OBJECTIVETo investigate the reproducibility of Gleason scores for prostate cancer.

METHODSBased on the revised Gleason Scoring System of the International Society of Urological Pathology ( ISUP) , we analyzed the reproducibility and difference of Gleason scores in 49 cases of prostate cancer using the methods of combination and grouping.

RESULTSThe total reproducibility of Gleason scores among 15 pathologists was good (κ = 0.642), 62.2% by the combination method, the highest in Gleason 5 + 5 (81.2%) and 5 +4 (73.3%), then in Gleason 4 + 4 (67.5%), 3 + 3 (64.0%), 4 +3 (61.3%), and 3 + 4 (44.0%), and the lowest in Gleason 4 + 5 (38.9%) and 3 + 5 (33.3%). The total reproducibility of Gleason scores by the grouping method was 71.4%, the highest in Gleason 9-10 (84.9%) , then in Gleason 7 (76.7%) and 6 (64.0%), and the lowest in Gleason 8 (60.7%).

CONCLUSIONThe reproducibility of Gleason scores remains to be further improved in prostate cancer, mainly concerning the understanding of Gleason 3 and 4 carcinoma.

Carcinoma ; diagnosis ; Humans ; Male ; Neoplasm Grading ; Prostatic Neoplasms ; diagnosis ; Reproducibility of Results

Aim To investigate JAKs/STATs signal transduction change in HL-60 cells differentiation induced by diallyl disulfide(DADS)and molecular mechanism regulating the differentiation.Methods After incubation of HL-60 cells with DADS or AG490(50 ?mol?L -1),the cell differentiation indexes were observed by cytomorphology, NBT reduction ability assay,cell myeloid differentiation antigen CD11b by flow cytometry. Kinase activity of JAKs/STATs was tested by western-blotting and expressions of nucleus transcription genes stats,c-myc,c-fos,c-jun were detected by immumocyte chemistry method.Results Cell differentiation index changes indicated that HL-60 cells were induced differentiation toward granulocytic lineage by DADS, Western blot test demonstrated that constitive phosphorylation of Jak1,stat3 kinase was suppressed. Stat3,c-myc gene expression decreased and c-fos, c-jun gene expression increased in HL-60 cells treated with DADS through immunocyte chemistry.Conclusion Inhibition of phosphative Jak1, Stat3 was involved in HL-60 cells differentiation induced by DADS, its molecular mechanism might be related to modulation of gene expression associated proliferation and differentiation,and inhibition of DNA systhesis, induction differentiation.

Aim To investigate the molecular mechanisms of differentiation in human leukemia HL-60 cells induced by diallyl disulfide(DADS)using suppression subtractive hybridization(SSH).Methods In our privious study,the subtractive cDNA library was constructed successfully and efficiently. 30 clones were randomly analyed with restriction enzyme.The inserts of cDNAs were analyzed by restrictive enzyme EcoR I.Positive clones were sequenced and the homology of resulting cDNA sequences were analyzed through bioinformatics software Blastn.Results 18 clones contained 100～600 bp cDNA inserts.10 differantiation genes were obtained and involved in cell cycle,signal transduction,metabolism and RNA binding.And 3 of 10 genes,p21,STAT1 and CAMTA1 were up-regulated and detected by RT-PCR,the results matched with SSH.Conclusion sThere are tight correlation between the differentiation induced by DADS and three-upregulated gene:p21,STAT1 and CAMTA1.

Objective To investigate the expression of paired box2(Pax2),proliferation cell nuclear antigen(PCNA) and cell apoptosis in steroid-sensitive and steroid-resistant groups with primary nephrotic syndrome(PNS) and to find out the action of Pax2 expression in PNS with steroid-resistance.Method The expressions of Pax2,PCNA were evaluated by immunohistochemistry and cell apoptosis by fluorescence micoscope.Results Pax2 expression in renal tubule had a positive correlation with PCNA expression in steroid-sensitive group.In steroid-resistant group,Pax2 expression had no correlation with PCNA.Pax2 had a negative correlation with cell apoptosis.Conclusions Pax2 proper expression facilitate PCNA expression and repair tubulointerstitial lesions in steroid-sensitive group.Renal tubular epithelial cell proliferation coordinated with cell apoptosis.Pax2 overexpression in steroid-resistant group lead to the decrease of cell proliferation and cell apoptosis and lead to the severe tubule lesions,which made to glucocorticoid resistance.

OBJECTIVETo investigate the clinical effects of 360 degree circular decompression and transpedicle screw fixation to treat the ossification of thoracic posterior longitudinal ligament by posterior approach.

METHODSFrom December 2009 to November 2013, 18 patients with ossification of thoracic posterior longitudinal ligament ossification were treated with 360 degree circle decompression and transpedicle screw fixation by posterior approach. There were 8 males and 10 females,ranging in age from 32 to 67 years old, with an average of 51 years old. Four patients were accompanied with ligamentum flavum ossification. Longitudinal ossifications in 5 cases were located in the upper thoracic vertebra and 13 cases in the middle and lower thoracic vertebra. Five cases were typical type, 4 cases were segmental type, 6 cases were continuous type and 3 cases were mixed type. All the patients had the posterior spinal canal decompression combined with longitudinal ligament resection, interbody fusion with bone graft and internal fixation surgeries. The operation time,blood loss and complications were recorded. JOA score were applied to evaluate the neurological function recovery pre-surgery, 2 days post-surgery and the latest follow-up. The surgery effects were evaluated by Epstein-Schwall method.

RESULTSThe operation time ranged from 3 to 6 hours (mean, 4.2 hours). The blood loss ranged from 800 to 4 000 ml (mean, 1 800 ml). All the patients were followed up, and the duration ranged from 6 months to 3 years, with a mean of 1.8 years. The JOA score increased from preoperative 4.30 ± 2.60 to 7.60 ± 2.40 2 days after surgery, and 7.80 ± 1.90 at the latest follow-up (t = 4.61, P < 0.001). The JOA scores between 2 days after surgery and the latest follow-up had no significant differences (t = 0.28,P = 0.78). The neurological recovery rate was 74% 2 days after surgery and 71% at the latest follow-up. There were 4 cases got an excellent result,10 good,3 fair and 1 poor according to Epstein-Schwall evaluation method. Four patients had cerebrospinal fluid leakage, 3 patients had intercostal nerve paralysis or pain, and 1 patient had superficial incision infection. The neurological function in 3 patients became worse in the second day posteratively , and among them, 2 patients were recovered at the latest follow-up and 1 patient had no changes. All the patients got fusion of bone graft and no internal fixation loosening and fractures occurred.

CONCLUSION360 degree circular decompression and transpedicle screw fixation can resect different types of thoracic longitudinal ligament ossification, and can achieve a good clinical effect.

Adult ; Aged ; Decompression, Surgical ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; surgery ; Thoracic Vertebrae ; surgery

OBJECTIVETo evaluate the clinical effects of posterior spinal transpedicular wedge osteotomy for kyphosis due to delayed osteoporotic vertebral fracture in elderly.

METHODSFrom July 2009 to February 2014,26 patients with kyphosis caused by delayed osteoporotic vertebral fracture were treated with transpedicular wedge osteotomy. There were 10 males and 16 females,aged from 55 to 75 years old with an average of 67 years. There were 1 osteotomy in thoracic vertebra,21 osteotomies in thoracolumbar vertebrae and 4 in lumbar vertebrae. Total 29 vertebrae were involved, 23 cases with single vertebral fracture and 3 cases with double vertebral fractures. Preoperative Cobb angles were 32°~51° with the mean of (42.00 ± 4.75) ° and VAS scores were 6 to 9 points with an average of (8.40 ± 0.75) points. According to the Frankel grade of spinal cord function, 4 cases were grade D and 22 cases were grade E. Intraoperative bleeding, operation time and perioperative complications were recorded, and improvements of Cobb angle were evaluated by X-rays. VAS score and Frankel grade were respectively used to evaluate the pain and nerve function.

RESULTSThe average operation time were 155 min (ranged, 120 to 175) and the mean intraoperative bleeding were 1 100 ml (ranged,800 to 1 500). Postoperative at 2 days, Cobb angle and VAS score were (9.60 ± 2.50) ° and (4.00 ± 1.00) points, respectively, ranged from 5° to 15° and 1 to 5 points. VAS score and Cobb angle improved obviously compared with preoperative (P < 0.05), and the improvement rate of Cobb angle was 76%. Frankel grade of 1 case changed from grade E to C, and the others did not become worse. The follow-up period ranged from 3 to 24 months with an average of 16.4 months. At the final follow-up, Cobb angles and VAS score were (11.00 ± 3.50)° and (4.40 ± 1.25) points, respectively, ranged from 5° to 19° and 1 to 6 points. The patient whose Frankel grade E changed to C at 2 days after surgery and changed to grade D at the latest follow-up. Vertebral body fracture below the fusion level happened in 1 case at 3 months after surgery, vertebral body fracture above the fusion level happened in 1 case at 5 months after surgery, and their chest pain symptoms were relieved after symptomatic treatment and anti osteoporosis treatment. All osteotomy levels obtained fusion which confirmed by X-ray and no internal fixation loosening and breakage were found.

CONCLUSIONThe clinical effect of posterior transpedicular wedge osteotomy for kyphosis due to delayed osteoporotic vertebral fracture was satisfactory, but manipulation during the operation should be cautious and prevent adjacent vertebral body fracture should be pay attention to prevent.

Aged ; Female ; Humans ; Kyphosis ; etiology ; surgery ; Male ; Middle Aged ; Osteoporotic Fractures ; complications ; surgery ; Osteotomy ; methods ; Spinal Fractures ; complications ; surgery ; Visual Analog Scale

Objective To construct a MigR1-CD19 recombinant vector which contains CD19 gene, and to establish a CD19-K562 cell line over-expressing stably CD19 gene and its subcutaneous xenograft model in NOD-SCID mouse.Methods The CD19 gene was inserted into the retroviral vector (MigR1) through recombinant DNA technology after transfection into Plat-A packaging cells, and viral supernatant was collected to transduce K562 cell line repeatedly to obtain stable transduction CD19-K562 cell line.Flow cytometry was used to determine the transduction efficiency and RT-PCR was used to confirmed CD19 gene expression.Cell proliferation and apoptosis were detected by cell count and Annexin V/PI, respectively.Then the subcutaneous xenograft subtype of CD19-K562-a cell line was constructed through subcutaneous inoculation and was cultured in vitro and in vivo.Then its subcutaneous xenograft model in NOD-SCID mouse was established.The characteristics of CD19-K562-a cells were detected by RT-PCR, Wright staining and immunohistochemistry.Results MigR1-CD19 recombinant vector was successfully constructed, and the CD19 positive efficiency of K562 cell line was (99.80±0.17) ％ through retrovirus centrifugation transduction.The transduction and passage had no effects on proliferation and apoptosis of CD19-K562 cells.The CD19-K562-a cell line was constructed after CD19-K562 cells were injected subcutaneously and were passaged in vitro and in vivo.The CD19 positive efficiency of the xenograft subtype CD19-K562-a cell line was (99.78± 0.04) ％.CD19-K562-a and CD19-K562 cells were in an undifferentiated state.NOD-SCID subcutaneous xenografts were established through subcutaneous inoculation of CD19-K562-a cells.CD19 in the CD19-K562-a subcutaneous xenografts was positive, while it was negative in its counterparts K562 cells.Conclusion The CD19-K562 cell line over-expressing CD19 gene and its subcutaneous xenograft model in NOD-SCID mouse are successfully established.

Aim Toresearchthemolecularmecha-nisms of DADS-induced apoptosis in human leukemia K562cells.Methods Cellviabilitywasmeasuredby MTT. Levels of DADS-induced ROS were measured by 2ˊ, 7ˊ-dichlorofluorescein diacetate ( DCFH-DA) fluo-rescence. DADS-induced mRNA levels of components of the NADPH oxidase were detected by Real-time PCR. The combination of protein Rac2 and p67phox was measured by immunoprecipitation assays. Flow cy-tometry methods were used to determine the percentage of apoptosis cells. DADS-induced Rac2 levels were measuredbyWesternblot.Results TheDADS-trea-ted K562 cells showed a dose-and time-dependent de-crease in cell viability and proliferation. There was sig-nificant up-regulation of the mRNA level of components of the NADPH oxidase complex in K562 cells after treatment with 6 mg·L-1 DADS for 6 h. Western blot results revealed that, compared with the control group, there was a significant up-regulation of Rac2 protein in K562 cells treated with 5. 0 and 10. 0 mg·L-1 DADS for 24h. And Rac2 combined with p67phox in DADS-induced apoptosis in K562 cells. PMA markedly in-creased the percentage of apoptotic cells, and DPI re-duced the percentage of apoptotic cells in DADS-in-duced K562 cells. Levels of DADS-induced ROS, also showed enhancement when exposed in PMA, but there was no DADS-induced ROS production evident when exposed in DPI in DADS induced K562 cells. Conclu-sions TheseresultsindicatethatNADPHoxidaseis the main source of DADS-induced ROS production. Diallyl disulfide induces apoptosis in human leukemia K562 cells through activation of NADPH oxidase.

AIMTo investigate whether DADS induce MGC803 cell apop tosis and cell cycle arrest. METHODSMGC803 cell growth inhibitio n was measured by MTT assay. Flow cytometry and acridine orange fluorescent stai ning method were used to determine the induction of apoptosis and the change of cell cycle. RESULTSMTT assay showed that adding 20,30,40 mg?L -1 DADS for 72 h suppressed MGC803 growth by 25 7%,58 6%,69 0% respective ly. Partial cells presented the characteristic morphological changes of apoptosi s under the fluorescent microscope. The apoptosis rate ncreased in time-depende nt manner. Flow cytometry analysis revealed that treating MGC803 cell with DADS significantly increased in the percentage of cells in the G 2/M phase. The proportion of cells in the G 2/M phase after treatment with 30 mg?L -1 DADS for 24 hours was comparable (46 0%), and more than four times that occu rring in untreated cells (9 9%). Furthermore, flow cytometry analysis also demonstrated that DADS induced apoptosis of MGC803 cell in time-dependent manner. T he pencentage of apoptotic cell was 3 53% after 0 h of 30 mg?L -1 DADS tr eatment. This pencentage of apoptotic cell rose steadily over time reaching 9 8 % after 24 h and 39 5% after 48 h. CONCLUSIONDADS could induce apoptosis of MGC803 cells and block the cell cycle at G 2/M phase.