Objective To explore the clinical efficacy of lnstaTrak3500 Plus with Fluoro Trak magnetic imaging guidance for spine surgery, Methods The construction and operation progress of the system were applied in thoracic-lumbar spine surgery, Results The electric-magnetic guided imaging system can improve success rate in thoracic-lumbar spine surgery,and all operative pedicle screw were inserted accurately with no related complications error. Conclusion The electric-magnetic imaging system can improve the accuracy of operations and guide mini-invasive surgery, which can significantly decrease X-ray irradiation dosage.

BACKGROUND:Spinocerebel ar ataxia type 3 (SCA3) is a typical genetic neurodegenerative disease. To establish patient-specific disease models of genetic background contributes to studying the pathogenesis and exploring therapeutic manners.
OBJECTIVE:To observe the effectiveness of neural differentiation of induced pluripotent stem cel lines induced by SCA3 and the stability of CAG copy number.
METHODS:Skin tissue of SCA3 patient was obtained clinical y, and specific skin flbroblasts were isolated and cultured. Reprogramming fibroblasts could obtain induced pluripotent stem cel s. Patient-specific induced pluripotent stem cel s, normal person induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10) were induced to differentiate. Flow cytometry was used to compare the efficiency of differentiation. Western blot assay was utilized to detect ataxin-3 protein expression in neurons. Polymerase chain reaction was applied to measure the CAG repeat number of SCA3/ATXN3 gene.
RESULTS AND CONCLUSION:Induced pluripotent stem cel s that had identical genetic background to fibroblasts were successful y obtained, and had similar morphology and multi-directional differentiation potential to human embryonic stem cel s. Each cel line could differentiate into neural stem cel s. The CAG number did not apparently alter before and after reprogramming as wel as induction of neuronal differentiation. The effectiveness of the differentiation of induced pluripotent stem cel s derived from SCA3 into neural stem cel s was lower than that of normal person-derived induced pluripotent stem cel s (NHF) and embryonic stem cel s (ES-10). These findings demonstrate that reprogramming can successful y establish human induced pluripotent stem cel s, and induced the differentiation of above cel s into neural stem cel s. In the whole process, CAG number did not obviously alter, which was consistent with body cel s of patients.

Objective To study the effects of verbascoside on the proliferation and osteogenic differentiation of rat bone marrow stromal cells (BMSCs) and its mechanism.Methods BMSCs were obtained and identified from rat bone marrow by density gradient centrifugation.BMSCs were randomly divided into high,medium,low dose test groups and control group,and treated with 100,50,25,0 μmol · L-1verbascoside.The cell counting kit-8 (CCK-8) method was used to evaluate the BMSCs proliferation at 24,48,72 h after administration the medication.The alkaline phosphatase(ALP) activities were detected at 24,48,96 h after medication.The ALP staining was performed after 72 h of medication.Western blot and real time-polymerase chain reaction (PCR) were used to detect the osteogenic related proteins and genes expression.Results After 72 h,the proliferation of BMSCs in high dose test group was 1.22 ± 0.05,lower than that in control group,which was 1.50 ± 0.09 (P ＜ 0.05).The ALP expression levels in high,medium and low dose test groups were higher than control group at 24,48,96 h.After 72 h,ALP staining showed that the color of culture dishes in high,medium and low dose test groups were deeper than that in control group.Western blot showed that osteogenic related proteins of bone morphogenetic protein-2 (BMP2),Smad1,Smad5,Smad8,Runx2,Osterix in high,medium and low dose test groups were higher than those in control group (P ＜0.05),but there was no significant difference among those three test groups (P ＞ 0.05).Real time-PCR showed that the osteogenic related genes of BMP2,Smad1,Smad5,Smad8,Runx2,Osterix expression levels in high,medium and low dose test groups were higher than those in control group (P ＜ 0.05).However,there was no significant difference among three test groups(P ＞ 0.05).Conclusion The verbascoside at different concentrations has no obvious effect on BMSCs proliferation and promote osteogenic differentiation of BMSCs through BMP2-Smads -Runx2-Osterix pathway.

OBJECTIVETo analyze the expression of DD3 mRNA in the prostate tissues.

METHODSDD3 mRNA was detected by realtime fluorescent quantitative reverse transcription polymerase chain reaction (FQ-RT-PCR) based on the Taqman technique in the tissues of 27 patients with non-prostate cancer( NPCa), 21 prostate cancer( PCa), 39 benign prostatic hyperplasia (BPH) and 15 normal prostate (NP). The ROC curve was used to evaluate the diagnostic value of DD3 mRNA.

RESULTSDD3 mRNA expression was not detected in the NPCa tissues. The median expressions of DD3 mRNA in PCa, BPH and NP tissues were 7. 2 x 10(6), 2. 5 x 10(4) and 1.5 x 10(4) copies/mg tissue, respectively. The DD3 mRNA expression levels were significantly different between nonmalignant and malignant tissues (P < 0.01). No significant differences in DD3 mRNA expression were detected between the NP and BPH tissues and no significant correlation was found between the DD3 mRNA expression and clinical pathological parameters. The AUC-ROC was 0.937 (95% CI: 0.879 - 0.995) at cutoff value 1.4 x 10(5) copies/mg tissue. The sensitivity, specificity, accuracy, positive predictive value, negative predictive value, positive likelihood ratio and negative likelihood ratio for DD3 were 90.5%, 85.0%, 86.7%, 76.0%, 94.3%, 6.03 and 0.11 respectively.

CONCLUSIONThe DD3 mRNA expression is confined to prostate tissues and highly upregulated in PCa tissues. It has a potential application value in the early diagnosis of prostate cancer and the follow-up of the patient.

Aged ; Aged, 80 and over ; Antigens, Neoplasm ; biosynthesis ; genetics ; Humans ; Male ; Middle Aged ; Neoplasm Staging ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; RNA, Messenger ; genetics ; ROC Curve ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo identify predictive markers of the long-term outcome for neo-adjuvant chemotherapy (NC) in locally advanced breast cancer (LABC) treated with intravenous vinorelbine (V) and epirubicin (E) combination regimen.

METHODSOne hundred and nineteen patients with LABC were treated from September 2001 to May 2006. All patients were diagnosed as invasive breast cancer by 14G core needle biopsy and treated with three cycles of VE regimen before the operation. The patients were subjected to surgery and subsequently were given other three cycles of VE or cyclophosphamide+epirubicin+fluorouracil (CEF) regimen according to the clinical responses. Local-regional radiotherapy was applied to all patients after the chemotherapy and followed by hormone-therapy according to hormone receptor status. The impact of clinical, pathological, and immunohistochemical features on disease free survival (DFS) and overall survival (OS) was evaluated.

RESULTSAll patients were evaluable for responses: clinical complete response was documented in 27 patients (22.7%), 78 patients (65.5%) obtained partial clinical response. The pathological complete response was found in 22 cases (18.5%). Of the patients, 115 cases (96.6%) were followed-up for a median time of 63.4 months (range, 9-76 months), the 5-year DFS rate and OS rate was 58.7% and 71.3%, respectively. On multivariate analysis, high pre-Ki-67 (P=0.012) and post-Ki-67 expression (P=0.045), no pathological complete response after NC (P=0.034) were associated with the higher risk of disease relapse; high pre-Ki-67 (P=0.017) and post-Ki-67 expression (P=0.001), negative pre-ER (P=0.002) and no pathological complete response after NC (P=0.034) were associated with a shorter survival.

CONCLUSIONPathological response in primary tumor, pre-Ki-67 and post-Ki-67 expression, pre-ER expression are important predictors of long-term outcome for LABC patients with three cycles of VE regimen before operation.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Breast Neoplasms ; drug therapy ; pathology ; surgery ; Chemotherapy, Adjuvant ; Epirubicin ; administration & dosage ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Middle Aged ; Prognosis ; Retrospective Studies ; Treatment Outcome ; Vinblastine ; administration & dosage ; analogs & derivatives

Background: Focal segmental glomerulosclerosis (FSGS) is a kidney disease that is commonly associated with proteinuria and the progressive loss of renal function, which is characterized by podocyte injury and the depletion and collapse of glomerular capillary segments. The pathogenesis of FSGS has not been completely elucidated; however, recent advances in molecular genetics have provided increasing evidence that podocyte structural and functional disruption is central to FSGS pathogenesis. Here, we identified a patient with FSGS and aimed to characterize the pathogenic gene and verify its mechanism. Methods: Using next-generation sequencing and Sanger sequencing, we screened the causative gene that was linked to FSGS in this study. The patient's total blood RNA was extracted to validate the messenger RNA (mRNA) expression of coenzyme Q monooxygenase 6 (COQ6) and validated it by immunohistochemistry. COQ6 knockdown in podocytes was performed in vitro with small interfering RNA, and then, F-actin was determined using immunofluorescence staining. Cell apoptosis was evaluated by flow cytometry, the expression of active caspase-3 was determined by Western blot, and mitochondrial function was detected by MitoSOX. Results: Using whole-exome sequencing and Sanger sequencing, we screened a new causative gene, COQ6, NM_182480: exon1: c.G41A: p.W14X. The mRNA expression of COQ6 in the proband showed decreased. Moreover, the expression of COQ6, which was validated by immunohistochemistry, also had the same change in the proband. Finally, we focused on the COQ6 gene to clarify the mechanism of podocyte injury. Flow cytometry showed significantly increased in apoptotic podocytes, and Western blotting showed increases in active caspase-3 in si-COQ6 podocytes. Meanwhile, reactive oxygen species (ROS) levels were increased and F-actin immunofluorescence was irregularly distributed in the si-COQ6 group. Conclusions This study reported a possible mechanism for FSGS and suggested that a new mutation in COQ6, which could cause respiratory chain defect, increase the generation of ROS, destroy the podocyte cytoskeleton, and induce apoptosis. It provides basic theoretical basis for the screening of FSGS in the future.
Adolescent ; Animals ; Apoptosis ; genetics ; physiology ; Cell Line ; Female ; Flow Cytometry ; Glomerulosclerosis, Focal Segmental ; genetics ; Humans ; Immunohistochemistry ; Mice ; Mutation ; genetics ; Podocytes ; metabolism ; pathology ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; metabolism ; Ubiquinone ; analogs & derivatives ; genetics ; metabolism