OBJECTIVETo explore the relationship between salivary proteome and dental caries and to promote the biomarker studies of dental caries susceptibility by comparing the salivary proteome of caries-active children and caries-free children with electrospray ionization ion-trap tandem mass spectrometry (ESI-MS/MS).

METHODSTen caries-active children and ten caries-free children were sampled. The salivary proteome of the two groups was studied, and the differential protein between the two groups was analyzed by ESI-MS/MS after sodium dodecyl sulfate polyacrylamide gel electrophoresis, filter-aided sample preparation, and liquid chromatography.

RESULTSThe concentration of salivary protein was higher in the caries-active group than in the caries-free group. The polypeptide counts of thecaries-active and caries-free groups were 602 and 481, which belonged to 286 and 227 proteins, respectively. The differential polypeptide count of the two groups was 361, and the differential protein count was 118. The detected proteins included matrix metalloproteinase-9 (MMP9), mucin-7 (MUC7), lactotransferrin (LTF), carbonic anhydrase 6 (CA6), azurocidin (AZU), and cold agglutinin.

CONCLUSIONThe total salivary protein was higher in the caries-active group than in the caries-free group. The preliminary detection of differential proteins (MMP9, MUC7, LTF, CA6, AZU, and cold agglutinin) may lay some foundation for biomarker research of dental caries susceptibility.

Carbonic Anhydrases ; Child ; Dental Caries ; Humans ; Matrix Metalloproteinase 9 ; Proteome ; Saliva ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

AIM: To investigate the pharmacokinetic properties and bioequivalence of nifedipine sustained-release tablets after multiple doses administration in healthy volunteers. METHODS: Twenty two male healthy volunteers were enrolled in a randomized two-way crossover design with multiple doses (20 mg·d-1×7 d) study. Nitrendipine was used as the internal standard and the concentrations of nifedipine in plasma were determined by HPLC-APCI-MS. The pharmacokinetic parameters were calculated and the bioequivalence were compared by DAS (ver 1.0) program. RESULTS: The pharmacokinetic parameters of test and reference preparations were as follows: Cmax (52.5±27.4) and (54.0±31.2) ng·ml-1;Cmin (5.4±4.1) and (6.2±5.9) ng·ml-1;Cav (16.8±9.2) and (19.3±12.4) ng·ml-1;Tmax (3.7±0.9) and (4.1±1.1) h;t1/2 (8.9±4.9) and (8.5±3.1) h;AUC0-τ (403.4±221.0) and (461.9±296.6) μg·h·L-1, AUC0-36h (444.4±256.1) and (503.1±330.9) ng·h·ml-1;AUC0-∞ (482.1±268.9) and (542.3±348.4) ng·h·ml-1;DF (299.8±117.7)% and (279.2±97.5)%, respectively. There were no significant differences (P>0.05) in Tmax, Cmax, Cmin, Cav, DF, AUC0-τ, AUC0-36h, AUC0-∞ and t1/2 between the two preparations. The relative bioavailability of test tablets was (100.6±38.6)%. CONCLUSION:The test and reference preparations were bioequivalence.

The molecular delivery vectors used in gene therapy need provide the features of safety,stability, efficiency and capacity. The current studies on the structure and action mechanism of pRNA, a packaging RNA of phage?29, showed that pRNA with multiple binding sites can through cell membrane easily and escort exogenous molecules to target cell, without inducing immune reaction. As an ideal nano-scale gene therapy vehicle, pRNA presents a promising application in delivering multiple therapeutic components to detect and treat human diseases.

OBJECTIVE Autosomal dominant polycystic kidney disease (ADPKD) is a common-monogenetic disease characterized by progressive development of renal cysts. Thereis still further need for effective therapy.Based on our precious study that Ganoderma triterpenes(GT),which is the major secondary metabolites of Ganoderma lucidum,is able to attenuate renal cyst development.The aim of this study was to investigate the effect of a monomer,Ganoderic acid A(GA-A)that was purified from the GT,which has been reported to exhibit antinociceptive,antioxidative,hepatoproctive and anti-cancer activities,to have a potent anti-cyst effect in ADPKD. METHODS We first evaluated the potential cytotoxicity of GA-A on MDCK cells using a CCK-8 assay.Then we used MDCK cyst model,cultivated MDCK cells in vitro to form fluid-filled cysts surrounded by monolayer cells.MDCK cells were co-incu-bated with 10 μmol·L-1FSK with or without GA-A(25 μg·mL-1)and equal concentration GT as positive control from day 0 to day 6 to investigate the inhibitory effect of GA-A on cyst formation.And to further investigate the inhibitory effect of GA-A on cyst enlargement, MDCK cysts were treated with different concentration of GA-A(6.25,25 and 100 μg·mL-1)from day 5 to day 12.Next we used an embryonic kidney cyst model, wile-type mice kidneys were taken out on embryonic day 13.5 to form renal cysts stimulated with 8-Br-cAMP to prove the renal cyst inhibition at organ level.Meanwhile,we explored the possible mechanisms underlying GA-A inhibition on renal cyst development using MDCK cells treated with 10 μmol·L-1FSK co-incubated with GA-A(25 μg·mL-1)and equal concentration GT.Several key components of Ras/MAPK pathway was evaluated by Western blot,the protein expression of H-ras,B-raf, p-ERK, Egr-1 and c-fos was evaluated. RESULTS MDCK cell viability was not affected by GA-A that were used ofincreasing concentrations up to 200 μg·mL-1. GA-A had no significant influence on cyst formation,but inhibited cyst enlargement dose-dependently and the inhibitory effect is significantly better than that of the same concentration of GT which proved that GA-A may be an effective monomer from GT.And after washing out GA-A on day 8,MDCK cysts regrew to a large size,suggesting that the inhibitory effect of GA-A on MDCK cyst enlargement was reversible. GA-A inhibited embryonic kidney cyst development significantly in a dose-dependent and reversible manner proving GA-A cyst inhibition at organ level,which is also more effective than equal concentration GT.Treatment of MDCK cells with FSK caused a significant elevation of H-ras,B-raf,p-ERK,Egr-1 and c-fos signaling molecules,while treatment with GA-A reduced the level of H-ras,B-raf,p-ERK,Egr-1 and c-fos expression.GA-A down-regulated Ras/MAPK signaling pathway could contribute to its inhibitory effect on cyst development. CONCLUSION Ganoderic acid A from Ganoderma lucidum retard ADPKD renal cyst development via down-regulating Ras/MAPK signaling pathway.

Objective To evaluate diagnostic value of the combined examination of ultrasound , contrast-enhanced CT and physical examination in axillary lymph node status in breast cancer .Methods 316 cases of breast cancer were studied from Jan .2012 to Dec.2013.The axillary lymph nodes were divided into negative group, suspicious negative group , suspicious positive group and positive group by physical examination , axillary ultrasound , and contrast-enhanced breast CT imaging .ROC curve was used to evaluate diagnostic value and the sensitivity,specificity,and accuracy were analyzed .Results The evaluation model presented the AUC of 0.859. The accuracy, sensitivity, specificity, positive and negative predictive value was 91.3%, 93.8%, 92.5%, 94.0%and 90.9%in negative group and positive group .Conclusion This evaluation method can accurately predict the axillary lymph node status and may be instructive to clinical operation mode selection .

Objective To study the efficacy and complications of reconstruction of failed urethro-plasty for hypospadias with pedicle flap , bladder mucosa , buccal mucosa , and biological patch . Methods 23 patients were enrolled from Jul .2005 to Dec.2011.8 patients, with good local skin condition , were performed with pedicle flap urethroplasty .The other 15 patients, with bad local skin condition or with long segment urethral stricture , were performed with free grafts , including 6 bladder mucosa , 7 buccal muco-sal and 2 biological patch. Results Of the 23 cases, 7 cases were cured by one phase operation .There were 16 (16/25) cases had complications.3 (3/16) cases were failed because of serious infection (2 pedi-cle flap, 1 bladder mucosa ) .The failed cases were implemented with urethroplasty 6 months later by the buccal mucosa installments operation .4 (4/16) cases had solitary urethral fistula (1 pedicle flap, 2 bladder mucosa, and 1 buccal mucosal), who were successfully treated with simple fistula repair 3 to 6 months later. 9 ( 9/16) cases had urethral stricture ( 2 pedicle flap , 3 bladder mucosa , 3 buccal mucosal , and 1 biologi-cal patch graft ) , who were treated with urethral sound and got good result .We had reconstructed the urethra using mucosa graft onlay urethroplasty .All of the 23 patients were followed up with an average of 14.5 ( 6-24) months.23 cases were satisfied with the stretched penis , urination and no need to expand the urethra more than 6 months.3 cases were not satisfied with penile appearance .After communication, these patients did not require a further penis orthopedic surgery . Conclusions Pedicle flap, bladder mucosa , buccal mucosa and biological patch can be used in urethral repair and construction of failed urethroplasty for hypos -padias.Urethral sound dilation plays an important role in hypospadias repair .

Objective:To observe the effects of different moxibustion time on cartilage morphology,tumor necrosis factor(TNF)-α and interleukin(IL)-10 of the knee joint in rats with knee osteoarthritis(KOA),and to explore the best treatment time of moxibustion for KOA.Methods:Healthy male Wistar rats were randomly divided into a blank group,a model group,a 15-minute-moxibustion group,a 30-minute-moxibustion group,and a 60-minute-moxibustion group,with 10 rats in each group.Except for the blank group,the KOA model was established in all groups by injecting sodium iodoacetate solution into the knee joint cavity of rats.Rats in the 15-minute-moxibustion group,the 30-minute-moxibustion group,and the 60-minute-moxibustion group were all treated with mild moxibustion intervention for 15 min,30 min,and 60 min,respectively at Neixiyan(EX-LE4)and Dubi(ST35)points near the patella,3 times a week for 4 weeks,12 times in total.Rats in the blank group and the model group were fixed for 30 min without moxibustion intervention.Macroscopic observation for the smoothness of knee cartilage surface was performed after the intervention.Hematoxylin-eosin staining,toluidine blue staining,and Mankin score were used to evaluate the pathological changes in the cartilage.The expression levels of TNF-α and IL-10 in the serum were detected by enzyme-linked immunosorbent assay.Results:Compared with the blank group,the articular cartilage surface in the model group was rough,the chondrocyte arrangement was irregular,the Mankin score and the serum TNF-α expression were significantly increased(P＜0.05),while the expression of serum IL-10 was significantly decreased(P＜0.05).Compared with the model group,the articular cartilage surface was smoother,the chondrocytes were arranged neatly,the Mankin score and serum TNF-α expression level were significantly lower in the three moxibustion intervention groups(P＜0.05);the serum IL-10 level in the 30-minute-moxibustion group and the 60-minute-moxibustion group was increased significantly(P＜0.05).Compared with the 15-minute-moxibustion group,the articular cartilage surface in the 30-minute-moxibustion group and the 60-minute-moxibustion group was smoother,the chondrocyte arrangement was more regular,the Mankin score and the serum TNF-α level were decreased significantly(P＜0.05),and the serum IL-10 level was increased(P＜0.05).There was no significant difference in the serum TNF-α or IL-10 level between the 30-minute-moxibustion group and the 60-minute-moxibustion group(P＞0.05).Conclusion:Moxibustion can obviously improve the morphology and structure of KOA articular cartilage,protect articular cartilage,inhibit cartilage inflammation,and delay KOA cartilage degeneration.Moxibustion's effect is closely related to moxibustion time;the therapeutic effect of the 30-minute-moxibustion and the 60-minute-moxibustion is better than that of the 15-minute-moxibustion.

OBJECTIVETo explore the clinical efficacy of moxibustion on rheumatoid arthritis (RA) in the patients and the impacts on the inflammatory indices.

METHODSForty RA patients were randomized into an observation group and a control group, 20 cases in each one. In the control group, diclofenac sodium sustained release tablets were used, 0.3 g each time, twice a day; methotrexate tablets were used, 10 mg each time, once a week; folic acid was used, 5 mg each time, once a week. In the observation group, on the basis of the treatment as the control group, moxibustion was added specially on the swelling and painful joints, once a day, 15-20 min each time till: the local skin turned slightly red. The treatment was given once a day, at the interval of 1 day after every 6 treatments. In the two groups, the efficacy was observed after 30-day treatment. Before and after treatment, the rheumatoid factor (RF), high sensitivity C-reactive protein (hs-CRP), erythrocyte sedimentation rate (ESR), disease activity score-28 (DAS-28) and the score of joint symptom and physical signs were observed.

RESULTSCompared with those before treatment, the levels of RF, hs-CRP, ESR, DAS-28 and the symptom scale score were all reduced in the RA patients of the two groups after treatment (all P < 0.05). Compared with the control group, the levels of RF, hs-CRP, ESR, DAS-28 and the symptom scale score in the observation group were improved much apparently (all P < 0.05). The total effective rate was 85.0% (17/20) in the observation group and was 80.0% (16/20) in the control group. The total effective rate in the observation group was apparently better than that in the control group (P < 0.05).

CONCLUSIONMoxibustion as supplement relieves the joint symptoms, reduces the inflammatory reactive indices and improves the clinical efficacy in the RA patients.

Adult ; Aged ; Arthritis, Rheumatoid ; blood ; therapy ; C-Reactive Protein ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Rheumatoid Factor ; blood ; Treatment Outcome ; Young Adult

Objective To investigate the relationship of TCRCα-575A/G polymorphism with anti-neutrophil antibody(ANCA) associated vasculitis in Chinese Han population. Methods 86 cases of ANCA associated vasculi-tis in Chinese Han population and 196 healthy subjects were enrolled. TCRCα-575A/G was genotyped by PCR-re-striction fragment length polymorphism (PCR-RFLP) assay. Case-control study was performed. Results No signifi-cant difference was found in either genotype distribution(AA,AG,GG) or allele frequencies between 86 patients and healthy subjects(P>0.05);But significant differences between AA group, AG group, and GG group in systolic pres-sure[(127.47±24.18)、(124.11±25.21)、(148.92±19.23) mm Hg],diastolic pressure [(75.35±14.12)、 (74.50±13.01)、(85.46±9.40) mm Hg],red blood cell count[(3.41±1.01)×109/L、(3.46±1.04)× 109/L、(2.68±0.67)×109/L] and hemoglobin [(90.45±20.69)、(100.66±29.80)、(77.61±15.81) g/L (P<0.05 for each) were found. The patients in GG group had higher blood pressure and more severe anaemia;By following the patients about (16.0±36.8) months,no statistics significance was found between groups with and without chronic renal failure in distributions and genetypes of TCRCα-575A/G (P>0.05 ). Conclusions In Chi-nese Han population,TCRCα-575A/G polymorphism might not be related to genetic susceptibility and chronic renal failure of ANCA associated vaseulitis;but G allele might be associated with more serious anaemia and hypertension.

Objective:To observe the influence of radiofrequency catheter ablation of the fat pad at the right pulmonary vein- atria junction(RPV fat pad)on Bezold-Jarisch reflex(BJR)induced by veratridine in canine.Methods:Ten mongrel dogs were anesthetized and their chest were opened through a right thoracotomy to expose the RPV fat pad.Veratridine was injected into the left ventricle at bolus doses of 10?g/kg through a catheter to induce BJR.The sinus cycle length(SCL),systolic arterial pressure(SAP),diastolic arterial pressure(DAP),mean arterial pressure(MAP),left ventricle systolic pressure(LVSP), left ventricle diastolic pressure(LVDP)and mean left ventricle pressure(MLVP)were measured under the baseline and after ablation of the RPV fat pad;the changes of the above parameters(?SCL,?SAP,?DAP,?MAP,?LVSP,?LVDP,and?MLVP)were calculated.Then the same dose of veratridine was injected under the pad after ablation and the changes of the a- bove parameters were observed.Results:BJR was successfully induced in the 8 animals;the animals has prolonged sinus cycle length(SCL)and decreased MAP and MLVP.After ablation,the prolongation of SCL after injection of veratridine was markedly reduced([229.2?92.3]ms vs[39.3?14.1]ms,P