Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Molar ; Root Canal Therapy ; methods

Objective To compare the main pharmacodynamic actions between Siji Sanhuang Capsules (SSC) and Siji Sanhuang Pills (SSP). Methods Antipyretic, anti-inflammatory and antibacterial effects of SSC and its effect on the intestinal movement in normal mice and constipation mice were observed. Results SSC could relieve fever in rats induced by the fresh beer yeast and 2,4-dinitrophenol, significantly inhibit the auricular swelling induced by xylol and the increased capillary permeability caused by histamine in mice, and promote the intestinal movement in normal mice and constipation mice. In-vitro antibacterial experiments showed that SSC had an obvious bacteriostatic effect on Staphylococcus aureus,typhoid bacillus, Bacillus coli, Shigella flexneri, pseudomonas aeruginosa and Bacillus proteus. The effects were in a dose-dependent manner and the effects of SSC were stronger than those of SSP. Conclusion SSC exerts a Strong antipyretic, anti-bacterial, anti-inflammatory and carthartic effect.

OBJECTIVEThe aim of this study is to establish a transgenic mouse model for cleft palate relevant gene thyroid transcription factor-2 (TTF-2), which can be used to study palatal shelf development when the expression pattern and regular activation of TTF-2 is altered.

METHODSThe C57BL/6J mouse TTF-2 gene was cloned through polymerase chain reaction (PCR) from the mouse genomic DNA. The TTF-2 gene was inserted into the expression vector pBROAD3-mcs to construct the recombinant expression vector pBROAD3-TTF-2. This expression vector was then microinjected into the male pronuclei of the fertilized mouse ovum. Thus, the TTF-2 transgenic mice model was established. The genotype of the transgenic mice was identified by PCR and Southern blot analysis. Immunohistochemistry identified the consistent expression of TTF-2 gene during its palatal shelf development.

RESULTSTTF-2 genes were microinjected into 982 fertilized ova. A total of 580 two-cell-stage embryos cultured and transplanted into the oviducts of 48 pseudopregnant female mice. Overall, 68 embryos were obtained for analysis. The genotype of the mice was determined through PCR and Southern blot analysis using genomic DNA extracted from tail biopsies of the transgenic fetus. A total of 13 TTF-2 transgenic mice were detected. The expression of TTF-2 gene during the palatal shelf development of the transgenic mice was consistently detected by immunohistochemistry.

CONCLUSIONThe recombinant expression vector pBROAD3-TTF-2 was integrated into mouse genome through microinjection. The transgenic mouse in the palatal shelf that consistently expressed TTF-2 was successfully established and displayed a cleft palate phenotype.

Animals ; Cleft Palate ; Disease Models, Animal ; Female ; Forkhead Transcription Factors ; Genotype ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Thyroid Gland

Objective: To further understand the role of folic acid supplements rivaling MTHFR gene silencing in pathogenesis of NCLP, RNA interference (RNAi) was applied to knock down MTHFR in mouse embryonic palatal mesenchymal (EPM) cells. Methods: MTHFR ShRNA expression vector were transfected into the primary cultured EPM cells. MTT was used to observe cell proliferation after MTHFR gene silencing. FCM was used to observe cell cycle after MTHFR gene silencing. Results: The results showed the cells proliferation had an inequality amelioration after using folic acid supplements in MEPM cells with MTHFR gene silencing. Using folic acid supplements rivaled the effect of MTHFR gene silencing had a dose-dependent manner. Using 20 μg/ml folic acid supplements could improve the cell proliferation to achieve normal level of cell proliferation. Conclusion: MTHFR gene is an important candidate gene of NCL/P. Using folic acid supplements could prevent teratogenic MTHFR gene silencing for embryonic palate development.

Objective To investigate the correlation between liver-type fatty acid binding protein (L-FABP) and nonalcoholic fatty liver disease (NAFLD) extent and other clinical parameters.Methods Ninety-six patients of NAFLD (NAFLD group) and 100 cases of healthy controls (control group) were selected.The levels of serum L-FABP and blood biochemical parameters were measured by enzyme-linked immunosorbent assay.The lesion degree was assessed by ultrasound.The body mass index (BMI),waist to hip ratio (WHR) and homeostasis model assessment insulin resistance index (HOMA-IR) were calculated.Results The WHR,BMI,fasting plasma glucose (FBG),triglycerides (TG),alanine aminotransferase (ALT),HOMA-IR and L-FABP in NAFLD group were higher than those in control group,there were statistical differences (P ＜ 0.05).The result of correlation analysis showed L-FABP level was positively related with ALT,TG,FBG,WHR,BMI,HOMA-IR (r =0.735,0.728,0.681,0.713,0.699,0.673 ;P ＜0.05),and negative correlation with HDL-C (r =-0.607,P ＜ 0.05).The L-FABP level in control group was (15.42 ± 2.51) g/L,mild NAFLD was (15.96 ± 2.92) g/L,moderate NAFLD was (17.48 ± 3.91) g/L,serious NAFLD was (25.14 ± 5.37) g/L.There was statistical difference in L-FABP level between serious NAFLD and control group,mild NAFLD,moderate NAFLD (P ＜ 0.05).Conclusion Serum L-FABP level of serious NAFLD patient significantly increases,and L-FABP level is related with biochemical parameters of liver function.

Objective To research the operation method for hypospadias by one-stage urethroplasty.Methods According to the length of urethra defect,18 cases were operated by using one-stage urethroplasty with perineal and preputial island flaps.Results 16 cases were successful in all patients(88.9%),except one case of urethrostenosis and urethral fistula respective.The case of urethrostenosis recovered by dilation of urethra after 5 months and the patient of urethral fistula was successfully repaired by operation after 8 months.All patients were followed up for 6～26 months with satisfactory penis and micturition.Conclusion The technique of one-stage urethroplasty by using perineal and preputial island flaps is the one of best choices for hypospadias.

Objective:To develop a highly efficacious and sensitive immunological reagent for further investigation on the retinoic acid-induced gene I (Rig-I) of mouse .Methods:The Helicase domain coding region (726-2 240 bp) of mRig-I-H was cloned into plasmid pET15b (+) to construct the recombinant plasmid pET15b(+)-mRig-I-H.Then the plasmid was transformed into E.coli BL21 for protein expression.Rabbits were immunized with electrophoresis-purified recombinant protein to obtain the polyclonal antibody against mRig-I-H.The titer of polyclonal antibody was detected by ELISA and the specificity was identified by Western blot and Immunofluorescence.Results:The recombinant protein was expressed successfully in E.coli.Western blot analysis showed that target protein was expressed with a molecular weight of 40 kD.Titer of the polyclonal antibody was about 1∶1?105 by ELISA assay.With this antibody,we could detect the expression of Rig-I in RAW 264.7 cell line by Western blot and Immunofluorescence.Conclusion:The high level expression of Rig-I Helicase domain is induced in E.coli expressing system.Anti-mRig-I-H polyclonal antibody with high titer and fine specificity could be a novel tool in future investigation of Rig-I.

Objective To evaluate the function of regulatory T (Treg)cells in peripheral blood from patients with psoriasis, and to explore the possible role of the STAT3 signaling pathway in Treg cell dysfunction. Methods Totally, 81 patients with psoriasis vulgaris, who all presented with chronic plaques and had a psoriasis area and severity index (PASI)score of 10 - 30, were enrolled into this study. Forty-six healthy blood donors served as the control group. Venous blood samples were collected from these subjects followed by isolation of Treg cells and responder T (Tresp)cells. Flow cytometry was performed to determine the proportion of Treg cells in peripheral blood as well as that of cells secreting phosphorylated-STAT3(p-STAT3), interferon γ(IFN-γ), tumor necrosis factor α(TNF-α)and interleukin 17(IL-17)in Treg cells, and quantitative real-time PCR (qRT-PCR)to measure the expression levels of IFN-γ, TNF-α and IL-17 mRNAs in Treg cells. Some Treg cells and Tresp cells were cultured in vitro alone or in combination, and flow cytometry was conducted to estimate cellular proliferative activity on day 7 after stimulation with IL-2. Some patient-derived Treg cells were classified into several groups to be cultured alone or in combination with Tresp cells with or without the presence of the STAT3 pathway inhibitor, Stattic V (10 or 50 μg/L), for 7 days. Subsequently, flow cytometry was performed to evaluate the proliferative activity of Tresp cells, and qRT-PCR to measure the expression levels of IFN-γ, TNF-α and IL-17 mRNAs in Treg cells. Results No significant differences were observed in the proportion of Treg cells in peripheral blood between the patient group and control group (6.437% ± 0.186% vs. 6.812% ± 0.241%, t = 1.224, P >0.05). Compared with control-derived Treg cells, the patient-derived Treg cells showed significantly decreased proliferative activity and inhibitory effects on Tresp cells, but increased proportion of cells secreting p-STAT3, IFN-γ, TNF-α and IL-17 (all P < 0.05). After the treatment with 50 μg/L Stattic V, a significant increase was observed in the inhibitory effect of patient-derived Treg cells on Tresp cells (inhibition rate: 61.670% ± 4.640% vs. 28.820% ± 11.490%, P < 0.05), but a significant decrease in the mRNA expressions of IFN-γ (2-△△C t: 1.654 ± 0.879 vs. 23.350 ± 6.721, P <0.05), TNF-α(0.850 ± 0.705 vs. 4.847 ± 1.525, P < 0.05)and IL-17(0.572 ± 0.135 vs. 3.095 ± 0.650, all P < 0.05)in patient-derived Treg cells compared with untreated patient-derived Treg cells. Conclusions The negative regulatory effect of Treg cells on Tresp cells is decreased in patients with psoriasis, which may be associated with abnormal activation of the STAT3 signaling pathway, and inhibition of the pathway may restore the function of Treg cells to a certain extent.

The aim of this study was to investigate the effect of a synthetic fifteen-residue peptide, Gly-Thr-Pro-Gly-Pro-Gln-Gly-Ile-Ala-Gly-Gln-Arg-Gly-Val-Val(P-15),on islet cell insulin secretion and insulin gene expression.Wistar rat islets,isolated by collagenase and trypsin digestion,were exposed to 0,0. 72,7.2,36?mol/L P-15 with 16.7mmol/L glucose.Insulin release increased approximately twofold in re- sponse to the concentration of 7.2?mol/L P-15 and Northern-blot analysis of islet mRNA with a rat pre- proinsulin cDNA probe showed a concomitant increase in mRNA level as compared with levels observed in islets treated with control.It is concluded that P-15 not only can stimulate the release of insulin but also in- crease the preproinsulin mRNA expression of islet cells.

· AIM: To study the in situ relative intraocular position of the iris-claw phakic intraocular lens (PIOL)for high myopia using an anterior chamber optical coherence tomography (AC OCT)prototype.· METHODS: Six PIOLs (11.50 to 22.00D lens powers) were implanted in phakic myopic eyes. Using AC OCT, tomography was taken in the anterior chamber to measure the preoperative anterior chamber depth, postoperative distance between the PIOL and the corneal endothelium (endothelial-optic distance), and the postoperative distance between the PIOL and the crystalline lens.· RESULTS: Preoperative anterior chamber depth ranged from 3.27 to 3.91 mm and the postoperative endothelial-optic distance measured 2,07 to 2,24 mm. The distance between the crystalline lens and the posterior surface of the IOL ranged from 0.82 to 1.32 mm. Several tomography revealed the position of the PIOL on the iris, The pigment layer of the iris did not seem to be disturbed by the presence of the PIOL.· CONCLUSION, The original anterior chamber depths were reduced by 36,1% to 44.6% after implantation. This study of 6 eyes revealed that tomography taken by AC OCT are useful in verifying the intraocular position of the PIOL within the anterior chamber. Adequate space was maintained between the iris-fixated phakic intraocular lens and the corneal endothelium, angle, and crystalline lens.