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Objective To explore the Helicobacter pylori (Hp) infection rate and antibody typing of 1111 physical examination people in plateau area, and to analyze the risk factors of Hp infection by logistics regression analysis. Methods 1111 healthy people with physical examination in plateau area from January 2022 to December 2022 were selected as the research subjects. The Hp infection rate and antibody typing were calculated, and the risk factors of Hp infection were analyzed by logistics regression analysis. Results The Hp infection rate of physical examination people in plateau area was 62.47% (694/1 111). The infection rate of type I HP in infected patients was higher than that of type Ⅱ HP(75.50% vs 24.50%) (χ2=361.141, P2=4.418, 8.708, 16.565, 32.583, P=0.036, 0.003, 2=5.153, P=0.023). Often eating pickled or barbecued foods [OR (95%CI)=2.038 (1.049-3.961)], history of chronic gastric disease [OR(95%CI)=1.706 (1.132-2.569)] and family members living together≥4 [OR (95%CI)=1.857 (1.135-3.037)] were risk factors of Hp infection, and regular garlic consumption [OR (95%CI)=0.559 (0.346-0.903)] was a protective factor (P=0.036, 0.011, 0.014, 0.018). Conclusion The Hp infection rate and antibody Ure positive rate are higher in physical examination people in plateau area, and chronic gastric disease history and often eating pickled or barbecued foods are risk factors of Hp infection.
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Dental fluorosis is a developmental disturbance of dental enamel caused by excessive fluoride intake during tooth development, leading to the changes in morphology, structure and function of tooth enamel, which can affect the aesthetics and function of teeth. There are many factors which may account for the occurrence of dental fluorosis. However, the pathogenesis mechanism underlying dental fluorosis has not been fully clarified.In recent years, researches in the fields of fluoride-induced stress response pathways, signaling pathways and apoptosis at the molecular and genetic level had provided extensive knowledge of dental fluorosis. This article focuses on the latest research progress in the mechanism of dental fluorosis, which include the effects of fluoride on ameloblasts and enamel matrix proteins, genetic polymorphism and dietary nutrients, in order to provide new references for the targeted prevention and treatment of dental fluorosis.
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Peri-implant disease, an important group of diseases that cause implant failure, are associated with metabolic abnormality. Metabolic syndrome (MetS) is a common metabolic disorder comprising abdominal obesity, hyperglycemia, systemic hypertension and atherogenic dyslipidemia. Previous studies had reported that MetS and its diversified clinical manifestations might be associated with peri-implant diseases, but the relationship and underlying mechanisms were unclear. This review aims to explore the relationship between MetS and peri-implant disease, in order to provide beneficial reference for the prevention and treatment of peri-implant disease in patients with MetS.
Subject(s)
Humans , Metabolic Syndrome/complications , Peri-Implantitis , Dental Implants/adverse effects , Hypertension/complications , Risk FactorsABSTRACT
OBJECTIVE@#To explore the association of single nucleotide polymorphisms (SNPs) of the vitamin D receptor gene ( VDR) with circulating lipids considering gender differences.@*METHODS@#Of the Han Chinese adults recruited from a health examination center for inclusion in the study, the circulating lipids, 25-hydroxyvitamin D (25OHD), and other parameters were measured. The VDR SNPs of Cdx2 (rs11568820), Fok1 (rs2228570), Apa1 (rs7975232), and Taq1 (rs731236) were genotyped with a qPCR test using blood DNA samples, and their associations with lipids were analyzed using logistic regression.@*RESULTS@#In the female participants ( n = 236 with dyslipidemia and 888 without dyslipidemia), multiple genotype models of Fok1 indicated a positive correlation of B (not A) alleles with LDLC level ( P < 0.05). In the male participants ( n = 299 with dyslipidemia and 564 without dyslipidemia), the recessive model of Cdx2 and the additive and recessive models of Fok1 differed ( P < 0.05) between the HDLC-classified subgroups, respectively, and Fok1 BB and Cdx2 TT presented interactions with 25OHD in the negative associations with HDLC ( P < 0.05).@*CONCLUSION@#In the Chinese Han adults included in the study, the Fok1 B-allele of VDR was associated with higher LDLC in females, and the Fok1 B-allele and the Cdx2 T-allele of VDR were associated with lower HDLC in males. The interaction of VD and Fok1 BB or Cdx2 TT in males synergistically decreased HDLC levels.
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Alleles , Asian People/genetics , China/ethnology , Dyslipidemias/genetics , Genetic Predisposition to Disease/genetics , Genotype , Lipids/blood , Polymorphism, Single Nucleotide , Receptors, Calcitriol/genetics , Sex Factors , Vitamin D/bloodABSTRACT
Objective:To explore the clinical efficacy of Chaihu Shugansan combined with saccharomyces boulardii in treating functional dyspepsia (FD) in children and the effect on serum cytokines and gastrointestinal hormones expression. Method:Totally 120 cases were divided into control group (60 cases) and observation group (60 cases).The control group was treated with Duopanlitong Pian 0.3 mg·kg-1, po, tid, and Saccharomyces boulardii 0.25 g,po,bid. The observation group was treated with Chaihu Shugansan, po,qd, and saccharomyces boulardii 0.25 g,po,bid. Four weeks was a course of treatment.The changes of serum Gastric Hunger Hormone and cytokines, Hamilton depression scale (HAMD) score, symptoms improvement time and clinical effect in the two groups were observed. And the recurrence rate was observed after 6 months follow-up. Result:The total effective rate was compared between control group (76.67%,46/60) and observation group (96.15%,57/60),with significant differences (P<0.01). After treatment, serum tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in both groups were decreased, while serum levels of ghrelin of both group were increased. However,the decreasing range and the increasing range of observation group were statistically obvious than those of control group (P<0.01). HAMD score and disappearance time of clinical symptoms in observation group was significantly different from those in control group (P<0.01). In adverse reactions during treatment, the control group was inferior to the observation group (P<0.05). After the 6-month follow-up, the recurrence rate in observation group was lower than that in control group (P<0.01). Conclusion:Chaihu Shugansan combined with Saccharomyces boulardii was effective in treatment of FD in children, with a low recurrence rate and good compliance. The mechanism may be related to the effective regulation of brain-intestinal axis, balance of intestinal flora, inhibition of inflammatory reaction, regulation of cytokines and gastrointestinal hormone levels.
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OBJECTIVE@#To explore the changes in bone height of the maxillary sinus floor at different sinus ridge heights after transcrestal sinus floor elevation (tSFE) with the simultaneous implantation of short implants.@*METHODS@#A total of 74 Bicon short implants were implanted into 37 patients during the same period of maxillary sinus elevation. The residual bone height (RBH)<4 mm group has 43 sites, and the RBH≥4 mm group has 31 sites. After 5 years of follow-up observation, the implant survival rate and the change in bone height achieved in the maxillary sinus over time were measured and analyzed via clinical examination and X-ray imaging.@*RESULTS@#In the 74 implantation sites, the elevation height of the sinus floor was (6.64±1.32) mm and the bone height of the sinus floor was (3.35±1.29) mm 5 years after loading. No statistical difference was observed in the bone resorption of the implant neck between the RBH<4 mm and RBH≥4 mm groups. Meanwhile, a statistical difference was noted in the bone height obtained in the maxillary sinus between the two groups.@*CONCLUSIONS@#When RBH in the maxillary posterior tooth area was <4 mm, the simultaneous implantation of Bicon short implants with tSFE can achieve a high implant survival rate and bone gain in the maxillary sinus, but does not increase the absorption of the alveolar ridge bone.
Subject(s)
Humans , Dental Implantation, Endosseous , Dental Implants , Maxilla , Maxillary Sinus/surgery , Retrospective Studies , Sinus Floor Augmentation , Treatment OutcomeABSTRACT
<p><b>OBJECTIVE</b>To express and purify the mouse endothelial cell-targeted recombinant Notch ligand protein mD1R, and to investigate its effect on hematopoiesis after carbon tetrachloride damage.</p><p><b>METHODS</b>PCR was performed to clone and construct the expression vector pET22b(+)-mD1R. The mD1R successfully transformed into E. coli was induced by IPTG, and purified with Ni-beads affinity chromatography. The target protein was detected by SDS-PAGE. The fluorescence-activated cell sorting analysis (FACS), cell adhesion test, immunofluorescence staining and quantitative real-time PCR were employed to detect the endothelial cell-targeted and Notch signaling-activated biological characteristics of mD1R. The carbon tetrachloride mouse model was established to observe the effects of mD1R on the hematopoietic stem cell (HSC), myeloid cells and lymphoid cells by flow cytometry. The LinScal-1c-Kit cells were sorted by magnetic bead, FACS was performed to analyze the cell cycle, and RT-PCR was employed to observe the expression of interleukin (IL)-10.</p><p><b>RESULTS</b>The prokaryotic expression vector was successfully cloned and constructed. The purity and the activity were confirmed in mD1R recombinant protein. The purified mD1R activated the Notch signaling pathway of hematopoietic stem cells in carbon tetrachloride damaged mouse, and internally elevated the number of HSC and long-term HSC to 2.96-fold and 6.18-fold. In addition, mD1R improved the amplification of the myeloid progenitor cells and the myeloid-derived suppressor cells, particularly the granulocyte/monocyte into blood. Mechanistically, the further analyses suggested that Notch pathway could increase the proliferation of HSC and enhance expression of IL-10 after stress injury.</p><p><b>CONCLUSIONS</b>A new and activated recombinant Notch ligand protein has been obtained successfully to communicate hematopoietic stem cells and hematopoietic microenvironment. The Notch- mediated intrinsic hematopoiesis has been regulated by the anti-inflammatory factor after stress injury.</p>
Subject(s)
Animals , Mice , Carbon Tetrachloride , Escherichia coli , Hematopoiesis , Hematopoietic Stem Cells , Ligands , Receptors, Notch , Signal TransductionABSTRACT
OBJECTIVE: To study the pharmacological function of the petroleum ether extract from Citrullus lanatus vine (PEECLV) in treating adjuvant arthritis(AA) and explore the mechanism. METHODS: AA in mice was made to observe the anti-inflammatory effect of PEECLV. Arthritis index was calculated by 5 grades evaluation. Thymus and spleen index were measured. The content of RF, PGE2, COX-1, COX-2 in serum were measured. The levels of TNF-α, IL-1β in serum were measured. RESULTS: PEECLV can suppress the foot swelling degree in model mice. It can increase thymus index, reduce spleen index and AI of AA in mice. High-dose group can significantly reduce serum PGE2, IL-1β, COX-2 and COX-1 levels in AA in mice. All groups can reduce TNF-α and RF levels. CONCLUSION: PEECLV has a certain degree therapeutic effect on AA, and the possible mechanism is to affect the production of inflammatory mediators and antioxidant.
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Objective To identify the success rate of peripheral indwelling needle puncture in pediatric patients, and analyze its influencing factors. Methods A survey was conducted in a sample of 902 pediatric patients.Personal information table of pediatric patients who receive infusionwas adopted to collect data. Results The success rate of first puncture was 85.37%(770/902), the success rate of two punctures was 95.34%(860/902). The success rate was affected by several factors, such as department, ages of the patients, condition of the veins, years of working as a nurse(OR=1.128, 2.308, 2.351, P <0.05). Conclusions Success rate of peripheral indwelling needle puncture in children still can be improved. When formulate management program, the influencing factors should be considered for management standard.
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<p><b>OBJECTIVE</b>To observe the effects of blood activating wind dissipating acupuncture (BAWDA) on blood pressure (BP) of prehypertension (PHT) patients.</p><p><b>METHODS</b>Totally 60 PHT patients were assigned to the control group and the acupuncture group according to random digit table, 30 in each group. All patients were intervened by life style. BAWDA was additionally performed in patients in the acupuncture group for 6 weeks (30 times). The improvement of BP after intervened by acupuncture was observed. BP success rates and the proportion of PHT progressing to hypertension (HT) were also observed after 6-week intervention of acupuncture and at 1-year follow-up.</p><p><b>RESULTS</b>Systolic blood pressure (SBP) and diastolic blood pressure (DBP) decreased after 6-week intervention in the acupuncture. The BP control rate was 56.7% (17/30 cases) in the acupuncture group vs.10.0% (3/30 cases) in the control group with statistical difference (chi2 = 14.70, P < 0.01). At 1-year follow-up BP success rate was 36.7% (11/30 cases) in the acupuncture group, remarkably higher than that of the control group [13.3%, (4/30 cases)] (chi2 = 4.36, P < 0.05).</p><p><b>CONCLUSIONS</b>BAWDA showed BP regulating roles in a gradually stable decreasing tendency. It also could elevate BP success rate of PHT, and reduce the risk of PHT progressing to HT.</p>
Subject(s)
Humans , Acupuncture , Methods , Acupuncture Therapy , Methods , Blood Pressure , Hypertension , Prehypertension , Therapeutics , WindABSTRACT
To analyze the effects of hyaluronidase and hirudoid treatment on drug extravasation in neonates. The medical records of 13 neonates with drug extravasation treated with hyaluronidase and hirudoid between August 1[st], 2010 and May 1[st], 2012 were analyzed retrospectively. The treatment procedure for drug extravasation adhered to the protocol in neonatal department The information including age, sex, weight, diagnosis, size of affected area, site of extravasation and treatment was collected. The extravasation injuries alleviated and the symptoms improved after treatment, no adverse drug effects were reported with use of hyaluronidase and hirudoid. The treatment appeared to be beneficial in the management of extravasations of various medications in neonates and may be useful in reducing the severity of cutaneous toxicosis. However, further studies with large samples are still needed to assess the effectiveness and safety of hyaluronidase and hirudoid
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OBJECTIVE: To investigate the hypoglycemic effects of different fractions from Citrullus lanatus vine on diabetic model induced by alloxan in mice.
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<p><b>OBJECTIVE</b>To study in vitro sperm damage caused by trichloroethylene in male rats.</p><p><b>METHODS</b>Sperms of Sprague-Dawley (SD) rats were collected 4 hours after being contaminated by trichloroethylene of 0, 2, 4, 6, 8, and 10 mmol/L in vitro. Giemsa staining was performed to observe the morphological changes of sperms, and flow cytometer was used to detect the changes in mitochondrial membrane potential.</p><p><b>RESULTS</b>The sperm motilities in 6, 8, and 10 mmol/L trichloroethylene groups decreased significantly compared with that in control group (P <0.01); the sperm aberration rates in 8 and 10 mmol/L trichloroethylene groups were significantly higher than that in control group (P<0.01). With the increase in exposure dose, the proportion of sperms with reduced mitochondrial membrane potential increased, and there were significant differences in sperm apoptosis rate between the 4, 6, 8, and 10 mmol/L trichloroethylene groups and control group (P<0.01).</p><p><b>CONCLUSION</b>In vitro exposure to trichloroethylene can reduce sperm motility and increase the aberration rate and apoptosis rate of sperms in male SD rats.</p>
Subject(s)
Animals , Male , Rats , Apoptosis , Membrane Potential, Mitochondrial , Rats, Sprague-Dawley , Sperm Motility , Spermatozoa , Cell Biology , Trichloroethylene , ToxicityABSTRACT
<p><b>OBJECTIVE</b>To explore the effects of bisphenol A (BPA) exposure on toxicity characteristic and OCT4 and SOX2 gene expression of mouse embryonic stem cells (mESC).</p><p><b>METHODS</b>mESC were cultured, and treated with the doses of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L respectively of BPA and DMSO (the solvent control group)for 24 hours, and three groups of cells were treated with the same method. The morphological changes of mESC in the control and exposure groups were observed through an inverted microscope. Cell counting kit 8 (CCK8) was used to detect the effects of BPA on proliferation of mESC, and based on the results, the half inhibitory concentration (IC50) was calculated. Real-time fluorescent quantitative polymerase chain reaction (RT-QPCR) and western blotting were used to detect the expression of OCT4 and SOX2.</p><p><b>RESULTS</b>BPA had certain toxicity on mESC, the treatment of BPA significantly increased cell toxicity in a concentration-dependent manner, and the IC50 was 4.3×10(-4) mol/L, combined with the BPA exposure concentration of the environment and the related literature, eventually taking the five concentrations of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L as the experimental groups. The mESC morphology were effected after the treatment of BPA for 24 h, compared with the control group, the number of cells decreased, appearing some floating cells, and the cell cloning became irregular and differentiation in the higher concentration groups. The OCT4 mRNA expression level in the 10(-7) mol/L (1.146 ± 0.087), 10(-6) mol/L (1.156 ± 0.030), 10(-5) mol/L (1.158 ± 0.103) and the 10(-4) mol/L (1.374 ± 0.053) dose group were all significantly higher than the control group (1.000 ± 0.000) (t values were -2.384, -2.953, -3.203, -4.021 respectively, P value all < 0.05). Meanwhile, the SOX2 mRNA expression level in the 10(-4) mol/L (1.113 ± 0.052) were higher than the control group (1.000 ± 0.000) (t value was -2.765, P value < 0.05). Moreover, the OCT4 protein expression level in the 10(-5) mol/L (1.360 ± 0.168) and 10(-4) mol/L (1.602 ± 0.151) were all significantly higher than the control group (1.000 ± 0.000) (t values were -3.538, -4.002 respectively, P value all < 0.05), while no obvious change of the SOX2 protein expression level was detected in all treated groups.</p><p><b>CONCLUSION</b>BPA in a certain dose range could upregulate the expression of OCT4 gene in mouse embryonic stem cells while had no significant effect on the expression of SOX2 gene.</p>
Subject(s)
Animals , Mice , Benzhydryl Compounds , Toxicity , Cells, Cultured , Embryonic Stem Cells , Metabolism , Gene Expression , Octamer Transcription Factor-3 , Genetics , Phenols , Toxicity , SOXB1 Transcription Factors , Genetics , Signal TransductionABSTRACT
The prevalence of human papilloma virus (HPV)-16 in patients with cervical cancer, the physical status of HPV-16 in patients with cervical lesions, and the role of HPV-16 integration in cervical carcinogenesis were investigated. HPV genotyping was performed by using PCR approach with the primer GP5+/GP6+ and type-specific primer on biopsy specimens taken operatively from 198 women. Multiple PCR was done to detect physical status of HPV-16 in a series of cervical liquid-based cytology samples and biopsy specimens obtained from different cervical lesions with HPV-16 infection, including 112 specimens with cervical cancer, 151 specimens with CIN I, 246 specimens with CIN and 120 specimens with CINIII. The results showed that there were 112 cervical cancer samples (56.57% of total cervical cancer patients) with HPV-16 infection. The frequency of HPV-16 pure integration was 65.18% (73/112), 56.57% (47/120), 23.58% (58/246) and 7.95% (12/151) in cervical cancer, CINIII, CINII and CINI patients respectively. In situ hybridization was performed on some paraffin-embedded sections of CINII, CINIII and cervical cancer to verify the physical status of HPV-16 infection. Significant difference was observed between cervical cancer and CIN I, CINII, CINIII in the frequency of HPV-16 integration (P<0.01). It is suggested that HPV-16 is the most prevalent type and is associated with cervical cancer. In the case of HPV-16 infection there are close associations between the severity of cervical lesions and the frequency of HPV-16 integration. The application of testing HPV genotyping and physical status based on detection of HC-II HPV DNA would be in favor of predicting the prognosis of cervical precancerosis and enhancing the screening accuracy of cervical cancer.
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<p><b>OBJECTIVE</b>To investigate the indications, safety and efficacy of catheter directed thrombolysis for early left lower extremity deep venous thrombosis (DVT) without vena cava filters protection.</p><p><b>METHODS</b>Clinical data of 54 cases of early left lower extremity DVT received catheter directed thrombolysis without vena cava filters from July 2008 to June 2010 were retrospectively analyzed. The thrombosis was entire without free floating clots and no thrombosis in vena cava detected with ultrasound scan. Twenty-five patients were male and 29 were female with the average age of 52.8 years. Fifty-one of which were iliofemoral and popliteal, the other 3 were iliofemoral. The course were ≤ 7 d in 45 cases and these were 8 to 30 d in 9 cases. Urokinase of 300 000 U was infused through catheters per 2 h twice a day. Meanwhile 4000 U of low weight heparin was administered subcutaneously per 12 h, or heparin infusion at dosage of 18 U×kg(-1)×h(-1).</p><p><b>RESULTS</b>The procedure technically succeeded in all patients. In total cases venous score decreased to 4.6 ± 2.1 post 6 to 10 d of thrombolysis from 10.8 ± 1.0 with thrombolysis rate of 58% ± 18% which was not significantly different between groups of ≤ 7 d and 8 to 30 d (t = 1.02, P = 0.34). On 14(th) day, 11 patients (20.4%) completely recovered, 35 cases (64.8%) experienced large improvement, 8 patients (14.8%) had mild improvement and nobody was failed, resulting in total efficacy of 100%. No patient developed clinical symptomatic pulmonary embolism. SpO2 did not alter markedly post thrombolysis [(91.0 ± 2.6)% vs. (90.8 ± 2.4)%, t = 2.03, P = 0.05]. No patients suffered from cerebral hemorrhage and haemoturia, and catheter induced inflammation occurred in 4 cases (7.41%). There was mild bleeding in puncture sites in 11 patients (20.4%) during the course. There were 36 patients (66.7%) had been followed up with the time of 6 to 21 months. In which 31 cases had no lower extremity edema or had mild edema after activities. Two patients developed serious edema after activities for deep venous insufficiency. Three cases combined with malignant tumor or renal failure recurred.</p><p><b>CONCLUSIONS</b>For early left extremity DVT which is entire without free floating clots and no thrombosis in vena cava, catheter directed thrombolysis without filter protection maybe administered with safety, efficiency and lower expense.</p>
Subject(s)
Female , Humans , Male , Middle Aged , Catheterization, Peripheral , Fibrinolytic Agents , Therapeutic Uses , Follow-Up Studies , Lower Extremity , Pulmonary Embolism , Retrospective Studies , Thrombolytic Therapy , Methods , Urokinase-Type Plasminogen Activator , Therapeutic Uses , Vena Cava Filters , Venous Thrombosis , TherapeuticsABSTRACT
Objective This study is to understand the interpersonal conflict management styles of head nurses and its related factors. Methods A cross- sectional survey was conducted in 440 head nurses in Shanghai Hospital, using"personal information table"and the "Rahim Organizational Conflict Inventory- Ⅱ"as the tools for data collection. Results When dealing with conflicts, "integrating"is the most commonly used approach of head nurses, while "dominating"the least used one. Age, education, work experience and other factors were statistically significant with the conflict management styles head nurses used. Conclusions The most commonly used conflict management style of head nurses tends to a winwin way. Age, education, work experience and other factors can affect the ways of managing conflict.
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<p><b>OBJECTIVE</b>To investigate the expression of macrophage migration-inhibitory factors (MIF) in clinically healthy and inflamed human pulp tissues and the effects of rhMIF on the proliferation of human dental pulp cells (HDPC).</p><p><b>METHODS</b>Immunohistochemistry was used to detect the localization of MIF expression in clinically healthy pulp and inflamed pulp tissues. Quantitative real-time polymerase chain reaction (PCR) was performed to evaluate the mRNA levels of MIF in pulp specimens. In addition, the culture supernatants of HDPC were collected after HDPC was stimulated by lipopolysaccharide (LPS) for 24 h, and then the MIF levels were assayed by quantitative sandwich enzyme-linked immunosorbent assay. Meanwhile, the effects of rhMIF on the proliferation of HDPC at different concentrations for 24 and 48 h were observed by cell counting kit-8 (CCK-8).</p><p><b>RESULTS</b>MIF was mainly distributed in odontoblasts of healthy pulp tissue, however, in inflamed pulp tissue, it was widely detected in fibroblasts, inflammatory infiltrates and endothelial cells as well as odontoblasts. Quantitative real-time PCR showed that there was no significant difference in MIF mRNA levels between inflamed pulps and healthy pulps (P > 0.05). Additionally, the secretion of MIF was significantly increased by stimulation with LPS at the concentration of 0.1 and 1.0 mg/L [(1772.58 ± 495.05), (1692.58 ± 337.45) ng/L] (P < 0.05), and the concentration was (1048.53 ± 161.81) ng/L in control group. rhMIF stimulated the HDPC's proliferation at the concentration of 10, 30, 60 µg/L for 24 and 48 h.</p><p><b>CONCLUSIONS</b>MIF was expressed in pulp tissue and its expression was increased after stimulation by LPS. rhMIF increased the proliferation of HDPC. These results suggest that MIF may be involved in the process of pulpal inflammation.</p>
Subject(s)
Humans , Cell Proliferation , Cells, Cultured , Dental Pulp , Cell Biology , Pathology , Dose-Response Relationship, Drug , Lipopolysaccharides , Pharmacology , Macrophage Migration-Inhibitory Factors , Genetics , Metabolism , Pulpitis , Metabolism , Pathology , RNA, Messenger , Metabolism , Reverse Transcriptase Polymerase Chain ReactionABSTRACT
<p><b>OBJECTIVE</b>To investigate DNA methylation variation in human cells induces by B(a)P, and to explore the role of PARP1 during this process.</p><p><b>METHODS</b>The changes of DNA methylation of 16HBE and its PARP1-deficient cells exposed to B(a)P (1.0, 2.0, 5.0, 10.0, 15.0, 30.0 µmol/L) were investigated by immunofluorescence and high performance capillary electrophoresis, and simultaneously, the expression level of PARP 1 and DNMT 1 were monitored dynamically.</p><p><b>RESULTS</b>The percentage of methylated DNA of overall genome (mCpG%) in 16HBE and 16HBE-shPARP1 cells were separately (4.04 ± 0.08)% and (9.69 ± 0.50)%. After being treated by 5-DAC for 72 hours, mCpG% decreased to (3.15 ± 0.14)% and (6.07 ± 0.54)%. After both being exposed to B(a)P for 72 hours, the mCpG% in 16HBE group (ascending rank) were separately (5.10 ± 0.13), (4.25 ± 0.10), (3.91 ± 0.10), (4.23 ± 0.27), (3.70 ± 0.15), (3.08 ± 0.07); while the figures in 16HBE-shPARP1 group (ascending rank) were respectively (10.63 ± 0.60), (13.08 ± 0.68), (9.75 ± 0.55), (7.32 ± 0.67), (6.90 ± 0.49) and (6.27 ± 0.21). The difference of the results was statistically significant (F values were 61.67 and 60.91, P < 0.01). For 16HBE group, expression of PARP 1 and DNMT 1 were 141.0%, 158.0%, 167.0%, 239.0%, 149.0%, 82.9% and 108.0%, 117.0%, 125.0%, 162.0%, 275.0%, 233.0% comparing with the control group, whose difference also has statistical significance (t values were 11.45, 17.32, 32.24, 33.44, 20.21 and 9.87, P < 0.01). For 16HBE-shPARP1 group, expression of PARP 1 and DNMT 1 were 169.0%, 217.0%, 259.0%, 323.0%, 321.0%, 256.0% and 86.0%, 135.0%, 151.0%, 180.0%, 229.0%, 186.0% comparing with the control group, with statistical significance (t values were 9.06, 15.92, 22.68, 26.23, 37.19 and 21.15, P < 0.01). When the dose of B(a)P reached 5.0 µmol/L, the mRNA expression of DNMT 1 in 16HBE group (ascending rank) were 125.0%, 162.0%, 275.0%, 233.0% times of it in control group, with statistical significance (t values were 12.74, 24.92, 55.11, 59.07, P < 0.01); while the dose of B(a)P reached 2.0 µmol/L, the mRNA expression of DNMT 1 in 16HBE-shPARP1 group were 135.0%, 151.0%, 180.0%, 229.0%, 186.0% of the results in control group, and the differences were statistically significant (t values were 23.82, 40.17, 32.69, 74.85, 46.76, P < 0.01).</p><p><b>CONCLUSION</b>The hypomethylation of 16HBE cells induced by B(a)P might be one important molecular phenomenon in its malignant transformation process. It suggests that PARP1 could regulate DNA methylation by inhibiting the enzyme activity of DNMT1, and this effect could be alleviated by PARP1-deficiency.</p>
Subject(s)
Humans , Benzo(a)pyrene , Cell Line , DNA (Cytosine-5-)-Methyltransferase 1 , DNA (Cytosine-5-)-Methyltransferases , Genetics , Metabolism , DNA Damage , DNA Methylation , Epithelial Cells , Metabolism , Poly (ADP-Ribose) Polymerase-1 , Poly(ADP-ribose) Polymerases , Genetics , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To construct DNA methyltransferase 1 (DNMT1) low expression 16HBE cell line and observe the variation of cell cycle and global genomic DNA methylation.</p><p><b>METHODS</b>The method of Lenti-virus induced RNA interference was applied to introduce four different shRNA fragment into 16HBE cells. Flow cytometry and 5-mC immunofluorescence methods were used to observe the cell cycle and global DNA methylation status of DNMT1 low expression 16HBE cells.</p><p><b>RESULTS</b>The DNMT1 protein relative expression level of 16HBE-shDNMT1-4 cell line was down regulated about 44% (P < 0.05) compared with the control. No obvious differences of cell cycle and global genome DNA methylation status were observed between the 16HBE and 16HBE-shDNMT1.</p><p><b>CONCLUSION</b>The DNMT1 gene low expression cell is successfully constructed, and there are no obvious changes happened on the cell cycle and global genomic DNA methylation.</p>