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.

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

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 investigate the association between HPV genotypes and cervical lesion in Hybrid Capture 2 (HC2) HPV test positive samples.

METHODS602 cervical samples randomly obtained detected as HPV positive by the HC2 high-risk probe cocktail were determined by polymerase chain reaction (PCR)-reverse dot blot (RDB), among them 344 participated Thinprep Cytology test.

RESULTS569 (94.5%) samples were successfully amplified. The most common HPV genotypes were HPV16 (31.6%), 52 (16.7%), 58 (15.1%), 56 (8.1%), 39 (7.9%); HPV16 or HPV16/18 infection was significantly related to ASCUS, LSIL and HSIL; 24 samples suggested single HPV infection with the genotypes not available for HC2.

CONCLUSIONThe most common HPV genotypes in random screening were HPV16, 52 and HPV58; Our data demonstrated the pseudo-positivity of HC2 test was 4.0%; HPV16 was a index for progression of HSIL.

Adult ; Aged ; Alphapapillomavirus ; classification ; genetics ; isolation & purification ; Cervix Uteri ; pathology ; virology ; China ; epidemiology ; Female ; Humans ; Middle Aged ; Papillomavirus Infections ; diagnosis ; epidemiology ; pathology ; virology ; Risk Factors ; Uterine Cervical Diseases ; diagnosis ; epidemiology ; pathology ; virology ; Vaginal Smears ; Young Adult

OBJECTIVETo observe the infection of the high risk human papillomavirus (HPV) in the specimen of the small cell neuroendocrine carcinoma of the uterine cervix (SCNEC).

METHODSWe extracted the nucleic acids in the formalin-fixed and paraffin-embedded specimen from a 33-year-old patient diagnosed as small cell neuroendocrine carcinoma of uterine cervix and detected the HPV genotype with the nested PCR.

RESULTSWe identified HPV18, a high-risk genotype, in the specimen.

CONCLUSIONThe HPV detection with the nested PCR was available for identification of the HPV genotype(s) in the paraffin-embedded specimens of small cell neuroendocrine carcinoma of the uterine cervix with a high accuracy and sensitivity.

Adult ; Base Sequence ; Carcinoma, Neuroendocrine ; virology ; Carcinoma, Small Cell ; virology ; Female ; Human papillomavirus 18 ; genetics ; isolation & purification ; Humans ; Molecular Sequence Data ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; virology

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 investigate the infection of the high risk human papillomavirus (HPV) in the specimen of the clear cell carcinoma of the cervix.

METHODSWe extracted the nucleic acids in the formalin-fixed and paraffin-embedded specimens from a 37-year-old patient with clear cell carcinoma of the cervix and detected the HPV genotype with the nested PCR.

RESULTSWe identified HPV18, a high-risk genotype, in the specimens.

CONCLUSIONThe HPV detection with the nested PCR was available for identification of the HPV genotype(s) in the paraffin-embedded specimens of clear cell carcinoma of the cervix (CCCC) with a high accuracy and sensitivity.

Adenocarcinoma, Clear Cell ; diagnosis ; virology ; Adult ; Female ; Genotype ; Human papillomavirus 18 ; genetics ; isolation & purification ; Humans ; Papillomavirus Infections ; diagnosis ; virology ; Paraffin Embedding ; Polymerase Chain Reaction ; Sensitivity and Specificity ; Uterine Cervical Neoplasms ; diagnosis ; virology

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