Objective:To investigate if transforming growth factor-?1 (TGF-?1) induces apoptosis in odontoblast cell line MDPC-23. Methods:MDPC-23 cells were treated with TGF-?1 at 0.5,2.5 or 10 ng/ml for 48 h,apoptosis of MDPC-23 cells was detected by annexin V and propidium iodide (PI) staining, cell death detection ELISA and DNA electrophoretic analysis. Results: Consistent results were obtained from three different methods.TGF-?1 at 0.5-10 ng/ml induced apoptosis of MDPC-23 cells in a dose-dependent manner. Conclusion:TGF-?1 may paly a role in apoptosis of odontoblasts.

Objective: Our purpose was to study the effect of L-arginine (L-Arg) on the kidneys of rats with obstructive jaundice and the underlying mechanism. Methods: Forty male Sprague-Dowley rats, weighting 200 to 230 g, were randomly assigned to four groups (n=10): sham-operation group, control group, L-Arg group, L-NG-nitroarginine methyl (L-NAME) group.Except the sham-operation group,the other groups rendered jaundiced by doubly ligating the common bile duct. Fourteen days later,we administrated L-Arg to L-Arg group,and L-NAME to L-NAME group for 7 days.On the 21st day after bile duct ligation, we measured the amount of nitric oxide (NO), malondrialdehyde (MDA), and superoxide dismutase (SOD) in the kidneys, and assessed the changes of the renal function.We also observed the morphological changes of the kidneys. Results: In the control group,the amount of NO and MDA increased while that of SOD decreased (P<0.01, P<0.01and P<0.01), and the renal function and the nephric tissue were impaired on the 21st day after bile duct ligation compared with the sham-operation group.In the L-Arg group, the amount of NO and SOD increased significantly (P<0.01, P<0.01), whereas that of MDA decreased (P<0.05), and the damage in the nephric tissue and renal function was alleviated,as compared with the control group.On the contrary, the amount of NO in the L-NAME group decreased significantly (P<0.05) compared with the control group.Although there was no significant difference in the levels of SOD, MDA, and the parameters of the renal function between the L-NAME group and control group,those indexes in the L-NAME group tended to be aggravated. Moreover, the damage in the nephric tissue of the L-NAME group was not alleviated. Conclusion: L-arginine may protect the kidney from impairment in rats with obstructive jaundice through L-Arg-NO pathway.

Objective To investigate the inhibitory effects and possible related mechanism of OTX008 [a selective inhibitor of galectin-1 (Galectin-1)] on retinal neovascularization (RNV) in mouse model of oxygeninduced retinopathy (OIR).Methods 7-day-old (P7) C57BL/6J mice were randomly (according to random number table) divided into 4 groups including normal group,OIR group,OIR-OTX008 group and OIRphosphate buffered saline (PBS) group.To establish the OIR mouse model,mice from all groups except normal group were expose to (75±2)％ oxygen for 5 days and then to room air.OIR-OTX008 group received an intravitreal injection of 1 μl (0.25 μg/μl) OTX008 at P12,OIR-PBS group received the equal volume (1 μl) of PBS injection.Mice from 4 groups were euthanized at P17,and retinas were collected for molecular biological analysis and morphological study.RNV was evaluated by counting the number ofpre-retinal neovascular nuclei and the whole-mount immunofluorescent staining of mouse retina.Cyrosections of retinas were imaged via confocal microscopy to observe the enrichment of staining of Galectin-1.Protein levels of Galectin-1,Neuropilin-1 and phosphorylation of vascular endothelial growth factor receptor 2 (pVEGFR2) were determined with Western blot.Results At P17,Galectin-1 expressed higher in retinal ganglion cell layer,inner plexiform layer and inner nuclear layer from OIR group and OIR-PBS group than normal group.Galectin-1 expressed less in cryosection retinas from OIR-OTX008 group than OIR group and OIR-PBS group.The numbers ofpre-retinal neovascular cell nuclei from OIR group and OIR-pBS group were obviously more than that from normal group (t=9.314,P＜0.05).The number of pre-retinal neovascular cell nuclei from OIR-OTX008 group were obviously lower than those from OIR group and OIR-PBS group (t=8.038,7.774;P＜0.05).The RNV tufts area (t=13.250,12.570),non-perfusion area (t=15.590,12.430) and hypoxic area (t=9.542,9.928) from OIR-OTX008 group were significantly smaller than those in OIR group and OIR-PBS group (P＜ 0.05).Protein levels of Galectin-1 (t=24.800,23.060),Neuropilin-1 (t=4.120,3.530) and pVEGFR2 (t=25.880,15.480) in the OIR-OTX008 group were significantly down-regulated than those from OIR group and OIR-PBS group (P＜0.05).Conclusion Intravitreal injection of OTX008 inhibits RNV and ameliorates retinal hypoxia in mice model of OIR possibly through down-regulating Galectin-1,Neurolinpin-1 and pVEGFR2.

Objective:To clone and sequence cDNA of MH2 domain of Smad1 gene from human dental papilla cells. Methods:Total RNA was isolated from primarily cultured human dental papilla cells and reversely transcribed into single stranded cDNA.The desired DNA product was obtained by PCR with two gene specific primers.The segment was inserted into PUC19 vector and the plasmid was transformed into E.coli JM109.The double stranded cDNA of positive clone was sequenced.Results:The sequence of MHz domain of Smad1 cloned from human dental papilla cells was consistent with that reported by Hoodless et al. Conclusion:Smad1 exists in human dental papilla cells,BMP signaling may be mediated by smad1 in human dental papilla cells.

Objective To evaluate the effects of dexmedetomidine on the cellular immune function during analgesia with morphine after radical resection for esophageal cancer in the patients.Methods Sixty patients of both sexes,of ASA physical status Ⅰ or Ⅱ,after radical resection for esophageal cancer under general anesthesia,were randomly divided into 2 groups (n =30 each) using a random number table:control group (group C) and dexmedetomidine group (group Dex).Patient-controlled intravenous analgesia (PCIA) was performed immediately after operation in the two groups.In group C,the PCIA solution (150 ml) contained morphine 0.48 mg/kg.In group Dex,the PCIA solution (150 ml) contained morphine 0.48 mg/kg and dexmedetomidine 1 μg/kg.The postoperative visual analogue scale (VAS) scores were maintained ≤ 3.The consumption of morphine was recorded within 24,48 and 72 h after operation.The adverse effects such as nausea,vomiting,pruritus,bradycardia,hypotension,oversedation and respiratory depression were also recorded after operation.Before induction of anesthesia (T0),immediately after extubation (T1),and at 24,48 and 72 h after operation (T2-4),venous blood samples were obtained for determination of the levels of T-lymphocyte subsets (CD3+,CD4+,CD8+) and natural killer (NK) cells by flow cytometry.CD4+/CD8+ ratio was calculated.Results Compared with group C,the consumption of morphine within 24,48 and 72 h after operation and incidence of nausea,vomiting and pruritus after operation were significantly decreased in group Dex.The levels of CD3+,CD4+,CD4+/CD8+ ratio and NK cells were significantly lower at T1-4 than at T0 in the two groups.The levels of CD3+,CD4+,CD4+/CD8+ ratio and NK cells were significantly higher at T1-4 in group Dex than in group C.Conclusion Dexmedetomidine can improve the cellular immune function during analgesia with morphine after radical resection for esophageal cancer in the patients.

Background and purpose:Aberrant DNA methylation that leads to the inactivation of tumor suppressor genes plays important roles in development and progression of breast cancer. Clinically, related gene methylation is considered to be a promising biomarker for tumor diagnosis and prognosis. This study aimed to investigate the methylation status ofSox17 gene in breast cancer tissue and its corresponding plasma circulating DNA, as well as to investigate its value in breast cancer early diagnosis and prognosis.Methods:TheSox17 gene promoter methylation status was detected by MSP in 86 cases of breast cancer, 36 normal breast tissues and its paired plasma DNA, the results were analyzed with corresponding clinical and pathological features.Results:The frequency ofSox17 gene methylation rate among 86 breast cancer tissues was 77.9%(67/86), and was 61.6%(53/86)in plasma circulating DNA, however, noSox17 gene methylation was found in normal breast tissues.Sox17 gene promoter methylation in plasma circulating DNA was signiifcantly associated with the methylation status in tumor tissues (r=0.502,P=0.000). In breast cancer tissue specimens,Sox17 methylation status was significantly correlated with tumor stage (χ2=6.18,P=0.041) and lymph node metastasis (χ2=13.54,P=0.001);Sox17 gene methylation rate was signiifcantly correlated with tumor stage (χ2=27.06,P=0.000), tumor size (χ2=9.65,P=0.007) and lymph node metastasis (χ2=20.80,P=0.000) in plasma samples, and there was no signiifcant difference ofSox17 gene methylation between patient age, histological grade and ER, PR, HER-2/neu status.Conclusion:Sox17 gene promoter methylation plays an important role in the carcinogenesis and development of breast cancer, and may be associated with the prognosis of breast cancer. Furthermore, methylatedSox17 gene may be a useful tumor biomarker in plasma circulating DNA for breast cancer detection and disease monitoring.

Objective To evaluate the efficacy of dexmedetomidine and dezocine used to supplement awake tracheal intubation assisted by fiberoptic bronchoscope (FOB) in elderly patients.Methods Sixty elderly patients aged 65-77 yr,of ASA physical status Ⅱ or Ⅲ (Mallampati grade Ⅰ or Ⅱ),scheduled for elective surgery under general anesthesia,were randomly divided into 3 groups (n =20 each) using a random number table:dezocine group (group DEZ),dexmedetomidine group (group DEX) and dezocine combined with dexmedetomidine group (group DEZ+DEX).Dezocine 0.1 mg/kg was injected intravenously in group DEZ.Dexmedetomidine 0.4 μg/kg was infused intravenously over 10-15 min in group DEX.In group DEZ+DEX,dexmedetomidine 0.4 μg/kg was infused intravenously over 10-15 min,and dezocine 0.1 mg/kg was injected simultaneously.Laryngeal mucous membrane was sprayed with 2％ lidocaine for topical anesthesia during infusion in all the three groups.In addition,1％ tetracaine 3 ml was injected into trachea through cricothyroid membrane.Awake tracheal intubation was performed and assisted by FOB after the end of administration in all the three groups.Cardiovascular response (MAP or HR＞30％ of baseline values) and respiratory depression (SpO2＜90％ and RR＜8 bpm) were recorded during the period between induction of anesthesia and 3 min after intubation was completed.The intubation time was recorded.The tolerance of tracheal tube was assessed in the patients.At the time of topical anesthesia,when epiglottis came into view,immediately after tracheal tube was successfully inserted into trachea,and at 3 min after successful intubation,perfusion index and Ramsay sedation score,and patients' satisfaction with the sedation (Ramsay sedation score 2-4) were recorded.Results Compared with group DEZ or DEX,the tolerance of tracheal tube was significantly enhanced,intubation time was shortened,the rate of satisfactory sedation was increased,perfusion index and the incidence of cardiovascular response were decreased in DEZ+DEX group.There was no significant difference in respiratory depression among the three groups.Conclusion Dexmedetomidine and dezocine can provide better condition for awake tracheal intubation assisted by FOB than dexmedetomidine or dezocine alone in elderly patients.

OBJECTIVE:To establish a method for the simultaneous determination of ornidazole and dexamethasone sodium phosphate in Compound ornidazole film. METHODS:HPLC was performed on the column of Inertsil ODS-3 with mobile phase of methanol-20 mmol/L phosphate buffer(pH was adjusted to 7.40 with glacial acetic acid)(55∶45,V/V)at a flow rate of 1.0 ml/min, column temperature was 30 ℃,detection wavelength was 242 nm and volume injection was 10 μl. RESULTS:The linear range were 1-100μg/ml for both ornidazole(r=0.999 7)and dexamethasone sodium phosphate(r=0.999 9);RSDs of precision,stabili-ty and reproducibility tests were lower than 2.0%;recoveries were 96.50%-99.80%(RSD=1.02%,n=9) and 96.50%-99.60%(RSD=0.99%,n=9). CONCLUSIONS:The method is specific with good precision and stability and high accuracy,and can be used for the simultaneous determination of ornidazole and dexamethasone sodium phosphate in Compound ornidazole film.

The differentiation of periodontal ligament (PDL) progenitor cells is important for maintaining the homeostasis of PDL tissue and alveolar bone. Vitamin C (VC), a water-soluble nutrient that cannot be biosynthesized by humans, is vital for mesenchymal stem cells differentiation and plays an important role in bone remodeling. Therefore, the objective of this study was to determine the function and mechanism of VC in PDL progenitor cells osteogenic differentiation at the molecular level. We demonstrated that VC could induce the osteogenic differentiation and maturation of PDL progenitor cell without other osteogenic agents. During the process, VC preferentially activated ERK1/2 but did not affect JNK or p38. Co-treatment with ERK inhibitor effectively decreased the Vitamin C-induced expression of Runx2. ERK inhibitor also abrogated Vitamin C-induced the minimized nodules formation. PELP1, a nuclear receptor co-regulator, was up-regulated under VC treatment. PELP1 knockdown inhibited ERK phosphorylation. The overexpression of PELP1 had a positive relationship with Runx2 expression. Taken together, we could make a conclude that VC induces the osteogenic differentiation of PDL progenitor cells via PELP1-ERK axis. Our finding implies that VC may have a potential in the regeneration medicine and application to periodontitis treatment.
Ascorbic Acid ; pharmacology ; Butadienes ; pharmacology ; Cell Differentiation ; drug effects ; Cells, Cultured ; Co-Repressor Proteins ; antagonists & inhibitors ; genetics ; metabolism ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Humans ; MAP Kinase Signaling System ; drug effects ; Mitogen-Activated Protein Kinase 1 ; antagonists & inhibitors ; metabolism ; Mitogen-Activated Protein Kinase 3 ; antagonists & inhibitors ; metabolism ; Nitriles ; pharmacology ; Periodontal Ligament ; cytology ; Phosphorylation ; drug effects ; RNA Interference ; RNA, Small Interfering ; metabolism ; Stem Cells ; cytology ; Transcription Factors ; antagonists & inhibitors ; genetics ; metabolism ; Up-Regulation ; drug effects

AIM:To investigate the expression of centromere protein-H(CENP-H)in adrenocortical carcino-ma(ACC)and its relationship with disease progression and prognosis,and to explore the impact of CENP-H gene knock-down on the viability and migration of ACC cells.METHODS:The mRNA expression level of CENP-H in 76 ACC pa-tients and 128 healthy controls,and its correlations with tumor stages and prognosis were analyzed by GEPIA2 database.The mRNA expression of CENP-H in different stages of ACC and its correlation with disease prognosis were further ana-lyzed by ULCAN database.The protein expression of CENP-H was examined by immunohistochemical staining of paraffin-embedded ACC and normal adrenal gland specimens.Knockdown of CENP-H by siRNA(siCENP-H)was performed in human ACC cell line H295R.The viabilty of H295R cells transfected with siCENP-H or siNC was measured by CCK-8 as-say,the cell migration was detected by wound-healing assay,and the protein levels of CENP-H,p-ERK1/2,t-ERK1/2,p-P38,t-P38,p-JNK1/2 and t-JNK1/2 were detected by Western blot.RESULTS:The mRNA level of CENP-H was signifi-cantly higher in ACC than that in normal controls,and was correlated with tumor stages and prognosis.The protein level of CENP-H was significantly higher in ACC specimens than that in normal adrenal gland.Knockdown of CENP-H in H295R cells resulted in decreased cell viability and migration.The protein levels of p-P38 and p-JNK1/2 were decreased in si-CENP-H group.CONCLUSION:CENP-H is highly expressed in ACC,and is correlated with tumor stages and poor prognosis.Knockdown of CENP-H can inhibit the viability and migration of ACC cells,and its mechanism may related to inactivation of P38 and JNK signaling pathways.