This research explored the synergistic effects and the potential mechanisms of RCE-4 and various nonsteroidal anti-inflammatory drugs (NSAIDs) on the proliferation of cervical cancer Ca Ski cells. The MTT assay and CalcuSyn V2.0 software were used to detect cell proliferation and calculate the combination index (CI); the expression levels of various proteins were analyzed using Western blot assay; mitochondrial membrane potential (MMP) was assessed using JC-1 staining; acridine orange/ethidium bromide (AO/EB) double-fluorescence staining was used to detect the apoptosis of Ca Ski cells; a co-immunoprecipitation (Co-IP) assay was used to analyze the relative content of Bcl-2-Beclin 1 complex in Ca Ski cells. The results demonstrate that the combination of RCE-4 and NSAIDs increases the inhibition of Ca Ski cells compared to the single-RCE-4 group, and celecoxib provided the best synergistic effect among the four NSAIDs tested, with a CI of 0.32. The combination of RCE-4 and celecoxib significantly down-regulated the expression of cyclooxygenase-2 (COX-2) and nuclear transcription factor-κB (NF-κB), and promoted the expression of non-steroidal anti-inflammatory drugs activity gene-1 (NAG-1). In addition, autophagy induced by RCE-4 was markedly inhibited in combination with celecoxib, which was associated with down-regulation of the expression of microtubule-associated protein 3 (LC3)-II, Beclin 1, p62 and autophagy-related gene (ATG) 3/4B/5/7/14. RCE-4-induced apoptosis was significantly enhanced by altering the depolarization of mitochondrial membrane potential and the expression of B cell lymphoma-2 (Bcl-2), B cell lymphoma-xl (Bcl-xl), Bcl-2 associated X protein (Bax), Bcl-2/Bcl-xl-associated death promoter (Bad) and cleaved cysteinyl aspartate specific proteinase (cleaved-caspase) 3/7/9. Furthermore, the formation of the Bcl-2-Beclin 1 complex was significantly inhibited in Ca Ski cells treated with RCE-4 in combination with celecoxib. Taken together, this research shows that the combination of RCE-4 and celecoxib has a significant synergistic effect on the proliferation of Ca Ski cells by promoting apoptosis, inhibiting autophagy and disturbing the formation of the Bcl-2-Beclin 1 complex, which may be a novel strategy to increase the sensitivity of anti-cervical cancer drugs.

Objective To study the effects of dexamethasone (DEX) and/or hyperbaric oxygen (HBO) on the subdermal vascular network (SVN).Methods SVNs were selected on dorsal skin flaps of 40 Wistar rats.The animals were divided randomly into a control group,a DEX group,a HBO group and a HBO+DEX group.Cranially based,2.5 cm?10 cm dorsal SVN skin flaps were sharply incised and elevated between the dartos and SVN,then sutured to their beds.On the 7th postoperative day,the surviving flap area was measured along with the number of new capillary,the thickness of meat tissue and the number of infiltrated neutrophilie granulocytes in the SVN skin flap.Results The mean surviving flap area for the control group was 7.90 cm~2,for the DEX group it was 10.48 cm~2,for the HBO group 15.53 cm~2,and for the DEX+HBO group it was 15.58 cm~2.The improvement in surviving flap area was highly statistically signifieant compared with the control.The improvement was also statistical- ly significant when the HBO group or HBO+DEX group was compared with the DEX group.However,no statistically significant difference was found between the HBO group and HBO+DEX group.Conclusion In a rat dorsal skin flap model,DEX or HBO improved skin flap viability,but DEX alone is not as efficacious as HBO or as DEX+ HBO.DEX plus HBO showed no additive beneficial effect over HBO alone.

Objective To analyze the effects of-γ-knife treatment with different dosages on level of prolactin (PRL) in patients with different sizes of functional pituitary prolactinomas, and determine an index to guide hormone replacement therapy and the prognosis of -γ-knife treatment in patients with functional pituitary prolactinomas through comparing the changes of tumor sizes and the levels of PRL before and after -γ-knife treatment. Methods A retrospective analysis of the clinical data of 248 patients with functional pituitary prolactinomas was performed; gamma knife treatment was performed on these patients from September 2004 to March 2008. We divided the patients into 3 groups: group Ⅰ (50 Gy≤central dose<60 Gy, 20 Gy＜marginal dose<30 Gy), group Ⅱ (40 Gy≤ central dose＜50 Gy, 15 Gy＜marginal dose<25 Gy) and group Ⅲ (30 Gy ≤ central dose<40 Gy, 12 Gy＜marginal dose<20 Gy). The irradiation dose on optic nerves in the 3 groups was under 9 Gy. Radioimmunoassay was employed to detect the serum PRL level before and 1, 3 and 12 months after γ-knife treatment. The changes of the tumor sizes were observed and compared with cranial MRI 1 and 2 years after -γ-knife treatment.Results Significant differences on the PRL level were noted before -γ-knife treatment between each 2 groups (P<0.05); the PRL level in group Ⅲ was lower as compared with that in group Ⅰ and Ⅱ before γ-knife treatment; however, the PRL level in group Ⅲ was higher as compared with that in group 112 months after -γ-knife treatment; the PRL level in all the 3 groups after γ-knife treatment was significantly lower as compared with that before γ-knife treatment (P<0.05). MRI showed that the tumor had 80% partial response rate (198/248) in the 1st year, 82% complete response rate (203/248) in the 2nd year, increased volume in 19 patients (7.7%) and no change in 26 patients (10.4%). Conclusion Different treatment doses of Gamma knife on functional pituitary prolactinomas has great influences on postoperative recovery of endocrine; the higher doses of the center and edge (especially center), the higher normal rate of postoperative PRL level. Whether it will cause long-term hypopituitarism needs continue follow-up.

BACKGROUNDPatients with pelvic fractures are often treated in hospitals without the capacity to implement an open reduction internal fixation (ORIF). This often leads to pelvic malunion in patients with unstable pelvic fracture, shock or even death due to uncontrollable pelvic hemorrhage and unstable hemodynamics. This study explored the role of early external fixation (within 7 days) for patients with unstable pelvic fractures.

METHODSA retrospective analysis was conducted on 32 patients with unstable pelvic fractures treated with early external fixation from January 2005 to January 2010 (Tile type B: 18 cases; C: 14 cases). The study comprised 28 males and 4 females, with a mean age of (32 ± 8) years (range, 21-56 years). Of these patients, 22 were treated with emergency pelvic external fixation and 10 were treated with external fixation within 1-7 days. Fifteen cases suffered traumatic hemorrhagic shock. A statistical analysis was conducted to compare fluid infusion and blood transfusion volumes within the first 24 hours of these shock patients with another cohort of patients treated without early external fixation from January 1993 to January 1998.

RESULTSThe average follow-up was (34.7 ± 14.6) months (range, 6-66 months). Six to eight weeks after external fixation, patients could walk with crutches; by 12 weeks, external fixation was removed and all fractures had healed. Seven patients presented with sequelae, including 3 patients with long-term lumbosacral pain, 3 patients with erectile dysfunction and 1 patient with Morel-Lavallee lesion and other complications. The 15 shock patients in this study (2005-2010 group) required significantly lower volumes of fluid infusion and blood transfusion (P(fluid) = 0.000; P(transfusion) = 0.000) as compared to the 1993-1998 cohort.

CONCLUSIONSThe early application of external fixation in unstable pelvic fracture patients positively affects hemodynamic stability, with outstanding efficacy as a final fixation option for unstable pelvic fractures.

Adult ; External Fixators ; Female ; Fracture Fixation ; methods ; Fractures, Bone ; physiopathology ; surgery ; Hemodynamics ; Humans ; Male ; Middle Aged ; Pelvic Bones ; injuries ; Retrospective Studies

OBJECTIVETo analyze the reasons of the complications,and the measures taken for the prevention and treatment by percutaneous vertebroplasty (PVP) for osteoporotic thoracolumbars vertebral compressive fractures.

METHODSFrom May 2004 to June 2008, 204 patients with 286 diseased vertebrae underwent PVP under the guidance of C-arm fluoroscopy, and 56 patients with complications included 18 males, 38 females, ranging in age from 58 to 93 years, with an average of 72 years. The condition of leakage of bone cement and complications was observed.

RESULTSThe complications related to the leakage of PMMA found in 60 vertebrae in 49 cases: vertebral canal in 5 cases, intervertebral foramina in 3 cases, soft tissue besides vertebrae in 20 cases,intervertebral discs in 15 cases, venous plexus besides vertebrae in 6 cases. The complications no related to the leakage of PMMA found in 7 cases: the nerve roots injuried in 3 cases; the pain of the patient worsened temporarily in 2 cases; the blood pressure of the patient descent temporarily in 2 cases.

CONCLUSIONThe main reasons of complications are no-integrated of the compressived vertebrae,unstandard skills of injecting,unsuitable opportunity and quantity of injecting PMMA, unsufficient monitoring of C-arm fluoroscopy, the toxicity of PMMA. The measures of prevention and treatment is strict indication, standard skills of injecting, sufficient monitoring of C-arm fluoroscopy, suitable opportunity and quantity of injecting PMMA, electrocardio-monitoring in operation.

Aged ; Aged, 80 and over ; Bone Cements ; adverse effects ; Female ; Fractures, Compression ; complications ; surgery ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Polymethyl Methacrylate ; adverse effects ; Postoperative Complications ; prevention & control ; Spinal Fractures ; complications ; surgery ; Vertebroplasty

Objective to explore the current situation and related influencing factors on the retention time of patients receiving methadone maintenance treatment (MMT). Methods Information on basic situation and daily treatment of the patients were collected from the 7 MMT clinics opened in the pro-two batch in Hunan province. Retention rate and influencing factors were analyzed. Results (1) The retention rates after 6 and 12 months of MMT became 72.06% and 49.65% respectively. (2) The retention rates of high-dosage group and low-dosage group were 85.03% and 68.03% after 6 months on MMT program while became 60.48% and 46.28% after 12 months of MMT respectively. (3) The mean retention time of HIV+ patients and HIV- patients were 9.46 months and 8.62 months respectively during the 12 months follow-up observation, showing a significant difference. (4) Patients who took large dose methadone, did not share needles, at older age or HIV+ , were prone to keep MMT at a long period. Conclusion The retention rates for 6 months and 12 months in the MMT program in Hunan province were similar to the national data. Dose, type of drug abuse, age and HIV status were related to the period of retention.

BACKGROUNDIt was reported that telomerase expression is closely associated with cellular immortality and cancer. This study was designed to investigate the relationship between telomerase expression and the carcinogenesis of cervical cancer, the possible use of telomerase as a marker of cervical intraepithelial neoplasia (CIN) progression or regression, and the natural history of CIN.

METHODSTelomeric repeat amplification protocol (TRAP) assay was used to measure telomerase activity in cervical scrapings and biopsy samples obtained from 105 cases affected with various cervical conditions, including chronic cervicitis (n = 20), CIN (n = 64, 16 cases of CIN I, 20 cases of CIN II, and 28 cases of CIN III), and invasive squamous cell carcinoma (n = 21).

RESULTSIn exfoliated cell samples, telomerase activity was detected in 5 of 20 (25.0%) cases of cervicitis, 10 of 16 (62.5%) cases of CIN I, 11 of 20 (55.0%) cases of CIN II, 23 of 28 (82.1%) cases of CIN III, and 13 of 21 (61.9%) cases of carcinoma. In cervical biopsy samples, telomerase activity was detected in 6 of 20 (30.0%) cases of cervicitis, 8 of 16 (50.0%) cases of CIN I, 9 of 20 (45.0%) cases of CIN II, 27 of 28 (96.4%) cases of CIN III, and 20 of 21 (95.2%) cases of carcinoma. Telomerase activation was significantly higher in CIN samples than in cervicitis samples. Telomerase activity was detected at similar frequency in samples from cervical scrapings and cervical biopsies.

CONCLUSIONThese results seem to suggest that telomerase expression may be associated with carcinogenesis of the cervix. TRAP assay of cervical scraping samples could be used to monitor and predict the development of CIN in clinical practice.

Adult ; Biomarkers, Tumor ; analysis ; Cervical Intraepithelial Neoplasia ; enzymology ; Disease Progression ; Female ; Humans ; Middle Aged ; Telomerase ; metabolism ; Uterine Cervical Neoplasms ; enzymology ; Uterine Cervicitis ; enzymology

OBJECTIVETo study the effect of ex vivo expansion on the adhesion activities of umbilical cord blood hematopoietic stem and progenitor cells (HSPC).

METHODSFresh UCB CD(34)(+) cells were cultured in a serum and stroma-free culture system. At day 7, day 10 and day 14, CD(34)(+) cells were re-selected from the expanded products. The expression of adhesion molecules (CAMs) such as VLA-4, VLA-5, LFA-1, ICAM-1, HCAM, L-selectin and PECAM-1, and the adhesion activity of the expanded CD(34)(+) cells were evaluated and compared with those of precultured fresh CD(34)(+) cells.

RESULTS(1) The CD(34)(+) cells expressing homing-related CAMs were increased (from 15-fold increase for CD(34)(+) CD(54)(+) subset to 72-fold increase for CD(34)(+) CD(49e)(+) subset at day 14). (2) The expressions of CD(49d), CD(44), CD(11a) and CD(49e) on the expanded CD(34)(+) cells were increased or sustained the same levels as those on fresh UCB CD(34)(+) cells, while the expression of CD(62L), CD(54) and CD(31) on expanded CD(34)(+) cells declined with the cultivating. (3) Spontaneous adhesion and SDF-1-induced adhesion tended to be increased in the course of the first 10 day's culture.

CONCLUSIONSThe culture system used in this study could substantially support the expansion of HSPCs expressing the above CAMs, and the expanded HSPCs would sustain their intrinsic adhesion potentials.

Antigens, CD ; analysis ; Antigens, CD34 ; analysis ; Cell Adhesion ; Cell Adhesion Molecules ; biosynthesis ; Cell Division ; Fetal Blood ; cytology ; immunology ; metabolism ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; immunology ; metabolism ; Humans ; Receptors, Lymphocyte Homing ; biosynthesis

OBJECTIVETo investigate the molecular mechanisms by which Qianliening Capsule (, QC) treats benign prostatic hyperplasia (BPH).

METHODSHuman prostate stromal cell line WPMY-1 was treated with 0, 1, 3 and 5 mg/mL of QC for 24, 48 and 72 h, respectively, in the presence of 10 ng/mL basic fibroblast growth factor (bFGF). The viability of WPMY-1 cells was determined by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell morphology was observed by phase-contrast microscopy. 4',6-diamidino-2-phenylindole (DAPI) staining and fluorescence activated cell sorting (FACS) analysis with Annexin-V/propidium iodide (PI) staining were performed to determine cell apoptosis. The loss of mitochondrial membrane potential was examined by FACS analysis with 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl-carbocyarine iodide (JC-1) staining. Activation of caspase-3 and -9 was evaluated by colorimetric assay. The mRNA and protein expression levels of Bcl-2 and Bax were measured by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting, respectively.

RESULTSUpon bFGF stimulation, the viability of WPMY-1 cells was increased to 122%-118% compared with the control cells (P <0.05). However, treatment with 1-5 mg/mL of QC for 24, 48 and 72 h decreased the viability of bFGF-stimulated cells to 80%-92%, 59%-82%, 36%-62% compared with the untreated cells (P <0.05). In addition, QC treatment reduced WPMY-1 cell density in a dose-dependent manner. Moreover, QC treatment dose-dependently induced the loss of plasma membrane asymmetry, the nuclear condensation and fragmentation, collapse of mitochondrial membrane potential, activation of caspase-9 and caspase-3, and increase of pro-apoptotic Bax/Bcl-2 ratio.

CONCLUSIONPromoting mitochondrion-dependent apoptosis of prostate stromal cells might be one of the mechanisms by which QC treats BPH.

Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Capsules ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Down-Regulation ; drug effects ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Humans ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria ; drug effects ; physiology ; Prostate ; cytology ; drug effects ; physiology ; Stromal Cells ; drug effects ; physiology

OBJECTIVETo investigate the role of mesenchymal stem cells (BMSCs) in multiple myeloma (MM) bone marrow (BM) microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.

METHODSBMSCs were derived from BM of untreated myeloma patients (MM-BMSCs) and healthy donors (HD-BMSCs), respectively. The phenotype, proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs. Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro. The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.

RESULTSMM-BMSCs and HD-BMSCs were isolated successfully. The phenotype of MM-BMSCs was similar to that of HD-BMSCs. Expressions of CD73, CD105, CD44 and CD29 were positive, but those of CD31, CD34, CD45 and HLA-DR (< 1%) negative. The proliferation time of MM-BMSCs was longer than that of HD-BMSCs (82 h vs 62 h, P < 0.05). Moreover, over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ± 9.4) pg/ml vs (115.0 ± 15.1) pg/ml and (1497.2 ± 39.7) pg/ml vs (1329.0 ± 21.1) pg/ml, respectively]. MM- BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.

CONCLUSIONSMM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib. Over-expression of IL-6 and VEGF maybe play a critical role in these effects.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Bortezomib ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Multiple Myeloma ; metabolism