Contrasting assembly mechanisms explain the biogeographic patterns of benthic bacterial and fungal communities on the Tibetan Plateau.

The Tibetan Plateau characterized by high altitude and low temperature, where a great number of lakes are located, is a hotspot of global biodiversity research. Both bacterial and fungal communities are vital participants of biogeochemical cycling in lake ecosystems. However, we know very little about the large-scale biogeographic patterns and the underlying assembly mechanisms of lake benthic microbial communities on the Tibetan Plateau. To investigate the biogeographic patterns and their underlying assembly mechanisms of benthic bacterial and fungal communities, we collected sediment samples from 11 lakes on the Tibetan Plateau (maximum geographic distance between lakes over 1100 km). Benthic community diversity and composition were determined using the high-throughput sequencing technique. Our results indicated that there were contrasting distance-decay relationships between benthic bacterial and fungal communities on a regional scale. Benthic bacterial communities showed a significant distance-decay relationship, whereas no significant relationship was observed for benthic fungal communities. Deterministic processes dominated the bacterial community assembly, whereas fungal community assembly was more stochastic. pH was a dominant factor in influencing the geographic distribution of benthic microbial communities. Co-occurrence network analysis revealed that bacterial communities showed higher complexity and greater stability than those of the fungal communities. Taken together, this study contributes to a novel understanding of the assembly mechanisms underlying the biogeographic distribution of plateau benthic bacterial and fungal communities at a large scale.

Ru2 and Ru encode mouse orthologs of the genes mutated in human Hermansky-Pudlak syndrome types 5 and 6.

Hermansky-Pudlak syndrome (HPS) is a genetically heterogeneous disease involving abnormalities of melanosomes, platelet dense granules and lysosomes. Here we have used positional candidate and transgenic rescue approaches to identify the genes mutated in ruby-eye 2 and ruby-eye mice (ru2 and ru, respectively), two 'mimic' mouse models of HPS. We also show that these genes are orthologs of the genes mutated in individuals with HPS types 5 and 6, respectively, and that their protein products directly interact. Both genes are previously unknown and are found only in higher eukaryotes, and together represent a new class of genes that have evolved in higher organisms to govern the synthesis of highly specialized lysosome-related organelles.

MiR-23b-3p functions as a positive factor for osteoporosis progression by targeting CCND1 in MC3T3-E1 cells.

MiRNAs have gained tremendous attention as studies have shown that miRNAs play important roles in osteoporosis (OP) progression. This study attempted to explore whether miR-23b-3p is involved in the pathogenesis of OP. We detected the miR-23b-3p and Cyclin D1 (CCND1) expressional patterns in the bone of patients with or without OP relying on the GEO database. ß-Glycerophosphate disodium salt and L-ascorbic acid were utilized to stimulate differentiation of MC3T3-E1 cells. Cell proliferative, apoptotic abilities, and cell cycle distribution were determined by CCK-8 and flow cytometry experiments. TargetScan and dual-luciferase reporter analysis were employed to predict and verify the targets of miR-23b-3p. Western blot was implemented to detect the expression of CCND1, apoptosis-related proteins, and cell osteogenesis-related proteins. ALP activity of MC3T3-E1 cells was measured using ALP kit. MiR-23b-3p was increased in OP specimens. Gain-/loss-of-function analysis indicated that the miR-23b-3p inhibited proliferation and differentiation and promoted apoptosis of MC3T3-E1 cells. The levels of Bax and cleaved caspase-3 were increased while those of Bcl-2 were decreased. ALP activity was reduced, and the levels of ALP, Runx2, Osterix, and OPN were declined in MC3T3-E1 cells relative to control. Further analyses demonstrated that CCND1 was a putative target gene of miR-23b-3p. Moreover, knockdown of CCND1 could reverse the impacts of miR-23b-3p inhibitor in MC3T3-E1 cells. MiR-23b-3p functioned as an O-positive factor through regulating cell cycle, proliferation, apoptosis, and differentiation via targeting CCND1.

ERK Inhibitor LY3214996 Targets ERK Pathway-Driven Cancers: A Therapeutic Approach Toward Precision Medicine.

The ERK pathway is critical in oncogenesis; aberrations in components of this pathway are common in approximately 30% of human cancers. ERK1/2 (ERK) regulates cell proliferation, differentiation, and survival and is the terminal node of the pathway. BRAF- and MEK-targeted therapies are effective in BRAF V600E/K metastatic melanoma and lung cancers; however, responses are short-lived due to emergence of resistance. Reactivation of ERK signaling is central to the mechanisms of acquired resistance. Therefore, ERK inhibition provides an opportunity to overcome resistance and leads to improved efficacy. In addition, KRAS-mutant cancers remain an unmet medical need in which ERK inhibitors may provide treatment options alone or in combination with other agents. Here, we report identification and activity of LY3214996, a potent, selective, ATP-competitive ERK inhibitor. LY3214996 treatment inhibited the pharmacodynamic biomarker, phospho-p90RSK1, in cells and tumors, and correlated with LY3214996 exposures and antitumor activities. In in vitro cell proliferation assays, sensitivity to LY3214996 correlated with ERK pathway aberrations. LY3214996 showed dose-dependent tumor growth inhibition and regression in xenograft models harboring ERK pathway alterations. Importantly, more than 50% target inhibition for up to 8 to 16 hours was sufficient for significant tumor growth inhibition as single agent in BRAF- and KRAS-mutant models. LY3214996 also exhibited synergistic combination benefit with a pan-RAF inhibitor in a KRAS-mutant colorectal cancer xenograft model. Furthermore, LY3214996 demonstrated antitumor activity in BRAF-mutant models with acquired resistance in vitro and in vivo. Based on these preclinical data, LY3214996 has advanced to an ongoing phase I clinical trial (NCT02857270).

[The anatomical study of safe zones of the screw fixations in acetabular revision surgery].

OBJECTIVE: To evaluate the safe zones of screw fixations of Jumbo cup and reinforcement rings in acetabular revision surgery. METHODS: From December 2006 to January 2008, 11 fresh frozen human cadaver pelvises were prepared by removing all soft tissues except the medial neurovascular structures, each specimen was held in supine position with clamps. The anteroposterior radiographs were taken after the Jumbo cups or reinforcement rings were fixed in places. Left sides of acetabulum were used to create the models to evaluate the safe zones of the high hip center and Jumbo cups. Right sides were used to evaluate the safe zones of 3 different designs of acetabular reinforcement rings (Müller, Ganz, Burch-Schneider cage). RESULTS: Jumbo cups: screws in the superoanterior and anteroinferior quadrants could cause neurovascular injuries. High hip center: neurovascular injuries could be expected in all quadrants except the inferoposterior and posterior half of superoposterior quadrant. Müller ring: screws placed in the intra-acetabular and extra-acetabular areas of the superoanterior quadrant could cause neurovascular injuries. Ganz ring:the screws placed in superoanterior quadrant and apex area were dangerous. The insults were confirmed in all of the specimens. Burch-Schneider cage: the medial nervous and vasculature structures were injured in all of the specimens if the screws placed in the superoanterior (intra-acetabular and extra-acetabular) quadrant. Screws for ischial fixation could cause sciatic nerve injury. CONCLUSIONS: In revision acetabular surgery, the superoanterior quadrant, the anterior half of the superoposterior quadrant and the implant's central area are unsafe for screw fixation, especially when the exposed thread is longer than 15 mm.

[A cadaveric study of relationships among rotational alignment reference axes of distal femur and tibial mechanical axis].

OBJECTIVES: To investigate the relationships among rotational alignment reference axes of distal femur and tibial mechanical axis, and determine the safest rotational alignment reference axis. METHODS: Digital photos were taken of 30 cadaveric lower extremities with knee in extension and flexion at 90 degrees , angles were measured among tibial mechanical axis and a line perpendicular to clinical epicondylar axis, a line perpendicular to surgical epicondylar axis, Whiteside's line and femoral mechanical axis. Statistical analysis of relationships among those axes were performed. RESULTS: The angles among the tibial mechanical axis and a line perpendicular to the clinical epicondylar axis, a line perpendicular to the surgical epicondylar axis, Whiteside's line and femoral mechanical axis were 0.6 degrees varus, 3.9 degrees varus, 0.2 degrees valgus and 3.0 degrees varus respectively. The angle between the femoral mechanical axis and the tibial mechanical axis was significantly larger than the angles among the tibial mechanical axis and a line perpendicular to the clinical epicondylar axis, the Whiteside's line (P < 0.05). There was no significant difference compared with the angle between a line perpendicular to the surgical epicondylar axis and the tibial mechanical axis. Angles of the clinical epicondylar axis, the surgical epicondylar axis and the Whiteside's line between knee extension and flexion were 2.3 degrees valgus, 0.9 degrees varus and 3.1 degrees valgus respectively. CONCLUSION: The surgical epicondylar axis rather than the clinical epicondylar axis or the Whiteside's line is the safest femoral rotational alignment reference axis intraoperatively.

Identification of 9 uterine genes that are regulated during mouse pregnancy and exhibit abnormal levels in the cyclooxygenase-1 knockout mouse.

BACKGROUND: Preterm birth is the leading cause of all infant mortality. In 2004, 12.5% of all births were preterm. In order to understand preterm labor, we must first understand normal labor. Since many of the myometrial changes that occur during pregnancy are similar in mice and humans and mouse gestation is short, we have studied the uterine genes that change in the mouse during pregnancy. Here, we used microarray analysis to identify uterine genes in the gravid mouse that are differentially regulated in the cyclooxygenase-1 knockout mouse model of delayed parturition. METHODS: Gestational d18.0 uteri (n = 4) were collected from pregnant wild-type and cyclooxygenase-1 knockout mice. Part of the uterus was used for frozen sections and RNA was isolated from the remainder. Microarray analysis was performed at the Indiana University School of Medicine Genomic Core and analyzed using the Microarray Data Portal. Northern analysis was performed to confirm microarray data and the genes localized in the gravid uterus by in situ hybridization. RESULTS: We identified 277 genes that are abnormally expressed in the gravid d18.0 cyclooxygenase-1 knockout mouse. Nine of these genes are also regulated in the normal murine uterus during the last half of gestation. Many of these genes are involved in the immune response, consistent with an important role of the immune system in parturition. Expression of 4 of these genes; arginase I, IgJ, Tnfrsf9 and troponin; was confirmed by Northern analysis to be mis-regulated during pregnancy in the knockout mouse. In situ hybridization of these genes demonstrated a similar location in the gravid wild-type and Cox-1 knockout mouse uteri. CONCLUSION: To our knowledge, this is the first work to demonstrate the uterine location of these 4 genes in the mouse during late pregnancy. There are several putative transcription factor binding sites that are shared by many of the 9 genes identified here including; estrogen and progesterone response elements and Ets binding sites. In summary, this work identifies 9 uterine murine genes that may play a role in parturition. The function of these genes is consistent with an important role of the immune system in parturition.

Preparation of hollow capsule-stabilized gold nanoparticles through the encapsulation of the dendrimer.

The narrow-dispersed dendrimer-encapsulated gold nanoparticles without agglomeration were prepared from the grafted-dendrimers on the surface of the silica microspheres, successively by complexation of the gold chloride anion with the nitrogen atom of the dendrimer and the reduction with sodium borohydride as reductant. The hollow capsule-stabilized gold nanoparticles were prepared through the encapsulation of dendrimer successively by LBL self-assembly process of polyelectrolytes with modified-silica as template and removal of the silica core by hydrofluoric acid. The size of gold nanoparticles was 2.3+/-0.8 nm. The catalytic activity of the capsule-stabilized gold nanoparticles was investigated in the reduction reaction of 4-nitrophenol to 4-aminophenol with sodium borohydride as reductant.

[Treatment of patellar instability of adolescence through arthroscopic reconstruction of medial patellofemoral ligament with trabs of medial soft tissues].

OBJECTIVE: To explore the clinical effectiveness of arthroscopic reconstruction of medial patellofemoral ligament (MPFL) with trabs of medial soft tissues in the treatment of patellar instability of adolescence. METHODS: From January 2005 to December 2006, 23 cases of patellar instability were treated, including 10 males and 13 females with an average age of 16 years old (13 to 20 years old). The locations were left knee in 11 cases, right knee in 12 cases. The disease course was 1-28 weeks (mean 15 weeks). All patients had patellar instability sense and knee arthralgia during strenuous exercise. Preoperative, the International Knee Documentation Committee (IKDC) score and Lysholm score were 48.30 +/- 5.77 and 50.80 +/- 7.61. The congruence angle, lateral patellar angle, and Q angle were (9.00 +/- 2.46), (2.94 +/- 2.55) and (19.10 +/- 4.16) degrees. All of the patients experienced the operation of reconstruction of MPFL with trabs of medial soft tissues (medial patellar retinaculum, joint capsule and vastus medialis oblique muscle fiber) through arthroscope. RESULTS: All the wounds healed by first intention, and no postoperative early complication occurred. All cases were followed up 19 months on average (12 to 24 months). Apprehensive test and patella tilt test were negative. The range of motion returned to normal. There was no recurrence of dislocation after operation. At 12 months after operation, the congruence angle, lateral patellar angle, and Q angle were (-7.03 +/- 0.60), (11.00 +/- 3.47) and (11.30 +/- 1.90) degrees; the IKDC score and Lysholm score were 93.20 +/- 3.51 and 94.10 +/- 4.26. There were statistically significant differences between preoperation and postoperation (P < 0.05). CONCLUSION: Arthroscopic reconstruction of MPFL with trabs of medial soft tissues can improved obviously the affected limb function in treatment of patellar instability of adolescence.

Identification of transcription factors at the site of implantation in the later stages of murine pregnancy.

Despite medical advances, preterm delivery continues to complicate 12% of all births in the United States and is a major cause of neonatal deaths. One of the reasons that preterm labor continues to be a significant problem is that very little is understood about the factors involved in normal labor. Many investigators have studied parturition in the mouse and defined essential pathways for normal labor. Prostaglandins play an essential role in mouse labor and are important in human labor as well. We examined the 23 transcription factors from pregnant mouse uterus that change expression after the induction of cyclooxygenase-1, the enzyme that catalyzes the first committed step in prostaglandin synthesis. Using in situ hybridization, we have identified three of these transcription factors, Hoxa10, Hoxa11 and GILZ as being expressed in the decidua and regulated at the end of pregnancy. Both Hoxa10 and Hoxa11 are known to be critical for implantation, but very little is known about their roles in late gestation. GILZ has not previously been identified in the gravid uterus. In summary, we have identified three transcription factors that are regulated in the decidua at the end of pregnancy, suggesting a role in detachment of the fetus and placenta.

The genomic sequence of the accidental pathogen Legionella pneumophila.

We present the genomic sequence of Legionella pneumophila, the bacterial agent of Legionnaires' disease, a potentially fatal pneumonia acquired from aerosolized contaminated fresh water. The genome includes a 45-kilobase pair element that can exist in chromosomal and episomal forms, selective expansions of important gene families, genes for unexpected metabolic pathways, and previously unknown candidate virulence determinants. We highlight the genes that may account for Legionella's ability to survive in protozoa, mammalian macrophages, and inhospitable environmental niches and that may define new therapeutic targets.