Fabrication and evaluation of porous coatings doped with bioactive elements on titanium surfaces.

OBJECTIVE: Although pure titanium (PT) and its alloys exhibit excellent mechanical properties, they lack biological activity as implants. The purpose of this study was to improve the biological activity of titanium implants through surface modification. MATERIALS AND METHODS: Titanium was processed into titanium discs, where the titanium discs served as anodes and stainless steel served as cathodes, and a copper- and cobalt-doped porous coating [pure titanium model (PTM)] was prepared on the surface of titanium via plasma electrolytic oxidation. The surface characteristics of the coating were evaluated using field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and profilometry. The corrosion resistance of PTM was evaluated with an electrochemical workstation. The biocompatibility and bioactivity of coated bone marrow mesenchymal stem cells (BMSCs) were evaluated through in vitro cell experiments. RESULTS: A copper- and cobalt-doped porous coating was successfully prepared on the surface of titanium, and the doping of copper and cobalt did not change the surface topography of the coating. The porous coating increased the surface roughness of titanium and improved its resistance to corrosion. In addition, the porous coating doped with copper and cobalt promoted the adhesion and spreading of BMSCs. CONCLUSIONS: A porous coating doped with copper and cobalt was prepared on the surface of titanium through plasma electrolytic oxidation. The coating not only improved the roughness and corrosion resistance of titanium but also exhibited good biological activity.

Macrophages promote cell proliferation in colorectal cancer via IL-1ß-mediated downregulation of miR-28-3p.

Infiltration of macrophages is associated with tumor progression and poor prognosis in multiple malignancies, but the underlying mechanisms by which macrophages contribute to colorectal cancer (CRC) have not yet been elucidated. The purpose of this study was to discuss the potential mechanisms of macrophages in CRC. The MTT assay was used to assess cell viability. The expression of the proliferation-related marker PCNA was detected by Western blot analysis. The 10 most important factors (PDGF, VEGF, TNFα, bFGF, IL-8, TGF-ß, IFN-γ, SPARC, IL-1ß and IL-6) secreted by macrophages were knocked down by RNA interference (RNAi), and the mRNA expression levels of these 10 factors were analyzed by qRT-PCR. The effect of these factors on cell proliferation was assessed by the MTT assay. The miRNAs regulated by IL-1ß in CRC cells were identified by miRNA microarray and qRT-PCR analyses. The proliferation ability of miR-28-3p inhibitor on CRC cells was detected by colony formation assay. The association of IL-1ß and miR-28-3p expression with the clinicopathological characteristics in patients with CRC was analyzed by TCGA RNA-seq data. As a result, macrophages promoted the proliferation of CRC cells in a time- and number-dependent manner, and these effects were associated with the upregulation of PCNA and the macrophage-secreted cytokine IL-1ß, which had the most significant effect on CRC cell proliferation. Furthermore, downregulation of miR-28-3p was induced by IL-1ß in CRC cells. The miR-28-3p inhibitor promoted the proliferation in CRC cells. Moreover, upregulation of IL-1ß expression or downregulation of miR-28-3p expression was associated with poor survival in patients with CRC. Therefore, these data demonstrated that macrophages promoted CRC cell proliferation via IL-1ß-mediated downregulation of miR-28-3p.

[Radio frequency catheter ablation of arrhythmias with low dose X-ray guided by three-dimensional mapping system in 14 children].

Objective: To assess the feasibility and efficacy of radiofrequency catheter ablation (RFCA) of arrhythmias with low dose X-ray in children guided by three-dimensional mapping system. Methods: Fourteen children with tachyarrhythmia who were transferred to Tianjin Medical University General Hospital after being hospitalized in Tianjin Children's Hospital and underwent RFCA from April 2017 to May 2018 were included.The results of intraoperative electrophysiological examination, intraoperative X-ray dosage,the immediate success rateand complications of RFCA,and the recurrence during the follow-up for at least 6 months were recorded. Results: Among the 14 children, 11 cases were diagnosed with supraventricular tachycardia.Five cases had atrioventricular reentrant tachycardia (AVRT) which included two cases with left accessory pathway and three cases with right accessory pathway, and four cases had atrioventricular nodular reentrant tachycardia (AVNRT), one case had atrial flutter (AFL), one case had both AFL and AVNRT. One case had focal atrial tachycardia, one case had ventricular tachycardia, and one case had premature ventricular contraction. Eleven cases underwent RFCA with zero radiation, and 3 cases underwent atrial septal puncture with low dose X-ray. The exposure amounts were 3.85, 3.23 and 4.67 mGy, respectively. No complications occurred except for one case with AVRT had atrioventricular block and recovered to normal in 25 days after operation.During the follow-up of 7 to 20 months, no arrhythmias had been found in 13 cases, and one case with AVRT and AFL recurred. Conclusion: Under the guidance of three-dimensional mapping system, RFCA of tachyarrhythmia with low dose X-ray in children is feasible.

The POU homeodomain protein Oct-1 binds Cis-regulatory element essential for the human GnRH upstream promoter activity in JEG-3 cells.

In previous studies, we have localized four specific nuclear protein-binding elements in the human GnRH upstream promoter. To test whether these four elements are reproductive tissue specific, we placed the four elements upstream to a thymidine kinase (TK) promoter/luciferase reporter gene, and transfected the constructs into human placental choriocarcinoma (JEG-3) cells. The 272-bp fragment (-994 to -723) containing the four elements can drive heterologous TK promoter expression in JEG-3 cells about 15 times more than that of basal TK promoter activity. Deletion of element 4 (E4, -987/-968) significantly decreased (4-fold) the luciferase activity. Further deletion of the other elements (E3 individual, -960/-940 or E3 and E2 in combination, -919/-896) only slightly decreased the luciferase activity. In contrast, deletion of element 1 (E1, -876/-851) caused a 2-fold loss of luciferase activity and elimination of E2 and E3 only lost less than 2-fold of the luciferase activity. Study performed with 5' end deletion of this region confirmed these observations. Furthermore, E4 DNA-protein complex can be supershifted by Oct-1 antibody, indicating that Oct-1 binds to E4. These results clearly demonstrated that all four elements are required to confer tissue-specific expression of the hGnRH gene in JEG-3 cells. However, the E4 is the most important for the tissue-specific expression of the hGnRH gene in JEG-3 cells. Oct-1 factor binds with E4 element and may be involved in the mediation of the human GnRH upstream promoter activity.

Estrogen receptor-mediated repression of gonadotropin-releasing hormone (gnRH) promoter activity in transfected CHO-K1 cells.

To examine the transcriptional regulation of gonadotropin-releasing hormone (GnRH) gene in reproductive tissues, the expression of human GnRH gene promoter in cultured human granulosa cells and a Chinese hamster ovary-derived CHO-K1 tumor cells was studied. Transfection of luciferase reporter gene construct containing either upstream (hU) or downstream (hD) human GnRH gene promoter into both human granulosa and CHO-K1 cells showed that the upstream promoter, hU, was more active than hD in directing luciferase expression in these ovarian tissues. CHO-K1 cells transfected with either hU or hD construct showed insignificant changes in luciferase activity in response to 17beta-estradiol and GnRH. However, cotransfection of hU construct with a vector expressing a human estrogen receptor-alpha (ER-alpha) cDNA results in dose-dependent decreases in luciferase activity in response to both 17beta-estradiol and a GnRH agonist. By functional analysis of a series of deletion constructs, the ER-mediated suppression of GnRH promoter activity by 17beta-estradiol was localized to a region between -169 and -548 bp 5' of the upstream transcription start site of the human GnRH gene. Results of this study demonstrated that estrogen receptor can mediate the negative feedback regulation of human GnRH upstream promoter activity by both estrogen and GnRH in the ovary.

Viral induction of low frequency interferon-alpha producing cells.

The human peripheral blood mononuclear cells responsible for IFN-alpha production in response to viral stimuli have been most often described as either monocytes (as typified by the response to Sendai virus) or as a light density, HLA-DR+ population which is negative for most cell surface markers characteristic of mature T cells, B cells, monocytes, or natural killer cells (as typified by the response to Herpes simplex virus (HSV)). The frequency of IFN-alpha-producing cells (IPC) responding to Sendai virus is typically 10-fold or more higher than those responding to HSV. In the current study, we have used ELISpot assays to determine the frequency of IPC responding to DNA and RNA viruses including HSV, Sendai, vesicular stomatitis virus, cytomegalovirus, adenovirus, SV40, influenza, measles, mumps, Newcastle disease virus (NDV) and human immunodeficiency virus (HIV). The enveloped viruses but not the nonenveloped viruses (adenovirus and SV40) elicited an IFN-alpha response. The frequency of IPC for each of the other viruses was more similar to the low frequency HSV-responding population than to the higher frequency Sendai virus response. These included several viruses in the same family as Sendai virus, namely the paramyxo viruses measles, mumps, and NDV. IPC were also tested for sensitivity to the lysosomotropic drug chloroquine, which diminishes IFN-alpha produced in response to HSV but not Sendai virus. With the exception of Sendai virus, chloroquine treatment abrogated the majority of IFN-alpha produced and IPC against each of the viruses. We conclude that low frequency, nonmonocytic NIPC account for the majority of IFN-alpha production in response to different viruses.

[Changes in serum FSH, LH and ovarian follicular growth during electroacupuncture for induction of ovulation].

Changes in serum FSH, LH and follicular sizes were observed in chronically anovulatory patients during electroacupuncture treatment (EAT) for induction of ovulation. 7 cases were diagnosed as PCOD, 3 as dysfunctional uterine bleeding, and 1 as hypogonadotropic amenorrhea. Among them 8 cases complained of infertility for 2.7 years on average. Ovulation was confirmed by pregnancy or the combination of biphasic BBT and ultrasonographic evidence. During one cycle with 3-day EAT on acupoints Ren 3, 4, Extra 16 and Sp 6, ovulation resulted in 5 patients (ovulatory group) and among the 5 cases, 3 of 4 infertile cases became pregnant. The other 5 cases remained in anovulation (anovulatory group); of them 3 cases got biphasic BBT, but no typical ovulatory signs were found on ultrasonography; 2 cases remained in monophasic BBT. Serum FSH, LH values were elevated in ovulatory group, and FSH pulsatile frequency increased significantly during EAT (from 2.10 +/- 0.42/4h to 3.70 +/- 1.64/4h), but not in anovulatory group. No apparent changes were found in serum LH pulsatile frequency and pulsatile amplitudes of FSH and LH in this study. In ovulatory group diameters of ovarian follicles increased markedly, while diameters of anovulatory group stopped to grow at 14-16 mm. It is suggested that ovulation may be induced by EAT via a regulation on hypothalamic-pituitary function leading to normal secretion of FSH and LH.

[Termination of pregnancy with 16,16-dimethyl-trans-delta2 PGE1 methyl ester vaginal suppositories: an analysis of 182 cases].

PIP: The results of pregnancy termination with ONO 802 as a vaginal suppository in 182 cases are presented. The drug was tested during the different phases of pregnancy, as well as in abnormal pregnancies (intrauterine fetal death and hydatidiform mole) and normal pregnancy. 1 suppository containing 1 mg of the drug was inserted in the posterior vaginal fornix every 3 hours, with 1 full course consisting of 5 suppositories. Cases which did not terminate spontaneously were administered the same treatment the next day. A success rate of 90.5% was obtained; 91.6% for the 1st trimester, 88.8% for the 2nd, 94.7% for intrauterine fetal death, and 100% for hydatidiform mole and 3rd trimester cases. A dosage of between 1-5 mg was successful in 83.9%. 75% terminated within 24 hours. In the 34 patients who were given 2 suppositories and whose surgical termination followed 6 hours later, the cervix was so soft and dilated that a No. 8 Hegar dilator was easily inserted and blood loss was reduced to an average of 12.5 ml. After termination, vaginal bleeding ceased within 2 weeks and menses resumed in 6 weeks. Few complications and side effects occurred. Of the 8 cases which later became pregnant, 2 have delivered normal fetuses spontaneously. In light of these results, the use of ONO 802 vaginal suppository is 1 of the better methods of pregnancy termiantion. (author's modified)^ieng