[Effect of different support angles on the fitness of removable partial denture framework fabricated using selective laser melting technique].

Objective: To evaluate the fitness of bilateral free-end dentition defect removable partial denture framework fabricated by selective laser melting (SLM) technique with different support angles. Methods: After the control group has been set to eliminate the system error, and according to the standard model of bilateral mandibular posterior teeth loss, eighteen titanium alloy removable partial denture frameworks fabricated by SLM technology were divided into 3 groups with support angles of 0° (horizontal group), 45°(45° group) and 90° (vertical group). Plaster cast with duplicated structure of tissue surface of the removable partial denture (RPD) framework was obtained. A three-dimensional scanner was used to scan original and duplicated plaster casts. The gaps between framework and the model in different parts were analyzed using Geomagic Qualify software to evaluate the fitness of the framework with visual method. Results: The framework fits on the plaster model completely, and its tissue surface fitted on the plaster model well. The deviation between frameworks and plaster casts was calculated as follow: the total deviations of the horizontal, 45°, and vertical group were (0.146±0.017), (0.182±0.015) and (0.185±0.022) mm respectively. The mean deviation of the horizontal group was significantly less than those of the 45° group and the vertical group (P<0.05). Moreover, there was no significant difference in the total deviation between the 45° group and the vertical group. The total deviation of occlusal rest of the horizontal group was significantly less than that of the 45° group (P<0.05). However, no significant difference was detected in the deviation of occlusal rest among the vertical group, the horizontal group, and the 45° group (P>0.05). There was no significant difference in the deviation of occlusal rest among the vertical group, the horizontal group, and the 45° group. The deviation of clasp of the horizontal group was significantly smaller than those of the 45° group and the vertical group (P<0.05). Whereas, there was no significant difference in the deviation of clasp between the 45° group and the 90° group (P>0.05). No significant difference was found in the deviation of lingual bar among the three groups (P>0.05). Conclusions: Among the three kinds of bilateral free-end dentition defect RPD framework fabricated by SLM in different support angles, horizontal printing was proved to reach the minimal deviation, even though the fitness of all three kinds of frameworks can fullfil clinical requirements according to previous studies.

Up-regulation of miR-190b promoted growth, invasion, migration and inhibited apoptosis of Wilms' tumor cells by repressing the PTEN expression.

OBJECTIVE: Wilms' tumor (WT) is the most common malignant tumor in the children's urogenital system. MiR-190b was found to participate in the development and progression of several cancers. However, the molecular mechanism of miR-190b in WT is still unclear. PATIENTS AND METHODS: We detected the miR-190b in WT tissue samples compared to adjacent normal samples as well as in WT patients' blood sample compared to normal volunteers using qRT-PCR. With over-expression and knockdown of miR-190b in WT-derived cell line SK-NEP-1, we next studied cell proliferation, cell circle, apoptosis, invasion and migration abilities change caused by miR-190b ectopic expression. Dual-luciferase assay and Western-blot analysis were used to explain the mechanism of miR-190b in WT. RESULTS: MiR-190b was over-expressed in WT tissue and blood samples compared to normal group, relatively. Up-regulation of miR-190b in SK-NEP-1 cells significantly increased the growth and decreased the apoptosis of cells, while its down-regulation reduced cell proliferation and promoted cell apoptosis of SK-NEP-1. Also, cell invasion and migration abilities were significantly improved after miR-190b over-expression. Moreover, PTEN was proved to be a direct target of miR-190b and its protein level was remarkably decreased after miR-190b up-regulation. CONCLUSIONS: miR-190b over-expressed in WT and promoted cell proliferation, invasion and migration while reduced cell apoptosis of WT cells by repressing PTEN repression, which might provide a potential target for WT diagnosis and therapy.

Seroprevalence of bluetongue virus infection in goats in the central China.

Seroprevalence of Bluetongue virus (BTV) in goats from Hubei was investigated by a commercial competitive enzyme-linked immunosorbent assay kits. Blood samples (n= 1157) were collected during the year 2014 and 2015. The results showed that 13.31% (CI 95% 11.4%-15.4%) serum samples were positive for BTV antibodies in goats in Hubei. The prevalence of BTV antibodies in each region ranged from 1.32% to 27.70%, and differences among the regions were statistically significant (p < 0.01). The prevalence of BTV in male and female goats was 14.23% (95% CI: 11.3, 17.6) and 12.58% (95% CI: 10.1, 15.4), respectively, no significant difference in genders (p > 0.05). In different seasons, the seroprevalence were 8.94% (95% CI: 5.6, 13.3) in spring; 18.31% (95% CI: 14.5, 22.7) in summer; 23.08% (95% CI: 17.0, 30.2) in autumn and 6.98% (95% CI: 4.6, 10.0) in winter, respectively with a significant difference of the prevalence in the different seasons (p < 0.01).

Autophagy protects chondrocytes from glucocorticoids-induced apoptosis via ROS/Akt/FOXO3 signaling.

OBJECTIVE: Glucocorticoids (GCs) have been widely used in the management of osteoarthritis (OA) and rheumatoid arthritis (RA). Nevertheless, there has been some concern about their ability of increasing reactive oxygen species (ROS) in the cartilage. Forkhead-box class O (FOXO) transcription factors have been proved to have a protective role in chondrocytes through regulation of autophagy and defending oxidative stress. The objective of this study was to investigate the role of FOXO3 in Dex-induce up-regulation of ROS. DESIGN: Healthy cartilages debris from six patients were used for chondrocytes culture. After the treatment of dexamethasone (Dex), the ROS levels, autophagic flux, the expression of FOXO3 in chondrocytes were measured. RNA interference technique was also used to determine the role of FOXO3 in Dex-induced autophagy. The metabolism of the extra-cellular matrix was also investigated. THE RESULTS: Dex increased intracellular ROS level, the expression of Akt, FOXO3 as well as autophagy flux in human chondrocytes. The expression of aggrecanases also increased after the treatment of Dex. Catalase, the ROS scavenger, suppressed Dex-induced up-regulation of autophagy flux and expression of aggrecanases and Akt. MK-2206 and LY294002, the PI3K/Akt inhibitors, repressed Dex-induced up-regulation of FOXO3. Silencing FOXO3 resulted in down-regulation of Dex-induced autophagy. Moreover, knockdown of FOXO3 increased Dex-induced apoptosis as well as ROS levels in chondrocytes. In addition, up-regulation of autophagy by Rapamycin resulted in decreasing ROS level in chondrocytes. CONCLUSION: Dex could advance the degenerative process in cartilage. Autophagy was induced in response to Dex-induced up-regulation of ROS via ROS/Akt/FOXO3 signal pathway.

IFN-γ induces aberrant CD49b⁺ NK cell recruitment through regulating CX3CL1: a novel mechanism by which IFN-γ provokes pregnancy failure.

Interferon-γ (IFN-γ), a pleiotropic lymphokine, has important regulatory effects on many cell types. Although IFN-γ is essential for the initiation of uterine vascular modifications and maintenance of decidual integrity, IFN-γ administration can also cause pregnancy failure in many species. However, little is known about the effector mechanisms involved. In this study, using an IFN-γ-induced abortion mouse model, we reported that no Dolichos biflorus agglutinin lectin-positive uterine natural killer (uNK) cells were observed in the uteri from IFN-γ-induced abortion mice. By contrast, the percentage of CD3(-)CD49b(+) NK cells in the uterus and blood from a foetal resorption group was significantly higher than that of the control group. Similarly, significantly upregulated expression of CD49b (a pan-NK cell marker), CX3CL1 and CX3CR1 (CX3CL1 receptor) was detected in the uteri of IFN-γ-induced abortion mice. Using isolated uterine stromal cells, we showed that upregulated expression of CX3CL1 by IFN-γ was dependent on a Janus family kinase 2-signal transducers and activators of transcription 1 (JAK2-STAT1) pathway. We further demonstrated the chemotactic activity of CX3CL1 in uterine stromal cell conditioned medium on primary splenic NK cells. Finally, we observed increased recruitment of CD49b(+) NK cells into the endometrium after exogenous CX3CL1 administration. Collectively, our findings indicate that IFN-γ can significantly increase uterine CX3CL1 expression via activation of the JAK2-STAT1 pathway, thus inducing CD49b(+) NK cell uterine homing, and eventually provoke foetal loss. Thus, we provide a new line of evidence correlating the deleterious effects of IFN-γ on pregnancy with the aberrant regulation of CX3CL1 and CD49b(+) NK cells.

The prognosis of hearing impairment complicating HIV-negative cryptococcal meningitis.

Eight patients who had sensorineural hearing loss (SNHL) associated with cryptococcal meningitis were studied. After a minimum 3-year follow-up, one had died. Among the seven survivors, three had improved, two stabilized, and two progressed. Predictive factors included visual disturbance, meningeal enhancements on MRI, and a CSF cryptococcal antigen titer of >1:1,024. SNHL accounted for 30.8% (8/26) of cryptococcal meningitis patients in our study.

Identification of beta subunit of the rhesus monkey chorionic gonadotropin (rmCG).

Chorionic gonadotropin (CG) is a placental derived hormone that plays a crucial role in successful implantation and establishment of early pregnancy in the primates. The rhesus monkey was chosen as a model to understand the feasibility of developing human DNA immuno-contraceptive. The coding region of rhesus monkey CG beta-subunit (rmCGbeta) was isolated by the TDRT-PCR method. The nucleotide sequence including the leader peptide was 499 nucleotide long and encoded 166 amino acids. In comparing with the previous known primates CG beta-subunits, the rmCGbeta was the highest degree of homology with baboon CG beta-subunit at the deduced amino acid sequence (94%), 79.5% homology with human CG beta-subunit and 70.4% homology with marmoset monkey CG beta-subunit. The eukaryotic expression vector pCMV4-rmCGbeta inserted full-coding cDNA sequence of rmCGbeta was constructed, and the expression of rmCG beta-subunit in HeLa cells transient expressing system in vitro and BALB/c mice in vivo was determined. The results demonstrated that the recombinant PCMV4-rmCGbeta eukaryotic expression vector could express rmCG beta-subunit in vitro and in vivo.

Auditory responses from the frontal cortex in the mustached bat, Pteronotus parnellii.

Response properties of neurons in an auditory field in the frontal cortex of the mustached bat, Pteronotus parnellii, have not been studied before. We recorded neural responses to constant frequency (CF) stimuli from the frontal auditory field in awake animals. The majority (75%) of neurons in this area responded well and often exhibited low thresholds to CF stimuli. Most CF-responsive neurons exhibited sharp tuning with values of > 180 for Q10db, a quality factor expressing the sharpness of tuning at 10dB above threshold. Neurons at 13 recording sites exhibited combination sensitivity in that their responses were facilitated by presenting combinations of either CF1/CF2 and/or CF1/CF3 components of the mustached bat's echolocation signal. Unlike the typical on-responses to a 30 ms tone, observed in the mustached bat's auditory cortex and at subcortical levels, many frontal auditory neurons exhibited loosely time locked firing patterns that lasted for > 100 ms.

Pathophysiology of thrombocytopenia and anemia in mice lacking transcription factor NF-E2.

Expression of the p45 subunit of transcription factor NF-E2 is restricted to selected blood cell lineages, including megakaryocytes and developing erythrocytes. Mice lacking p45 NF-E2 show profound thrombocytopenia, resulting from a late arrest in megakaryocyte differentiation, and a number of red blood cell defects, including anisocytosis and hypochromia. Here we report results of studies aimed to explore the pathophysiology of these abnormalities. Mice lacking NF-E2 produce very few platelet-like particles that display highly disorganized ultrastructure and respond poorly to platelet agonists, features consistent with the usually lethal hemorrhage in these animals. Thrombocytopenia was evident during fetal life and was not corrected by splenectomy in adults. Surprisingly, fetal NF-E2-deficient megakaryocyte progenitors showed reduced proliferation potential in vitro. Thus, NF-E2 is required for regulated megakaryocyte growth as well as for differentiation into platelets. All the erythroid abnormalities were reproduced in lethally irradiated wild-type recipients of hematopoietic cells derived from NF-E2-null fetuses. Whole blood from mice lacking p45 NF-E2 showed numerous small red blood cell fragments; however, survival of intact erythrocytes in vivo was indistinguishable from control mice. Considered together, these observations indicate a requirement for NF-E2 in generating normal erythrocytes. Despite impressive splenomegaly at baseline, mice lacking p45 NF-E2 survived splenectomy, which resulted in increased reticulocyte numbers. This reveals considerable erythroid reserve within extra-splenic sites of hematopoiesis and suggests a role for the spleen in clearing abnormal erythrocytes. Our findings address distinct aspects of the requirements for NF-E2 in blood cell homeostasis and establish its roles in proper differentiation of megakaryocytes and erythrocytes.

Hypothalamic neurons preferentially respond to female nest coo stimulation: demonstration of direct acoustic stimulation of luteinizing hormone release.

Avian vocalizations are generally understood to play a pivotal role in reproductive functions. The role of the hypothalamus in gonadotropin release in higher vertebrates including birds is well established. To date, however, a direct linkage between the neuronal processing of vocal input and the contingent luteinizing hormone (LH) response has not been demonstrated. In this study, using female ring doves, we recorded neuronal activity from hypothalamic nuclei that, as we have shown previously, receive acoustic inputs from the auditory thalamic relay. Concurrently with recording single-unit responses to stimulation with species-specific coo vocalizations, we sampled LH levels in blood from the pituitary veins. LH concentration in the plasma was significantly elevated in birds hearing species-typical coos but not in birds exposed to experimentally altered coos or white noise or in birds that received no vocal stimulation. We found two types of neurons in the preoptic and anterior hypothalamus that selectively responded to the female nest coo: excitatory units and inhibitory units. Among the excitatory neurons are units characterized by two bursts separated by a period of slow spiking or complete silence, in a pattern approximately corresponding temporally to the two-note coo. We designate them as female-nest-coo-specific units. Most neurons in the posterior hypothalamus were nonselective in their response. Female nest coo and male nest coo stimulation evoked an equal magnitude of discharge changes from responsive units in the preoptic-anterior hypothalamic area. We found, however, that the LH increment was three times greater for birds hearing female nest coos than for birds hearing male nest coos. These observations suggest that feature-detecting neurons such as the female-nest-coo-specific units are involved in gonadotropin-releasing hormone output. The present findings are consistent with the well established behavioral evidence that female nest coos mediate ovarian growth.

Reciprocal talk between the auditory thalamus and the hypothalamus: an antidromic study.

In this study we sought to validate physiologically the hypothalamus afferent projections from the auditory thalamus previously identified with tract tracing techniques in the ring dove. In total, we recorded the responses of 628 units in the nucleus ovoidalis (Ov) and its shell region to electrical stimulation applied to anterior hypothalamus and ventromedial nucleus. Ninety-six acoustic units in the shell region displayed good antidromic responses, confirming this region's axonal projections into these nuclei of the hypothalamus. Orthodromic responses (143 units) recorded in the Ov-Ov shell region suggest on the other hand reciprocal projections from the hypothalamus back to the auditory thalamus.

Furostanol glycosides from bulbs of Allium chinense.

Two new furostanol saponins, named chinenosides II and III, were isolated along with seven known compounds, from the bulbs of Allium chinese G. Don by a combination of silica gel, Diaion HP-20, and octadecylsilanized (ODS) silica gel column chromatographies and preparative HPLC. On the basis of chemical and spectroscopic evidence, the structures of chinenosides II and III were determined to be 26-O-beta-glucopyranosyl 3 beta,26-dihydroxy-(25R)-5 alpha-furost-20(22)-en-6-one 3-O-beta-xylopyranosyl-(1 --> 4)-[alpha-arbinopyranosyl(1 --> 6) ]-beta-glucopyranoside and 26-O-beta-glucopyranosyl 3 beta,26-dihydroxy-(25R)-5 alpha-furost-20 (22)-en-6-one 3-O-alpha-arabinopyranosyl (1 --> 6)-beta-glucopyranoside, respectively.

[Two new steroidal saponins from Allium sativum and their inhibitory effects on blood coagulability].

Six compunds were isolated from the fresh bulbs of Allium sativum L (garlic). Their structures were elucidated as proto-iso-eruboside-B (I), eruboside-B (II), iso-eruboside-B (III), sativioside C (IV), adenosine (V) and tryptophan (VI). I and III are new steroidal saponins. This paper deals with the structural determination of I and III and their effects on platelet aggregation, blood coagulation and fibrinolysis in vitro.

[Studies on two new furostanol glycosides from Allium macrostemon Bunge].

Further studies on the active constituents in the bulbs of Allium macrostemon Bunge led to the isolation and structural determination of two new furostanol saponin macrostemonoside E and F. On the basis of chemical evidences and spectral analysis (UV, IR, 1H-NMR, 13C-NMR and FAB-MS), the structure of macrostemonoside E(I) was elucidated as (25R)-26-O-beta-D-glucopyranosyl-5 alpha-furost-20(22)-ene-3 beta,26-diol-3-O- beta-D-glucopyranosyl (1-->2) [beta-D-glucopyranosyl (1-->3)]-beta-D-glucopyranosyl (1-->4)-beta-D-galactopyranoside; macrostemonoside F(II) was established to be (25R)-26-O-beta-D-glucopyranosyl-5 beta-furost-20(22)-ene-3 beta,26-diol-3-O- beta-D-glucopyranosyl (1-->2)-beta-D-galactoside. Preliminary pharmacological tests showed that both macrostemonoside E and F could strongly inhibit ADP-induced human platelet aggregation in vitro. The IC50 of the former was 0.417 mM and that of the latter was 0.020 mM.

[Two new steroidal saponins from Allium macrostemon].

Two new steroidal saponins, macrostemonoside A and D, were isolated from the bulbs of Allium macrostemon Bung (Chinese name as Xie bai), a Traditional Chinese Medicine used for the treatment of myocardial infarction. Their structures were established as tigogenin-3-O-beta-D-glucopyranosyl(1-->2) [beta-D-glucopyranosyl(1-->3)1]-beta-D-glucopyranosyl(1-->4)-beta-D- galactopyranoside (1) and tigogenin-3-O-beta-D-glucopyranosyl(1-->2)[beta-D-glucopyranosyl (1-->3)(6-O-acetyl-beta-D-glucopyranosyl)] (1-->4)-beta-D-galactopyranoside (2) by spectral and chemical evidences. 1 showed remarkable inhibitory effect on rabbit platelet aggregation induced by ADP in vitro (IC50 = 0.065 mmol).