Female ; Human papillomavirus 18 ; physiology ; Humans ; Papillomavirus Infections ; complications ; Risk Factors ; Uterine Cervical Neoplasms ; epidemiology ; etiology ; pathology ; virology ; Virus Integration ; genetics ; physiology

Objective To study the effect of bel-2 siRNA on apoptosis of HL-60 cells.Methods bcl-2 siRNA was synthesized in vitro transcription with silencer siRNA construction kit.The synthesized siRNA was transfected into HL-60 cells with Amine siPORT transfection.We used MTT flow cytometer and hoechst 33258 flourescence stainning t0 evaluate cell proliferation and apoptosis. Results.Bcl-2 siRNA could partially inhibit the growth of HL-60 cells.After incubated with bcl-2 siRNAl for 48 hours,HL-60 cells exhibited morphologic characteristic of apoptosis including chromatin condensation,crescents formation and nuclear fragmentation.Conclusion Effective bcl-2 siRNA can induce apoptosis and inhibit cell proliferation.

A 31P-quantitative nuclear magnetic resonance(qNMR)method was established for simultaneous determination of sodium α-glycerophosphate,sodium β-glycerophosphate and phosphoric acid in concentrated divitamins and sodium phosphate syrup.The qNMR experimental conditions were optimized,including hexamethylphosphoramide as internal standard,20%deuterium oxide solution as solvent,zgig pulse sequence,delay time of 30 s,and scan number of 64.The 31P-NMR peaks atδ29.80 of hexamethylphosphoramide,δ0.69 of sodiumα-glycerophosphate,δ0.17 of sodiumβ-glycerophosphate andδ0.03 of phosphoric acid were chosen as the quantitative peaks(pH of the test solution was around 4.8).Method validation was performed in terms of precision(Relative standard deviation less than 0.6%),linearity(Correlative coefficient greater than 0.999),limit of detection(23.58 μg/mL for sodium glycerophosphate and 11.61 μg/mL for phosphoric acid)and limit of quantitation(78.60 μg/mL for sodium glycerophosphate and 38.70 μg/mL for phosphoric acid).The recoveries were 99.8%?103.2%,and relative standard deviations were 0.41%?1.98%.The results showed that the reliability of 31P-qNMR method were suitable for its intended use.Seven batches of concentrated divitamins and sodium phosphate syrups were tested by the established method,of which the total phosphorus content was consistent with that of colorimetry method,but the content of sodium glycerophosphate(Sum ofαtype andβtype)was relatively low,about 82%of the labeled amount.The content of phosphoric acid was high.This method simplified sample pretreatment and had high specificity,and was more suitable for determination and quality control of concentrated divitamins and sodium phosphate syrup.

To establish single- and double-transfected transgenic cells stably expressing hMATE1, hMATE1 cDNA was cloned by RT-PCR from human cryopreserved kidney tissue, and subcloned into pcDNA3.1(+) plasmid by virtue of both HindIII and Kpn I restriction enzyme sites. Subsequently, the recombined pcDNA3.1(+)- hMATE1 plasmid was transfected into MDCK, MDCK-hOCT1 or MDCK-hOCT2 cells using Lipofectamine 2000 Reagent. After a 14-day-cultivation with hygromycin B at the concentration of 400 µg · mL(-1), all clones were screened with DAPI and MPP+ as substrates to identify the best candidate. The mRNA content of hMATE1, the cellular accumulation of metformin with or without cimetidine as inhibitor, or transportation of cimetidine was further valuated. The results showed that all of the three cell models over expressed hMATE1 mRNA. The cellular accumulation of metformin in MDCK-hMATE1 was 17.6 folds of the control cell, which was significantly inhibited by 100 µmol · L(-1) cimetidine. The transcellular transport parameter net efflux ratios of cimetidine across MDCK-hOCT1/hMATE1 and MDCK-hOCT2/hMATE1 monolayer were 17.5 and 3.65, respectively. In conclusion, cell models with good hMATE1 function have been established successfully, which can be applied to study the drug transport or drug-drug interaction involving hMATE1 alone or together with hOCT1/2 in vitro.
Animals ; Biological Transport ; Cimetidine ; pharmacology ; DNA, Complementary ; Dogs ; Drug Interactions ; Humans ; Madin Darby Canine Kidney Cells ; Metformin ; pharmacology ; Organic Cation Transport Proteins ; genetics ; metabolism ; Transfection

Objective To determine the effect of ostecytic TGF-β/Smad4 signaling on osteoblastic and osteoclastic differentiation in bone marrow stromal cells (BMSCs).Methods Mice with osteocytic TGF-β/Smad4 conditional knock down (Smad4ot CKD) were generated as previously by crossing DMP1-8kb-Cre mice with Smad4lox(ex8)/lox(ex8) mice.The osteocytes were isolated from tibial and femoral diaphysis and co-cultured with wild-type BMSCs.ALP staining, Alizarin red staining and TRAP staining were performed to show osteoblastic and osteoclastic differentiation.Then, their marker genes were detected by qPCR and proteins measured by Western blot.ResultsThe expression of Runx2 and Osterix were reduced in smad4 CKDot co-cultured with BMSCs compared with controls(P<0.01).Similarly, the specific markers of osteoblastic differentiation were decreased (P<0.01).Additionally, the expression of RANKL was not significantly changed in with BMSCs.However, OPG was highly expressed incontrol group compared with smad4 CKD in co-cultured group (P<0.05).Thus, the radio of RANKL/OPG was significantly reduced (P<0.05).Furthermore, the expression of RANK was inhibited.Conclusions The terminally-differentiated osteocytes are the cells regulating bone metabolism, while down-regulation of osteocytic-TGF-β/Smad4 inhibits BMSC osteoblastic and osteoclastic differentiation.

BACKGROUNDConnexin43 (Cx43) is the predominant gap junction protein in heart and is involved in the control of cell-to-cell communication to modulate the contractility and the electrical coupling of cardiac myocytes. Left ventricular (LV) hypertrophy is accompanied by changes of Cx43 expression. Recent studies have demonstrated that statins reduced cardiac hypertrophy. However, it is unknown whether statins can affect Cx43 expression in hypertrophied left ventricular myocardium. This study was designed to assess the effects of atorvastatin on LV hypertrophy and Cx43 expression in spontaneously hypertensive rats (SHR).

METHODSNine-week old SHRs were randomly divided into two groups. Some received atorvastatin at 30 mg/kg by oral gavage once daily for 8 weeks (SHR-A); others received vehicle. Age-matched Wistar-Kyoto rats (WKY) received atorvastatin or vehicle for 8 weeks were used as controls. At the end of the experiment, we investigated LV hypertrophy and the expression of Cx43 in LV myocardium in four groups. Cx43 expression was investigated by the methods of Western blotting, immunohistochemistry, and transmission electron microscope. LV hypertrophy was accessed by pathological analysis and plasma brain natriuretic peptide (BNP) level.

RESULTSLV hypertrophy was prominent in untreated SHR. In SHR, LV myocardium Cx43 level was upregulated, and the distribution of Cx43 was displaced from their usual locations to other sites at various distances away from the intercalated disks. After atorvastatin treatment, myocardium Cx43 level was reduced in SHR-A, and the distribution of Cx43 gap junction became much regular and confined to intercalated disk. Statins also prevented LV hypertrophy in SHR.

CONCLUSIONSThese results provide novel in vivo evidence for the key role of Cx43 gap junctions in LV hypertrophy and the possible mechanism in anti-hypertrophic effect of statins. Atorvastatin treatment may have beneficial effects on LV hypertrophy in spontaneously hypertensive rats.

Animals ; Anticholesteremic Agents ; pharmacology ; Atorvastatin Calcium ; Blood Pressure ; drug effects ; Blotting, Western ; Connexin 43 ; metabolism ; Heart ; drug effects ; physiopathology ; Heptanoic Acids ; pharmacology ; Hydroxymethylglutaryl-CoA Reductase Inhibitors ; pharmacology ; Hypertrophy, Left Ventricular ; blood ; drug therapy ; metabolism ; pathology ; Immunohistochemistry ; Lipid Metabolism ; drug effects ; Lipids ; blood ; Male ; Microscopy, Electron, Transmission ; Myocardium ; metabolism ; pathology ; ultrastructure ; Natriuretic Peptide, Brain ; blood ; Pyrroles ; pharmacology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

OBJECTIVETo observe the immune response after the transplantation of a deproteinized heterogeneous bone scaffold and provides the theoretic reference for clinical practice.

METHODSThe fresh pig bone and deproteinized bone were transplanted respectively to establish BABL/C thigh muscle pouches model of male mice and take the samples for detection at 1, 2, 4, 6 weeks after operation. Lymphocyte stimulation index, subset analysis, serum specific antibody IgG, cytokine detection and topographic histologic reaction after implantation were investigated.

RESULTSAfter the transplantation of deproteinized bone, lymphocyte stimulation index, CD(4)(+) and CD(8)(+) T-lymphocyte subsets, serum specific antibody IgG and cytokines in deproteinized bone group were significantly lower than those in fresh pig bone group at each time point (P<0.05). The histological examination found that in fresh bone group at each time point, a large quantity of inflammatory cells infiltrated in the surrounding of bone graft, and they were mainly lymphocytes, including macrophages and monocytes. In deproteinized bone group, there were few inflammatory cells infiltration around bone graft one week after operation. The lymphocytes were decreased as time went by. At 6 weeks, fibroblasts and fibrous tissue grew into the graft, and osteoclasts and osteoprogenitor cells appeared on the verge.

CONCLUSIONSThe established heterogeneous deproteinized bone has low immunogenicity and is a potentially ideal scaffold material for bone tissue engineering.

Animals ; Bone Transplantation ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Swine ; Tissue Engineering ; methods ; Tissue Scaffolds ; Transplantation, Heterologous

OBJECTIVETo study the clinicopathological features of spindle cell rhabdomyosarcoma (SCRMS) in order to differentiate it from other myosarcomas.

METHODSThe clinical features, morphologic and immunohistochemical phenotypes of 8 SCRMSs were analyzed.

RESULTSSCRMS cells were found to be arranged in a fascicular or storiform pattern, in which a number of enlarged plump or polygonal shaped rhabdomyoblasts containing abundant eosinophilic cytoplasm with eccentrically placed enlarged hyperchromatic nuclei were mixed. Immunohistochemical staining results showed that vimentin, MyoD1, desmin, actin, myoglobin were positive in tumor cells, but S-100, plap, AE1/AE3, CK, CD117 negative. The follow-up data showed that four cases had died of the recurrent disease, one still alive and the remain three patients lost follow-up.

CONCLUSIONSpindle cell rhabdomyosarcoma is a rare embryonal rhabdomyosarcoma which occurs in the childhood or adulthood with a poor prognosis, and is frequently presented as a painless mass most frequently involveing the head and neck or cervical area or para-testis site. A combination of MyoD1, desmin and myoglobin immunohistochemical staining is helpful in differential diagnosis.

Adolescent ; Adult ; Child ; Combined Modality Therapy ; Desmin ; metabolism ; Diagnosis, Differential ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; MyoD Protein ; metabolism ; Myoglobin ; metabolism ; Neoplasm Recurrence, Local ; Retrospective Studies ; Rhabdomyosarcoma, Embryonal ; metabolism ; pathology ; surgery ; Soft Tissue Neoplasms ; metabolism ; pathology ; surgery ; Young Adult

OBJECTIVETo analyze the early clinical and radiographic outcomes of Hoffa fractures treated by a standard protocol of open reduction and internal fixation using headless compression screws combined with back buttress plate in a consecutive series of 8 Chinese patients.

METHODSOpen reduction and internal fixation was performed on all patients. The fractures were anatomically reduced and held temporarily by K-wire. If the ends of fractures were atrophic, autologous bone graft from the ipsilateral iliac crest was packed between the ends. Then the fracture fragments were fixed with AO 6.5 mm headless compression cannulated screws. At least two screws were used to provide rotational stability. One pre-contoured reconstruction plate was placed on the nonarticular surface posteromedially or posterolaterally as back buttress plate.

RESULTSAll the patients were followed up for at least 12 months (range 12-25 months). All fractures achieved anatomical reduction and healed clinically and radiographically. At recent follow-up, the mean flexion degree was 120.6° (range 110°-135°) and the mean extension degree was 2.5° (range 0°-5°). The average visual analogue scale score was 1.6 points (range 0-3). Six patients were assessed as excellent and 2 as good according to the hospital for special surgery knee score system. There were no superficial or deep infections, or hardware breakages. No patient had giving way or locking of the knee, though some had intermittent pain and swelling after strenuous exercise. Injury mechanism had significant influence on the functional outcome (P=0.046).

CONCLUSIONHeadless compression screws combined with back buttress plate and/or autologous bone grafting to treat old Hoffa fracture is one of effective measures. It would be conducive to not only fracture healing but also early exercise and functional recovery.

Adult ; Bone Plates ; Bone Screws ; Female ; Femoral Fractures ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

OBJECTIVE: To report the prospective efficacy of 45 patients intracranial germinoma treated by radiotherapy and discuss its treatment. METHODS: From February 1998 to October 2007, a total of 45 intracranial germinoma patients were performed radiotherapy, including 15 combined chemotherapy in the Department of Oncology. Of them 23 were pathologically diagnosed while 22 cases were clinical diagnosed. Life table method showed the 5-year and 10-year survival rate. RESULTS: Forty patients were followed-up. Most symptoms of the patients were significantly reduced or disappeared completely. The 5-year and 10-year survival rate of all patients were 84% and 74%. CONCLUSION Radiotherapy is the main treatment for intracranial germinoma. Craniospinal irradiation, whole brain irradiation and partial brain irradiation are the main treatments. Radiotherapy combined with chemotherapy, which can reduce the radiation range and dose will be the trend.
Adolescent ; Adult ; Antineoplastic Agents ; therapeutic use ; Brain Neoplasms ; drug therapy ; mortality ; radiotherapy ; Child ; Female ; Follow-Up Studies ; Germinoma ; drug therapy ; radiotherapy ; Humans ; Male ; Young Adult