Objective To compare the effects of combination of maternal hydration and hands and knees posture with simple hands and knees posture on correction of breech pregnancy. Methods One hundred patients who were diagnosed breech pregnancy from November 2015 to November 2016 were assigned to experimental group and control group with 50 cases each according to individual willingness. The patients in experimental group received the combination of maternal hydration and hands and knees posture;while the patients in control group received simple hands and knees posture. Results Amniotic fluid index was (18.94±2.44)cm in experimental group and (15.97±2.33)cm in control group, the difference had statistic significance (t=6.239, P<0.05). The effective transfer rate and successfully used time was 92%(46/50) , (2.10 ± 0.75) weeks in experimental group and 64%(32/50), (4.27 ± 0.98) weeks in control group, the difference had statistic significance (χ2=11.422, P<0.05; t=-12.463, P<0.05). For primipara and multipara, the effective transfer rate was 90.91%(30/33), 16/17 in experimental group, and 68.75%(22/32), 10/18 in control group, the difference had statistic significance (χ2=4.986, 6.806, all P<0.05). The rate of vaginal and cesarean section deliveries was 86%(43/50), 14%(7/50) in experimental group, and 56%(28/50), 44%(22/50) in control group, the difference had statistic significance(χ2=10.928, P=0.001). Patients who had fetal movement or who didn′t have time to do correction or missed was 17.10%(46/269), 1.49%(4/269) in experimental group, and 12.55% (22/255), 7.06% (18/255) in control group. Conclusions @Therapic effects of combination of maternal hydration and hands and knees posture is much better than that of the simple hands and knees posture therapy on correction of breech pregnancy, which is worthy of popularization and application in clinic.

BACKGROUND:Induced pluripotent stem cel technology have solved the contradiction between the ethics and immune rejection, and this high-efficient and safe technique is becoming the mainstream of today’s research. OBJECTIVE:To comprehensively review the safety and application of induced pluripotent stem cel s.METHODS:A computer-based online retrieval of PubMed and CNKI was performed to search relevant papers published from January 2006 to April 2016, with the key words of“induced pluripotent stem cel , reprogramming, clinical application, safety, transcription factor, disease mode”in English and Chinese, respectively. RESULTS AND CONCLUSION:In recent years, research on induced pluripotent stem cel s has attracted much attention from the scientific community and the medical community, and this technique has successful y gained induced pluripotent stem cel s and overcome the problems of immunity and ethics. However, it is limited to the theoretical and laboratory research due to the inability to solve the safety, efficiency and re-differentiation mechanism of induced pluripotent stem cel s. Therefore, we are faced with enormous difficulties and chal enges, which involve al aspects of basic research, including how to safely and effectively induce the differentiation of induced pluripotent stem cel s into the desired cel type and how to establish a suitable disease model as wel as a high-throughput drug screening platform.

Objective To study the effects of different transport protein on the transport of 7,4'-dihydroxyflavone (7,4'-DHF) and its metabolite (7,4'-DHF-S) in Caco-2 cell model.Methods Ultra performance liquid chromatography was employed to determinethe content of 7,4'-DHF and 7,4'-DHF-S incubation buffer,their structures were identified by LC-MS/MS.Bidirectional transport of Caco-2 cells model was used to investigate the influence of ko143 (the inhibitor of BCRP) and MK571 (the inhibitor of MRP2) on the transport of 7,4'-DHF and 7,4'-DHF-S,respectively.Results Metabolic product of 7,4'-DHF in Caco-2 monolayer cell was identified as one monosulfate;PDR of 7,4'-DHF was (1.43 ± 0.11),PDR of ko143 and MK571 on the apparent permeability of 7,4'-DHF was (1.59 ± 0.04) and (1.48 ± 0.07) (P ＞ 0.05);PDR of 7,4'-DHF-S was (1.60 ± 0.06);ko143 could significantly reduce the apparent permeability of 7,4'-DHF-S,and the PDR was (0.23 ±0.03) (P ＜ 0.01);MK571 had no significant effect on the apparent permeability of the 7,4'-DHF-S,and the PDR was (1.51±0.04) (P ＞ 0.05).Conclusion Caco-2 cells can mediate the suffonated reaction of 7,4'-DHF;7,4'-dihydroxyflavone sulfonated combination product may be a substrate for BCRP.

Objective:To construct the human DcR3 expression vector and verify its expression in vitro. Methods: 915 bp human DcR3 gene CDS was amplified from porcine lung tissues,and was cloned into eukaryotic expression vector pEF1a-IRES-DsRed-Express2 which show red fluorescence. And then pEF1a-IRES-DsRed-Express2-DcR3 was transfected into LX-2 cells by FuGene HD. Expression of mRNA and protein lever of Human DcR3 were detected by RT-PCR and Western blot. Results:The levels of DcR3 gene transcription and translation in the hepatic stellate cells were significantly increased after transfection with pEF1a-IRES-DsRed-Ex-press2-DcR3 by RT-PCR and Western blot analysis. Conclusion: DcR3 expression vector was successfully constructed and highly expressed in LX-2 cells.

Objective:To investigate the mRNA expression of the WIF-1 gene and the methylation of its promoter in breast can-cer, and to determine the correlation between the epigenetic aberrant WIF-1 DNA methylation and the clinicopathological significance of WIF-1 in breast cancer . Methods:RT-PCR and sensitive methylation-specific-PCR (MSP) were used to detect WIF-1 mRNA ex-pression and the methylation of the WIF-1 promoter in 30 breast cancer samples as well as in tumor-adjacent tissue samples and 9 be-nign breast tissues. Results:The WIF-1 mRNA expression in 30 breast cancer samples significantly decreased compared with those of the other two groups. In addition, WIF-1 methylation was more frequent in breast-tumor tissues compared with those in tumor-free tis-sues. Meanwhile, WIF-1 mRNA expression in breast cancer tissues involved the abnormal methylation of its promoter. Clinicopatholog-ical correlation analysis showed that the abnormal methylation of the WIF-1 gene promoter was not associated with age, TNM stage, histotype, or lymph node metastasis. Conclusion:WIF-1 mRNA expression loss due to abnormal methylation may be a crucial factor in breast cancer development and can thus be used in the prognosis and progression of the disease.

Objective To investigate the effects of granulocyte macrophage colony-stimulating factor (GM-CSF) combined with interleukin-12 (IL-12) genes on apoptosis of hepatoma cells.Methods The hepatoma cell lines were cultured in vitro and were divided into four groups: GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,negative control group (empty load group),respectively.The PIB-CMV3-GM-CSF and PIB-CMV3-IL-12 eukayotic expression vector was built,and 36 h after transfection,fluorescence microscope was used to detect the transfection effect;the expression level of IL-12,GM-CSF,p53,p38 and C-JUN mRNA were detected by RT-PCR,and Western blot was used to examine the expression level of IL-12,GM-CSF,p53,p38 and C-JUN protein.In addition,the flow cytometry was applied to detect cell apoptosis.Results Through fluorescence microscope,green fluorescence was observed in cells of GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group,indicating that the plasmid has successfully transferred into cells.In addition,the expression of p53mRNA in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were 1.2±0.10,4.3±0.98,4.2±0.34,9.2±0.87,and the protein expression were 1.0±0.10,3.6±0.34,3.8±0.30,5.0±0.60.Compared with the empty load group,the expression level of p53 mRNA and protein were significantly increased in the three plasmid transfection groups (P<0.01).The expression of p53 mRNA and protein were significantly increased in co-transfection group than GM-CSF group and IL-12 group (P<0.01),while in the comparison with GM-CSF transfection group and IL-12 transfection group,the expression level of p53mRNA and protein in the co-transfection group could be improved to a higher degree(P<0.01).Meanwhile,p38 C-JUN mRNA expression levels in empty load group,GM-CSF transfection group,IL-12 transfection group,GM-CSF and IL-12 co-transfection group were as follows: 7.5± 0.9,3.5±0.45,3.7±0.25,1.0±0.11,while p38protein expression levels were 10.1±1.03,6.1± 0.67,7.1 ± 0.61,1.0 ± 0.12,respectively,C-JUN mRNA expression levels were 11.2 ± 1.20,4.1 ± 0.19,3.3 ± 0.30,1.0 ± 0.01,separately,C-JUN protein expression levels were 2.25 ± 0.2,1.8 ± 0.13,1.4 ± 0.12,1.0 ± 0.09.P38, C-JUN mRNA and protein levels were significantly reduced in the three plasmid transfection groups compared with the empty load group (P<0.01).The expression of p38,C-JUN mRNA and protein were reduced to a lower degree in co-transfection group than in GM-CSF transfection group and IL-12 transfection group (P<0.01).Flow cytometer showed that the hepatoma cell apoptosis rate of the empty load group,GM-CSF transfection group,IL-12 transfection group,co-transfection group were (3.43±0.9)%,(5.87±1.02)%,(7.32±1.1)%,(17.47±2.11)%,the rates of the three plasmid transfection groups were significantly higher than that of the empty load group (P<0.01).And the apoptosis rate was significantly increased in the co-transfected group compared with other plasmid groups (P<0.01). Conclusion The combination of GM-CSF and IL-12 could significantly accelerate the apoptosis of hepatoma cells by up-regulating the expression of p53,and down-regulating the expression of p38 and C-JUN.

Objective To investigate the kinectics characteristics of sulfation of apigenin mediated by SULTIA3.Methods After incubation of apigenin using in vitro SULT1A3 system, high-performance liquid chromatography was utilized to determine the sulfates of apigenin.Mass spectrum(MS) were employed to elucidate the structure of metabolite.The program GraphPad Prism 5 was used to perform the kinetic characterization of SULT1A3 catalyzed metabolism of apigenin.Results A liner calibration curve for the assay of apigenin was validated in the range of 0.15625 ~30 μM with the recoveries of at least 80% and intra-day and inter-day RSD of less than 15%.Metabolic product of apigenin and SULT1A3 in the incubated system was identified one monosulfate.The metabolic behavior of apigenin in SULT1A3 was followed substrate inhibition kinetics.Apparent kinetic parameters of metabolism of apigenin by SULT1A3, Kmwas(0.355 ±1.04) μM and Ksi was(23.62 ±0.06) μM,Vmax was(65.71 ±1.30) nmol/(min? mg),Vmax/Km was 185.10 mL/(min? mg).Conclusion SULT1A3 can mediate the binding of apigenin sulfonated reaction, and the character of enzymatic kinetics shows substrate inhibition.Sulfation of apigenin mediated by SULTIA3 may play an important role in phaseⅡmetabolic in vivo.

ObjectiveTo investigate the effect of decoy receptor 3 (DcR3) gene on hepatocyte apoptosis, as well as the possible mechanism of action of DcR3 in this process. MethodsThe human liver cell lines were cultured in vitro, and pEF1α-DcR3 transfection group, pEF1α-IRES transfection group, and negative control group were established. The pEF1α-IRES-DsRed-Express2-DcR3 eukaryotic expression vector was constructed and transfected into human liver cell lines for 36 hours. qRT-PCR was used to measure the mRNA expression of DcR3, Fas ligand (FasL), α-smooth muscle actin (α-SMA), and transforming growth factor-β1 (TGF-β1), Western blot was used to measure the change in the protein expression of DcR3, and flow cytometry was used to measure apoptosis. An analysis of variance was used for comparison of continuous data between groups, and the least significant difference t-test was used for comparison between any two groups. ResultsAfter human liver cell lines were transfected with pEF1α-DcR3 for 36 hours, the pEF1α-DcR3 transfection group showed significant increases in the mRNA and protein expression of DcR3 compared with the pEF1α-IRES transfection group and negative control group (F=33 1695 and 14154, all P＜0.01). Compared with the other two groups, the pEF1α-DcR3 transfection group showed significant reductions in the mRNA expression of FasL, α-SMA, and TGF-β1 (F=269 4518, 20 7904, and 80678, all P＜0.01), which suggested that DcR3 inhibited the expression of FasL, α-SMA, and TGF-β1. Compared with the pEF1α-IRES transfection group and negative control group, the pEF1α-DcR3 transfection group showed a significant reduction in apoptosis rate (F=55863, all P＜0.01). ConclusionDcR3 can inhibit hepatocyte apoptosis and downregulate the mRNA expression of FasL, α-SMA, and TGF-β1.

OBJECTIVE: To observe the morphology of eroded auditory ossicles obtained in middle ear surgery for cholesteatoma and to investigate the mechanism of bone erosion in cholesteatoma. METHOD: The morphology of eroded auditory ossicles in 8 cholesteatoma cases and 2 normal cases were observed with light microscopy. The ultrastructure of eroded auditory ossicles in 5 cholesteatoma cases and the ultrastructure of control bones in external ear canal of 2 cases were observed and compared with transmission electron microscopy. RESULT: Osteomyelitis and multinucleate osteoclasts with ruffled borders were observed in the eroded auditory ossicles of cholesteatoma. Intramembranous and endochondral ossification were both observed. The obvious bone destruction and remodeling were observed consistently. CONCLUSION Osteoclasts and Osteomyelitis are both responsible for bone destruction in cholesteatoma. Intramembranous and endochondral ossification may co-participate in bone remodeling. Osteogenesis is also a basic pathologic phenomena in cholesteatoma. The obvious bone destruction and remodeling can coexist in cholesteatoma cases.
Case-Control Studies ; Cholesteatoma, Middle Ear ; pathology ; Ear Ossicles ; pathology ; ultrastructure ; Humans ; Microscopy, Electron, Transmission ; Osteoclasts ; pathology ; ultrastructure

OBJECTIVE: To observe the expression of receptor activator nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) in cholesteatoma and to determine their roles in the destruction of bone. To observe the role of osteoclasts in bone destruction. METHOD: Immunohistochemical technique and computer image analysis were used to examine the expression of RANKL and OPG in 27 cholesteatoma and 11 normal tissue respectively. RESULT: RANKL was expressed mainly in promatrix cells, including fibroblasts and foam cells. OPG was expressed both in epithelium and promatrix cells. The results of the computer image quantitative analysis showed that the mean optical density of RANKL and the ratio of RANKL/OPG in cholesteatoma were higher than that in normal skin epithelial tissue (P<0.05). There was no significant difference between the mean optical density of OPG in cholesteatoma and that in normal skin epithelial tissue (P>0.05). CONCLUSION The overexpression of RANKL and the increased ratio of RANKL/OPG in cholesteatoma are associated with the destruction of bone. Osteoclasts is possible mediator of osteolysis.
Adolescent ; Adult ; Aged ; Cholesteatoma ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Osteoclasts ; metabolism ; pathology ; Osteoprotegerin ; metabolism ; RANK Ligand ; metabolism ; Receptor Activator of Nuclear Factor-kappa B ; metabolism ; Young Adult