Objective To evaluate the diagnostic approaches and treatment choices for priapism,and to upgrade the diagnostic and therapeutic efficacy. Methods Thirteen patients with priapism were evaluated by physical examinations,cavernous blood gas analysis,color duplex ultrasonography and cavernosography.Among them 9 cases of priapism resulted from intracavernosal injection of vaso-active agents,1 from perineal trauma and 3 from unknown cause.All the 13 patients were treated with intracavernosal injection of aramine and cavernosal decompression. Results Among these cases of priapism,12 were of low-flow and 1 was of high-flow.After the treatment,all the patients with priapism achieved remission.During the follow-up of 3 to 43 months,4 cases experienced decreased erection;9 had no significant change of erection.No cavernous fibrosis of penis occurred in them. Conclusions Cavernous blood gas analysis and color duplex ultrasonography are helpful with the accurate and timely dignosis of priapism.Cavernosal decompression and intracavernosal injection of arimine can be applied to most of the patients,and in the course of the treatment systemic adverse events should be avoided.

Objective To study the physical characteristics of the OPEN stereotactic body radiotherapy system for the clinical application. Methods The 0. 125cc ioniztion chamber, 160 mm polystyrene sphere model,Gafchromic EBT2 films and IBA film analysis software were used to evaluated the focus position tolerance,dose rate,repeatability,linear relation,penumbra and composite error of the OPEN stereotactic body gamma knife. We used the DTA method to verify the accuracy of dose distribution between the plans and measured value. Resualts The focus error was 0. 36 mm,max dose rate tolerance 3%,linear relation error 2%,repeatability error 0. 3%,composited error 2. 5 mm. There was 90% pass rate when the distance away from test point was less than 2 mm and the dose error was set less than 5 % . Conclusions Parts of the test resualts were similar to the head gamma knife national protocal of OPEN stereotatic body gamma knife. The deliver dose distribution can meet the clinic need.

ObjectiveTo investigate the injury mechanism,clinical characteristics and surgical treatments of Segond fracture combined with anterior cruciate ligament (ACL) injury.MethodsNine patients suffering from Segond fracture combined with ACL injury were treated between January 2008 and December 2010.All the patients revealed ACL and medial collateral ligament (MCL) breakage under arthroscopy.Furthermore,one patient was associated with lateral collateral ligament (LCL) breakage and medial meniscus injury,four with medial meniscus tear and two with lateral meniscus tear.All the patients underwent arthroscopic tendon allotransplantation,ACL reconstruction and MCL repair.Besides,synchronous LCL reconstruction was performed in one patient,meniscus suture in three and meniscus plasty in four.Six patients with large Segond fracture fragments were fixed with two hollow lag crews and three with relatively small fracture fragments fixed with one hollow lag crew.ResultsThe mean followup period was 12 months,which showed average postoperative Lysholm score of 59 points and satisfactory clinical outcome.ConclusionsSegond fracture is often combined with ACL injury and is predictive for ACL injury.In ACL reconstruction,large Segond fracture blocks should be reduced and fixed and the combined injuries also should be treated in the same period.

Objective To investigate the inhibitive effect of siRNA(small interfering RNA,siRNA) on the phosphodiesterase type 5(PDE5) in smooth muscle cells of human corpus cavernosum,and provide experimental groundwork for the gene therapy of erectile dysfunction (ED). Methods Small interfering RNAs targeting PDE5 gene were systhesized by using web design software provided by Ambion,there siRNAs and control siRNA were systhesized by Ambion. SiRNAs were transfected into smooth muscle cells of human corpus cavernosum by using siPORTTM Lipid reagent;down-regulation of PDE5 mRNA was detected by RT-PCR;the inhibitive effect of PDE5 was detected by Western Blotting. Results The results of RT-PCR indicated siRNA1、siRNA2 and siRNA3 made down-regulations of PDE5 mRNA expression in the transfected groups 58.2%、14.9% and 11.8%;the PDE5 expression decreased 70.5%、19.8% and 17.3%;however the expression did not have different in control siRNA and frank group. Conclusions The synthesized siRNAs in vitro were able to down-regulate the expression of PDE5.There were different capabilities of the specific siRNAs down-regulation.It was suggested that the siRNA technique provide not only an extremely powerful tool for the functional analysis of genome but also a new method for ED gene therapy.

BACKGROUND: Glucocorticoid-induced osteoporosis has relationship with the down-regulation of osteoprotegedn expression. Osteoprotegerin could inhibit bone resorption in the animal experiment and clinical application for treating oestrogenic hormone deficiency osteoporosis. OBJECTIVE: To investigate the effects of exogenous recombinant osteoprotegerin fusion protein on glucocorticoid-induced osteoporosis in rats. DESIGN, TIME AND SETTING: Randomized grouping, controlled animal expenment was performed in the Institute of Orthopedics, Chinese PLA General Hospital between January 2006 and June 2008. MATERIALS: Sixty healthy male Wistar rats of clean grade; Dexamethasone was produced by Tianjin Jinyao Amino Acid Co., Ltd (Licenca No. H12020515). METHODS: Sixty rats were divided into 3 groups randomly with 20 rats in each. Control group: the rats were administrated with 0.9％ sodium chloride. Dexamethasone group: the rats were administrated with dexamethasone intramuscularly. Osteoprotegedn group: the rats were administrated with dexamethasone and recombinant osteoprotegerin intramuscularly. MAIN OUTCOME MEASURES: All rats were sacrificed at 12 weeks after administration. The urine calcium, phosphor, creatinine, bone mineral density, biomechanics tests of femur and vertebral body, were measured. Immunohistochemistry staining were performed to observe osteoprotegerin expression.RESULTS: Sixty rats were all involved in the final analysis. ①Compared with control group, udne calcium increased in the Dexamethasone group (P < 0.05); the bone mineral density of lumbar vertebra and femur decreased significantly (P < 0.05), especially lumbar vertebra (P < 0.01); biomechanics tests of femur and vertebral body (maximum load, maximum stress, elasticity load, elasticity stress, elastic modulus) decreased significantly (P < 0.05); immunohistochemistry staining showed that endogenous osteoprotegerin expressions were reduced significantly in bone marrow of Dexamethasone group (P < 0.01). ②Compared with Dexamethasone group, urine calcium decreased in the osteoprotegerin group (P < 0.01 ); the bone mineral density of lumbar vertebra and femur increased (P < 0.05); the parameters of biomechanics testa of femur and vertebral body increased (P < 0.05); the osteoprotegerin expression was not changed between Dexamethasone group and osteoprotegerin group.CONCLUSION: Glucocorticoid could inhibit osteoprotegerin expression in the bone followed by progressive bone loss and induce osteoporosis. Recombinant osteoprotegerin works effectively in inhibiting bone resorption after administrated with glucocorticoid, reduce bone resorption index, increase bone mineral index and bone strength, thus improving the osteoporosis which is induced by glucocorticoid.

We investigated the effects of dendritic cell (DC) pulsed with acute leukemia cell frozen-thawed antigen on inducing the cytotoxic T lymphocyte (CTL) to get specific anti-tumor activity in vitro. DC was generated from healthy human bone marrow mononuclear cell (BMMC) in the presence of granulocyte/macrophage-colony stimulating factor(GM-CSF), interleukin-4 (IL-4) in vitro. DC pulsed with acute leukemia cell frozen-thawed antigen was co-cultured to induce T cell into specific CTL. Then we observed the effects of CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen killing acute leukemia cell specially and the influence of dendritic cell affecting the function and CD expression on CTL. The levels of CD1a, CD86, HLA-DR expression on DC pulsed with acute leukemia cell frozen-thawed antigen were obviously higher than those before culture (P<0.01). There were more CD3+CD8+ T cells in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen, compared with those in the T cell uncultured group (P<0.01). The CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen significantly had higher activity in killing acute leukemia cell than in killing k562 cell (P<0.01), and the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen was also more effective for killing acute leukemia cell as compared with the CTL induced by DC simply, T cell co-cultured with IL-2 and T cell simply (P<0.01). The DC generated from human bone marrow mononuclear cell (BMMC) in the presence of granulocyte/macrophage-colony stimulating factor (GM-CSF), interleukin-4 (IL-4) was CD14- CD1a+CD83+DC, and it could also induce the cytotoxic T lymphocyte (CTL) to get specific anti-tumor activity in vitro. Otherwise,the increasing of CD3+CD8+ T cells in the CTL induced by DC pulsed with acute leukemia cell frozen-thawed antigen implied the main role of the CD3+CD8+ T cells in the anti-tumor immunity.
Antigens, Neoplasm ; immunology ; Cells, Cultured ; Coculture Techniques ; Cytotoxicity, Immunologic ; Dendritic Cells ; cytology ; immunology ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Interleukin-4 ; pharmacology ; Leukemia, Myeloid, Acute ; immunology ; pathology ; Lymphocyte Activation ; T-Lymphocytes, Cytotoxic ; immunology

Objective To investigate the effect of simvastatin (SV) in combination with cytosine arabinoside (ARA-C) on the proliferation and apoptosis of K562 cells. Methods Human K562 cells were incubated with SV and cytosine arabinoside alone or in combination and K562 cells without any treatment were taken as normal control. Cells in different groups were collected at 24, 48 and 72 h after incubation for further detections. Morphological changes by Wright stain were performed. MTT method was used to assay the growth inhibition rate and cytoflowmetry was used to detect the early stage apoptosis ratio and cell necrosis ratio. Results Compared with Ara-C group and SV group, cells in the group treated with SV combined with Ara-C showed obvious karyopyknosis,apoptosis bodies formation and significant cell growth inhibition, which were positively correlated with culture time. Combination of 15 μmol/L SV and Ara-C showed the most significant cell growth inhibition with a inhibition rate of (72±1) % at 72 h of culture, as was significantly higher than that of 15 μmol/L SV group (45±2) % and 20 μmol/L Ara-C group (44±0) % (P ＜0.01),furthermore, combination of 15 μmol/L simvastatin and Ara-C showed synergistic inhibition with Q value of 1.24 and 1.19 at 24 h and 48 h in each. The apoptosis rates at early stage (AnnexinV) detected by flow cytometry in 20 μmol/L, 15 μmol/L and 10 μmol/L SV treated K562 cells were significantly higher than that in normal K562 cells (P ＜0.01), as were positively correlated with culture time and SV dose (P ＜0.05). There were no significant difference of early apoptosis rate between the 20 μmol/L SV and 15 μmol/L SV groups (P ＞0.05), yet the very two were both higher than that of 10 μmol/L SV group (P ＜0.05). There were no statistic differences of late apoptosis rate (PI) amongdifferent treated groups (P ＞0.05). Conclusion SV inhibited K562 cell proliferation and induced cell apoptosis in vitro, and combination of SV and Ara-C exhibited obvious synergistic inhibition and apoptosis, which may increase the sensitivity of K562 cell to chemotherapy. SV at 15 μmol/L may be the best concentration for K562 cells in vitro.

Objective To investigate the effects of simvastatin (SV) on the proliferation,differentiation and apoptosis of human promyelocytic leukemia cell line NB4.Methods NB4 cells were incubated with SV at different concentration with or without all-trans retinoic acid (ATRA),and NB4 cells without any treatment were taken as normal control.Cells of different groups were collected at 24 h,48 h and 72 h after incubation for further detection.Morphological changes by Wright stain were performed.MTT method was used to assay the growth inhibition rate and flow cytometry was used to detect the surface CD11b expression levels,the early stage apoptosis ratio and cell necrosis ratio.Results Treated with 15 μ mol/L SV,10 μ mol/L SV and 5 μ mol/L SV respectively,with the NB4 cells growth,the cell inhibition rates gradually increased (F =7.15,P =0.000),as well as CD11b expression levels (F =3.41,P =0.014) and AnnexinVexpression levels (F =43.38,P =0.000).Furthermore the NB4 cells treated with 15 μ mol/L SV exhibited the most significant changes with cell inhibition rate of 0.96±0.02,CD11b expression level increased to (62.41±6.37) ％ and AnnexinV expression level increased to (87.38±2.94) ％ after 72 h incubation.Combination of 15 μmol/L SV with 0.5 μmol/L ATRA displayed obvious interaction for increasing CD11b expression levels (F =4.093,P =0.025),while no significant interaction for cell inhibition rates and Annexin V expression levels were observed.After 72 h incubation,the CD11b expression levels (89.46±9.13) ％ in NB4 cells treated with 15 μ mol/L SV in combination with 0.5 μ mol/L ATRA were significantly higher than those treated with ATRA (71.27±7.27) ％ and SV (62.41±6.37) ％ (t =2.71,P =0.054; t =4.37,P =0.017)' solely.Conclusion Simvastatin in vitro inhibits NB4 cell proliferation,promotes cell apoptosis,and synergistically induces cell differentiation with ATRA dose-dependently in vitro,which indicates that SV may have the effect of synergistic anti-promyelocytic potency with ATRA.

OBJECTIVETo detect putative suppressor loci involved in tumor progressing or metastases.

METHODSThirty microsatellite marker primers were employed to amply the corresponding loci of the genome DNA from 83 patients with sporadic colorectal cancer. The PCR products were electrophoresed on a 377 PRISM sequencer and the fluorescent signals were analyzed by Genotyper and Genescan software.

RESULTSThe data were obtained from 24 loci, with an average LOH frequency of 15.16%. The LOH at D2S206 and D2S364 was more frequent than 30%, and was less than 20% at the rest loci. Significant difference was observed between the percentage of LOH and tumor staging or differentiation at D2S142 (2q24.1), D2S126 (2q35), D2S2211 (2p24.2), D2S305 (2p23.3). Occarrence of deletion at the later two loci was correlative.

CONCLUSIONSFrequent LOH was not observed at the loci around known mismatch repair genes on chr. 2. The region between D2S305 (2p23.3) and D2S2211 (2p24.2) deleted holistically, and was correlated to the stage and differentiation of tumor attended by D2S142 (2q24.1) and D2S126 (2q35) on 2q. It is suggested that unknown genes associated with tumor progressing or metastases reside in the two loci on 2q or the region on 2p.

Adult ; Aged ; Aged, 80 and over ; Cell Differentiation ; Chromosome Mapping ; Colorectal Neoplasms ; genetics ; pathology ; Female ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Middle Aged ; Neoplasm Metastasis

BACKGROUNDTo investigate the expression of the SOCS3 gene and its effect on proliferation of A549 cells.

METHODSA549 cells were cotransfected with pEFSOCS3 and pSV2neo by liposome, then G418 was used to screen the positive cells. Expression of SOCS3 mRNA and protein was detected by RT-PCR and immunocytochemistry respectively before and after transfection. MTT assay was used to detect the cell growth. Flow cytometric DNA analysis was used to determine the cell cycle.

RESULTSRT-PCR and immunocytochemistry showed that no expression of SOCS3 mRNA and protein was detected in A549 cells before transfection, but a stable expression of SOCS3 gene was observed after transfection with SOCS3 gene. Compared with control group, growth of A549 cells transfected with SOCS3 gene was significantly suppressed, with a suppressive rate of 41.07%. The cells at G₀/G₁ cell phases increased, and those at S and G₂/M phases decreased significantly after transfection.

CONCLUSIONSSOCS3 protein might inhibit the proliferation of A549 cells by negatively regulating cellular signal pathways.