting, Western ; Cadherins ; Cytokines ; Down-Regulation ; Enzyme-Linked Immunosorbent Assay ; Epithelial-Mesenchymal Transition ; Hirudins ; Interleukin-6 ; Interleukins ; Matrix Metalloproteinases ; Neoplasms, Glandular and Epithelial ; Ovarian Neoplasms ; Phenotype ; Polymerase Chain Reaction ; Prognosis ; Proteins ; RNA, Messenger ; Thrombin ; Up-Regulation ; Vimentin

OBJECTIVETo observe the clinical effect of a self-designed posterior STB thoracolumbar transpedicular screw-plate system in the treatment of spondylolysis and spondylolisthesis.

METHODSThis STB screw-plate system is developed with titanium alloy (TC4, Ti6Al4V). During December, 1999 and January, 2001, this system was applied in 51 cases, including lumbar spondylolysis and spondylolisthesis (36 cases), degenerative lumbar instability (15 cases). The patients were aged 22 - 78 years, mean age: 47.5 - years; among them there were 14 male cases, 37 female cases; There were 15 cases degenerative spondylolisthesis with grade I-II slip, 36 cases of spondylolysis and spondylolisthesis, including 34 cases with grade I-II slips and 2 cases with grade III slips; single level of 33 cases, two levels 3 cases. Decompression was performed for the 43 cases with interbody fusion using iliac crest or bilateral-lateral fusion using demineralized bone matrix and 8 cases were fixed with demineralized bone matrix fusion without decompression.

RESULTSAll the operations heeded about 60 - 120 minutes, and during the operations bleeding amounts were 200 - 500 ml. The cases completely recovered to work or normal action after 3 months of surgeries. 46 of all cases were achieved to satisfactorily reduction and clinical effect, 5 cases of grade II-III remained incomplete reduction (grade I slip).

CONCLUSIONThis STB system has advantages of biomechanical stability and reduction capability and its effect in clinical application is undoubtedly positive.

Adult ; Biomechanical Phenomena ; Bone Plates ; Bone Screws ; Humans ; Joint Dislocations ; surgery ; Lumbar Vertebrae ; Male ; Spinal Fusion ; instrumentation ; methods ; Spondylolisthesis ; physiopathology ; surgery ; Spondylolysis ; physiopathology ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVESTo investigate the role of glycyrrhizin on TGFbeta1 stimulated signaling transduction in rat hepatic stellate cells (HSCs).

METHODSThe mice HSCs were isolated and cultured with or without glycyrrhizin (1 micromol/L-1000 micromol/L) in vitro after TGFbeta1 stimulation. The mRNA level of Smad2, 3, 7 were measured with RT-PCR; protein expression level of Smad2, 3, 7 and collagen I, III were analyzed with Western blot.

RESULTSTGFbeta1 increased the mRNA level and protein expression of Smad2, 3, 7 in HSC; it also increased protein expression of collagen I and III. 1 micromol/L-1000 micromol/L glycyrrhizin decreased the mRNA level and protein expression of Smad2, 3, 7; it also inhibited protein expression of collagen I and III gradually.

CONCLUSIONInterventing the TGFbeta signaling pathway and decreasing the synthesis of collagen, might be involved in the anti-fibrosis mechanism of glycyrrhizin.

Animals ; Glycyrrhizic Acid ; pharmacology ; Hepatocytes ; metabolism ; Male ; Rats ; Signal Transduction ; Smad Proteins ; metabolism ; Transforming Growth Factor beta ; pharmacology

OBJECTIVETo investigate how to select an appropriate surgical approach in the management of fracture and dislocation of lower cervical spine combined spinal cord injury.

METHODSThe clinical data of 54 patients of lower cervical spine fracture and dislocation were retrospectively analyzed. There were 29 cases with vertebral body compressive fracture and dislocation, 7 cases with vertebral body bursting fracture and dislocation, 3 cases with unilateral facet dislocation, 15 cases with bilateral facet dislocation. All cases were associated with spinal cord injury. According to American Spinal Injury Association (ASIA) grades, 21 cases were in A grade, 5 cases in B grade, 22 cases in C grade and 6 cases in D grade. All patients had surgical reduction, decompression, stabilization and fusion, 43 cases in anterior approach and 11 cases in posterior approach.

RESULTSAll patients were followed up in 12 to 36 months, the mean follow-up time was 18 months. There were no great vessels, trachea, esophagus or spinal cord iatrogenic injury. There were no pull-out and breakage of screws or plates. Fusion was achieved in all patients at an average of 12 weeks postoperatively. There were no pseudarthrosis or bone nonunion. Of all the patients, 96.3% were acquired completely reduction and the normal intervertebral height and lordosis were maintained. Patients with complete spinal cord had no neurologic recovery, but they felt relief from upper limb pain or numb. Incomplete spinal cord lesions improved on average 1-2 Frankel grade after surgery.

CONCLUSIONSFor lower cervical spine fracture and dislocation, an ideal anatomy reduction can be obtained with either anterior or posterior approach surgery. It is important to select a suitable surgical approach according to different types of cervical fracture and dislocation.

Adolescent ; Adult ; Aged ; Cervical Vertebrae ; injuries ; surgery ; Diskectomy ; methods ; Female ; Humans ; Joint Dislocations ; complications ; surgery ; Laminectomy ; methods ; Male ; Middle Aged ; Retrospective Studies ; Spinal Cord Injuries ; complications ; Spinal Fractures ; complications ; surgery ; Spinal Fusion ; methods ; Treatment Outcome

OBJECTIVETo study the clinicopathologic features, immunophenotype and prognosis of primary cutaneous anaplastic large cell lymphoma (C-ALCL).

METHODSEight cases of C-ALCL were enrolled into the study. The clinicopathologic features, immunohistochemical findings and results of in-situ hybridization for EBER 1/2 were analyzed.

RESULTSThree of the 8 patients were males and 5 were females. The median age was 49.5 years. C-ALCL often presented with solitary skin nodule, without systemic symptoms. Histologically, the lymphoma cells infiltrated the dermis and subcutis in a sheet-like pattern. They were of large size and showed conspicuous nuclear atypia. Immunohistochemical study showed that more than 75% of the lymphoma cells were positive for CD30. All cases expressed one to three T cell markers (CD3, CD5 or CD45RO) and cytotoxic granule-associated antigens (TIA-1, granzyme B or perforin). The staining for leukocyte common antigen was positive in all cases, while the expression of CD5, CD8, ALK-1 and epithelial membrane antigen was noted in 5, 1, 1 and 3 cases, respectively. The staining for CD15, CD20, CK and HMB45 was negative. In-situ hybridization for EBER 1/2 was also negative in all the cases studied. Follow-up information was available in 6 patients. Five of them were still alive and 1 died of unclear cause.

CONCLUSIONSC-ALCL has distinctive clinicopathologic and immunophenotypic features. It is not Epstein-Barr virus-related and often carries a favorable prognosis.

Adult ; Aged ; CD5 Antigens ; metabolism ; Child ; Combined Modality Therapy ; Female ; Follow-Up Studies ; Humans ; Immunophenotyping ; In Situ Hybridization ; Ki-1 Antigen ; metabolism ; Leukocyte Common Antigens ; metabolism ; Lymphoma, Primary Cutaneous Anaplastic Large Cell ; immunology ; metabolism ; pathology ; therapy ; Male ; Middle Aged ; Prognosis ; RNA, Viral ; metabolism ; Skin Neoplasms ; immunology ; metabolism ; pathology ; therapy ; Young Adult

OBJECTIVETo investigate the long-term efficacy of anterior approach surgery on cervical spondylotic myelopathy and factors affecting prognosis.

METHODSThe data in 116 patients suffered from cervical spondylosis from January 1992 to December 2000 were reviewed, including 80 male cases and 36 female cases, whose age ranged from 36 to 76 years (mean, 51 years). The preoperative course of disease was 2 months to 20 years (mean, 19 months). There were 65 cases (56.0%) with single segments involved, 44 cases (37.9%) with two segments, 7 cases (6.0%) with three segments. Ninety-eight cases were onset slowly, 18 cases with no remote cause and aggravating quickly. Three kinds of surgeries were performed: anterior cervical decompression and autoiliac bone interbody fusion, anterior cervical decompression and fusion with threaded fusion cage, anterior cervical decompression and autoiliac bone interbody fusion with anterior screw-plate system. Improvement in spinal cord function was assessed using the Japanese Orthopaedic Association (JOA) scoring system, the long-term efficacy and influential factors were also analyzed.

RESULTSThe mean follow-up time was 7 years and three months (5 - 12 years). The mean preoperative JOA score was 9.34 +/- 1.81. The mean postoperative JOA score was 10.35 +/- 1.85. At the final follow-up, the JOA score was 14.09 +/- 1.90 and the recovery rate was 63.2%. Among the total patients, 27 cases were excellent, 47 cases were fine, 23 cases were good, 19 cases were poor, the fineness rate was 63.8%. The long-term efficacy of anterior approach surgery has close correlations with time of course, age of onset, preoperative spinal cord function and the number of affected segments, but has no correlations with modes of fusion and internal fixation.

CONCLUSIONSThe patients will be attentively observed while having a definite diagnosis of cervical spondylotic myelopathy. The good long-term results will be obtained after early anterior cervical decompression and fusion.

Adult ; Aged ; Cervical Vertebrae ; pathology ; surgery ; Decompression, Surgical ; methods ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Spinal Fusion ; methods ; Spinal Osteophytosis ; surgery ; Time Factors ; Treatment Outcome

OBJECTIVETo analyze the causes of perioperative complications of single-stage surgical management for spinal tuberculosis.

METHODSOne hundred and twenty patients with thoracic, lumbar and lumbosacral spinal tuberculosis were treated by single-stage surgical management from January 1997 to January 2006 in our unit, including seventy-five males and forty-five females. The mean age was 34.5 (range 17 to 68) years old. The lesion ranged from T(6) to S(1). The anterior procedures of anterior debridement, interbody fusion and anterior fixation were carried out in sixty-five cases, posterior procedures in twenty-six cases, and combined anterior and posterior procedures in twenty-nine cases, respectively. The complications that occurred during surgical procedure and 1 month after operation were recorded. Underlying causes were analysed.

RESULTSThere were 10 cases (8.3%) were recorded of mild to severe complications during perioperative period in 120 patients. The complications and underlying causes were as follows: (1) A patient died from liver failure and blood coagulation dysfunction after operation due to inappropriate surgical timing (n = 1), in which case the patient with lumbosacral spinal tuberculosis also suffered from alcoholic liver sclerosis and dysfunction. (2) False diabetes insipidus (n = 1) and deep vein thrombosis of lower limbs (n = 1) occurred as result of surgical trauma. (3) Tear of iliac vein (n = 1) occurred with lumbosacral spinal tuberculosis because of unclear anatomical relationships when anterior debridement was performed. Injury of lumbar nerve roots (n = 3) and hemothorax (n = 1) also occurred due to mispractice of surgical procedures. (4) Paralysis intestinal obstruction and hypokalemia (n = 2) occurred after anterior procedures for lumbar spinal tuberculosis as a result of other reasons.

CONCLUSIONSImproper perioperative care will lead to complications of single-stage surgical procedures for spinal tuberculosis. Emphasis should be put on preoperative evaluation, surgical planning, and postoperative caring for prevention of complications.

Adolescent ; Adult ; Aged ; Female ; Humans ; Intraoperative Complications ; etiology ; prevention & control ; Male ; Middle Aged ; Perioperative Care ; adverse effects ; methods ; Postoperative Complications ; etiology ; prevention & control ; Retrospective Studies ; Tuberculosis, Spinal ; surgery

OBJECTIVE: To compare the consistency of programmed cell death 1-ligand 1 (PD-L1, clone E1L3N, 22C3, SP263) in different immunohistochemical staining methods. METHODS: The first step was to select the optimal process: The PD-L1(clone E1L3N) antibody recommended process, self-built process ①, self-built process ② and self-built process ③ were used to perform immunohistochemical staining in 5 cases of tonsil tissue. The quality of all slides was scored by expert pathologists (0-6 points). The process with the highest score was selected. The second step was to compare the consistency between the optimal procedure and the two standard procedures. Thirty-two cases of lung non-small cell carcinoma diagnosed by pathology in Peking University First Hospital in the past two years were randomly selected. The 32 cases were stained in parallel with the SP263 and 22C3 standard procedures, and all stained slides were scored by specialized pathologists for tumor proportion score (TPS). The scoring results were grouped according to < 1%, ≥1% to < 10%, ≥10% to < 50%, and ≥50%. The consistency of PD-L1 detection antibody clone E1L3N and 22C3, E1L3N and SP263 staining results was analyzed. RESULTS: Tonsil stained slides scores (0-6 points) were as follows: The recommended protocol was 5, 5, 5, 5 and 5. The self-built process ① was 5, 6, 6, 5 and 6. The self-built process ② was 4, 4, 4, 4 and 4.The self-built process ③ was 3, 3, 3, 3 and 3. The self-built process ① was the best with the highest score. The TPSs of 32 non small cell lung carcinoma (NSCLC) cases were as follows: Of self-built process ①, 6 cases were lower than 1%, 5 cases were from 1% to 10%, 10 cases were from 10% to 50%, and 11 cases were higher than 50%; of 22C3 standard procedure, 5 cases were lower than 1%, 3 cases were from 1% to 10%, 13 cases were from 10% to 50%, 11 cases were higher than 50%; of SP263 standard procedure, 7 cases were lower than 1%, 4 cases were from 1% to 10%, 11 cases were from 10% to 50%, 10 cases were higher than 50%. The results of the consistency test were as follows: The κ value for self-built process ① and 22C3 standard procedure was 0.736 (P < 0.001), the agreement was good; the κ value for self-built process ① and SP263 standard procedure was 0.914 (P < 0.001), the agreement was very good. CONCLUSION The immunostaining using PD-L1(E1L3N) with validated self-built staining protocol ① by Ventana Benchmark GX platform can obtain high quality of slides, and the TPSs based on these slides are in good agreement with 22C3 and SP263 standard procedures.
Humans ; Carcinoma, Non-Small-Cell Lung ; Lung Neoplasms/pathology* ; Immunohistochemistry ; B7-H1 Antigen/metabolism* ; Ligands ; Antibodies ; Staining and Labeling ; Apoptosis

Animals ; Cellular Reprogramming ; Dentate Gyrus ; cytology ; Fibroblasts ; cytology ; Mice ; Neural Stem Cells ; cytology ; transplantation ; Swine